TEST METHODS FOR EVALUATING
SOLID WASTE, PHYSICAL/CHEMICAL
METHODS, SW-846, 3RD EDITION,
PROPOSED UPDATE II
\) Printed on Recycled Paper
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Test Methods for Evaluating Solid Waste, Physical/Chemical Methods
Third Edition
Promulgated Update Package
Instructions
Enclosed is the proposed Update 2 package for "Test Methods for Evaluating Solid
Waste, Physical/Chemical Methods", SW-846, Third Edition. Attached is a list
of methods included in the proposed update, indicating whether the method is a
new method, a partially revised method, or a totally revised method.
Do not discard or replace any of the current pages in the SW-846 manual until
the proposed Update 2 package is promulgated. Until promulgation of the update
package, the methods in the update package are not officially part of the SW-846
manual, and thus do not carry the status of EPA approved methods.
Enclosure
Revised methods are designated by the letter "A" (Revision 1) or the
letter "B" (Revision 2) in the method number. In order to properly
document the method revision used, the entire method number, Including
the letter designation, must be referenced.
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TABLE OF CONTENTS
VOLUME ONE
SECTION A
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
ACKNOWLEDGEMENTS
PART I METHODS FOR ANALYTES AND PROPERTIES
CHAPTER ONE -- QUALITY CONTROL
1.0 Introduction
2.0 Quality Assurance Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitons
6.0 References
CHAPTER TWO -- CHOOSING THE CORRECT PROCEDURE
2.1 Purpose
2.2 Required Information
2.3 Implementing the Guidance
2.4 Characteristics
2.5 Ground Water
2.6 References
CHAPTER THREE -- METALLIC ANALYTES
3.1 Sampling Considerations
3.2 Sample Preparation Methods
Method 3005: Acid Digestion of Waters for Total Recoverable or Dissolved
Metals for Analysis by Flame Atomic Absorption Spectroscopy
or Inductively Coupled Plasma Spectroscopy
Method 3010: Acid Digestion of Aqueous Samples and Extracts for Total Metals
for Analysis by Flame Atomic Absorption Spectroscopy or
Inductively Coupled Plasma Spectroscopy
Method 3015: Microwave Assisted Acid Digestion of Aqueous Samples and
Extracts
Method 3020: Acid Digestion of Aqueous Samples and Extracts for Total Metals
for Analysis by Graphite Furnace Atomic Absorption Spectroscopy
Method 3040: Dissolution Procedure for Oils, Greases, or Waxes
Method 3050: Acid Digestion of Sediments, Sludges, and Soils
CONTENTS - 1
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Method 3051:
Method 5050:
Microwave Assisted Acid Digestion of Sludges, Soils, and Oils
Bomb Combustion Method for Solid Waste
3.3 Methods for Determination of Metals
Method 6010: Inductively Coupled Plasma Atomic Emission Spectroscopy
Method 6020: Inductively Coupled Plasma Mass Spectrometry
Method 7000: Atomic Absorption Methods
Method 7020: Aluminum (AA, Direct Aspiration)
Method 7040: Antimony (AA, Direct Aspiration)
Method 7041: Antimony (AA, Furnace Technique)
Method 7060: Arsenic (AA, Furnace Technique)
Method 7061: Arsenic (AA, Gaseous Hydride)
Method 7062: Antimony and Arsenic (AA, Gaseous Borohydride)
Method 7080: Barium (AA, Direct Aspiration)
Method 7081: Barium (AA, Furnace Technique)
Method 7090: Beryllium (AA, Direct Aspiration)
Method 7091: Beryllium (AA, Furnace Technique)
Method 7130: Cadmium (AA, Direct Aspiration)
Method 7131: Cadmium (AA, Furnace Technique)
Method 7140: Calcium (AA, Direct Aspiration)
Method 7190: Chromium (AA, Direct Aspiration)
Method 7191: Chromium (AA, Furnace Technique)
Method 7195: Chromium, Hexavalent (Coprecipitation)
Method 7196: Chromium, Hexavalent (Colorimetric)
Method 7197: Chromium, Hexavalent (Chelation/Extraction)
Method 7198: Chromium, Hexavalent (Differential Pulse Polarography)
Method 7200: Cobalt (AA, Direct Aspiration)
Method 7201: Cobalt (AA, Furnace Technique)
Method 7210: Copper (AA, Direct Aspiration)
Method 7211: Copper (AA, Furnace Technique)
Method 7380: Iron (AA, Direct Aspiration)
Method 7381: Iron (AA, Furnace Technique)
Method 7420: Lead (AA, Direct Aspiration)
Method 7421: Lead (AA, Furnace Technique)
Method 7430: Lithium (AA, Direct Aspiration)
Method 7450: Magnesium (AA, Direct Aspiration)
Method 7460: Manganese (AA, Direct Aspiration)
Method 7461: Manganese (AA, Furnace Technique)
Method 7470: Mercury in Liquid Waste (Manual Cold-Vapor Technique)
Method 7471: Mercury in Solid or Semi sol id Waste (Manual Cold-Vapor
Technique)
Method 7480: Molybdenum (AA, Direct Aspiration)
Method 7481: Molybdenum (AA, Furnace Technique)
Method 7520: Nickel (AA, Direct Aspiration)
Method 7550: Osmium (AA, Direct Aspiration)
Method 7610: Potassium (AA, Direct Aspiration)
Method 7740: Selenium (AA, Furnace Technique)
Method 7741: Selenium (AA, Gaseous Hydride)
Method 7742: Selenium (AA, Gaseous Borohydride)
Method 7760: Silver (AA, Direct Aspiration)
Method 7761: Silver (AA, Furnace Technique)
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Method 7770: Sodium (AA, Direct Aspiration)
Method 7780: Strontium (AA, Direct Aspiration)
Method 7840: Thallium (AA, Direct Aspiration)
Method 7841: Thallium (AA, Furnace Technique)
Method 7870: Tin (AA, Direct Aspiration)
Method 7910: Vanadium (AA, Direct Aspiration)
Method 7911: Vanadium (AA, Furnace Technique)
Method 7950: Zinc (AA, Direct Aspiration)
Method 7951: Zinc (AA, Furnace Technique)
APPENDIX -- COMPANY REFERENCES
CONTENTS - 3
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VOLUME ONE
SECTION B
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
ACKNOWLEDGEMENTS
CHAPTER ONE. REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 Quality Assurance Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitons
6.0 References
CHAPTER FOUR -- ORGANIC ANALYTES
4.1 Sampling Considerations
4.2 Sample Preparation Methods
4.2.1 Extractions and Preparations
Method
Method
Method
Method
Method
Method
Method
Method
Method
3500:
3510:
3520:
3540:
3541:
3550:
3580:
5030:
5040:
Method 5041:
Method 5100:
Method 5110:
4.2.2 Cleanup
Organic Extraction and Sample Preparation
Separatory Funnel Liquid-Liquid Extraction
Continuous Liquid-Liquid Extraction
Soxhlet Extraction
Automated Soxhlet Extraction
Ultrasonic Extraction
Waste Dilution
Purge-and-Trap
Analysis of Sorbent Cartridges from Volatile Organic
Sampling Train (VOST): Gas Chromatography/Mass
Spectrometry Technique
Analysis of Sorbent Cartridges from Volatile Organic
Sampling Train (VOST): Wide-bore Capillary Gas
Chromatography/Mass Spectrometry Technique
Determination of the Volatile Organic Content of Waste
Samples
Determination of Organic Phase Vapor Pressure in Waste
Samples
Method 3600: Cleanup
Method 3610: Alumina Column Cleanup
Method 3611: Alumina Column Cleanup and Separation of Petroleum Wastes
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Method 3620: Florisil Column Cleanup
Method 3630: Silica Gel Cleanup
Method 3640: Gel-Permeation Chromatography (GPC) Cleanup
Method 3650: Acid-Base Partition Cleanup
Method 3660: Sulfur Cleanup
Method 3665: Sulfuric Acid/Permanganate Cleanup
4.3 Determination of Organic Analytes
4.3.1 Gas Chromatographic Methods
Method 8000:
Method 8010:
Method 8011:
Method 8015:
Method 8020:
Method 8021:
Method 8030:
Method 8031:
Method 8032:
Method 8040:
Method 8060:
Method 8061:
Method 8070:
Method 8080:
Method 8081:
Method 8090:
Method 8100:
Method 8110:
Method 8120:
Method 8121:
Method 8140:
Method 8141:
Method 8150:
Method 8151:
Gas Chromatography
Halogenated Volatile Organics
1,2-Dibromoethane and l,2-Dibromo-3-chloropropane by Gas
Chromatography
Nonhalogenated Volatile Organics by Gas Chromatography
Aromatic Volatile Organics by Gas Chromatography
Halogenated and Aromatic Volatiles by Gas Chromatography
using Electrolytic Conductivity and Photoionization
Detectors in Series: Capillary Column Technique
by
Gas
Acrolein, Acrylonitrile, Acetonitrile
Chromatography
Acrylonitrile by Gas Chromatography
Acrylamide by Gas Chromatography
Phenols by Gas Chromatography
Phthalate Esters by Gas Chromatography
Phthalate Esters by Gas Chromatography: Capillary
Technique
Nitrosamines by Gas Chromatography
Organochlorine Pesticides and Polychlorinated Biphenyls
by Gas Chromatography
Organochlorine Pesticides and Polychlorinated Biphenyls
by Gas Chromatography: Capillary Column Technique
Nitroaromatics and Cyclic Ketones by Gas Chromatography
Polynuclear Aromatic Hydrocarbons by Gas Chromatography
Haloethers by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography: Capillary
Column Technique
Organophosphorus Pesticides by Gas Chromatography
Organophosphorus Compounds by Gas Chromatography:
Capillary Column Technique
Chlorinated Herbicides by Gas Chromatography
Chlorinated Herbicides by Gas Chromatography: Capillary
Column Technique
4.3.2 Gas Chromatographic/Mass Spectroscopic Methods
Method 8240:
Method 8250:
Volatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS): Packed Column Technique
Semivolatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS): Packed Column Technique
CONTENTS - 5
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Method 8260:
Method 8270:
Method 8280:
Method 8290:
Volatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS): Capillary Technique
Semivolatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS): Capillary Technique
The Analysis of Polychlorinated Dibenzo-p-dioxins and
Polychlorinated Dibenzofurans
Appendix A: Signal-to-Noise Determination Methods
Appendix B: Recommended Safety and Handling Procedures
for PCDDs/PCDFs
Polychlorinated Dibenzodioxins (PCDDs) and Polychlorinated
Dibenzofurans (PCDFs) by High Resolution Gas
Chromatography/High Resolution Mass Spectrometry
(HRGC/HRMS)
4.3.3 High Performance Liquid Chromatographic Methods
Method 8310:
Method 8315:
Method 8316:
Method 8318:
Method 8321:
Method 8330:
Method 8331:
Polynuclear Aromatic Hydrocarbons
Formaldehyde by High Performance Liquid Chromatography
Acrylamide, Acrylonitrile and Acrolein by High Performance
Liquid Chromatography (HPLC)
N-Methyl Carbamates by High Performance Liquid
Chromatography (HPLC)
Reverse Phase High Performance Liquid Chromatography with
Thermospray/Mass Spectrometry (HPLC/TSP/MS) Detection
Nitroaromatics and Nitramines by High Performance Liquid
Chromatography (HPLC)
Tetrazene by High Performance Liquid Chromatography (HPLC)
4.3.4 Fourier Transform Infrared Methods
Method 8410: Semivolatile Organics by Gas Chromatography/Fourier
Transform Infrared Spectroscopy (GC/FTIR): Capillary
Column Technique
4.4 Miscellaneous Screening Methods
Method 3810:
Method 3820:
Method 8275:
Headspace
Hexadecane Extraction and Screening of Purgeable Organics
Semivolatile Organic Compounds by Thermal Chromatography/Mass
Spectrometry (TC/MS): Screening Technique
APPENDIX -- COMPANY REFERENCES
CONTENTS - 6
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VOLUME ONE
SECTION C
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
CHAPTER ONE. REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 Quality Assurance Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitons
6.0 References
CHAPTER FIVE -- MISCELLANEOUS TEST METHODS
Method 9010: Total and Amenable Cyanide (Colorimetric, Manual)
Method 9011: Cyanide Extraction Procedure for Solids and Oils
Method 9012: Total and Amenable Cyanide (Colorimetric, Automated UV)
Method 9020: Total Organic Hal ides (TOX)
Method 9021: Purgeable Organic Hal ides (POX)
Method 9022: Total Organic Hal ides (TOX) by Neutron Activation Analysis
Method 9030: Acid-Soluble and Acid-Insoluble Sulfides
Method 9031: Extractable Sulfides
Method 9035: Sulfate (Colorimetric, Automated, Chloranilate)
Method 9036: Sulfate (Colorimetric, Automated, Methylthymol Blue, AA II)
Method 9038: Sulfate (Turbidimetric)
Method 9056: Ion Chromatography Method
Method 9060: Total Organic Carbon
Method 9065: Phenolics (Spectrophotometric, Manual 4-AAP with Distillation)
Method 9066: Phenolics (Colorimetric, Automated 4-AAP with Distillation)
Method 9067: Phenolics (Spectrophotometric, MBTH with Distllation)
Method 9070: Total Recoverable Oil & Grease (Gravimetric, Separatory Funnel
Extraction)
Method 9071: Oil & Grease Extraction Method for Sludge Samples
Method 9073: Total Recoverable Hydrocarbons by Infrared Spectroscopy
Method 9075: Test Method for Total Chlorine in Used Oil by X-ray Fluorescence
spectrometry (XRF)
Test Method for Total Chlorine in New and Used Petroleum Products by
Oxidative Combustion and Microcoulometry
Test Methods for Total Chlorine in New and Used Petroleum Products
(Field Test Kit Methods)
Method 9131: Total Coliform: Multiple Tube Fermentation Technique
Method 9132: Total Coliform: Membrane Filter Technique
Method 9200: Nitrate
Method 9250: Chloride (Colorimetric, Automated Ferricyanide AAI)
Method 9076:
Method 9077:
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Method 9251: Chloride (Colorimetric, Automated Ferricyanide AAII)
Method 9252: Chloride (Titrimetric, Mercuric Nitrate)
Method 9253: Chloride (Titrimetric, Silver Nitrate)
Method 9320: Radium-228
CHAPTER SIX -- PROPERTIES
Method 1320: Multiple Extraction Procedure
Method 1330: Extraction Procedure for Oily Wastes
Method 9040: pH Electrometric Measurement
Method 9041: pH Paper Method
Method 9045: Solid and Waste pH
Method 9050: Specific Conductance
Method 9080: Cation-Exchange Capacity of Soils
Method 9081: Cation-Exchange Capacity of Soils
Method 9090: Compatibility Test for Wastes and
Method 9095: Paint Filter Liquids Test
Method 9096: Liquid Release Test (LRT) Procedure
Method 9100: Saturated Hydraulic Conductivity, Saturated Leachate Conductivity,
and Intrinsic Permeability
Method 9310: Gross Alpha & Gross Beta
Method 9311: Determination of Gross Alpha Activity in
Coprecipitation
Method 9312: Method for Gross Alpha in Solid Samples
Method 9315: Alpha-Emitting Radium Isotopes
(Ammonium Acetate)
(Sodium Acetate)
Membrane Liners
Drinking Water by
PART II CHARACTERISTICS
CHAPTER SEVEN -- INTRODUCTION AND REGULATORY DEFINITIONS
7.1
7.2
7.3
Ignitability
Corrositivity
Reactivity
Test Method to Determine Hydrogen Cyanide Released from Wastes
Test Method to Determine Hydrogen Sulfide Released from Wastes
7.4 Extraction Procedure Toxicity
CHAPTER EIGHT -- METHODS FOR DETERMINING CHARACTERISTICS
8.1 Ignitability
Method 1010: Pensky-Martens Closed-Cup Method for Determining Ignitability
Method 1020: Setaflash Closed-Cup Method for Determining Ignitability
8.2 Corrosivity
Method 1110: Corrosivity Toward Steel
8.3 Reactivity
CONTENTS - 8
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8.4 Toxicity
Method 1310: Extraction Procedure (EP) Toxicity Test Method and Structural
Integrity Test
Method 1311: Toxicity Characteristic Leaching Procedure
Method 1312: Synthetic Precipitation Leaching Procedure
APPENDIX -- COMPANY REFERENCES
CONTENTS - 9
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VOLUME TWO
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
CHAPTER ONE. REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 Quality Assurance Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitons
6.0 References
PART III SAMPLING
CHAPTER NINE -- SAMPLING PLAN
9.1 Design and Development
9.2 Implementation
CHAPTER TEN -- SAMPLING METHODS
Method 0010: Modified Method 5 Sampling Train
Appendix A: Preparation of XAD-2 Sorbent Resin
Appendix B: Total Chromatographable Organic Material Analysis
Method 0020: Source Assessment Sampling System (SASS)
Method 0030: Volatile Organic Sampling Train
PART IV MONITORING
CHAPTER ELEVEN -- GROUND WATER MONITORING
11.1 Background and Objectives
11.2 Relationship to the Regulations and to Other Documents
11.3 Revisions and Additions
11.4 Acceptable Designs and Practices
11.5 Unacceptable Designs and Practices
CHAPTER TWELVE -- LAND TREATMENT MONITORING
12.1 Background
12.2 Treatment Zone
12.3 Regulatory Definition
12.4 Monitoring and Sampling Strategy
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12.5 Analysis
12.6 References and Bibliography
CHAPTER THIRTEEN - INCINERATION
13.1 Introduction
13.2 Regulatory Definition
13.3 Waste Characterization Strategy
13.4 Stack-Gas Effluent Characterization Strategy
13.5 Additional Effluent Characterization Strategy
13.6 Selection of Specific Sampling and Analysis Methods
13.7 References
APPENDIX -- COMPANY REFERENCES
CONTENTS - 11
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CHAPTER TWO
CHOOSING THE CORRECT PROCEDURE
2.1 PURPOSE
This chapter aids the analyst in choosing the appropriate methods for
samples, based upon sample matrix and the analytes to be determined.
2.1.1 Trace Analysis vs. Macroanalvsis
The methods presented in SW-846 were designed through sample sizing and
concentration procedures to address the problem of "trace" analyses (<1000 ppm),
and have been developed for an optimized working range. These methods are also
applicable to "minor" (1000 ppm - 10,000 ppm) and "major" (>10,000 ppm) analyses,
as well as to "trace" analyses, through use of appropriate sample preparation
techniques that result in analyte concentration within that optimized range. Such
sample preparation techniques include:
1) adjustment of size of sample prepared for analysis,
2) adjustment of injection volumes,
3) dilution or concentration of sample,
4) elimination of concentration steps prescribed for "trace" analyses.
5) direct injection (of samples to be analyzed for volatile constituents)
The performance data presented in each of these methods were generated
from "trace" analyses, and may not be applicable to "minor" and "major"
analyses." Generally, extraction efficiency improves as concentration
increases.
Caution: Care should be taken when analyzing samples for trace analyses
subsequent to analysis of concentrated samples due to the possibility
of contamination.
2.1.2 Choice of Apparatus and Preparation of Reagents
Since many types and sizes of glassware and supplies are commercially
available, and since it is possible to prepare reagents and standards in many
different ways, those given in these methods may be replaced by any similar
types as long as this substitution does not affect the overall quality of the
analyses.
2.2 REQUIRED INFORMATION
In order to choose the correct combination of methods to form the
appropriate analytical procedure, some basic information is required.
2.2.1 Physical State(s) of Sample
The phase characteristics of the sample must be known. There are several
general categories of phases in which the sample may be categorized:
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o Aqueous o Oil and Organic Liquid
o Sludges o Solids
o Multiphase Samples o EP and TCLP Extracts
o Ground Water
2.2.2 Analvtes
Analytes are divided into classes based on the determinative methods which
are used to identify and quantify them. The organic compounds are divided into
different groups as indicated by Tables 2-1 through 2-28. Some of the analytes
appear on more than one table, as they may be determined using any of several
methods.
2.2.3 Detection Limits Required
Regulations may require a specific sensitivity or detection limit for an
analysis,-as in the determination of analytes for the Extraction Procedure (EP)
or for delisting petitions. Drinking water detection limits, for those specific
organic and metallic analytes covered by the National Interim Primary Drinking
Water Standards, are desired in the analysis of ground water.
2.2.4 Analytical Objective
Knowledge of the analytical objective will be useful in the choice of
aliquoting procedures and in the selection of a determinative method. This is
especially true when the sample has more than one phase. Knowledge of the
analytical objective may not be possible or desirable at all management levels,
but that information should be transmitted to the analytical laboratory
management to ensure that the correct techniques are being applied to the
analytical effort.
2.2.5 Detection and Monitoring
The strategy for detection of compounds in environmental or process samples
may be contrasted with the strategy for monitoring samples. Detection samples
define initial conditions. When there is little information available about the
composition of the sample source, e.g., a well or process stream, mass spectral
identification of organic analytes leads to fewer false positive results. Thus,
the most practical form of detection for organic analytes, given the analytical
requirements, is mass spectral identification. The choice of technique for
metals is governed by the detection limit requirements and potential
interferents.
Monitoring samples, on the other hand, are analyzed to confirm existing
and on-going conditions, tracking the presence or absence of constituents in an
environmental or process matrix. A less compound(s)-specific detection mode may
be used because the matrix and the analytical conditions are well defined and
stable.
2.2.6 Sample Containers. Preservations, and Holding Times
Appropriate sample containers, sample preservation techniques, and sample
holding times for aqueous matrices are listed in Table 2-29, at the end of this
chapter. Similar information for solid matrices may be found in Table 3-1
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(inorganic analytes) and Table 4-1 (organic analytes). Samples must be
extracted/analyzed within the specified holding times for the results to be
considered reflective of total concentrations. Analytical data generated outside
of the specified holding times must be considered to be minimum values only.
2.3 IMPLEMENTING THE GUIDANCE
The choice of the appropriate sequence of methods depends on the
information required and on the experience of the analyst. Figure 2-1 summarizes
the organic analysis options available. Appropriate selection is confirmed by
the quality control results. The use of the recommended procedures, whether they
are approved or mandatory, does not release the analyst from demonstrating the
correct execution of the method.
2.3.1 Determinative Procedures
The determinative methods for organic analytes have been divided into
three categories, shown in Figure 2-2: gas chromatography (GC); gas
chromatography/mass spectrometry (GC/MS); and high pressure liquid chromatography
(HPLC). This division is intended to help an analyst choose which determinative
method will apply. Under each analyte column, SW-846 method numbers have been
indicated, if appropriate, for the determination of the analyte. A blank has
been left if no chromatographic determinative method is available.
Generally, the MS procedures are more specific but less sensitive than
the appropriate gas chromatographic/specific detection method.
Method 8000 gives a general description of the method of gas
chromatography. This method should be consulted prior to application of any of
the gas chromatographic methods.
Method 8140 and 8141, for organophosphorus pesticides, and Methods 8150
and 8151, for chlorinated herbicides, are preferred to GC/MS because of the
combination of selectivity and sensitivity of the flame photometric, nitrogen-
phosphorus, and electron capture detectors.
Methods 8240 and 8260 are both GC/MS methods for volatile analytes. Method
8240 uses a packed column whereas Method 8260 employs a capillary column. Better
chromatographic separation of the volatile compounds may be obtained by using
Method 8260 rather than 8240. Performance criteria will be based on Method 8260.
Method 5030 has been combined with both Method 8240 and 8260, with which it was
used exclusively. A GC with a selective detector is also useful for the
determination of volatile organic compounds in a monitoring scenario, described
in Section 2.2.5.
Methods 8250 and 8270 are both GC/MS methods for semi volatile analytes.
Method 8250 uses a packed column whereas Method 8270 employs a capillary column.
Better chromatographic separation of the semivolatile compounds may be obtained
by using Method 8270 rather than 8250. Performance criteria will be based on
Method 8270.
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2.3.2 Cleanup Procedures
Each category in Figure 2-3, Cleanup of Organic Analyte Extracts,
corresponds to one of the possible determinative methods available in the manual.
Cleanups employed are determined by the analytes of interest within the extract.
However, the necessity of performing cleanup may also depend upon the matrix from
which the extract was developed. Cleanup of a sample may be done exactly as
instructed in the cleanup method for some of the analytes. There are some
instances when cleanup using one of the methods may only proceed after the
procedure is modified to optimize recovery and separation. Several cleanup
techniques may be possible for each analyte category. The information provided
is not meant to imply that any or all of these methods must be used for the
analysis to be acceptable. Extracts with components which interfere with
spectral or chromatographic determinations are expected to be subjected to
cleanup procedures.
The analyst's discretion must determine the necessity for cleanup
procedures, as there are no clear cut criteria for indicating their use. Method
3600 and associated methods should be consulted for further details on extract
cleanup.
2.3.3 Extraction and Sample Preparation Procedures
Methods for preparing organic analytes are shown in Figure 2-4. Method
3500 and associated methods should be consulted for further details on preparing
the sample for analysis.
2.3.3.1 Aoueous Samples
/ '
The choice of a preparative method depends on the sample. Methods
3510 and 3520 may be used for extraction of the semi volatile organic
compounds. Method 3510, a separatory funnel extraction, is appropriate
for samples which will not form a persistent emulsion interphase between
the sample and the extraction solvent. The formation of an emulsion that
can not be broken up by mechanical techniques will prevent proper
extraction of the sample. Method 3520, a liquid-liquid continuous
extraction, may be used for any aqueous sample; this method will minimize
emulsion formation.
2.3.3.1.1 Basic or Neutral Extraction of Semivolatiles
The solvent extract obtained by performing either Method 3510
or 3520 at a neutral or basic pH will contain the compounds of
interest. Refer to Table 1 in the extraction methods (3510 and/or
3520) for guidance on the pH requirements for extraction prior to
analysis.
2.3.3.1.2 Acidic Extraction of Phenols and Acids
The extract obtained by performing either Method 3510 or 3520
at pH 2 will contain the phenols and acid extractables.
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2.3.3.2 Solid Samples
Soxhlet (Method 3540) and ultrasonic extraction (Method 3550)
extraction are used with solid samples. Consolidated samples should be
ground finely enough to pass through a 9.5 mm sieve. In limited
applications, waste dilution (Method 3580) may be used if the entire sample
is soluble in the specified solvent.
Method 3540 and 3550 are neutral-pH extraction techniques and
therefore, depending on the analysis requirements, acid-base partition
cleanup (Method 3650) may be necessary. Method 3650 will only be needed
if chromatographic interferences are severe enough to prevent detection
of the analytes of interest. This separation will be most important if
a GC method is chosen for analysis of the sample. If GC/MS is used, the
ion selectivity of the technique may compensate for chromatographic
interferences.
2.3.3.3 Oils and Organic Liquids
Method 3580, waste dilution, may be used and the resultant sample
analyzed directly by GC or GC/MS. To avoid overloading the analytical
detection system, care must be exercised to ensure that proper dilutions
are made. Method 3580 gives guidance on performing waste dilutions.
To remove interferences, Method 3611 may be performed on an oil
sample directly, without prior sample preparation.
Method 3650 is the only other preparative procedure for oils and
other organic liquids. This procedure is a back extraction into an aqueous
phase. It is generally introduced as a cleanup procedure for extracts
rather than as a preparative procedure. Oils generally have a high
concentration of semivolatile compounds and, therefore, preparation by
Method 3650 should be done on a relatively small aliquot of the sample.
Generally, extraction of 1 ml of oil will be sufficient to obtain a
saturated aqueous phase and avoid emulsions.
2.3.3.4 Sludge Samples
There is no set ratio of liquid to solid which enables the analyst
to determine which of the three extraction methods cited is the most
appropriate. If the sludge is an organic sludge (solid material and
organic liquid, as opposed to an aqueous sludge), the sample should be
handled as a multiphase sample.
Determining the appropriate methods for analysis of sludges is
complicated because of the lack of precise definition of sludges with
respect to the relative percent of liquid and solid components. They may
be classified into three categories but with appreciable overlap.
2.3.3.4.1 Liquids
Use of Method 3510 or Method 3520 may be applicable to sludges
that behave like and have the consistency of aqueous liquids.
Ultrasonic extraction (Method 3550) and Soxhlet (Method 3540)
TWO - 5 Revision 2
November 1990
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procedures will, most likely, be ineffective because of the
overwhelming presence of the liquid aqueous phase.
2.3.3.4.2 Solids
Soxhlet (Method 3540) and ultrasonic extraction (Method 3550)
will be more effective when applied to sludge samples that resemble
solids. Samples may be dried or centrifuged to form solid materials
for subsequent determination of semivolatile compounds.
Using Method 3650, Acid-Base Partition Cleanup, on the extract
may be necessary, depending on whether chromatographic interferences
prevent determination of the analytes of interest.
2.3.3.4.3 Emulsions
Attempts should be made to break up and separate the phases
of an emulsion. Several techniques are effective in breaking
emulsions or separating the phases of emulsions.
1. Freezing/thawing: Certain emulsions will separate if exposed
to temperatures below 0°C.
2. Salting out: Addition of a salt to make the aqueous phase of
an emulsion too polar to support a less polar phase promotes
separation.
3. Centrifugation: Centrifugal force may separate emulsion
components by density.
4. Addition of water or ethanol: Emulsion polymers may be
destabilized when a preponderance of the aqueous phase is added.
If techniques for breaking emulsions fail, use Method 3520.
If the emulsion can be broken, the different phases (aqueous, solid,
or organic liquid) may then be analyzed separately.
2.3.3.5 Multiphase Samples
Choice of the procedure for aliquoting multiphase samples is very
dependent on the objective of the analysis. With a sample in which some
of the phases tend to separate rapidly, the percent weight or volume of
each phase should be calculated and each phase should be individually
analyzed for the required analytes.
An alternate approach is to obtain a homogeneous sample and attempt
a single analysis on the combination of phases. This approach will give
no information on the abundance of the analytes in the individual phases
other than what can be implied by solubility.
A third alternative is to select phases of interest and to analyze
only those selected phases. This tactic must be consistent with the
sampling/analysis objectives or it will yield insufficient information for
the time and resources expended. The phases selected should be compared
TWO - 6 Revision 2
November 1990
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with Figures 2-1 through 2-4 for further guidance. Figure 2-5 outlines
the testing sequence for determining if a waste exhibits one or more of
the characteristics of a hazardous waste.
2.4 CHARACTERISTICS
2.4.1 EP and TCLP extracts
The leachate obtained from using either the EP (Figure 2-6A) or the TCLP
(Figure 2-6B) is an aqueous sample and, therefore, requires further solvent
extraction prior to the analysis of semivolatile compounds. Figure 3 gives
further information on aqueous sample extraction.
The TCLP leachate is solvent extracted with methylene chloride at a pH > 11
by either Method 3510 or 3520. Method 3510 should be used unless the formation
of emulsions between the sample and the solvent prevent proper extraction. If
this problem is encountered, Method 3520 should be employed.
The solvent extract obtained by performing either Method 3510 or 3520 at
a basic or neutral pH will contain the base/neutral compounds of interest.
Refer to the specific determinative method for guidance on the pH requirements
for extraction prior to analysis.
Due to the high concentration of acetate in the TCLP extract, it is
recommended that purge-and-trap/GC/MS, Methods 8240 or 8260, be used to introduce
the volatile sample into the gas chromatograph.
2.5 GROUND WATER
Appropriate analysis schemes for the determination of analytes in ground
water are presented in Figures 2-7A, 2-7B, and 2-7C. Quantitation limits for
the metallic analytes should correspond to the drinking water limits which are
available.
2.5.1 Special Techniques for Metal Analvtes
All atomic absorption analyses must be performed using background
correction (i.e.. Zeeman, Smith-Hieftje or deuterium arc). Background correction
by the deuterium arc technique may not adequately compensate for high
concentrations of certain interferants in arsenic and selenium analyses (i.e.,
Al, Fe). Zeeman or Smith-Hieftje background correction, or appropriate matrix
modification, may allow analysis for low concentrations of selenium in the
presence of high concentrations of iron, and low concentrations of arsenic in
the presence of high concentrations of aluminum. If significant interference
is suspected, the analyst must switch to an alternate wavelength, or take other
appropriate actions to compensate for the interference effects.
To reduce matrix interferences, all graphite furnace atomic absorption
(GFAA) analyses should be performed using techniques which maximize an isothermal
environment within the furnace cell. Data indicate that two such techniques,
L'vov platform and the Delayed Atomization Cuvette (DAC), are equivalent in this
respect, and produce high quality results.
TWO - 7 Revision 2
November 1990
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Cadmium and antimony should be determined by GFAA. These two elements are
analyzed by GFAA to achieve lower detection limits. Typical GFAA detection limits
for antimony and cadmium are 3 jzg/L and 0.1 ng/l, compared to 60 M9/L and 3 M9/L
by ICP.
All furnace atomic absorption analysis should be carried out using the
exact matrix modifiers listed below. (See also the appropriate methods.)
Element(s) Modifier
As and Se Nickel Nitrate
Pb Phosphoric Acid
Cd Ammonium Phosphate
Sb Ammonium Nitrate
Tl Platinum/Palladium
The ICP calibration standards must match the acid composition and strength
of the acids contained in the samples. Acid strengths in the ICP calibration
standards should be stated in the raw data.
2.5.2 Special Techniques for Indicated Analvtes and Anions
If an Auto-Analyzer is used to read the cyanide distillates, the
spectrophotometer must be used with a 50 mm path length cell. If a sample is
found to contain cyanide, the sample must be redistilled a second time and
analyzed to confirm the presence of the cyanide. The second distillation must
fall within the 14 day holding time.
2.6 REFERENCES
1. Barcelona, M.J. "TOC Determinations in Ground Water"; Ground Water 1984,
22(1). 18-24.
2. Riggin, R.; et al. Development and Evaluation of Methods for Total Organic
Halide and Purgeable Organic Halide in Wastewater; U.S. Environmental
Protection Agency. Office of Research and Development. Environmental
Monitoring and Support Laboratory. ORD Publication Offices of Center for
Environmental Research Information: Cincinnati, OH, 1984; EPA-600/4-84-008.
3. McKee, G.; et al. Determination of Inorganic Anions in Water by Ion
Chromatoqraphv; (Technical addition to Methods for Chemical Analysis of
Water and Wastewater, EPA 600/4-79-020), U.S. Environmental Protection
Agency. Environmental Monitoring and Support Laboratory. ORD Publication
Offices of Center for Environmental Research Information: Cincinnati, OH,
1984; EPA-600/4-84-017.
4. Jarrell Ash Corporation, 590 Lincoln Street, Box 9036, Waltham, MA
02254-9036.
TWO - 8 Revision 2
November 1990
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TABLE 2-1.
ORGANIC COMPOUND CLASSIFICATIONS
CompoundTable(s)
Acenaphthene
Acenaphthylene
Acetaldehyde
Acetone
Acetonitrile
Acetophenone
2-Acetylaminofluorene
l-Acetyl-2-thiourea
Acifluorfen
Acrolein (Propenal)
Aery! amide
Acrylonitrile
Aldicarb (Temik)
Aldicarb Sulfone
Aldrin
Ally! alcohol
Allyl chloride
4-Aminobiphenyl
2-Aminoanthraquinone
Aminoazobenzene
3-Amino-9-ethylcarbazole
Aniline
Anilazine
o-Anisidine
Anthracene
Aramite
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
Asulam
Azinphos ethyl
Azinphos methyl
Barban
Bentazon
Benzal chloride
Benz(a)anthracene
Benzene
Benzidine
Benzoic acid
Benzo(b)fluoranthene
Benzo ( j ) f 1 uoranthene
Benzo(k)fluoranthene
Benzo(g,h,i)perylene
2-14, 2-19
2-14, 2-19
2-23
2-13
2-13, 2-16
2-14
2-14
2-14
2-10
2-13, 2-16, 2-24
2-21, 2-24
2-13, 2-16, 2-24
2-25
2-25
2-9, 2-14, 2-22
2-13
2-13, 2-11
2-14
2-14
2-14
2-14
2-14
2-14
2-14
2-14, 2-19
2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-26
2-8
2-8, 2-14
2-14
2-10
2-18
2-7, 2-14, 2-19
2-12, 2-13, 2-17
2-14
2-2, 2-14
2-7, 2-14, 2-19
2-19
2-14, 2-19, 2-22
2-14, 2-19
TWO - 9
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Benzo(a)pyrene
p-Benzoquinone
Benzotrichloride
Benzyl alcohol
Benzyl benzoate
Butyl benzyl phthalate
Benzyl chloride
BHC (Hexachlorocyclohexane)
•y-BHC (Lindane, gamma-Hexachlorocyclohexane)
a-BHC (al pha-Hexachl orocycl ohexane)
/3-BHC (beta-Hexachl orocycl ohexane)
6-BHC (del ta-Hexachl orocycl ohexane)
Bis(2-n-butoxyethyl) phthalate (BBEP)
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethoxyethyl) phthalate (BEEP)
Bis(2-ethylhexyl) phthalate
Bis(2-methoxyethyl) phthalate (BMEP)
Bis(4-methyl-2-pentyl) phthalate (BMPP)
Bolstar (Sulprofos)
Bromoacetone
Bromobenzene
Bromochl oromethane
Bromodichloromethane
4-Bromof 1 uorobenzene
Bromoform
Bromomethane
4-Bromophenyl phenyl ether
Bromoxynil
2-Butanone (Methyl ethyl ketone)
2-sec-Butyl -4,6-dinitrophenol (DNBP)
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Caffeine
Captafol
Captan
Carbofenthion
Carbaryl
Carbazole
Carbofuran
Carbophenothion
Carbon disulfide
Carbon tetrachloride
Chloramben
Table(s)
2-7, 2-14,
2-14
2-18
2-14
2-3
2-3, 2-14
2-11, 2-13,
2-18
2-9, 2-14,
2-9, 2-14,
2-9, 2-14,
2-9, 2-14,
2-3
2-6, 2-11,
2-6, 2-14
2-6, 2-14
2-3
2-3, 2-14
2-3
2-3
2-8
2-11, 2-13
2-11, 2-12,
2-12, 2-13
2-11, 2-12,
2-13
2-11, 2-12,
2-11, 2-12,
2-6, 2-14
2-14
2-13
2-2, 2-14
2-12, 2-13
2-12, 2-13
2-12, 2-13
2-26
2-14
2-14
2-8
2-14, 2-25
2-22
2-14, 2-25
2-14
2-13
2-11, 2-12,
2-10
2-19, 2-22
2-18
2-18
2-18
2-18
2-18
2-14
2-13
2-13
2-13
2-13
2-13
TWO - 10
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Table(s)
Chlordane
Chlorfenvinphos
Chlorinated dibenzodioxins
4-Chloro-3-methyl phenol
Chloroacetaldehyde
4-Chloroaniline
Chlorobenzene
Chlorobenzilate
Chlorodibromomethane
Chloroethane
2-Chloroethanol
Bi s (2-chl oroethoxy)methane
Bis(2-chloroethyl) sulfide*
2-Chloroethyl vinyl ether
Chloroform
1-Chlorohexane
Bis(2-chloroisopropyl) ether
Chloromethane
5-Chloro-2-methylaniline
Chloromethyl methyl ether
4-Chloro-3-methyl phenol
1-Chloronaphthalene
2-Chl oronaphthal ene
2-Chlorophenol
4-Chloro-l,2-phenylenediamine
4-Chl oro- 1 , 3-phenyl enedi ami ne
4-Chlorophenyl phenyl ether
Chloroprene
3-Chloropropiom'trile
Chlorotoluene(s)
2-Chlorotoluene
4-Chlorotoluene
5-Chloro-o-toluidine
3- (Chloromethyl Jpyridine hydrochloride
Chlorpyrifos
Chrysene
Coumaphos
Coumarin Dyes
Creosote
p-Cresidine
Cresols (methyl phenols) (Cresylic acids)
o-Cresol (2-methyl phenol)
m-Cresol (3-methyl phenol)
p-Cresol (4-methylphenol)
Crotoxyphos
2-Cyclohexyl-4,6-dinitrophenol
2,4-D
2-7, 2-9, 2-14
2-8, 2-14
2-7
2-2, 2-14
2-11
2-14
2-11, 2-12, 2-13, 2-17
2-14
2-13
2-11, 2-12, 2-13
2-11, 2-13
2-11
2-13
2-11, 2-13
2-11, 2-12, 2-13
2-11
2-11
2-11, 2-12, 2-13
2-14
2-11
2-22
2-14, 2-22
2-14, 2-18
2-2, 2-14, 2-22
2-14
2-14
2-6, 2-14
2-11, 2-13
2-13
2-11
2-12, 2-13
2-11, 2-12, 2-13
2-14
2-14
2-8
2-7, 2-14, 2-19
2-8, 2-14
2-26
2-7
2-14
2-2, 2-7
2-14
2-14
2-14
2-14
2-2, 2-14
2-10
TWO - 11
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Dalapon
2,4-DB
DCPA diacid
4,4'-DDD
4,4'-DDE
4,4'-DDT
Demeton-o,s
Diallate (cis, trans)
2,4-Diaminotoluene
Diamyl phthalate (DAP)
Diazinon
Dibenz(a,h)acridine
Dibenz (a, h) anthracene
Dibenz(a,j)acridine
Dibenzo(a,e)pyrene
Dibenzo(a,h)pyrene
Dibenzo(a,i)pyrene
7H-Dibenzo(c,g)carbazole
Dibenzofuran
Dibenzothiophene
Di bromochl oromethane
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Dibromomethane
Di-n-butyl phthalate
Dicamba
Di chl one
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Dichlorobenzene(s)
3,3'-Dichlorobenzidine
3,5-Dichlorobenzoic acid
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene (Vinylidene chloride)
cis-1, 2-Di chl oroethene
trans -1, 2-Di chl oroethene
Di chl oromethane (Methylene chloride)
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorophenoxyacetic acid
Dichloroprop
1, 2-Di chl oropropane
1,3-Dichloropropane
Table(s)
2-10
2-10
2-10
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-8, 2-14
2-14
2-14
2-3
2-8
2-19
2-14, 2-19
2-14, 2-19
2-19, 2-14
2-19
2-19
2-19
2-14
2-22
2-11, 2-12, 2-13
2-11, 2-13, 2-14
2-12, 2-13
2-11, 2-12, 2-13
2-3, 2-14
2-10
2-14
2-11, 2-12, 2-13,
2-11, 2-12, 2-13,
2-11, 2-12, 2-13,
2-7, 2-18
2-14
2-10
2-11, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-2, 2-14, 2-22
2-2, 2-14
2-7
2-10
2-11, 2-12, 2-13
2-12, 2-13
2-14, 2-17, 2-18
2-14, 2-17, 2-18
2-14, 2-17, 2-18
TWO - 12
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
2,2-Dichloropropane
l,3-Dichloro-2-propanol
1,1-Dichloropropene
cis-l,3-Dichloropropene
trans - 1 , 3-Di chl oropropene
Dichlorvos
Dicrotophos
Dicyclohexyl phthalate (DCP)
Dieldrin
l,2:3,4-Diepoxybutane
Diethyl ether
Diethylstilbestrol
Diethyl sulfate
Diethyl phthalate
1,4-Difluorobenzene
Dihexyl phthalate (DHP)
Dihydrosaffrole
Diisobutyl phthalate (DIBP)
Dimethoate
3,3'-Dimethoxybenzidine
3, 3' -Dimethyl benzidine
Dimethyl phthalate
p-Dimethylaminoazobenzene
7,12-Dimethylbenz(a)anthracene
a-,a-Dimethylphenethylamine
2,4-Dimethylphenol
4, 6-Dinitro-2-methyl phenol
Dinitrobenzene
1,2-Dinitrobenzene
1,3-Dinitrobenzene (DNB)
1,4-Dinitrobenzene
4,6-Dinitro-o-cresol
2,4-Dinitrophenol
2,4-Dinitrotoluene (24DNT)
2,6-Dinitrotoluene (26DNT)
Dinocap
Dinonyl phthalate
Dinoseb
Di-n-octyl phthalate
Dioxacarb
1,4-Dioxane
Dioxathion
Diphenylamine
1,2-Diphenylhydantoin
1,2-Diphenylhydrazine
Disperse Blue 3
Table(s)
2-12, 2-13
2-11, 2-13
2-12, 2-13
2-11, 2-13
2-11, 2-13
2-8, 2-14, 2-26
2-14
2-3
2-9, 2-14
2-13
2-15
2-14
2-14
2-3, 2-14
2-13
2-3
2-14
2-3
2-8, 2-14, 2-26
2-14
2-14
2-3, 2-14
2-14
2-14
2-14
2-2, 2-14
2-14
2-5, 2-7
2-14
2-14, 2-27
2-14
2-7
2-2, 2-14
2-5, 2-7, 2-14, 2-22, 2-27
2-5, 2-14, 2-27
2-14
2-3
2-10, 2-14
2-3, 2-14
2-25
2-13
2-8, 2-14
2-14, 2-22
2-14
2-14
2-26
TWO - 13
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Table(s)
Disperse Blue 14
Disperse Brown 1
Disperse Orange 3
Disperse Orange 30
Disperse Red 1
Disperse Red 5
Disperse Red 13
Disperse Red 60
Disperse Yellow 5
Disulfoton
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Epichlorohydrin
EPN
Ethanol
Ethion
Ethoprop
Ethyl benzene
Ethyl carbamate
Ethyl methacrylate
Ethyl methanesulfonate
Ethylene dibromide
Ethyl ene oxide
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
Fluorescent Brightener 61
Fluorescent Brightener 236
2-Fluorobiphenyl
2-Fluorophenol
Formaldehyde
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl orophene
Hexachl oropropene
Hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX)
2-26
2-26
2-26
2-26
2-26
2-26
2-26
2-26
2-26
2-8, 2-14, 2-26
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-9, 2-14
2-14
2-11, 2-13
2-8, 2-14
2-13, 2-15
2-8, 2-14
2-8
2-12, 2-13, 2-17
2-14
2-13
2-14
2-11
2-13
2-8, 2-14, 2-26
2-8, 2-14, 2-26
2-8, 2-14
2-14
2-14, 2-19
2-14, 2-19, 2-22
2-26
2-26
2-14
2-14
2-23
2-7, 2-9, 2-14
2-9, 2-14
2-7, 2-14, 2-18, 2-22
2-7, 2-12, 2-13, 2-14, 2-18
2-7, 2-14, 2-18
2-7, 2-14, 2-18
2-14
2-14
2-27
TWO - 14
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Hexamethyl phosphoramide
2-Hexanone
Hexyl 2-ethylhexyl phthalate (HEHP)
HMPA
1,2,3,4,6,7,8-HpCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,7,8-HxCDF
3-Hydroxycarbofuran
5-Hydroxydicamba
Hydroquinone
2-Hydroxypropi oni tri 1 e
Indeno(l,2,3-cd)pyrene
lodomethane
Isobutyl alcohol
Isodrin
Isophorone
Isopropyl benzene
p- 1 sopropyl toluene
Isosafrole
Leptophos
Malathion
Malononitrile
MCPA
MCPP
Merphos
Mestranol
Methacrylonitrile
Methapyrilene
Methiocarb (Mesurol)
Methomyl (Lannate)
Methoxychlor
3-Methylcholanthrene
2-Methyl -4,6-dinitrophenol
4,4'-Methylenebis(2-chloroaniline)
4, 4' -Methyl enebis(N,N-dimethyl aniline)
Methyl ethyl ketone (MEK)
Methyl iodide
Methyl isobutyl ketone (MIBK)
Methyl methacrylate
Methyl methanesulfonate
2-Methyl naphthalene
2-Methyl -5-nitroaniline
Methyl Parathion
4-Methyl -2-pentanone
4-Methyl phenol
2-Methyl pyridine
Methyl -2,4,6-trinitrophenylnitramine (Tetryl )
Table(s)
2-14
2-13
2-3
2-8
2-20
2-20
2-20
2-25
2-10
2-14
2-13
2-14, 2-19
2-13
2-13
2-14
2-5, 2-14
2-12, 2-13
2-12, 2-13
2-14
2-8, 2-14
2-8, 2-14
2-13
2-10
2-10
2-8, 2-26
2-14
2-13
2-14
2-25
2-25, 2-26
2-9, 2-14
2-14, 2-19
2-2
2-14
2-14
2-15
2-11, 2-13
2-15
2-13
2-14
2-14
2-14
2-14, 2-26
2-13
2-22
2-14
2-27
TWO - 15
Revision 2
November 1990
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TABLE 2-1.
(Continued)
Compound
Table(s)
Mevinphos 2-8,
Mexacarbate ' 2-14
Mirex 2-14
Monochrotophos 2-8,
Naled 2-8,
Naphthalene 2-7,
Naphthoquinone 2-5
1,4-Naphthoquinone 2-14
1-Naphthylamine 2-14
2-Naphthylamine 2-14
Nicotine 2-14
5-Nitroacenaphthene 2-14
2-Nitroaniline 2-14
3-Nitroaniline 2-14
4-Nitroaniline 2-14
5-Nitro-o-anisidine 2-14
Nitrobenzene (NB) 2-5,
4-Nitrobiphenyl 2-14
Nitrofen 2-14
2-Nitrophenol 2-2,
4-Nitrophenol 2-2,
Nitroquinoline-1-oxide 2-14
N-Nitrosodibutylamine 2-14
N-Nitrosodiethylamine 2-14
N-Nitrosodimethylamine 2-4,
N-Nitrosodiphenylamine 2-4,
N-Nitrosodi-n-propylamine 2-4,
N-Nitrosomethylethyl amine 2-14
N-Nitrosomorpholine 2-14
N-Nitrosopiperidine 2-14
N-Nitrosopyrrolidine 2-14
o-Nitrotoluene (2NT) 2-27
m-Nitrotoluene (3NT) 2-27
p-Nitrotoluene (4NT) 2-27
5-Nitro-o-toluidine 2-14
OCDF 2-20
Octamethyl pyrophosphoramide 2-14
Octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine (HMX) 2-27
4,4'-Oxydianiline 2-14
Parathion 2-14
Parathion ethyl 2-8
Parathion methyl 2-8
PCB-1016 (Aroclor-1016) 2-9
PCB-1221 (Aroclor-1221) 2-9
PCB-1232 (Aroclor-1232) 2-9
PCB-1242 (Aroclor-1242) 2-9
PCB-1248 (Aroclor-1248) 2-9
TWO - 16
2-14
2-14, 2-26
2-14, 2-26
2-12, 2-13, 2-14, 2-19, 2-22
2-7, 2-14, 2-27
2-14
2-10, 2-14
2-14
2-14
2-14
Revision 2
November 1990
-------
TABLE 2-1.
(Continued)
Compound
Table(s)
PCB-1254 (Aroclor-1254)
PCB-1260 (Aroclor-1260)
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
Pentachlorobenzene
Pentachloroethane
Pentachlorohexane
Pentachloronitrobenzene
Pentachlorophenol
Phenacetin
Phenanthrene
Phenobarbital
Phenol
Phenylenediamine
Phorate
Phosalone
Phosmet
Phosphamidion
Phthalic anhydride
2-Picoline
Picloram
Piperonyl sulfoxide
B-Priopiolactone
Promecarb
Pronamide
Propargyl alcohol
Propionitrile
Propoxur (Baygon)
n-Propylamine
n-Propylbenzene
Propylthiouracil
Pyrene
Pyridine
Resorcinol
Ronnel
Safrole
Solvent Red 3
Solvent Red 23
Strychnine
Styrene
Sulfall ate
Sulfotep
2,4,5-T
2X7.8-TCDD
1,2,3,4-TCDD
2-9
2-9
2-20
2-20
2-20
2-14, 2-18
2-13
2-18
2-14
2-2, 2-10, 2-14
2-14
2-14, 2-19, 2-22
2-14
2-2, 2-14
2-14
2-7, 2-8, 2-14
2-14, 2-26
2-8, 2-14
2-8, 2-14
2-14
2-7, 2-13, 2-14, 2-17
2-10
2-14
2-13
2-25
2-14
2-13
2-13
2-25
2-13
2-12, 2-13
2-14
2-14, 2-19, 2-22
2-7, 2-13, 2-14, 2-17
2-14
2-8
2-14
2-26
2-26
2-14, 2-26
2-12, 2-13, 2-17
2-14
2-8
2-10
2-20
2-20
TWO - 17
Revision 2
November 1990
-------
TABLE 2-1.
(Continued)
Compound
Table(s)
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7,8-TCDD
1,2,8,9-TCDD
1,2,7,8-TCDF
TEPP
Terbuphos
Terphenyl
Tetrachlorobenzene(s)
1,2,3,4-Tetrachlorobenzene
1,2,3,5-Tetrachlorobenzene
1,2,4,5-Tetrachlorobenzene
1,1,2,2-Tetrachloroethane
1,1,1,2-Tetrachloroethane
Tetrachloroethene
2,3,4,6-Tetrachlorophenol
Tetrachlorophenol (s)
Tetrachlorvinphos (Stirophos)
Tetraethyl dithiopyrophosphate
Tetraethyl pyrophosphate
Tetrazene
Thiofanox
Thionazine
Thiophenol (Benzenethiol)
TOCP
Tokuthion (Prothiofos)
Toluene
Toluene diisocyanate
o-Toluidine
Toxaphene
2,4,5-TP (Silvex)
2,4,6-Tri bromophenol
1,2,3-Trichlorobenzene
1,2,4-Tri chlorobenzene
1,3,5-Trichlorobenzene
1,1,1-Tri chloroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chlorof1uoromethane
Trichlorfon
Trichloronate
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Trichlorophenol(s)
1,2,3-Tri chloropropane
Trichloropropane(s)
2-20
2-20
2-20
2-20
2-20
2-20
2-8
2-8, 2-14
2-14
2-7, 2-18
2-18
2-18
2-14, 2-18
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-14
2-2
2-8, 2-14
2-14
2-14
2-28
2-26
2-14
2-14, 2-17
2-8
2-8
2-12, 2-13, 2-17
2-14
2-14
2-7, 2-9, 2-14
2-7, 2-10
2-14
2-12, 2-13, 2-18
2-12, 2-13, 2-14, 2-18
2-18
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-11, 2-12, 2-13
2-26
2-8
2-14
2-2, 2-14
2-2
2-12, 2-13
2-11
TWO - 18
Revision 2
November 1990
-------
TABLE 2-1.
(Continued)
Compound Table(s)
Trifluralin 2-14
2,4,5-Trimethylaniline 2-14
1,2,4-Trimethylbenzene 2-12, 2-13
1,3,5-Trimethylbenzene 2-12, 2-13
Trimethyl phosphate 2-14
1,3,5-Trinitrobenzene (TNB) 2-14, 2-27
2,4,6-Trinitrotoluene (TNT) 2-27
Tris(2,3-dibromopropyl) phosphate (Tris-BP) 2-14, 2-26
Tri-p-tolyl phosphate 2-14
0,0,0-Triethyl phosphorothioate 2-14
Vinyl acetate 2-13
Vinyl chloride 2-11, 2-12, 2-13
o-Xylene 2-12, 2-17
m-Xylene 2-12, 2-17
p-Xylene 2-12, 2-17
Xylene(s) 2-13
TABLE 2-2.
METHOD 8040 - PHENOLS
2-sec-Butyl-4,6-dinitrophenol (DNBP) 2,4-Dimethylphenol
4-Chloro-3-methylphenol 2,4-Dinitrophenol
2-Chlorophenol 2-Methyl-4,6-dinitrophenol
Cresol(s) (methyl phenols) 2-Nitrophenol
2-Cyclohexyl-4,6-dinitrophenol 4-Nitrophenol
2,4-Dichlorophenol Pentachlorophenol
2,6-Dichlorophenol Phenol
Trichlorophenol(s) 2,4,6-Trichlorophenol
Tetrachlorophenol(s)
TWO - 19 Revision 2
November 1990
-------
TABLE 2-3. TABLE 2-4.
METHODS 8060/8061 - PHTHALATE ESTERS METHOD 8070 - NITROSAMINES
Benzyl benzoate* N-Nitrosodimethyl amine
Benzyl butyl phthalate N-Nitrosodiphenylamine
Bis(2-n-butoxyethyl) phthalate (BBEP)" N-Nitrosodi-n-propylamine
Bis(2-ethylhexyl) phthalate
Bis(2-ethoxyethyl) phthalate (BEEP)*
Bis(4-methyl-2-pentyl) phthalate (BMPP)*
Bis(2-methoxyethyl) phthalate (BMEP)*
Diamyl phthalate (DAP)*
Di-n-butyl phthalate
Dicyclohexyl phthalate (DCP)*
Diethyl phthalate
Dihexyl phthalate (DHP)"
Diisobutyl phthalate (DIBP)*
Dimethyl phthalate
Dinonyl phthalate*
Di-n-octyl phthalate
Hexyl 2-ethylhexyl phthalate (HEHP)*
Target analyte of Method 8061 only.
TABLE 2-5.
METHOD 8090 - NITROAROMATICS AND TABLE 2-6.
CYCLIC KETONES METHOD 8110 - HALOETHERS
Dinitrobenzene Bis(2-chloroethyl) ether
2,4-Dinitrotoluene Bis(2-chloroethoxy)methane
2,6-Dinitrotoluene Bis(2-chloroisopropyl) ether
Isophorone 4-Bromophenyl phenyl ether
Naphthoquinone 4-Chlorophenyl phenyl ether
Nitrobenzene
TABLE 2-7a.
METHOD 3650 - BASE/NEUTRAL FRACTION
Benz(a)anthracene Hexachlorobenzene
Benzo(a)pyrene Hexachlorobutadiene
Benzo(b)fluoranthene Hexachloroethane
Chlordane Hexachlorocyclopentadiene
Chlorinated dibenzodioxins Naphthalene
Chrysene Nitrobenzene
Creosote Phorate
Dichlorobenzene(s) 2-Picoline
Dinitrobenzene Pyridine
2,4-Dinitrotoluene Tetrachlorobenzene(s)
Heptachlor Toxaphene
TWO - 20 Revision 2
November 1990
-------
TABLE 2-7b.
METHOD 3650 - ACID FRACTION
2-Chlorophenol
Cresol(s)
Dichlorophenoxyacetic acid
2,4-Dimethylphenol
4,6-Dinitro-o-cresol
4-Nitrophenol
Pentachlorophenol
Phenol
Tetrachlorophenol(s)
Trichlorophenol(s)
2,4,5-TP (Silvex)
TABLE 2-8.
METHODS 8140/8141 - ORGANOPHOSPHORUS COMPOUNDS
(PACKED AND CAPILLARY COLUMNS)
Azinphos methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton, o,s
Diazinon
Dichlorvos^
Dimethoate*
Disulfoton
EPN*
Ethoprop
Fensulfothion
Fenthion
Trichloronate
Target analyte of Method 8141 only.
Malathion*
Merphos
Mevinphos
Monochrotophos"
Naled
Parathion ethyl"
Parathion methyl
Phorate
Ronnel
Stirophos (Tetrachlorvinphos)
Sulfotep*
TEPP"
TOCP*
Tokuthion (Prothiofos)
TABLE 2-9.
METHODS 8080/8081 - ORGANOCHLORINE PESTICIDES AND PCBs
Aldrin
a-BHC
0-BHC
6-BHC
•y-BHC (Lindane)
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Toxaphene
PCB-1016 (Arocl
PCB-1221 (Arocl
PCB-1232 (Arocl
PCB-1242 (Arocl
PCB-1248 (Arocl
PCB-1254 (Arocl
PCB-1260 (Arocl
or-1016)
or-1221)
or-1232)
or-1242)
or-1248)
or-1254)
or-1260)
TWO - 21
Revision 2
November 1990
-------
TABLE 2-10.
METHODS 8150/8151 - CHLORINATED HERBICIDES
Acifluorfen*
Bentazon*
Chloramben"
2,4-D
Dalapon
2,4-DB
DCPA diacid*
Dicamba
3,5-Dichlorobenzoic acid"
Dichlorprop
Dinoseb
5-Hydroxydicamba*
Target analyte of Method 8151 only.
MCPA
MCPP
4-Nitrophenol*
Pentachlprophenol"
Picloram"
2,4,5-TP (Silvex)
2,4,5-T
TABLE 2-11.
METHOD 8010 - HALOGENATED VOLATILES
Allyl chloride
Benzyl chloride
Bromoacetone
Bromobenzene
Bromodichloromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chloroacetaldehyde
Chlorobenzene
Chloroethane
Bis(2-chloroethoxy)methane
2-Chloroethanol
2-Chloroethyl vinyl ether
Chloroform
1-Chlorohexane
Bis(2-chloroisopropyl) ether
Chloromethane
Chloromethyl methyl ether
Chloroprene
4-Chlorotoluene
Di bromochloromethane
1,2-Di bromo-3-chloropropane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichloro-2-butene
Di chlorodi f1uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene (Vinylidene chloride)
trans-1,2-Dichloroethene
Dichloromethane (Methylene Chloride)
1,2-Dichloropropane
1,3-Dichloro-2-propanol
cis-l,3-Dichloropropene
trans-1,3-Dichloropropene
Epichlorohydrin
Ethylene dibromide
Methyl iodide
1,1,2,2-Tetrachloroethane
1,1,1,2-Tetrachloroethane
Tetrachloroethene
1,1,1-Tri chloroethane
1,1,2-Trichloroethane
Trichloroethene
Trichlorofluoromethane
Trichloropropane
Vinyl chloride
TWO - 22
Revision 2
November 1990
-------
TABLE 2-12.
METHOD 8021 (METHOD 8011") - HALOGENATED AND AROMATIC VOLATILES
Benzene
Bromobenzene
Bromochloromethane
Bromodi chloromethane
Bromoform
Bromomethane
n-Butylbenzene
sec-Butyl benzene
tert-Butylbenzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chloromethane
2-Chlorotoluene
4-Chlorotoluene
Di bromochloromethane
1,2-Di bromo-3-chlpropropane*
1,2-Dibromoethane*
Dlbromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Di chlorobenzene
Di chlorodi f1uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1,-Dichloroethene
cis-1,2-Dichloroethene
trans-1,2-Dichloroethene
Target analyte of Method 8011
1,2-Dichloropropane
1,3-Dichloropropane
2,2-Di chloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadi ene
Isopropylbenzene
p-Isopropyltoluene
Methylene chloride
Naphthalene
n-Propylbenzene
Styrene
1,1,1,2-Tetrachloroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1,1,2-Tri chloroethane
Trichloroethene
Tri chlorof1uoromethane
1,2,3-Trichloropropane
1,2,4-Tri methyl benzene
1,3,5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
TWO - 23
Revision 2
November 1990
-------
TABLE 2-13.
METHODS 8240/8260 - VOLATILES
Acetone"
Acetonitrile*
Acrolein (Propenal)*
Acrylonitrile
Allyl alcohol".
Allyl chloride*
Benzene
Benzyl chloride*
Bromobenzene*
Bromoacetone*
Bromochloromethane
Bromodi chloromethane
l-Bromo-4-fluorobenzene
Bromoform
Bromomethane
2-Butanone (Methyl ethyl ketone)*
n-Butylbenzene*
sec-Butyl benzene*
tert-Butylbenzene*
Carbon disulfide"
Carbon tetrachloride
Chlorobenzene
Chlorodi bromomethane*
Chloroethane
2-Chloroethanol*
Bis(2-chloroethyl) sulfide*
2-Chloroethyl vinyl ether*
Chloroform
Chloromethane
Chloroprene*
3-Chloropropionitrile*
2-Chlorotoluene*
4-Chlorotoluene*
Di bromochloromethane*
1,2-Di bromo-3-chloropropane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene*
1,3-Di chlorobenzene*
1,4-Dichlorobenzene*
l,4-Dichloro-2-butene*
Di chlorodi f1uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1,2-Dichloroethene*
trans-1,2-Dichloroethene
1,2-Dichloropropane
1,3-Dichloropropane*
2,2-Dichloropropane*
1,3-Dichloro-2-propanol*
1,1-Dichloropropene*
cis-1,3-Dichloropropene*
trans-1,3-Di chloropropene*
1,2:3,4-Di epoxybutane
1,4-Difluorpbenzene*
1,4-Dioxane"
Epichlorohydrin*
Ethanol*
Ethyl benzene
Ethylene oxide*
Ethyl methacrylate*
Hexachlorobutadi ene*
2-Hexanone*
2-Hydroxyprppionitrile*
lodomethane"
Isobutyl alcohol*
Isopropylbenzene*
p-Isopropylto]uene*
Malononitrile*
Methacrylonitrile*
Methylene chloride
Methyl iodide"
Methyl methacrylate*^
4-Methyl-2-pentanone*
Naphthalene*
Pentachlorpethane*
2-Picoline*
Propargyl alcohol*
b-Propiolactone"
Propionitrile]
n-Propylamine*
n-Propylbenzene*
Pyridine*
Styrene
1,1,1,2-Tetrachloroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene*
1,2,4-Trichlorobenzene*
1,1,1-Trichloroethane
1,1,2-Tri chloroethane
Trichloroethene
Tri chlorof1uoromethane
1,2,3-Trichloropropane
1,2,4-Trimethylbenzene*
1,3,5-Trimethylbenzene*
Vinyl acetate
Vinyl chloride
Xylene(s)
* Target analyte of Method 8240. All Method 8240 analytes should be analyzable by Method
8260.
* Target analyte of Method 8260 only.
TWO - 24
Revision 2
November 1990
-------
TABLE 2-14.
METHODS 8250/8270 - SEMIVOLATILES
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetylami nof1uorene"
1-Acetyl-2-thiourea"
Aldrin
2-Aminoanthraquinone"
Aminoazobenzene*
4-Aminobiphenyl
3-Ami no-9-ethylcarbazole*
Anilazine"
Aniline
o-Anisidine*
Anthracene
Aramite*
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
Azinphps-methyl*
Barban"
Benz(a)anthracene
Benzidine
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzoic acid
Benzo(g,h,i)perylene
Benzo(a)pyrenet
p-Benzoquinone*
Benzyl alcohol
a-BHC
/3-BHC
5-BHC
7-BHC (Lindane)
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Bromoxynil*
Butyl benzyl phthalate
2-sec-Butyl-4,6-dinitrophenoV
Captafpl*
Captan*
Carbaryl*
Carbofuran"
Carbofenthion*
Chlordane
Chlorfenvinphos"
4-Chloroaniline
Chiorobenzilate*
5-Chloro-2-methylani1i ne"
4-Chloro-3-methylphenol
3-(Chloromethyl) pyridine hydrochloride"
1-Chloronaphthalene
2-Chloronaphthalene
2-Chlorophenol
4-Chloro-l,2-phenylenediamine]
4-Chloro-l,3-phenylenediamine*
4-Chlorophenyl phenyl ether
5-Chloro-o-toluidine"
Chrysene
Coumaphos"
p-Cresidine]
Crotoxyphos"
2-Cyclohexyl-4,6-dinitrophenol*
4,4'-DDD
4,4'-DDE*
4,4'-DDT
Demeton-o]
Demeton-s*
Diallate (cis or trans)"
2,4-Diaminotoluene*
Dibenz(a,j)acridine
Dibenz(a,h)anthracene
Dibenzofuran
Dibenzo(a,e)pyrene*
1,2-Di bromo-3-chloropropane*
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorpphenol
Dichlorovos]
Dicrotophos"
Dieldrin
Diethyl phthalate
Diethylstilbestrol*
Diethyl sulfate"
Dihydrosaffrole"
Dimethoate"
3,3'-Dimethoxybenzidine*
TWO - 25
Revision 2
November 1990
-------
TABLE 2-14.
METHODS 8250/8270 - SEMIVOLATILES (CONTINUED)
p-Dimethylaminoazobenzene
7,12-Dimethylbenz(a)anthracene
3,3'-Dimethylbenzidine*
a-,a-Dimethylphenethylamine
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene*
1,3-Dinitrobenzene*
1,4-Dinitrobenzene*
4,6-Dinitro-2-methylphenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap*
Dinoseb*
Dioxathion"
Diphenylamine
5,5-Diphenylhydantoin*
1,2-Di phenylhydrazi ne
Di-n-octyl^phthalate
Disulfoton*
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
EPN*
Ethion*
Ethyl carbamate"
Ethyl methanesulfonate
Famphur*
Fensulfothion*
Fenthion*
Fluchloralin"
Fluoranthene
Fluorene
2-Fluorobiphenyl
2-Fluorophenol
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachlorocyclopentadi ene
Hexachloroethane
Hexachlorophene*
Hexachloropropene*
Hexamethyl phosphoramide*
Hydroquinone"
Indeno(l,2,3-cd)pyrene
Isodrin*
Isophorone
Isosafrole*
Kepone*
Leptophos*
Malathion*
Maleic Anhydride*
Mestranol*
Methapyrilene*
Methoxychlor
3-Methylcholanthrene
4,4'-Methylenebis(2-chloroaniline)*
4,4'-Methylenebis(N,N-dimethylaniline)"
Methyl methanesulfonate
2-Methylnaphthalene
2-Methyl-5-nitroaniline"
Methyl parathion*
2-Methylphenol (o-cresol)
3-Methylphenol (m-cresol)*
4-Methylphenol (p-cresol)
2-Methylpyridine
Mevinphos
Mexacarbate*
Mi rex*
Monocrotophos*
Naled*
Naphthalene
1,4-Naphthoquinone"
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene*
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitroanisidine*
Nitrobenzene
4-Nitrobiphenyl*
Nitrofen*
2-Nitrophenol
4-Nitrophenol
Nitroquinoline-1-oxide*
N-Ni trosodi butyl ami ne^
N-Nitrosodiethyl amine"
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi-n-propylamine
N-Ni trosomethylethyl ami ne*
N-Nitrosomorpholine*
N-Nitrosopiperidine
TWO - 26
Revision 2
November 1990
-------
TABLE 2-14.
METHODS 8250/8270 - SEMIVOLATILES (CONTINUED)
N-Nitrosopyrrolidine*
5-Nitro-o-toluidine*
Octamethyl pyrophpsphoramide*
4,4'-Oxydianiline*
Parathion*
Pentachlorobenzene
Pentachloron i trobenzene
Pentachlorophenol
Phenacetin
Phenanthrene
Phenobarbital*
Phenol
Phenylenediamine*
Phorate"
Phosalone*
Phosmet*
Phosphamidion*
Phthalic anhydride*
2-Picoline
Piperonyl sulfoxide"
Pronamide
Propylthiouracil"
Pyrene
Pyridine"
Resorcinol*
Target analyte of Method 8270 only.
Safrole*
Strychnine*
Sulfal1 ate*
Terbuphos*
Terphenyl
1,2,4,5-Tetrachlorobenzene
2,3,4,6-Tetrachlorophenol
Tetrachlorvinphos (Stirophos)*
Tetraethyl dithiopyrophosphate*
Tetraethyl^ pyrophosphate*
Thionazine*
Thiophenol (Benzenethiol)*
Toluene diisocyanate*
o-Toluidine*
Toxaphene
2,4,6-Tri bromophenol
1,2,4-Tri chlorobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Trifluralin"
2,4,5-Trimethylaniline*
Trimethyl phosphate*
1,3,5-Trinitrobenzene*
Tris(2,3-dibromopropyl) phosphate"
Tri-p-tolyl phosphate*
0,0,0-Triethyl phosphorothioate*
TABLE 2-15.
METHOD 8015 - NON-HALOGENATED VOLATILES
Diethyl ether
Ethanol
Methyl ethyl ketone (MEK)
Methyl isobutyl ketone (MIBK)
TABLE 2-16.
METHODS 8030/8031 - ACETONITRILE,
ACROLEIN, ACRYLONITRILE
Acetonitrile*
Acrolein (Propenal)"
Acrylonitrile
Target analyte of Method 8030 only.
TWO - 27
Revision 2
November 1990
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TABLE 2-17.
METHOD 8020 - AROMATIC VOLATILES
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Ethyl benzene
2-Picoline
Pyridine
Styrene
Toluene
Thiophenol (Benzenethiol)
o-Xylene
m-Xylene
p-Xylene
TABLE 2-18.
METHODS 8120/8121 - CHLORINATED HYDROCARBONS
Benzal chloride*<
Benzotrichloride"
Benzyl chloride*
2-Chloronaphthalene
Dichlorobenzene(s)*
1,2-Dichlorobenzene"
1,3-Dichlorobenzene]
1,4-Di chlorobenzene*
Hexachlorobenzene
Hexachlorobutadiene
Hexachlorocyclohexane*
alpha-Hexachlorocyclohexane (alpha-BHC)*
beta-Hexachlorocyclohexane (beta-BHC)*
gamma-Hexachlorocyclohexane (gamma-BHC)]
delta-Hexachlorocyclohexane (delta-BHC)*
Hexachlorocyclopentadi ene
Hexachloroethane
Pentachlorobenzene*
Pentachlorohexane*
Tetrachlorobenzenets)*
1,2,3,4-Tetrachlorobenzene"
1,2,3,5-Tetrachlorobenzene]
1,2,4,5-Tetrachlorobenzene*
1,2,3-Tri chlorobenzene"
1,2,4-Tri chlorobenzene^
1,3,5-Tri chlorobenzene"
Target analyte of Method 8121 only.
* Target analyte of Method 8121 only.
TABLE 2-19.
METHODS 8100/8310 - POLYNUCLEAR AROMATIC HYDROCARBONS
Acenaphthene
Acenaphthylene
Anthracene
Benz(a)anthracene
Benzo(a)pyrene
Benzo(b)fl uoranthene
Benzo(g,h,i)perylene
Benzo(k)f1uoranthene
Chrysene
Dibenz(a,h)acridine
Fluoranthene
Fluorene
Indeno(l,2,3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
TWO - 28
Revision 2
November 1990
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TABLE 2-20.
METHODS 8280/8290 - DIOXINS AND DIBENZOFURANS
2,3,7,8-TCDD
1,2,3,4-TCDD
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7,8-TCDD
1,2,8,9-TCDD
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
1,2,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
OCDF
TABLE 2-21.
METHOD 8032 - ACRYLAMIDE
Acrylamide
TABLE 2-23.
METHOD 8315 - FORMALDEHYDE
Formaldehyde
Acetaldehyde
TABLE 2-22.
METHOD 8275 - SEMIVOLATILES (SCREENING)
2-Chlorophenol
4-Methylphenol
2,4-Dichlorophenol
Naphthalene
4-Chloro-3-methyl-phenol
1-Chioronaphthalene
2,4-Dinitrotoluene
Fluorene
Diphenylamine
Hexachlorobenzene
Dibenzothiophene
Phenanthrene
Carbazole
Aldrin
Pyrene
Benzo(k)fluoranthene
Benzo(a)pyrene
TABLE 2-24.
METHOD 8316 - ACRYLAMIDE,
ACRYLONITRILE AND ACROLEIN
Acrylamide
Acrylonitrile
Acrolein
TABLE 2-25.
METHOD 8318 - N-METHYL CARBAMATES
Aldicarb (Temik)
Carbaryl (Sevin)
Carbofuran (Furadan)
Dioxacarb
3-Hydroxycarbofuran
Methiocarb (Mesurol)
Methomyl (Lannate)
Promecarb
Propoxur (Baygon)
TWO - 29
Revision 2
November 1990
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TABLE 2-26.
MEfriOD 8321 - NONVOLATILES
Azo Dyes
Disperse Red 1
Disperse Red 5
Disperse Red 13
Disperse Yellow 5
Disperse Orange 3
Disperse Orange 30
Disperse Brown 1
Solvent Red 3
Solvent Red 23
Anthraquinone Dves
Disperse Blue 3
Disperse Blue 14
Disperse Red 60
Coumarin Dyes
(Fluorescent Brighteners)
Fluorescent Brightener 61
Fluorescent Brightener 236
Alkaloids
Caffeine
Strychnine
Orqanophosphorus Compounds
Methomyl
Thiofanox
Famphur
Asulam
Dichlorvos
Dimethoate
Disulfoton
Fensulfothion
Merphos
Methyl parathion
Monocrotophos
Naled
Phorate
Trichlorfon
Tris-(2,3-Dibromopropyl) phosphate, (Tris-BP)
TABLE 2-27.
METHOD 8330 - NITROAROMATICS AND NITRAMINES
Octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine (HMX)
Hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX)
1,3,5-Trinitrobenzene (TNB)
1,3-Dinitrobenzene (DNB)
Methyl -2,4,6-tri ni trophenylni trami ne (Tetryl)
Nitrobenzene (NB)
2,4,6-Trinitrotoluene (TNT)
2,4-Dinitrotoluene (24DNT)
2,6-Dinitrotoluene (26DNT)
o-Nitrotoluene (2NT)
m-Nitrotoluene (3NT)
p-Nitrotoluene (4NT)
TABLE 2-28.
METHOD 8331 - TETRAZENE
Tetrazene
TWO - 30
Revision 2
November 1990
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TABLE 2-29.
REQUIRED CONTAINERS. PRESERVATION TECHNIQUES, AND HOLDING TIMES FOR AQUEOUS MATRICES
Name
Bacterial Tests:
Collform, total
Inorganic Tests:
Chloride
Cyanide, total and amenable
to chlorlnation
Hydrogen Ion (pH)
Nitrate
Sulfate
Sulfide
Metals:
Chromium VI
Mercury
Metals, except chromium VI
and mercury
Organic Tests:
Oil and grease
Organic carbon, total (TOC)
Purgeable Halocarbons
Purgeable aromatic
hydrocarbons
Acrolein and acrylonitrile
Phenol s
Benzi dines
Phthalate esters
Nitrosamines
PCBs
Nitroaromatics and
cyclic ketones
Polynuclear aromatic
hydrocarbons
Haloethers
Chlorinated hydrocarbons
Dioxins and Furans
Total organic halides (TOX)
Pesticides
Radiological Tests:
Alpha, beta and radium
Container1
P, 6
P, 6
P, 6
P. G
P, G
P, G
P, G
P, G
P, 6
P, G
G
P. G
G, Teflon-lined
septum
G, Teflon-lined
septum
G, Teflon-lined
septum
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap .
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
G, Teflon-lined cap
P, G
Preservation
Cool, 4°C. 0.008% Na2S20,
None required
Cool, 4°C; if oxidizing
agents present add 5 ml
0.1N NaAsO, per L or 0.06 g
of ascorbic acid per L;
adjust pH>12 with 50% NaOH.
None required
Cool, 4°C
Cool, 4°C
Cool, 4°C, add zinc acetate
Cool, 4°C
HNO, to pH<2
HNO, to pH<2
Cool. 4°C2
Cool, 4°C2
Cool, 4°C3
Cool, 4°C, 0.008% Na2S2032'3
Cool, 4°C, 0.008% Na2S20,.
Adjust pH to 4-5
Cool, 4°C, 0.008% Na2S203
Cool, 4°C. 0.008% Na,S203
Cool. 4°C
Cool , 4"C, store in dark.
0.008% Na2S203
Cool, 4°C
Cool. 4°C. 0.008% Na,S20,
store in dark
Cool. 4°C. 0.008% Na2S203
store in dark
Cool, 4"C. 0.008% Na8S203
Cool. 4°C, 0.008% Na2S203
Cool, 4°C, 0.008% Na.SA
Cool, 4°C2
Cool, 4°C, pH 5-9
HN03 to pH<2
Maximum holding time
6 hours
28 days
14 days
Analyze immediately
48 hours
28 days
7 days
24 hours
28 days
6 months
28 days
28 days
14 days
14 days
14 days
7 days until extraction,
after extraction
7 days until extraction.
after extraction
7 days until extraction,
after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
7 days until extraction,
40 days after extraction
8 days
7 days until extraction,
40 days after extraction
6 months
40 days
40 days
40 days
1 Polyethylene (P) or Glass (G)
2Adjust to pH<2 with H2SO,. HC1 or solid NaHSO,
3Free chlorine must be removed prior to addition of HC1 by exact addition of Na2S203
TWO - 31
Revision 2
November 1990
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co
ro
Z 30
O*
o
?o
C7>
-
vo
o
-------
FIGURE 2-2.
DETERMINATION OF ORGANIC ANALYTES
Semi volatile Organic Compounds
GC/MS
Determination
Methods
Specific
Detection
Methods
HPLC
Phenol s
8270
8250
8040
Acids
8270
8250
Phthalate
Esters
8270
8250
8060
8061
Nitro-
soami nes
8270
8250
8070
Nitro-
& Cyclic
Ketones
8270
8250
8090
Poly-
nuclear
Hydro-
carbons
8270
8250
8100
8310
Haloethers
8270
8250
8110
Semi volatile Organic Compounds (Continued)
GC/MS
Determination
Methods
Specific GC
Detection
Methods
HPLC
Chlorinated
Hydro-
carbons
8270
8250
8120
8121
Base/
Neutral
8270
8250
Organo-
phosphorus
Pesticides
8270'
8140
8141
8321
Organo-
chlorine
Pesticides
& PCBs
8270*
8080
8081
Chlorinated
Herbicides
8270'
8150
8151
Carbamates
8318
Explosives
8330
8331
'This method is an alternative confirmation method. It is not the method of choice.
TWO - 33
Revision 2
November 1990
-------
FIGURE 2-2.
(Continued)
Volatile Organic Compounds
GC/MS
Determination
Methods
Specific GC
Detection
Methods
HPLC
Halogenated
Volatiles
8240
8260
8010
8011
8021
Non-
halogenated
Volatiles
8240
8015
Aromatic
Volatiles
8240
8260
8020
8021
Acrolein
Acryl o-
nitrile
Acetonitrile
8240
8030
8031
8316
Volatile
Organics
8240
8260
8021
Formal dehyde
8315
Acryl amide
8032
8316
FIGURE 2-3.
CLEANUP OF ORGANIC ANALYTE EXTRACTS
Phenols
3630
3640
3650
Acids
3650
Phthalate
Esters
3610
3620
3640
Nitro-
aromatics
& Cyclic
Ketones
3620
3640
Polynuclear
Aromatic
Hydrocarbons
3611
3630
3640
Chlorinated
Hydrocarbons
3620
3640
Base/Neutral
3650
Organo-
phosphorus
Pesticides
3620
Organo-
chlorine
Pesticides
& PCBs
3620
3640
3660
3665
Chlorinated
Herbicides
8150
TWO - 34
Revision 2
November 1990
-------
FIGURE 2-4.
PREPARATION METHODS FOR ORGANIC ANALYTES
Aqueous
PH3
Solids
Aqueous
Sludges
Emulsions'
PH3
Solids
Oils
Phenols
3510
3520
<2
3540
3550
35802
3520
<2
3650
35802
Acids
3510
3520
<2
3540
3550
35801
3520
<2
3650
35802
Phthalate
Esters
3510
3520
Neutral
3540
3550
35802
Sec
3520
Neutral
Sec
35802
Nitro-
aromatics
& Cyclic
Ketones
3510
3520
5-9
3540
3550
35802
Aqueous Ai
3520
5-9
Solids Abe
35802
Poly-
nuclear
Aromatic
Hydro-
carbons
3510
3520
Neutral
3540
3550
35802
3520
Neutral
3560
35802
Chlori-
nated
Hydro-
carbons
3510
3520
Neutral
3540
3550
35802
3520
Neutral
35802
Base/
Neutral
3510
3520
>11
3540
3550
35802
3520
>11
3650
35802
'If attempts to break up emulsions are unsuccessful, this method may be used.
2Waste dilution. Method 3580, is only appropriate if the sample is soluble in the
specified solvent.
3pH at which extraction should be performed.
TWO - 35
Revision 2
November 1990
-------
FIGURE 2-4.
(Continued)
Aqueous
PH3
Solids
Aqueous
Sludges
Emulsions'
PH'
Sol ids
Oils
Organo-
phosph-
orus
Pesti-
cides
3510
3520
6-8
3540
3550
35802
3520
6-8
35801
Organo-
chlorine
Pesti-
cides
& PCBs
3510
3520
3665
5-9
3540
3550
35802
3665
3541*
3520
5-9
3580*
Chlori-
nated
Herbi-
cides
8150
<2
8150
3580'
8150
<2
See
35801
Halo-
genated
Volatiles
5030
5030
Aqueous Al
5030
Non-
halo-
genated
Volatiles
5030
5030
5030
Aromatic
Volatiles
5030
5030
5030
Acrolein
Acrylo-
nitrlle
Aceto-
nitrile
5030
5030
5030
Volatile
Organics
5030
5030
5030
5030
5030
5030
5030
5030
'If attempts to break up emulsions are unsuccessful, this method may be used.
'Waste dilution. Method 3580, is only appropriate if the sample is soluble in the
specified solvent.
3pH at which extraction should be performed.
'Method 3541 Is appropriate if the sample is to be analyzed for PCBs only.
TWO - 36
Revision 2
November 1990
-------
(
FIGURE 2-5.
SCHEMATIC OF SEQUENCE OF TESTING TO DETERMINE
IF A WASTE IS HAZARDOUS BY CHARACTERISTICS
Method 1020
DOT 149 CFR 173 131)
A
• i r «r
«* ti
\
No
ir? / V
No
C~* ~"
lonhaurdout by
ignitibility
.
(N*nh«t«rd*u* \ f
(•r CP tonoilf I (
°h
-------
FIGURE 2-5.
(Continued)
DOT (49 CFR 173 30C;
at \
ical \
does J >
s te /
ve' /
Solid
Mixture
/ \
/ Is uaste >v Yes /^~ ' *
No
Paint
Filter Test
— ' I Nonhazardous \
•* 1 for ignitability 1
1
Liquid
Methods 1113 and 8240
Nonhaza rdous
for corrosivity
chrac teris tic
Method 1010, Method 1020
Yei
[ Hazardous V-
Nonhazardous
for ignitability
characteristic
TWO - 38
Revision 2
November 1990
-------
FIGURE 2-6A.
EP
Cr
Ag
TWO - 39
Revision 2
November 1990
-------
FIGURE 2-6B.
TCLP
Ba
Cr
Ag
As
Cd
Pb
Se
TWO - 40
Revision 2
November 1990
-------
FIGURE 2-7A.
GROUND WATER ANALYSIS
(Found In Easyflow, titled: ch2f1g7a)
VOA
8240
8260
Semivolatile
Org.
inie
lie
Pes ticides
3510 or 3S20
8270
82SO
Herbicides Oioxini
3510 or 3520
Neutral
1
3620, 3640,
and/or 3660
8080 or 8081
81SO or 81S1 8280 or 8290
Optional: Cleanup required only if interference* prevent analytia.
TWO - 41
Revision 2
November 1990
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METHOD 3051
MICROWAVE ASSISTED ACID DIGESTION OF SEDIMENTS.
SLUDGES. SOILS. AND OILS
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the microwave assisted acid digestion
of sludges, sediments, soils, and oils for the following elements:
Aluminum Cadmium Iron Molybdenum Sodium
Antimony Calcium Lead Nickel Strontium
Arsenic Chromium Magnesium Potassium Thallium
Boron Cobalt Manganese Selenium Vanadium
Barium Copper Mercury Silver Zinc
Beryllium
1.2 This method is provided as an alternative to Method 3050A. It is
intended to provide a rapid multielement acid leach digestion prior to
analysis so that decisions can be made about site cleanup levels, the need for
TCLP testing of a waste and whether a BOAT process is providing acceptable
performance. If a decomposition including hydrochloric acid is required for
certain elements, it is recommended that Method 3050A be used. Digests
produced by the method are suitable for analysis by flame atomic absorption
(FLAA), graphite furnace atomic absorption (GFAA), inductively coupled plasma
emission spectroscopy (ICP-ES) and inductively coupled plasma mass
spectrometry (ICP-MS).
2.0 SUMMARY OF METHOD
2.1 A representative sample of up to 0.5 g is digested in 10 mL of
concentrated nitric acid for 10 min using microwave heating with a suitable
laboratory microwave unit. The sample is placed in a Teflon PFA vessel with
10 mL of concentrated nitric acid. The vessel is capped and heated in the
microwave unit. After cooling, the vessel contents are diluted to volume and
analyzed by the appropriate SW-846 method (Ref. 1).
3.0 INTERFERENCES
3.1 Very reactive or volatile materials that may create high pressures
when heated may cause venting of the vessels with potential loss of sample and
analytes. The complete decomposition of either carbonates, or carbon based
samples, may cause enough pressure to vent the vessel if the sample size is
greater than 0.25 g when used in the 120 mL vessels with a pressure relief
device that has an upper limit of 7.5+ 0.7 atm (110 ± 10 psi).
3051 - 1 Revision 0
November 1990
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4.0 APPARATUS AND MATERIALS
4.1 Microwave apparatus requirements.
4.1.1 The microwave unit provides programmable power with a
minimum of 574 W and can be programmed to within ± 10 W of the required
power.
4.1.2 The microwave unit cavity is corrosion resistant as well as
ventilated.
4.1.3
operation.
All electronics are protected against corrosion for safe
4.1.4 The system requires Teflon PFA digestion vessels (120 ml
capacity) capable of withstanding pressures up to 7.5 ± 0.7 atm (110 ±
10 psi) and capable of controlled pressure relief at pressures exceeding
7.5 ± 0.7 atm (110 ± 10 psi).
4.1.5 A rotating turntable is employed to insure homogeneous
distribution of microwave radiation within the unit. The speed of the
turntable should be a minimum of 3 rpm.
4.1.6 Those laboratories now using or contemplating the use of
kitchen type microwave ovens for this method should be aware of several
significant safety issues. First, when an acid such as nitric is used
to assist sample digestion in microwave units in open vessels, or sealed
vessels equipped with venting features, there is the potential for the
acid gases released to corrode the safety devices that prevent the
microwave magnetron from shutting off when the door is opened. This can
result in operator exposure to microwave energy. Use of a unit with
corrosion resistant safety devices prevents this from occurring.
The second safety concern relates to the use of sealed containers
without pressure relief valves in the unit. Temperature is the
important variable controlling the reaction. Pressure is needed to
attain elevated temperatures but must be safely contained. However,
many digestion vessels constructed from certain Teflons may crack,
burst, or explode in the unit under certain pressures. Only unlined PFA
Teflon containers with pressure relief mechanisms are considered
acceptable at present.
Users are therefore advised not to use kitchen type microwave ovens or
to use sealed containers without pressure relief valves for microwave
acid digestions by this method. Use of laboratory-grade microwave
equipment is required to prevent safety hazards. For further details
consult reference 2.
3051 - 2
Revision 0
November 1990
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4.2 Polymeric volumetric ware in plastic (Teflon or polyethylene)
50 or 100 ml capacity.
4.3 Whatman No. 41 filter paper (or equivalent).
4.4 Disposable polypropylene filter funnel.
4.5 Analytical balance, 300 g capacity, and minimum ± 0.001 g.
5.0 REAGENTS
5.1 All acids should be sub-boiling distilled where possible to
minimize the blank levels due to metallic contamination. Other grades may be
used, provided it is first ascertained that the reagent is of sufficient
purity to permit its use without lessening the accuracy of the determination.
5.1.1 Concentrated nitric acid, HN03. Acid should be analyzed to
determine levels of impurity.
5.2 Reagent Water. Reagent water shall be interference free. All
references to water in the method refer to reagent water unless otherwise
specified (Ref. 3).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All sample containers must be prewashed with detergents, acids and
water. Plastic and glass containers are both suitable. See Chapter Three,
Step 3.1.3 of this manual, for further information.
6.3 Samples must be refrigerated upon receipt and analyzed as soon as
possible.
7.0 PROCEDURE
7.1 Calibration of Microwave Equipment
7.1.1 Measurement of the available power for heating is evaluated
so that absolute power in watts may be transferred from one microwave
unit to another. For cavity type microwave equipment, this is
accomplished by measuring the temperature rise in 1 kg of water exposed
to microwave radiation for a fixed period of time. The analyst can
relate power in watts to the partial power setting of the unit. The
calibration format required for laboratory microwave units depends on
the type of electronic system used by the manufacturer to provide
3051 - 3 Revision 0
November 1990
-------
partial microwave power. Few units have an accurate and precise linear
relationship between percent power settings and absorbed power. Where
linear circuits have been utilized, the calibration curve can be
determined by a three-point calibration method (7.1.3), otherwise, the
analyst must use the multiple point calibration method (7.1.2).
7.1.2 The multiple point calibration involves the measurement of
absorbed power over a large range of power settings. Typically, for a
600 W unit, the following power settings are measured;
100,99,98,97,95,90,80,70,60,50, and 40%using the procedure described in
section 7.1.4. This data is clustered about the customary working power
ranges. Nonlinearity has been commonly encountered at the upper end of
the calibration. If the unit's electronics are known to have nonlinear
deviations in any region of proportional power control, it will be
necessary to make a set of measurements that bracket the power to be
used. The final calibration point should be at the partial power
setting that will be used in the test. This setting should be checked
periodically to evaluate the integrity of the calibration. If a
significant change is detected (±10 W), then the entire calibration
should be reevaluated.
7.1.3 The three-point calibration involves the measurement of
absorbed power at three different power settings. Measure the power at
100% and 50% using athe procedure described in section 7.1.4. From the
2-point line calculate the power setting corresponding to the required
power in watts specified in the procedure. Measure the absorbed power
at that partial power setting. If the measured absorbed power does not
correspond to the specified power within ±10 W, use the multiple point
calibration in 7.1.2. This point should also be used to periodically
verify the integrity of the calibration.
7.1.4 Equilibrate a large volume of water to room temperature
(23 ± 2eC). One kg of reagent water is weighed (1,000.0 g + 0.1 g) into
a Teflon beaker or a beaker made of some other material that does not
significantly absorb microwave energy (glass absorbs microwave energy
and is not recommended). The initial temperature of the water should be
23 ± 2 °C measured to ± 0.05 °C. The covered beaker is circulated
continuously (in the normal sample path) through the microwave field for
2 minutes at the desired partial power setting with the unit's exhaust
fan on maximum (as it will be during normal operation). The beaker is
removed and the water vigorously stirred. Use a magnetic stirring bar
inserted immediately after microwave irradiation and record the maximum
temperature within the first 30 seconds to ± 0.05 8C. Use a new sample
for each additional measurement. If the water is reused both the water
and the beaker must have returned to 23 ± 2'C. Three measurements at
each power setting should be made.
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The absorbed power is determined by the following relationship
P = (K) (Cp) (m) (AT)
Eq. 1
P = the apparent power absorbed by the sample in watts (W).
(W=joule sec'1)
K = the conversion factor for thermochemical calories sec'1 to watts
(=4.184)
Cp = the heat capacity, thermal capacity, or specific heat
(cal g~1°C~1), of water, m = the mass of the water sample in grams (g).
AT = the final temperature minus the initial tempera ture (°C), and
t = the time in seconds (s).
Using the experimental conditions of 2 minutes and 1 kg of distilled
water (heat capacity at 25 °C is 0.9997 cal g"1 °C"1) the calibration
equation simplifies to:
Eq. 2 P = (AT) (34.85)
NOTE: Stable line voltage is necessary for accurate and reproducible
calibration and operation. The line voltage should be within
manufacturer's specification, and during measurement and operation
should not vary by more than ±2 V. A constant power supply may be
necessary for microwave use if the source of the line voltage is
unstable.
Electronic components in most microwave units are matched to
the units' function and output. When any part of the high voltage
circuit, power source, or control components in the unit have been
serviced or replaced, it will be necessary to recheck the units'
calibration. If the power output has changed significantly (±10 W),
then the entire calibration should be reevaluated.
7.2 All digestion vessels and volumetric ware must be carefully acid
washed and rinsed with reagent water. When switching between high concen-
tration samples and low concentration samples, all digestion vessels should be
cleaned by leaching with hot (1:1) hydrochloric acid for a minimum of two
hours followed with hot (1:1) nitric acid for a minimum of two hours and
rinsed with reagent water and dried in a clean environment. This cleaning
procedure should also be used whenever the prior use of the digestion vessels
is unknown or cross contamination from vessels is suspected. Polymeric
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volumetric ware and storage containers should be cleaned by leaching with more
dilute acids appropriate for the specific plastics used and then rinsed with
reagent water and dried in a clean environment.
7.3 Sample Digestion
7.3.1 Weigh the Teflon PFA digestion vessel, valve and cap
assembly to 0.001 g prior to use.
7.3.2 Weigh a well-mixed sample to the nearest 0.001 g into the
Teflon PFA sample vessel equipped with a single-ported cap and a
pressure relief valve. For soils, sediments, and sludges use no more
than 0.500 g. For oils use no more than 0.250 g.
7.3.3 Add 10 ± 0.1 ml concentrated nitric acid in a fume hood.
If a vigorous reaction occurs, allow the reaction to stop before capping
the vessel. Cap the vessel and torque the cap to 12 ft-lb (16 N-m)
according to the unit manufacturer's directions. The sample vessel may
be connected to an overflow vessel using Teflon PFA connecting tubes.
Weigh the vessels to the nearest 0.001 g. Place the vessels in the
microwave carousel. Connect the overflow vessels to the center well of
the unit.
CAUTION: When digesting samples containing volatile or easily oxidized
organic compounds, initially weigh no more than 0.10 g and observe the
reaction before capping the vessel. If a vigorous reaction occurs,
allow the reaction to cease before capping the vessel. If no
appreciable reaction occurs, a sample weight up to 0.25 g can be used.
7.3.4 Place the vessels evenly distributed in the turntable of
the microwave unit using groups of 2 sample vessels or 6 sample vessels.
Any vessels containing 10 ml of nitric acid for analytical blank
purposes are counted as sample vessels. When fewer than the recommended
number of samples are to be digested, i.e., 3 samples plus 1 blank, the
remaining vessels should be filled with 10 mL of nitric acid to achieve
the full complement of vessels. This provides an energy balance since
the microwave power absorbed is proportional to the total mass in the
cavity (Ref. 4). Irradiate each group of 2 sample vessels at 344 W for
10 minutes and each group of 6 sample vessels at 574 W for 10 minutes.
The temperature of each sample should rise to 175 °C in less than 5.5
minutes and remain between 170-180 CC for the balance of the 10 minute
irradiation period. The pressure should peak at less than 6 atm for
most soil, sludge, and sediment samples (Ref. 5). The pressure will
exceed these limits in the case of high concentrations of carbonate or
organic compounds. In these cases the pressure will be limited by the
relief pressure of the vessel to 7.5 ± 0.7 atm (110 ± 10 psi).
7.3.4.1 Newer microwave units may be capable of higher
power (W) that permits digestion of a larger number of samples per
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batch. If the analyst wishes to digest other that two or six
samples at a time, the analyst may use different values of power
as long as they result in the same time and temperature conditions
defined in 7.3.4. That is, any sequence of power that brings the
samples to 175°C in 5.5 minutes and permits a slow rise to 175 -
180°C during the remaining 4.5 minutes (Ref. 5).
Issues of safety, structural integrity (both temperature and
pressure limitations), heat loss, chemical compatibility,
microwave absorption of vessel material, and energy transport will
be considerations made in choosing alternative vessels. If all of
the considerations are met and the appropriate power settings
provided to reproduce the reaction conditions defined in 7.3.4,
then these alternative vessels may be used (Ref. 1,2).
7.3.5 At the end of the microwave program, allow the vessels to
cool for a minimum of 5 minutes before removing them from the microwave
unit. When the vessels have cooled to room temperature, weigh and
record the weight of each vessel assembly. If the weight of acid plus
sample has decreased by more than 10 percent from the original
weight, discard the sample. Determine the reason for the weight loss.
These are typically attributed to loss of vessel seal integrity, use of
a digestion time longer than 10 minutes, too large a sample, or improper
heating conditions. Once the source of the loss has been corrected,
prepare a new sample or set of samples for digestion beginning at 7.3.1.
7.3.6 Complete the preparation of the sample by carefully
uncapping and venting each vessel in a fume hood. Transfer the sample
to an acid-cleaned polyethylene bottle. If the digested sample contains
particulates which may clog nebulizers or interfere with injection of
the sample into the instrument, the sample may be centrifuged, allowed
to settle, or filtered.
7.3.6.1 Centrifugation: Centrifugation at 2,000-3,000 rpm
for 10 minutes is usually sufficient to clear the supernatant.
7.3.6.2 Settling: Allow the sample to stand until the
supernatant is clear. Allowing a sample to stand overnight will
usually accomplish this. If it does not, centrifuge or filter the
sample.
7.3.6.3 Filtering: The filtering apparatus must be
thoroughly cleaned and prerinsed with dilute nitric acid. Filter
the sample through quantitative filter paper into a second
acid-cleaned container.
7.3.7 The diluted digest has an approximate acid concentration
of 20 percent (v/v) HN03. The digest is now ready for analysis for
elements of interest using the appropriate SW-846 method.
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7.4 Calculations: The concentrations determined are to be reported on
the basis of the actual weight of the original sample.
8.0 QUALITY CONTROL
8.1 All quality control data must be maintained and available for
reference or inspection for a period of three years. This method is
restricted to use by, or under supervision of, experienced analysts. Refer to
the appropriate section of Chapter One for additional quality control
requirements.
8.2 Replicate samples should be processed on a routine basis. A
replicate sample is a sample brought through the whole sample preparation and
analytical process. A replicate sample should be processed with each
analytical batch or every 20 samples, whichever is the greater number. A
replicate sample should be prepared for each matrix type (i.e., soil, sludge,
etc.).
8.3 Spiked samples or standard reference materials should be included
with each group of samples processed or every 20 samples, whichever is the
greater number. A spiked sample should also be included whenever a new sample
matrix is being analyzed.
9.0 METHOD PERFORMANCE
9.1 Precision: The precision of Method 3051, as determined by the
statistical examination of interlaboratory test results is as follows:
9.2 Repeatability: If successive results are obtained by the same
analyst with the same apparatus under constant operating conditions on
identical test material, then the difference between these successive results
will not, with 95% probability, exceed the repeatability value. For example,
in the case of lead, an average of only 1 case in 20 would exceed
0.206 x
in the long run, where x is one result in g/g (Ref. 6).
9.3 Reproducibility: If two successive measurements are made indepen-
dently by each of two different analysts working in different laboratories on
identical test material, then the difference between the average result for
each analyst will not, with 95% probability, exceed the reproducibility value.
For example, in the case of lead, an average of only 1 case in 20 would exceed
0.303 x
in the long run, where x is the average of two successive measurements in g/g
(Ref. 2).
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As can be seen in Table 1, repeatability and reproducibility differ
between elements, and usually depend on that element's concentration. Table 2
provides an example of how users of the method can determine expected values
for repeatability and reproducibility; nominal values of lead have been used
for this model (Ref. 6).
9.4 Bias: In the case of SRM 1085 - Wear Metals in Oil, the bias of
this test method is different for each element. An estimate of bias, as shown
in Table 3, is:
Bias = Amount found - Amount expected.
However, the bias estimate inherits both the uncertainty in the
measurements made using Method 3051 and the uncertainty on the certificate, so
whether the bias is real or only due to measurement error must also be con-
sidered. The concentrations found for Al, Cr, and Cu using Method 3051 fall
within their certified ranges on SRM 1085, and 95% confidence intervals for Fe
and Ni overlap with their respective certified ranges; therefore, the observed
biases for these elements are probably due to chance and should be considered
insignificant. Biases should not be estimated at all for Ag and Pb because
these elements were not certified. Therefore, the only two elements
considered in this table for which the bias estimates are significant are Mg
and Mo.
10.0 REFERENCES
1. Test Methods for Evaluating Solid Waste. Physical/Chemical Methods, 3rd
ed; U.S. Environmental Protection Agency, Office of Solid Waste and
Emergency Response. U.S. Government Printing Office: Washington, DC,
1986; SW-846.
2. Kingston, H. M. and L. B. Jassie, "Safety Guidelines for Microwave
Systems in the Analytical Laboratory". In Introduction to Microwave
Acid Decomposition; Theory and Practice; Kingston, H. M. and Jassie, L.
B., eds.; ACS Professional Reference Book Series; American Chemical
Society: Washington, DC, 1988.
3. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water ; ASTM, Philadelphia, PA, 1985, D1193-77.
4. Introduction to Microwave Sample Preparation: Theory and Practice.
Kingston, H. M. and Jassie, L. B., Eds.; ACS Professional Reference Book
Series; American Chemical Society: Washington, DC, 1988.
3051 - 9 Revision 0
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Kingston, H. M. EPA IAG #DWI-393254-01-0 January 1-March 31, 1988,
quarterly Report.
Binstock, D. A., Yeager, W. M., Grohse, P. M. and Gaskill, A. Valida-
tion of a Method for Determining Elements in Solid Waste by Microwave
Digestion. Research Triangle Institute Technical Report Draft, RTI
Project Number 321U-3579-24, November, 1989, prepared for the Office of
Solid Waste, U.S. Environmental Protection Agency, Washington, DC 20460.
3051 - 10
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TABLE 1.
EQUATIONS RELATING REPEATABILITY AND REPRODUCIBILITY TO MEAN
CONCENTRATION OF DUPLICATE DETERMINATION WITH 95 PERCENT CONFIDENCE
Element Repeatability Reproducibilitv
Ag 0.195X* 0.314X
Al 0.232X 0.444X
B 12.9b 22.6b
Ba 0.238X 0.421X
Be 0.082b 0.082b
Ca 0.356X 1.27X
Cd 0.385X 0.571X
Co 0.291X 0.529X
Cr 0.187X 0.195X
Cu 0.212X 0.322X
Fe 0.257X 0.348X
Mg 0.238X 0.399X
Mn 1.96X1/2° 4.02X1/2
Mo 0.701X 0.857X
Ni 0.212X 0.390X
Pb 0.206X 0.303X
Sr 0.283X 0.368X
V 1.03X1/2 2.23X1/2
Zn 3.82X1/2 7.69X1/2
aLog transformed variable based on one-way analysis of variance.
"Repeatability and reproducibility were independent of concentratio .
cSquare root transformed variable based on one-way analysis of variance.
TABLE 2.
REPEATABILITY AND REPRODUCIBILITY FOR LEAD
BY METHOD 3051
Average Value Repeatability Reproducibility
50 10.3 15.2
100 20.6 30.3
200 41.2 60.6
300 61.8 90.9
400 82.4 121
500 103 152
all results are in mg/Kg
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TABLE 3.
RECOVERY AND BIAS DATA FOR SRM 1085 - WEAR METALS IN OIL
Element
Ag
Al
Cr
Cu
Fe
Mg
Mo
N1
Pb
Amount
Expected
(Certified
Range)
(291)**
296+4
298±5
295+10
300+4
297±3
292±11
303±7
(305)**
all values in mg/Kg
Amount
Found*
(95% Conf
Interval)
234±16
295±12
293110
289±9
311±14
270±11
238±11
293+9
27918
Absolute
Bias
( g/g)
-1
-5
-6
+11
-27
-54
-10
Relative
Bias
(Percent)
0
-2
-2
+4
-9
-18
-3
Significant
(due to more
than chance)
No
No
No
No
Yes
Yes
No
*Results taken from table 4-7, Ref. 2.
**Value not certified, so should not be used in bias detection and
estimation.
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Start
METHOD 3051
MICROWAVE ASSISTED ACID DIGESTION OF SEDIMENTS
SLUDGES, SOILS, AND OILS
7.3.1 W.igh
aliquot into
the digestion
ve*»el
7.3.2 Add
concentrated
HNO.,cap after
reaction
•topped
7.3.3 Place 6
•ample veaaeli
in oven, heat
according to
power program
7.3.4 Allow
•ample* to
cool to room
temperature
7.3.S Heigh
each venel
a»»embly
7.3.7 Uie the
appropriate
SH-846 method
to analyze
7.4 Calculate
concontration*
ba*ad on
original »ac
apl«
weight
Stop
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METHOD 3015
MICROWAVE ASSISTED ACID DIGESTION OF AQUEOUS
SAMPLES AND EXTRACTS
1.0 SCOPE AND APPLICATION
1.1 This digestion procedure is used for the preparation of aqueous
samples, mobility-procedure extracts, and wastes that contain suspended solids
for analysis, by flame atomic absorption spectroscopy (FLAA), graphite furnace
absorption spectroscopy (GFAA), inductively coupled argon plasma spectroscopy
(ICP), or inductively coupled argon plasma mass spectrometry (ICP-MS). The
procedure is a hot acid leach for determining available metals.
1.2 Samples prepared by Method 3015 using nitric acid digestion may
be analyzed by FLAA, GFAA, ICP, or ICP-MS for the following:
Aluminum Lead
Antimony Magnesium
*Arsenic Manganese
Barium Molybdenum
Beryllium Nickel
Cadmium Potassium
Calcium *Selenium
Chromium Silver
Cobalt Sodium
Copper Thallium
Iron Vanadium
Zinc
''Cannot be analyzed by FLAA
2.0 SUMMARY OF METHOD
2.1 Nitric acid is added to an aqueous sample in a 120 mL Teflon
digestion vessel. The vessel is capped and heated in a microwave unit. After
cooling, the vessel contents are filtered, centrifuged, or allowed to settle
in a clean sample bottle for analysis.
3.0 INTERFERENCES
3.1 Very reactive or volatile materials that may create high pres-
sures when heated may cause venting of the vessels with potential loss of
sample and analytes. Samples that contain carbonates or other carbon dioxide
generating compounds may cause enough pressure to vent the vessel. If this
situation is anticipated the analyst may wish to use a smaller sample.
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4.0 APPARATUS AND MATERIALS
4.1 Microwave apparatus requirements
4.1.1 The microwave unit provides programmable power with a
minimum of 574 W and can be programmed to within ± 10 W of the
required power.
4.1.2 The microwave unit cavity is corrosion resistant and
well ventilated.
4.1.3 All electronics are protected against corrosion for safe
operation.
4.1.4 The system requires Teflon PFA digestion vessels (120 ml
capacity) capable of withstanding pressures up to 7.5 ± 0.7 atm (110
+ 10 psi) and capable of controlled pressure relief at pressures
exceeding 7.5 + 0.7 atm (110 ± 10 psi).
4.1.5 A rotating turntable is employed to insure homogeneous
distribution of microwave radiation within the unit. The speed of
the turntable should be a minimum of 3 rpm.
4.1.6 Those laboratories now using or contemplating the use of
kitchen type microwave ovens for this method should be aware of
several significant safety issues. First, when an acid such as
nitric is used to assist sample digestion in microwave units in open
vessels, or sealed vessels equipped with venting features, there is
the potential for the acid gases released to corrode the safety
devices that prevent the microwave magnetron from shutting off when
the door is opened. This can result in operator exposure to micro-
wave energy. Use of a unit with corrosion resistant safety devices
prevents this from occurring.
The second safety concern relates to the use of sealed containers
without pressure relief valves in the unit. Temperature is the
important variable controlling the reaction. Pressure is needed to
attain elevated temperatures but must be safely contained. However,
many digestion vessels constructed from certain Teflons may crack,
burst, or explode in the oven under certain pressures. Only unlined
PFA Teflon containers with pressure relief mechanisms are considered
acceptable at present.
Users are therefore advised not to use kitchen type microwave ovens
or to use sealed containers without pressure relief valves for
microwave acid digestions by this method. Use of laboratory grade
microwave equipment is required to prevent safety hazards. For
further information consult reference 1.
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4.2 Plastic ware graduated cylinder, 50 or 100 ml capacity.
4.3 Quantitative filter paper, Whatman No. 41 or S&S White label or
equivalent.
4.4 Analytical balance, 300 g capacity, minimum ± 0.01 g.
4.5 Disposable polypropylene filter funnel.
4.6 Polyethylene bottles, 125 ml, with caps
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. All acids should
be sub-boiling distilled where possible to minimize the blank levels due to
metallic contamination. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Reagent Water. Reagent water shall be interference free. All
references to water in the method refer to reagent water unless otherwise
specified (Ref. 2).
5.3 Concentrated Nitric acid, HN03. Acid should be analyzed to
determine levels of impurities.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All sample containers must be prewashed with detergents, acids,
and water. Plastic containers are preferable. See Chapter Three, Step 3.1.3
of this manual, for further information.
6.3 Aqueous waste waters must be acidified to a pH of < 2 with HN03.
7.0 PROCEDURE
7.1 Calibration of Microwave Equipment
7.1.1 Measurement of the available power for heating is
evaluated so that absolute power in watts may be transferred from one
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microwave unit to another. For cavity type microwave equipment, this
is accomplished by measuring the temperature rise in 1 kg of water
exposed to microwave radiation for a fixed period of time. The
analyst can relate power in watts to the partial power setting of the
unit. The calibration format required for laboratory microwave units
depends on the type of electronic system used by the manufacturer to
provide partial microwave power. Few units have an accurate and
precise linear relationship between percent power settings and
absorbed power. Where linear circuits have been utilized, the
calibration curve can be determined by a three-point calibration
method (7.1.3), otherwise, the analyst must use the multiple point
calibration method (7.1.2).
7.1.2 The multiple point calibration involves the measurement
of absorbed power over a large range of power settings. Typically,
for a 600 W unit, the following power settings are measured; 100,99,-
98,97,95,90,80,70,60,50, and 40% using the procedure described in
section 7.1.4. This data is clustered about the customary working
power ranges. Nonlinearity has been commonly encountered at the
upper end of the calibration. If the unit's electronics are known to
have nonlinear deviations in any region of proportional power
control, it will be necessary to make a set of measurements that
bracket the power to be used. The final calibration point should be
at the partial power setting that will be used in the test. This
setting should be checked periodically to evaluate the integrity of
the calibration. If a significant change is detected (±10 W), then
the entire calibration should be reevaluated.
7.1.3 The three-point calibration involves the measurement of
absorbed power at three different power settings. Measure the power
at 100% and 50% using the procedure described in section 7.1.4, and
calculate the power setting corresponding to the required power in
watts specified in the procedure from the (2-point) line. Measure
the absorbed power at that partial power setting. If the measured
absorbed power does not correspond to the specified power within ±10
W, use the multiple point calibration in 7.1.2. This point should
also be used to periodically verify the integrity of the calibration.
7.1.4 Equilibrate a large volume of water to room temperature
(23 ± 2 °C). One kg of reagent water is weighed (1,000.0 g ± 0.1 g)
into a Teflon beaker or a beaker made of some other material that
does not significantly absorb microwave energy (glass absorbs
microwave energy and is not recommended). The initial temperature of
the water should be 23 ± 2 *C measured to ± 0.05 °C. The covered
beaker is circulated continuously (in the normal sample path) through
the microwave field for 2 minutes at the desired partial power
setting with the unit's exhaust fan on maximum (as it will be during
normal operation). The beaker is removed and the water vigorously
stirred. Use a magnetic stirring bar inserted immediately after
microwave irradiation and record the maximum temperature within the
3015 - 4 Revision 0
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first 30 seconds to ± 0.05 °C. Use a new sample for each additional
measurement. If the water is reused both the water and the beaker
must have returned to 23 ± 2 °C. Three measurements at each power
setting should be made.
The absorbed power is determined by the following relationship
P - (K) (Cp) (m) (AT)
Eq. 1
P = the apparent power absorbed by the sample in watts (W).
(W=joule sec"1)
K = the conversion factor for thermochemical calories sec"1 to watts
(=4.184)
Cp = the heat capacity, thermal capacity, or specific heat
(cal g"1°C"1), of water
m = the mass of the water sample in grams (g),
AT = the final temperature minus the initial temperature ( °C), and
t = the time in seconds (s).
Using the experimental conditions of 2 minutes and 1 kg of distilled
water (heat capacity at 25 "C is 0.9997 cal g"1 °C"1) the calibration
equation simplifies to:
P = (AT) (34.86) Eq. 2
NOTE: Stable line voltage is necessary for accurate and reproducible
calibration and operation. The line voltage should be within
manufacturer's specification, and during measurement and operation
not vary by more than ±2 V. A constant power supply may be necessary
for microwave use if the source of the line voltage is unstable.
Electronic components in most microwave units are matched to
the units' function and output. When any part of the high
voltage circuit, power source, or control components in the
unit have been serviced or replaced, it will be necessary to
recheck the units' calibration power. If the power output has
changed significantly (±10 W), then the entire calibration
should be reevaluated.
7.2 All digestion vessels and volumetric ware must be carefully acid
washed and rinsed with reagent water. When switching between high solids
3015 - 5 Revision 0
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(concentrated) samples and low solids (low concentration) samples all diges-
tion vessels should be cleaned by leaching with hot (1:1) hydrochloric acid
for a minimum of two hours followed with hot (1:1) nitric acid for a minimum
of two hours and rinsed with reagent water and dried in a clean environment.
This cleaning procedure should also be used whenever the prior use of the
digestion vessels is unknown or cross contamination from vessels is suspected.
Polymeric volumetric ware and storage containers should be cleaned by leaching
with more dilute acids appropriate for the specific plastics used and then
rinsed with reagent water and dried in a clean environment.
7.3 Sample Digestion
7.3.1 Weigh the Teflon PFA digestion vessel, valve and cap
assembly to 0.01 g prior to use.
7.3.2 A 45 ml aliquot of a well shaken sample is measured in a
graduated cylinder. This aliquot is poured into the Teflon digestion
vessel with the number of the vessel recorded on the preparation
sheet.
7.3.3 A blank sample of reagent water is treated in the same
manner along with spikes and duplicates.
7.3.4 Add 5 ml of concentrated nitric acid to each vessel that
will be used. Check to make sure the pressure relief disks are in
the caps with the smooth side toward the sample and start the caps a
few turns on the vessels. Finish tightening the caps in the capping
station which will tighten them to a uniform torque pressure of 12
ft.lbs. (16-N m). Weigh each capped vessel to the nearest 0.01 g.
7.3.5 Place five vessels evenly distributed in the carousel.
Blanks are treated as samples for the purpose of balancing the power
input. When fewer than the recommended number of samples are
digested, the remaining vessels should be filled with 45 ml of
reagent water and 5 mL of nitric acid to achieve the full compliment
of vessels. This provides an energy balance since the microwave
power absorbed is proportional to the total mass in the cavity (Ref.
1).
7.3.6 Place the carousel in the unit; be sure to seat it
carefully on the turntable. Program the microwave unit for the
first-stage of the power program to give 545 W for 10 minutes and the
second-stage program to give 344 W for 10 minutes. This sequence
brings the samples to 160°C ± 4"C in 10 minutes and permits a slow
rise to 165-170 °C during the second 10 minutes (Ref. 3). Start the
turntable motor and be sure the vent fan is running on high and the
turntable is turning. Start the microwave generator.
7.3.6.1 Newer microwave units may be capable of higher
power that permit digestion of a larger number of samples per
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batch. If the analyst wishes to digest more than 5 samples at
a time, the analyst may use different power settings as long as
they result in the same time and temperature conditions defined
in 7.3.6. That is, any sequence of power that brings the
samples to 160*C ± 4"C in 10 minutes and permits a slow rise to
165-170°C during the second 10 minutes (Ref. 2).
Issues of safety, structural integrity (both temperature and
pressure limitations), heat loss, chemical compatibility,
microwave absorption of vessel material, and energy transport
will be considerations made in choosing alternative vessels.
If all of the considerations are met and the appropriate power
settings are provided to reproduce the reaction conditions
defined in 7.3.6, then these alternative vessels may be used
(Ref. 1,2)
7.3.7 At the end of the microwave program, allow the vessels
to cool for at least 5 minutes in the unit before removal to avoid
possible injury if a vessel vents immediately after microwave
heating. The samples may be cooled outside the unit by removing the
carousel and allowing the samples to cool on the bench or in a water
bath. When the vessels have cooled to room temperature, weigh and
record the weight of each vessel assembly. If the weight of the
sample plus acid has decreased by more than 10% discard the sample.
7.3.8 Rinse virgin or acid-cleaned polyethylene 125 ml bottles
(or other suitable size) and caps with reagent water and shake out
the large water drops. Label the bottles.
7.3.9 Complete the preparation of the sample by carefully
uncapping and venting each vessel in a fume hood. Transfer the
sample to an acid-cleaned polyethylene bottle. If the digested
sample contains particulates which may clog nebulizers or interfere
with injection of the sample into the instrument, the sample may be
centrifuged, allowed to settle or filtered.
7.3.9.1 Centrifugation: Centrifugation at 2,000-3,000 rpm
for 10 minutes is usually sufficient to clear the supernatant.
7.3.9.2 Settling: Allow the sample to stand until the
supernatant is clear. Allowing a sample to stand overnight
will usually accomplish this. If it does not, centrifuge or
filter the sample.
7.3.9.3 Filtering: The filtering apparatus must be
thoroughly cleaned and prerinsed with dilute nitric acid.
Filter the sample through quantitative filter paper into a
second acid-cleaned container.
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7.3.10 The concentration values obtained from analysis must be
corrected for the dilution factor from the acid addition. If the
sample will be analyzed by ICP-MS additional dilution will generally
be necessary. For example, the sample may be diluted by a factor of
20 with reagent water and the acid strength adjusted back to 10%
prior to analysis. The dilutions used should be recorded and the
measured concentrations adjusted accordingly.
8.0 QUALITY CONTROL
8.1 All quality control measures described in Chapter One, of this
Manual, should be followed.
8.2 For each analytical batch of samples processed, analytical
reagent blanks (also field blanks if they were taken) should be carried
throughout the entire sample preparation and analytical process. These blanks
will be useful in determining if samples are being contaminated.
8.3 Replicate samples should be processed on a routine basis. A
replicate sample is a real sample brought through the whole sample preparation
and analytical process. A replicate sample should be processed with each
analytical batch or every 20 samples, whichever is the greater number.
8.4 Spiked samples or standard reference materials should be
employed to determine accuracy. A spiked sample should be included with each
group of samples processed and whenever a new sample matrix is being analyzed.
8.5 The method of standard addition shall be used for the analysis
of all EP extracts (see Method 7000, Step 8.7).
9.0 METHOD PERFORMANCE
9.1 Refer to Reference 4.
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10.0 REFERENCES
1. Introduction to Microwave Sample Preparation: Theory and Practice.
Kingston, H. M.; Jassie, L. B., Eds.; ACS Professional Reference Book
Series: American Chemical Society, Washington, DC, 1988; Ch 6 & 11.
2. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specifica-
tion for Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
3. Kingston, H. M., Final Report EPA IAG #DWI3932541-01-I, September 30,
1988, Appendix A.
4. Shannon, M., Alternate Test Procedure Application, USEPA Region Y,
Central Regional Laboratory, 536 S. Clark Street, Chicago, IL 60606,
1989.
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METHOD 3015
MICROWAVE ASSISTED ACID DIGESTION
OF AQUEOUS SAMPLES AND EXTRACTS
7.1 Calibrate
th* microwav*
•quipnant
7.2 Acid waah
and HtO rinaa
7.3.1 Moa.ur.
45 ml aliquot
into th«
digavtion
vmmmml
all dis* tion
gs*
enal* and
glaiaware
7.3.7 Rin..
virgin
bottU. with
rvagvnt water
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METHOD 6020
INDUCTIVELY COUPLED PLASMA - MASS SPECTROMETRY
1.0 SCOPE AND APPLICATION
1.1 Inductively coupled plasma-mass spectrometry (ICP-MS) is a technique
which is applicable to jxg/L concentrations of a large number of elements in
water and wastes after appropriate sample preparation steps are taken [1,2].
When dissolved constituents are required, samples must be filtered and acid-
preserved prior to analysis. No further digestion is required prior to analysis
for dissolved elements. Acid-digestion prior to filtration and analysis is
required for groundwater, aqueous samples, industrial wastes, soils, sludges,
sediments, and other solid wastes for which total (acid-leachable) elements are
required.
1.2 Elements for which Method 6020 has shown acceptable performance in a
multi-laboratory study are listed in TABLE 1. Acceptability of the method for
an element was based upon the multi-laboratory performance compared with that of
either furnace atomic absorption spectroscopy or inductively coupled plasma-
atomic emission spectroscopy. Other elements may be added to Table 1 as more
information becomes available. Multi-laboratory performance data for the listed
elements (and others) are provided in Section 9. Instrument detection limits,
sensitivities, and linear ranges for these elements will vary with the matrices,
instrumentation, and operating conditions.
1.3 Use of this method is restricted to spectroscopists who are
knowledgeable in the recognition and the correction of spectral, chemical, and
physical interferences in ICP-MS.
1.4 An appropriate internal standard is required for each analyte
determined by ICP-MS. Recommended internal standards are Li, Sc, Y, Rh,
mln, 159Tb, 1<5Ho, and 209Bi. The lithium internal standard should have an
enriched abundance of Li, so that interference from lithium native to the sample
is minimized. Other elements may need to be used as internal standards when
samples contain significant amounts of the recommended internal standards.
2.0 SUMMARY OF METHOD
2.1 Prior to analysis, samples which require total values must be digested
using appropriate sample preparation methods (such as Methods 3005 - 3051).
2.2 Method 6020 describes the multi-elemental determination of analytes
by ICP-MS. The method measures ions produced by a radio-frequency inductively
coupled plasma. Analyte species originating in a liquid are nebulized and the
resulting aerosol transported by argon gas into the plasma torch. The ions
produced are entrained in the plasma gas and introduced, by means of a water-
cooled interface, into a quadrupole mass spectrometer. The ions produced in the
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plasma are sorted according to their mass-to-charge ratios and quantified with
a channel electron multiplier. Interferences must be assessed and valid
corrections applied or the data flagged to indicate problems. Interference
correction must include compensation for background ions contributed by the
plasma gas, reagents, and constituents of the sample matrix.
3.0 INTERFERENCES
3.1 Isobaric elemental interferences in ICP-MS are caused by isotopes of
different elements forming atomic ions with the same nominal mass-to-charge ratio
(m/z). A data system must be used to correct for these interferences. This
involves determining the signal for another isotope of the interfering element
and subtracting the appropriate signal from the analyte isotope signal. Since
commercial ICP-MS instruments nominally provide unit resolution at 10% of the
peak height, very high ion currents at adjacent masses can also contribute to ion
signals at the mass of interest. Although this type of interference is uncommon,
it is not easily corrected, and samples exhibiting a significant problem of this
type could require resolution improvement, matrix separation, or use of another
method.
3.2 Isobaric molecular and doubly-charged ion interferences in ICP-MS are
caused by ions consisting of more than one atom or charge, respectively. Most
isobaric interferences that could affect ICP-MS determinations have been
identified in the literature [3,4]. Examples include ArCl* ions on the As
signal and MoO* on the cadmium isotopes. Since the Cl natural abundance of
75.8 percent is 3.13 times the Cl abundance of 24.2 percent, the choride
corrections can be calculated as follows (where the Ar Cl* contribution at m/z
75 is a negligible 0.06 percent of the 40Ar35cr signal):
corrected arsenic signal = (m/z 75 signal) - (3.13) (m/z 77 signal) +
(2.53) (m/z 82 signal), (where the final term adjusts for any selenium
contribution at 77 m/z),
Similarly,
corrected cadmium signal = (m/z 114 signal) - (0.027)(m/z 118 signal) -
(1.63)(m/z 108 signal), (where last 2 terms adjust for any tin or MoO*
contributions at m/z 114).
The above equations are based upon the constancy of the isotopic ratios for the
interfering species. Corrections that presume a constant fraction of a molecular
ion relative to the "parent" ion have not been found [5] to be reliable, e.g.,
oxide levels can vary. If a correction for an oxide ion is based upon the ratio
of parent-to-oxide ion intensities, the correction must be adjusted for the
degree of oxide formation by the use of an appropriate oxide internal standard
previously demonstrated to form a similar level of oxide as the interferant.
This type of correction has been reported [5] for oxide-ion corrections using
ThO*/Th* for the determination of rare earth elements.
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3.3 Physical interferences are associated with the sample nebulization and
transport processes as well as with ion-transmission efficiencies. Nebulization
and transport processes can be affected if a matrix component causes a change in
surface tension or viscosity. Changes in matrix composition can cause
significant signal suppression or enhancement [6]. Dissolved solids can deposit
on the nebulizer tip of a pneumatic nebulizer and on the interface skimmers
(reducing the orifice size and the instrument performance). Total solid levels
below 0.2% (2,000 mg/L) have been recommended [7] to minimize solid deposition.
An internal standard can be used to correct for physical interferences, if it is
carefully matched to the analyte so that the two elements are similarly affected
by matrix changes [8]. When the intensity level of an internal standard is less
than 30 percent or greater than 120 percent of the intensity of the first
standard used during calibration, the sample must be reanalyzed after a fivefold
(1+4) dilution has been performed.
3.4 Memory interferences can occur when there are large concentration
differences between samples or standards which are analyzed sequentially. Sample
deposition on the sampler and skimmer cones, spray chamber design, and the type
of nebulizer affect the extent of the memory interferences which are observed.
The rinse period between samples must be long enough to eliminate significant
memory interference.
4.0 APPARATUS AND MATERIALS
4.1 Inductively coupled plasma-mass spectrometer:
4.1.1 A system capable of providing resolution, better than or
equal to 1 amu at 10% peak height is required. The system must have a
mass range from at least 6 to 240 amu and a data system that allows
corrections for isobaric interferences and the application of the internal
standard technique. Use of a mass-flow controller for the nebulizer argon
and a peristaltic pump for the sample solution are recommended.
4.1.2 Argon gas supply: high-purity grade (99.99%).
5.0 REAGENTS
5.1 Acids used in the preparation of standards and for sample processing
must be of high purity. Redistilled acids are recommended because of the high
sensitivity of ICP-MS. Nitric acid at less than 2 per cent (v/v) is required for
ICP-MS to minimize damage to the interface and to minimize isobaric molecular-ion
interferences with the analytes. Many more molecular-ion interferences are
observed on the analytes when hydrochloric and sulfuric acids are used [3,4].
Concentrations of antimony and silver above 300 /xg/L require 1% (v/v) HC1 for
stability. If HC1 is added as a stabilizer, then corrections for the chloride
molecular-ion interferences must be applied to all data generated.
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5.2 Reagent water: Reagent water will be interference free. All
references to water in the method refer to reagent water unless otherwise
specified. Refer to Chapter One for a definition of reagent water.
5.3 Standard stock solutions may be purchased or prepared from ultra-high
purity grade chemicals or metals (99.99 to 99.999% pure). See Method 6010A,
Section 5.3, for instructions on preparing standard solutions from solids.
5.3.1 Bismuth internal standard solution, stock, 1 mL = 100 /xg Bi:
Dissolve 0.1115 g Bi203 in a minimum amount of dilute HN03. Add 10 ml
cone. HN03 and dilute to 1,000 ml with reagent water.
5.3.2 Holmium internal standard solution, stock, 1 mL = 100 jug Ho:
Dissolve 0.1757 g Ho2(C03)2-5H20 in 10 ml reagent water and 10 mL HN03.
After dissolution is complete, warm the solution to degas. Add 10 mL
cone. HN03 and dilute to 1,000 mL with reagent water.
5.3.3 Indium internal standard solution, stock, 1 mL = 100 M9 In:
Dissolve 0.1000 g indium metal in 10 mL cone. HN03. Dilute to 1,000 mL
with reagent water.
5.3.4 Lithium internal standard solution, stock, 1 mL = 100 jug 6Li:
Dissolve 0.6312 g 95-atom-% Li, Li2C03 in 10 mL of reagent water and 10 mL
HN03. After dissolution is complete, warm the solution to degas. Add
10 mL cone. HN03 and dilute to 1,000 mL with reagent water.
5.3.5 Rhodium internal standard solution, stock, 1 mL = 100 jug Rh:
Dissolve 0.3593 g ammonium hexachlororhodate (III) (NH,)3RhCl, in 10 mL
reagent water. Add 100 mL cone. HC1 and dilute to 1,000 mL with reagent
water.
5.3.6 Scandium internal standard solution, stock, 1 mL = 100 jug Sc:
Dissolve 0.15343 g Sc203 in 10 mL (1+1) hot HN03. Add 5 ml cone. HN03 and
dilute to 1,000 mL with reagent water.
5.3.7 Terbium internal standard solution, stock, 1 mL = 100 M9 Tb:
. Dissolve 0.1828 g Tb2(CO,)3-5H20 in 10 mL (1+1) HN03. After dissolution is
complete, warm the solution to degas. Add 5 ml cone. HN03 and dilute to
1,000 mL with reagent water.
5.3.8 Yttrium internal standard solution, stock, 1 mL = 100 /xg Y:
Dissolve 0.2316 g Y2(C03),.3H20 in 10 mL (1+1) HN03. Add 5 ml cone. HN03
and dilute to 1,000 mL with reagent water.
5.3.9 Titanium solution, stock, 1 mL = 100 ng Ti: Dissolve 0.4133 g
(NH,)2TiF6 in reagent water. Add 2 drops cone. HF and dilute to 1,000 mL
with reagent water.
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5.3.10 Molybdenum solution, stock, 1 ml = 100 M9 Mo: Dissolve
0.2043 g (NH4)2Mo04 in reagent water. Dilute to 1,000 ml with reagent
water.
5.4 Mixed calibration standard solutions -- Dilute the stock-standard
solutions to levels in the linear range for the instrument in a solvent
consisting of 1 percent (v/v) HNO, in reagent water. The calibration standard
solutions must contain a suitable concentration of an appropriate internal
standard for each analyte. Generally, an internal standard should be no more
than 50 amu removed from the analyte. Recommended internal standards include
*Li, 45Sc, ®Y, 1(53Rh, 115In, 159Te, 1*9Ho, and 209Bi. Prior to preparing the mixed
standards, each stock solution must be analyzed separately to determine possible
spectral interferences or the presence of impurities. Care must be taken when
preparing the mixed standards that the elements are compatible and stable.
Transfer the mixed standard solutions to freshly acid-cleaned FEP fluorocarbon
bottles for storage. Fresh mixed standards must be prepared as needed with the
realization that concentrations can change on aging. Calibration standards must
be initially verified using a quality control sample (see Section 5.8) and
monitored weekly for stability.
5.5 Blanks: Three types of blanks are required for the analysis. The
calibration blank is used in establishing the calibration curve. The reagent
blank is used to monitor for possible contamination resulting from the sample
preparation procedure. The rinse blank is used to flush the system between all
samples and standards.
5.5.1 The calibration blank consists of 1 percent HN03 (v/v) in
reagent water along with the selected concentrations of internal standards
such that there is an appropriate internal standard element for each of
the analytes.
5.5.2 The reagent blank must contain all the reagents in the same
volumes as used in processing the samples. The reagent blank must be
carried through the complete procedure and contain the same acid
concentration in the final solution as the sample solutions used for
analysis.
5.5.3 The rinse blank consists of 1 to 2 percent HN03 (v/v) in
reagent water. Prepare a sufficient quantity to flush the system between
standards and samples.
5.6 The instrument check standard is prepared by the analyst by combining
compatible elements at concentrations equivalent to the midpoint of their
respective calibration ranges.
5.7 The interference check solution(s) (ICS) is prepared to contain known
concentrations of interfering elements that will demonstrate the magnitude of
interferences and provide an adequate test of any corrections. Chloride in the
ICS provides a means to evaluate software corrections for chloride-related
interferences such as 35ClV on 3V and "Ar^Cl* on ^As*. Iron is used to
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demonstrate adequate resolution of the spectrometer for the determination of
manganese. Molybdenum serves to indicate oxide effects on cadmium isotopes. The
other components are present to evaluate the ability of the measurement system
to correct for various molecular-ion isobaric interferences. The ICS is used to
verify that the interference levels are corrected by the data system within
quality control limits.
5.7.1 The final concentrations of elements in ICS A and ICS AB are
shown in Table 2. These solutions must be prepared from ultra-pure
reagents. They can be obtained commercially or prepared by the following
procedure.
5.7.1.1 Mixed ICS solution I may be prepared by adding
13.903 g A1(NO,)3-9H,0, 2.498 g CaCO, (dried at 180 C for 1 h before
weighing), 1.000 g Fe, 1.658 g MgO, 2.305 g Na2C03, and 1.767 g K2C03
to 25 ml of reagent water. Slowly add 40 ml of (1+1) HN03. After
dissolution is complete, warm the solution to degas. Cool and
dilute to 1,000 ml with reagent water.
5.7.1.2 Mixed ICS solution II may be prepared by slowly
adding 7.444 g 85 % H,P04, 6.373 g 96% H,S04, 40.024 g 37% HC1, and
10.664 g citric acid t60,H8 to 100 mL of reagent water. Dilute to
1,000 ml with reagent water.
5.7.1.3 Mixed ICS solution III may be prepared by adding 5 ml
each of 100 M9/ml arsenic stock solution, chromium stock solution,
copper stock solution, manganese stock solution, selenium stock
solution, silver stock solution, and zinc stock solution, 10 ml each
of 100 /K|/ml cobalt stock solution, nickel stock solution, and
vanadium stock solution, and 2.5 ml of 100 ng/m\ cadmium stock
solution. Dilute to 100 ml with 2% HN03.
5.7.1.4 Working ICS Solutions
5.7.1.4.1 ICS A may be prepared by adding 50 mL of
mixed ICS solution I (5.7.1.1), 10 ml each of 100 /zg/ml
titanium stock solution (5.3.9) and molybdenum stock solution
(5.3.10), and 25 ml of mixed ICS solution II (5.7.1.2).
Dilute to 100 ml with reagent water. ICS solution A must be
prepared fresh weekly.
5.7.1.4.2 ICS AB may be prepared by adding 50 ml of
mixed ICS solution I (5.7.1.1), 10 ml each of 100 /ug/ml
titanium stock solution (5.3.9) and molybdenum stock solution
(5.3.10), 25 ml of mixed ICS solution II (5.7.1.2), and 2 ml
of Mixed ICS solution III (5.7.1.3). Dilute to 100 mL with
reagent water. ICS solution AB must be prepared fresh weekly.
5.8 The quality control sample is the initial calibration verification
solution, which must be prepared in the same acid matrix as the calibration
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standards. This solution must be an independent standard near the midpoint of
the linear range at a concentration other than that used for instrument
calibration. An independent standard is defined as a standard composed of the
analytes from a source different from those used in the standards for instrument
calibration.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Sample collection procedures should address the considerations
described in Chapter Nine of this Manual.
6.2 See the introductory material in Chapter Three, Inorganic Analytes,
Sections 3.1.3 for information on sample handling and preservation. Only
polyethylene or Teflon containers are recommended for use in Method 6020.
7.0 PROCEDURE
7.1 Solubilization and digestion procedures are presented in the Sample
Preparation Methods (e.g. Methods 3005 - 3050).
7.2 Initiate appropriate operating configuration of the instrument
computer.
7.3 Set up the instrument with the proper operating parameters.
7.4 Operating conditions: In general, the analyst should follow the
instructions provided by the instrument manufacturer. The following is a
suggested listing of operating conditions which may be useful.
Perkin-Elmer Sciex
Elan 500 VG Plasmaauad
Plasma Gas (1pm) 12 13
Aux. Gas (1pm) 1.2 0.65
Neb. Gas (1pm) 0.95 0.69
Forward power (kW) 1.2 1.30
Reflected power (W) < 5 < 5
Sampling Height 18 12
(mm above load coil)
Note: Addition of nitrogen to the plasma argon has been reported to
decrease many molecular interferences [9].
Allow at least 30 minutes for the instrument to equilibrate before analyzing
any samples. This must be verified by analyzing a tuning solution (such as 100
jiig/L Li, Co, In, and Tl) at least four times with relative standard deviations
of less than 10% for the analytes contained in the tuning solution.
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Note: Precautions must be taken to protect the channel electron multiplier
from high ion currents. The channel electron multiplier suffers from fatigue
after being exposed to high ion currents. This fatigue can last from several
seconds to hours depending on the extent of exposure. During this time period,
response factors are constantly changing, which invalidates the calibration
curve, causes instability, and invalidates sample analyses.
7.5 Conduct mass calibration and resolution checks in the mass regions of
interest. The mass calibration and resolution parameters are required criteria
which must be met prior to any samples being analyzed. If the mass calibration
exceeds a difference of more than 0.1 amu from the actual value, then the mass
calibration must be adjusted to the correct values. The resolution must also be
verified to be less than 1.0 amu full width at 10 percent peak height.
7.6 Calibrate the instrument for the analytes of interest for the isotopes
shown in Table 3 using the calibration blank and at least a single standard
according to the manufacturer's recommended procedure. Flush the system with the
rinse blank (5.5.3) between each standard solution. Use the average of the
multiple integrations for both standardization and sample analysis.
7.7 Some elements (such as Hg, W, and Mo) require extended flushing times
which need to be determined for each instrumental system.
7.8 All masses which could affect data quality should be monitored to
determine potential effects from matrix components on the analyte peaks. These
masses must be monitored either simultaneously in a separate scan or at the same
time quantification occurs.
7.9 Immediately after the calibration has been established, the
••libration must be verified and documented for every analyte by the analysis of
tne initial calibration verification solution (Section 5.8). When measurements
exceed ± 10% of the accepted value the analysis must be terminated, the problem
corrected, the instrument recalibrated, and the calibration reverified. Any
samples analyzed under an out-of-control calibration must be reanalyzed.
7.10 Flush the system with the rinse blank solution (5.5.3) for at least
30 seconds before the analysis of each sample (see Section 7.7). Aspirate each
sample for at least 30 seconds before collecting data. Analyze the instrument
check standard (Section 5.6) and the calibration blank (Section 5.5.1) at a
frequency of at least once every 10 analytical samples.
7.11 Dilute and reanalyze samples that are more concentrated than the
linear range for an analyte (or species needed for a correction) or measure an
alternate less-abundant isotope.
7.12 Calculations: The quantitative values shall be reported in units of
micrograms per liter (M9/L) for aqueous samples and milligrams per kilogram
(mg/kg) for solid samples. If dilutions were performed, the appropriate
corrections must be applied to the sample values.
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7.12.1 Results for solids must be reported on a dry-weight basis as
follows:
(1) A separate determination of percent solids must be
performed.
(2) The concentrations determined in the digest are to be
reported on the basis of the dry weight of the sample.
Concentration (dry weight)(mg/kg) = {* x ^
H A O
Where,
C = Digest Concentration (mg/L)
V = Final volume in liters after sample preparation
W = Weight in kg of wet sample
% Sol ids
100
Calculations should include appropriate interference corrections (see
Section 3.2 for examples), internal standard normalization, and the
summation of signals at 206, 207, and 208 m/z for lead to compensate for
any differences in the abundances of these isotopes between samples and
standards.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and be available for
easy reference or inspection.
8.2 Instrument Detection Limits (IDL's) (in M9/L) can be estimated by
multiplying by three the average of the standard deviations obtained on three
nonconsecutive days from the analysis of a standard solution (each analyte in
reagent water) at a concentration 3x-25x IDL, with seven consecutive measurements
per day. Each measurement must be performed as though it were a separate
analytical sample (i.e., each measurement must be followed by a rinse and/or any
other procedure normally performed between the analysis of separate samples).
IDL's must be determined at least every three months and kept with the instrument
log book.
8.3 The intensities of all internal standards must be monitored for every
analysis. When the intensity of any internal standard fails to fall between 30
and 120 percent of the intensity of that internal standard in the initial
calibration standard, the following procedure is followed. The sample must be
diluted fivefold (1+4) and reanalyzed with the addition of appropriate amounts
of internal standard. This procedure must be repeated until the internal
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standard intensities fall within the prescribed window. The intensity levels of
the internal standards for the calibration blank (Section 5.5.1) and instrument
check standard (Section 5.6) must agree within ± 20 percent of the intensity
level of the internal standard of the original calibration blank solution. If
they do not agree, terminate the analysis, correct the problem, recalibrate, and
reanalyze the affected samples.
8.4 To obtain analyte data of known quality, it is necessary to measure
more than the analytes of interest in order to determine whether interference
corrections are necessary. If the concentrations of interference sources (such
as C, Cl, Mo, Zr, W) are below the levels that show an effect on the analyte
level, uncorrected equations may be used provided all QC criteria are met. Note
that monitoring the interference sources does not necessarily require monitoring
the interferant itself, but that a molecular species may be monitored to indicate
the presence of the interferent. When corrected equations are used, all QC
criteria must also be met. Extensive QC for interference corrections are
required at all times. The monitored masses must include those elements whose
hydrogen, oxygen, hydroxyl, chlorine, nitrogen, carbon and sulfur molecular ions
could impact the analytes of interest. Unsuspected interferences may be detected
by adding pure major matrix components to a sample to observe any impact on the
analyte signals. When an interference source is present, the sample elements
impacted must be flagged to indicate (a) the percentage interference correction
applied to the data or (b) an uncorrected interference by virtue of the elemental
equation used for quantitation. The isotope proportions for an element or
molecular-ion cluster provide information useful for quality assurance.
NOTE: Only isobaric elemental, molecular, and doubly charged interference
corrections which use established isotopic response ratios or parent-to-oxide
ratios (provided an oxide internal standard is used as described in Section 3.2)
are acceptable corrections for use in Method 6020.
8.5 Serial dilution: If the analyte concentration is within the linear
dynamic range of the instrument and sufficiently high (minimally, a factor of 100
above the instrumental detection limit), an analysis of a fivefold dilution must
agree within ± 10% of the original determination. If not, an interference effect
must be suspected. One serial dilution must be analyzed for each twenty samples
or less of each matrix in a batch.
8.6 Matrix spike addition: An analyte spike added to a portion of a
prepared sample, or its dilution, should be recovered to within 75 to 125 percent
of the known value. The spike addition should produce a minimum signal level of
10 times and a maximum of 100 times the instrumental detection limit. If the
spike is not recovered within the specified limits, a matrix effect should be
suspected. The use of a standard-addition analysis procedure can usually
compensate for this effect. See Section 8.5.3 of Method 6010 for information on
standard additions.
8.7 A Laboratory Control Sample (LCS) should be analyzed for each analyte
using the same sample preparations, analytical methods and QA/QC procedures
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employed for the test samples. One LCS should be prepared and analyzed for each
sample batch at a frequency of one LCS for each 20 samples or less.
8.8 Check the instrument standardization by analyzing appropriate quality
control solutions as follows:
8.8.1 Check instrument calibration using a calibration blank
(Section 5.5.1) and the initial calibration verification solution
(Sections 5.8 and 7.9).
8.8.2 Verify calibration at a frequency of every 10 analytical
samples with the instrument check standard (Section 5.6) and the
calibration blank (Section 5.5.1). These solutions must also be analyzed
for each analyte at the beginning of the analysis and after the last
sample.
8.8.3 The results of the initial calibration verification solution
and the instrument check standard must agree within ± 10% of the expected
value. If not, terminate the analysis, correct the problem, and
recalibrate the instrument. Any sample analyzed under an out-of-control
calibration must be reanalyzed at no additional cost to the government.
8.8.4 The results of the calibration blank must be less than 3
times the current IDL for each element. If this is not the case, the
reason for the out-of-control condition must be found and corrected, and
affected samples must be reanalyzed.
8.9 Verify the magnitude of elemental and molecular-ion isobaric
interferences and the adequacy of any corrections at the beginning of an
analytical run or once every 12 hours, whichever is more frequent. Do this by
analyzing the interference check solutions A and AB
8.10 Analyze one duplicate sample for every matrix
frequency of one matrix duplicate for every 20 samples.
in a batch at a
8.10.1 The relative percent difference (RPD) between duplicate
determinations must be calculated as follows:
ID, - D2 |
RPD = x 100
(D, + D2)/2
where:
RPD = relative percent difference.
D1 = first sample value.
D2 = second sample value (duplicate)
A control limit of 20% RPD should not be exceeded for analyte values
greater than 100 times the instrumental detection limit. If this limit is
exceeded, the reason for the out-of-control situation must be found and
6020-11 Revision 0
November 1990
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corrected, and any samples analyzed during the out-of-control condition
must be reanalyzed.
9.0 METHOD PERFORMANCE
9.1 In an EPA multi-laboratory study, 10 laboratories applied the
ICP-MS technique to both aqueous and solid samples. TABLE 5 summarizes the
method performance data for aqueous samples. Performance data for solid samples
is provided in TABLE 6.
10.0 REFERENCES
1. Horlick, G., et al., Spectrochim. Acta 40B, 1555 (1985).
2. Gray, A.L., Spectrochim. Acta 40B, 1525 (1985); 41B, 151 (1986).
3. Tan, S.H., and Horlick, G., Appl. Spectrosc. 40, 445 (1986).
4. Vaughan, M.A., and Horlick, G., Appl. Spectrosc. 40, 434 (1986).
5. Lichte, F.E., et al., Anal. Chem. 59, 1150 (1987).
6. Beauchemin, D., et al., Spectrochim. Acta 42B, 467 (1987).
7. Houk, R.S., Anal. Chem. 58, 97A (1986).
8. Thompson, J.J., and Houk, R.S., Appl. Spectrosc. 41, 801 (1987).
9. Evans, E.H., and Ebdon, L., J. Anal. At. Spectrom. 4, 299 (1989).
6020-12 Revision 0
• November 1990
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TABLE 1. ELEMENTS APPROVED FOR ICP-MS DETERMINATION
Element
CAS* *
Estimated Detection
Limit (/ig/L)
Aluminum
Antimony
Arsenic
Barium
Beryl 1i urn
Cadmium
Chromium
Cobalt
Copper
Lead
Manganese
Nickel
Silver
Thallium
Zinc
7429-90-5
7440-36-0
7440-38-2
7440-39-3
7440-41-7
7440-43-9
7440-47-3
7440-48-4
7440-50-8
7439-92-1
7439-96-5
7440-02-0
7440-22-4
7440-28-0
7440-66-6
,1
0.1
0.02
0.4
0.02
0.
0.07
0.02
0.01
0.03
0.02
0.04
0.03
0.04
0.05
0.08
6020-13
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November 1990
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TABLE 2. RECOMMENDED INTERFERENCE CHECK SAMPLE COMPONENTS AND CONCENTRATIONS.
Interference Solution A Solution AB
component Concentration (mg/L) Concentration (mg/L)
Al
Ca
Fe
Mg
Na
P
K
S
C
Cl
Mo
Ti
As
Cd
Cr
Co
Cu
Mn
Ni
Ag
Zn
500.0
500.0
500.0
500.0
500.0
500.0
500.0
500.0
1000.0
3600.0
10.0
10.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
500.0
500.0
500.0
500.0
500.0
500.0
500.0
500.0
1000.0
3600.0
10.0
10.0
0.100
0.050
0.100
0.200
0.100
0.100
0.200
0.100
0.100
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November 1990
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TABLE 3. RECOMMENDED ISOTOPES FOR SELECTED ELEMENTS
Mass Element of interest
27 Aluminum
121. 123 Antimony
75 Arsenic
138, 137, 136, 135. 134 Barium
9 Beryl 1i urn
209 Bismuth (IS)
114. 112, 111. 110, 113, 116, 106 Cadmium
42, 43, 44, 46, 48 Calcium (I)
35, 37, (77, 82)a Chlorine (I)
52, 53, 50, 54 Chromium
59 Cobalt
63, 65 Copper
165 Hoi mi urn (IS)
115. 113 Indium (IS)
56, 54, 57, 58 Iron (I)
139 Lanthanum (I)
208, 207. 206. 204 Lead
6^7 Lithium (IS)
24, 25, 26 Magnesium (I)
55 Manganese
98, 96, 92, 97, 94, (108)a Molybdenum (I)
58, 60, 62, 61, 64 Nickel
39 Potassium (I)
103 Rhodium (IS)
45 Scandium (IS)
107. 109 Silver
23 Sodium (I)
159 Terbium (IS)
205. 203 Thallium
120, 118 Tin (I)
89 Yttrium (IS)
64, 66, 68, 67, 70 Zinc
NOTE: Method 6020 is recommended for only those analytes listed in Table
1. Other elements are included in this table because they are potential
interferents (labeled I) in the determination of recommended analytes, or because
they are commonly used internal standards (labeled IS). Isotopes are listed in
descending order of natural abundance. The most generally useful isotopes are
underlined and in boldface, although certain matrices may require the use of
alternative isotopes. a These masses are also useful for interference correction
(Section 3.2). Internal standard must be enriched in the Li isotope. This
minimizes interference from indigenous lithium.
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TABLE 4. SPIKING LEVELS FOR ICP-MS ANALYSIS (/zg/L)
Element Water Soil
Aluminum 500 *
Antimony 100 100
Arsenic 100 100
Barium 200 200
Beryl 1i urn 50 50
Cadmium 50 50
Chromium 50 50
Cobalt 100 100
Copper 50 50
Lead 50 50
Manganese 50 50
Nickel 100 100
Silver 50 50
Thallium 50 50
Vanadium 100 100
Zinc 100 100
* No spike required.
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TABLE 5. ICP-MS MULTI-LABORATORY PRECISION AND ACCURACY DATA FOR AQUEOUS
SOLUTIONS
Element
Comparability8
Range
%RSD
Range
Nb Sc
Aluminum 95 - 100 11 - 14 14-14 4
Antimony d 5.0 - 7.6 16-16 3
Arsenic 97 - 114 7.1 - 48 12-14 4
Barium 91 - 99 4.3 - 9.0 16-16 5
Beryllium 103 - 107 8.6 - 14 13-14 3
Cadmium 98 - 102 4.6 - 7.2 18-20 3
Calcium 99 - 107 5.7 - 23 17-18 5
Chromium 95 - 105 13 - 27 16-18 4
Cobalt 101 - 104 8.2 - 8.5 18-18 3
Copper 85 - 101 6.1-27 17-18 5
Iron 91 - 900 11 - 150 10-12 5
Lead 71 - 137 11 - 23 17-18 6
Magnesium 98 - 102 10 - 15 16-16 5
Manganese 95 - 101 8.8 - 15 18-18 4
Nickel 98 - 101 6.1 - 6.7 18-18 2
Potassium 101 - 114 9.9 - 19 11-12 5
Selenium 102 - 107 15 - 25 12-12 3
Silver 104 - 105 5.2 - 7.7 13-16 2
Sodium 82 - 104 24-43 9-10 5
Thallium 88 - 97 9.7 - 12 18-18 3
Vanadium 107 - 142 23-68 8-13 3
Zinc 93 - 102 6.8 - 17 16-18 5
8 Comparability refers to the percent agreement of mean ICP-MS values to those
of the reference technique. N is the range of the number of ICP-MS
measurements where the analyte values exceed the limit of quantitation (3.3 times
the average IDL value). c S is the number of samples with results greater
than the limit of quantitation. No comparability values are provided for
antimony because of evidence that the reference data is affected by an
interference.
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TABLE 6. ICP-MS MULT I-LABORATORY PRECISION AND ACCURACY DATA FOR SOLID MATRICES
Element
Comparability8
Range
%RSD
Range Nb
Sc
Aluminum 83 - 101 11 - 39 13-14 7
Antimony d 12-21 15-16 2
Arsenic 79 - 102 12 - 23 16-16 7
Barium 100 - 102 4.3 - 17 15-16 7
Beryllium 50 - 87 19 - 34 12 - 14 5
Cadmium 93 - 100 6.2 - 25 19-20 5
Calcium 95 - 109 4.1 - 27 15-17 7
Chromium 77 - 98 11 - 32 17-18 7
Cobalt 43 - 102 15 - 30 17-18 6
Copper 90 - 109 9.0 - 25 18-18 7
Iron 87 - 99 6.7 - 21 12-12 7
Lead 90 - 104 5.9 - 28 15-18 7
Magnesium 89 - 111 7.6 - 37 15-16 7
Manganese 80 - 108 11 - 40 16-18 7
Nickel 87 - 117 9.2 - 29 16-18 7
Potassium 97 - 137 11 - 62 10-12 5
Selenium 81 39 12 1
Silver 43 - 112 12 - 33 15-15 3
Sodium 100 - 146 14-77 8-10 5
Thallium 91 33 18 1
Vanadium 83 - 147 20-70 6-14 7
Zinc 84 - 124 14 - 42 18-18 7
a Comparability refers to the percent agreement of mean ICP-MS values to those
of the reference technique. N is the range of the number of ICP-MS
measurements where the analyte values exceed the limit of quantitation (3.3 times
the average IDL value). c S is the number of samples with results greater
than the limit of quantitation. No comparability values are provided for
antimony because of evidence that the reference data is affected by an
interference.
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METHOD 6020
INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY
Start
J
7 . 1 Analyia
by Mothod
7000 or
Method 6010
7.1 U..
Method 3040
!• aanpla
oil«,graa*a«
waxa«?
tha invtruamnt
I* H,0
acidified,
pr«-filt«r«d?
!• •anpla
vat«r?
I.
••mpla
•nalyiad by
FLAA/ICP or
CFAA?
7.8 Monitor all
!• ••nplv
•quaoui or
•olid?
instrument par
data quality a*
raooamandation*
calibration and
6020-19
Revision 0
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METHOD 7062
ANTIMONY AND ARSENIC (ATOMIC ABSORPTION. GASEOUS BOROHYDRIDE1
1.0 SCOPE AND APPLICATION
1.1 Method 7062 is an atomic absorption procedure for determining 1 jig/L
to 400 ng/L concentrations of antimony and arsenic in wastes, mobility procedure
extracts, soils, and ground water. Method 7062 is approved for sample matrices
that contain up to 4000 mg/L concentrations of cobalt, copper, iron, mercury, and
nickel. A solid sample can contain up to 40% by weight of the interferents
before exceeding 4000 mg/L in a digested sample. All samples including aqueous
matrices must be subjected to an appropriate dissolution step prior to analysis.
Spiked samples and relevant standard reference materials are used to determine
the applicability of the method to a given waste.
2.0 SUMMARY OF METHOD
2.1 Samples are prepared according to the nitric acid digestion procedure
described in Method 3010 for aqueous and extract samples and the
nitric/peroxide/hydrochloric acid digestion procedure described in Method 3050
(furnace AA option) for sediments, soils, and sludges. Excess peroxide is
removed by evaporating samples to near dryness at the end of the digestion
followed by degassing the samples upon addition of urea. L-cystine is then added
as a masking agent. Next, the antimony and arsenic in the digest are reduced to
the trivalent forms with potassium iodide. The trivalent antimony and arsenic
are then converted to volatile hydrides using hydrogen produced from the reaction
of the acidified sample with sodium borohydride in a continuous-flow hydride
generator.
2.2 The volatile hydrides are swept into an air-acetylene flame heated
quartz absorption cell located in the optical path of an atomic absorption
spectrophotometer. The resulting absorption of the lamp radiation is
proportional to the arsenic or antimony concentration.
2.3 The typical detection limit for this method is 1.0 [ig/L.
3.0 INTERFERENCES
3.1 Very high (>4000 mg/L) concentrations of cobalt, copper, iron,
mercury, and nickel can cause analytical interferences through precipitation as
reduced metals and associated blockage of transfer lines and fittings.
3.2 Traces of peroxides left following the sample work-up can result in
analytical interferences. Peroxides must be removed by evaporating each sample
to near dryness followed by reaction with urea and allowing sufficient time for
degassing before analysis (see Sections 7.1 and 7.2).
7062-1 Revision 0
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4.0 APPARATUS AND MATERIALS
4.1 Electric hot plate: Large enough to hold at least several 100 mL
Pyrex digestion beakers.
4.2 A continuous-flow hydride generator: A commercially available
continuous-flow sodium borohydride/HCl hydride generator or a generator
constructed similarly to that shown in Figure 1 (P. S. Analytical or equivalent).
4.2.1 Peristaltic Pump: A four-channel, variable-speed peristaltic
pump to permit regulation of liquid-stream flow rates (Ismatec Reglo-100
or equivalent). Pump speed and tubing diameters should be adjusted to
provide the following flow rates: sample/blank flow = 4.2 mL/min;
borohydride flow =2.1 mL/min; and potassium iodide flow = 0.5 mL/min.
4.2.2 Sampling Valve (optional): A sampling valve (found in the
P. S. Analytical Hydride Generation System or equivalent) that allows
switching between samples and blanks (rinse solution) without introduction
of air into the system will provide more signal stability.
4.2.3 Transfer Tubing and Connectors: Transfer tubing (1 mm I.D.),
mixing T's, and connectors are made of teflon and are of compatible sizes
to form tight, leak-proof connections (Latchat, Technicon, etc. flow
injection apparatus accessories or equivalent).
4.2.4 Mixing Coil: A 20-turn coil made by wrapping transfer tubing
around a 1-cm diameter by 5-cm long plastic or glass rod (see Figure 1).
4.2.5 Mixing Coil Heater: A 250-mL Erlenmeyer flask containing 100
mL of water heated to boiling on a dedicated one-beaker hotplate (Corning
PC-35 or equivalent). The mixing coil in 4.2.4 is immersed in the boiling
water to speed kinetics of the hydride forming reactions and increase
solubility of interfering reduced metal precipitates.
4.2.6 Gas-Liquid Separator: A glass apparatus for collecting
liquid and gaseous products (P.T. Analytical accessory or equivalent)
which allows the liquid fraction to drain to waste and gaseous products
above the liquid to be swept by a regulated carrier gas (argon) out of the
cell for analysis. To avoid undue carrier gas dilution, the gas volume
above the liquid should not exceed 20 mL. See Figure 1 for an acceptable
separator shape.
4.2.7 Condenser: Moisture picked up by the carrier gas must be
removed before encountering the hot absorbance cell. The moist carrier
gas with the hydrides is dried by passing the gasses through a small (< 25
mL) volume condenser coil (Ace Glass Model 6020-02 or equivalent) that is
cooled to 5°C by a water chiller (Neslab RTE-110 or equivalent). Cool tap-
water in place of a chiller is acceptable.
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November 1990
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4.2.8 Flow Meter: A meter capable of regulating up to 1 L/min of
argon carrier gas is recommended.
4.3 Absorbance Cell: A 17 cm or longer quartz tube T-cell (windowless is
strongly suggested) is recommended, as shown in Figure 1 (Varian Model VGA-76
accessory or equivalent). The cell is held in place by a holder that positions
the cell about 1 cm over a conventional AA air-acetylene burner head. In
operation, the cell is heated to around 900°C by an air-acetylene flame.
4.4 Atomic absorption spectrophotometer: Single or dual channel, single-
or double-beam instrument having a grating monochromator, photomultiplier
detector, adjustable slits, a wavelength range of 190 to 800 nm, and provisions
for interfacing with a strip-chart recorder.
4.5 Burner: As recommended by the particular instrument manufacturer for
an air-acetylene flame. An appropriate mounting bracket attached to the burner
that suspends the quartz absorbance cell between 1 and 2 cm above the burner slot
is required.
4.6 Antimony and arsenic hollow cathode lamps or antimony and arsenic
electrodeless discharge lamps and power supply. Super-charged hollow-cathode
lamps or EDL lamps are recommended for maximum sensitivity.
4.7 Strip-chart recorder (optional): Connect to output of
spectrophotometer.
5.0 REAGENTS
5.1 Reagent water: Water must be monitored for impurities. Refer to
Chapter 1 for definition of Reagent water.
5.2 Concentrated nitric acid (HN03): Acid must be analyzed to determine
levels of impurities. If a method blank is
-------
QUARTZ CELL
A A OURNER
'»0 ISCOHHECTEff
UIIRINO S*/Sn
ftHALVStS
20 TURN COIL
(TEFLON)
HOTPLATE
VALUE
(•LANK)
Figure 1. Continuous-flow sodium borohydride/hydride generator apparatus set-
up and an AAS sample introduction system.
7062-4
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November 1990
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5.5 Diluent solution: A 3% HC1 solution in reagent water must be prepared
as a diluent solution if excessive levels of analytes or interfering metals are
found in the undiluted samples.
5.6 Urea (H2NCONH2): A 5.00-g portion of reagent grade urea must be added
to a 25-mL aliquot of each sample for removal of excess peroxide through
degassing (see Section 7.2).
5.7 L-cystine (C6H12N204S2): A 1.00-g portion of reagent grade L-cystine
must be added to a 25-mL aliquot of each sample for masking the effects of
suppressing transition metals (see Section 7.2).
5.8 20% Potassium iodide (KI): A 20% KI solution (20 g reagent-grade KI
dissolved and brought to volume in 100 ml reagent water) must be prepared for
reduction of antimony and arsenic to their +3 valence states.
5.9 4% Sodium borohydride (NaBHJ: A 4% sodium borohydride solution (20
g reagent-grade NaBH4 plus 2 g sodium hydroxide dissolved in 500 ml of reagent
water) must be prepared for conversion of the antimony and arsenic to their
hydrides.
5.10 Analyte solutions:
5.10.1 Antimony and arsenic stock standard solution (1,000 mg/L):
Either procure certified aqueous standards from a supplier (Spex,
Inorganic Ventures, or equivalent) and verify by comparison with a second
standard, or dissolve 1.197 g of antimony trioxide Sbp03 and 1.320 g of
arsenic trioxide As203 in 100 ml of reagent water containing 4 g NaOH.
Acidify the solution with 20 ml concentrated HN03 and dilute to 1 liter.
5.10.2 Intermediate antimony and arsenic solution: Pipet 1 ml
stock antimony and arsenic solution into a 100-mL volumetric flask and
bring to volume with reagent water containing 1.5 mL concentrated
HN03/liter (1 ml = 10 ^g each of Sb and As).
5.10.3 Standard antimony and arsenic solution: Pipet 10 ml
intermediate antimony and arsenic solution into a 100-mL volumetric flask
and bring to volume with reagent water containing 1.5 ml concentrated
HN03/liter (1 ml = 1 jig each of Sb and As).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All sample containers must be prewashed with detergents, acids, and
reagent water. Plastic and glass containers are both suitable.
7062-5 Revision 0
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6.3 Special containers (e.g., containers used for volatile organic
analysis) may have to be used if very volatile antimony and arsenic compounds are
suspected to be present in the samples.
6.4 Aqueous samples must be acidified to a pH of <2 with nitric acid.
6.5 Nonaqueous samples shall be refrigerated, when possible, and analyzed
as soon as possible.
7.0 PROCEDURE
7.1 Place a 100-mL portion of an aqueous sample or extract or 1.000 g of
a dried solid sample in a 250-mL digestion beaker. Digest aqueous samples and
extracts according to Method 3010. Digest solid samples according to Method 3050
(furnace AA option) with the following modifications: add 5 ml of concentrated
hydrochloric acid just prior to the final volume reduction stage to aid in
antimony recovery; the final volume reduction should be to less than 5 ml but not
to dryness to adequately remove excess hydrogen peroxide (see note). After
dilution to volume, further dilution with diluent may be necessary if analytes
are known to exceed 400 ng/L or if interferents are expected to exceed 5000 mg/L
in the digest.
Note: For solid digestions, the volume reduction stage is critical to obtain
accurate data, especially for arsenic. Close monitoring of each sample is
necessary when this critical stage is reached.
7.2 Prepare samples for hydride analysis by adding 5.00 g urea, 1.00 g L-
cystine, and 20 ml concentrated HC1 to a 25-mL aliquot of digested sample in a
50-mL volumetric flask. Heat in a water bath until the L-cystine has dissolved
and effervescence has subsided (At least 30 minutes is suggested. If
effervescense is still seen, repeat step 7.1 with more volume reduction.). Bring
flask to volume with reagent water before analyzing. A 1:1 dilution correction
must be made in the final concentration calculations.
7.3 Prepare working standards from the standard antimony and arsenic
solution. Transfer 0, 0.5, 1.0, 1.5, 2.0, and 2.5 ml of standard to 100-mL
volumetric flasks and bring to volume with diluent. These concentrations will
be 0, 5, 10, 15, 20, and 25 jig Sb and As/liter.
7.4 If EP extracts (Method 1310) are being analyzed for arsenic, the
method of standard additions must be used. Spike appropriate amounts of
intermediate or standard antimony and arsenic solution to three 25 mL aliquots
of each unknown. Spiking volumes should be kept less than 0.250 ml to avoid
excessive spiking dilution errors.
7.5 Set up instrumentation and hydride generation apparatus and fill
reagent containers. The sample and blank flows should be set around 4.2 mL/min,
the borohydride flow around 2.1 mL/min, and the potassium iodide flow around 0.5
mL/min. The argon carrier gas flow is adjusted to about 200 mL/min. For the AA,
7062-6 Revision 0
November 1990
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use the 217.6-nm wavelength and 0.7-nm slit width without background correction
if analyzing for antimony. Use the 193.7-nm wavelength and 0.7-nm slit width
without background correction for the analysis of arsenic. Begin all flows and
allow 10 minutes for warm-up.
7.6 Place sample feed line into a prepared sample solution and start pump
to begin hydride generation. Wait for a maximum steady-state signal on the
strip-chart recorder or output meter. Switch to blank sample and watch for
signal to decline to baseline before switching to the next sample and beginning
the next analysis. Run standards first (low to high), then unknowns. Include
appropriate QA/QC solutions, as required. Prepare calibration curves and convert
absorbances to concentration. See following analytical flowchart.
CAUTION: The hydrides of antimony and arsenic are very toxic.
Precautions must be taken to avoid inhaling the gas.
7.7 If the method of standard additions was employed, plot the measured
concentration of the spiked samples and unspiked sample versus the spiked
concentrations. The spiked concentration axis intercept will be the method of
standard additions concentration. If the plot does not result in a straight
line, a nonlinear interference is present. This problem can sometimes be
overcome by dilution or addition of other reagents if there is some knowledge
about the waste. If the method of standard additions was not required, then the
concentration is determined from a standard calibration curve.
8.0 QUALITY CONTROL
8.1 See section 8.0 of Method 7000A.
9.0 METHOD PERFORMANCE
9.1 The relative standard deviations obtained by a single laboratory for
7 replicates of a contaminated soil were 18% for antimony at 9.1 ug/L in solution
and 4.6% for arsenic at 68 ug/L in solution.
10.0 REFERENCES
1. Methods for Chemical Analysis of Water and Wastes, EPA-600/4-82-055,
December 1982, Method 206.3.
2. "Evaluation of Hydride Atomic Absorption Methods for Antimony, Arsenic,
Selenium, and Tin", an EMSL-LV internal report under Contract 68-03-3249,
Job Order 70.16, prepared for T. A. Hinners by D. E. Dobb, and J. D.
Lindner of Lockheed Engineering and Sciences Co., and L. V. Beach of the
Varian Corporation.
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METHOD 7062
ANTIMONY AND ARSENIC (ATOMIC ABSORPTION, GASEOUS BOROHYDRIDE)
7.1 U». M.thod
3050 (furnace
AA option) to
dig..t 1.0 9
(ample
7.1-7.4
Digeat with
H.O. .>
deaoribed in
Method 3050
7.5 Add
concentrated
HC1
7.6 Do final
volume
reduction and
dilution, aa
described
No
7.1 Further
dilute with
diluent
7.1 Uae
Method 3010
to digeat 100
•1 «ample
7.2 Add to
aliquot urea,
L-cyatine, HC1;
heet H.O bath;
bring to volume
7.3 Prepare
itandarda from
itandard atook
•olutiona of Sb
and Aa
7.4 U.. the
method of
atandard
additiona on
•xtracti, only
7062-8
Revision 0
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METHOD 7742
SELENIUM (ATOMIC ABSORPTION. GASEOUS BOROHYDRIDE)
1.0 SCOPE AND APPLICATION
1.1 Method 7742 is an atomic absorption procedure for determining 3 |ig/L
to 750 ng/L concentrations of selenium in wastes, mobility procedure extracts,
soils, and ground water. Method 7742 is approved for sample matrices that
contain up to 1000 mg/L concentrations of cobalt, copper, iron, mercury, and
nickel. A solid sample can contain up to 10% by weight of the interferents before
exceeding 1000 mg/L in a digested sample. All samples including aqueous matrices
must be subjected to an appropriate dissolution step prior to analysis. Spiked
samples and relevant standard reference materials are employed to determine the
applicability of the method to a given waste.
2.0 SUMMARY OF METHOD
2.1 Samples are prepared according to the nitric acid digestion procedure
described in Method 3010 for aqueous and extract samples and the
nitric/peroxide/hydrochloric acid digestion procedure described in Method 3050
(furnace AA option) for sediments, soils, and sludges. Excess peroxide is
removed by evaporating samples to near-dryness at the end of the digestion
followed by dilution to volume and degassing the samples upon addition of urea.
The selenium is converted to the +4 oxidation state during digestion in HC1.
After a 1:10 dilution, selenium is then converted to its volatile hydride using
hydrogen produced from the reaction of the acidified sample with sodium
borohydride in a continuous-flow hydride generator.
2.2 The volatile hydride is swept into an air-acetylene flame heated
quartz absorption cell located in the optical path of an atomic absorption
spectrophotometer. The resulting absorption of the lamp radiation is
proportional to the selenium concentration.
2.3 The typical detection limit for this method is 3 ng/L.
3.0 INTERFERENCES
3.1 Very high (>1000 mg/L) concentrations of cobalt, copper, iron,
mercury, and, nickel can cause analytical interferences through precipitation as
reduced metals and associated blockage of transfer lines and fittings.
3.2 Traces of peroxides left following the sample work-up can result in
analytical interferences. Peroxides must be removed by evaporating each sample
to near-dryness followed by reacting each sample with urea and allowing
sufficient time for degassing before analysis (see Sections 7.1 and 7.2).
7742-1 Revision 0
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4.0 APPARATUS AND MATERIALS
4.1 Electric hot plate: Large enough to hold at least several 100 mL
Pyrex digestion beakers.
4.2 A continuous-flow hydride generator: A commercially available
continuous-flow sodium borohydride/HCl hydride generator or a generator
constructed similarly to that shown in Figure 1 (P. S. Analytical or equivalent).
4.2.1 Peristaltic Pump: A four-channel, variable-speed peristaltic
pump to permit regulation of liquid-stream flow rates (Ismatec Reglo-100
or equivalent). Pump speed and tubing diameters should be adjusted to
provide the following flow rates: sample/blank flow = 4.2 mL/min;
borohydride flow = 2.1 mL/min.
4.2.2 Sampling Valve (optional): A sampling valve (found in the
P. S. Analytical Hydride Generation System or equivalent) that allows
switching between samples and blanks (rinse solution) without introduction
of air into the system will provide more signal stability.
4.2.3 Transfer Tubing and Connectors: Transfer tubing (1 mm I.D.),
mixing T's, and connectors are made of teflon and are of compatible sizes
to form tight, leak-proof connections (Latchat, Technicon, etc. flow
injection apparatus accessories or equivalent).
4.2.4 Mixing Coil: A 20-turn coil made by wrapping transfer tubing
around a 1-cm diameter by 5-cm long plastic or glass rod (see Figure 1).
4.2.5 Mixing Coil Heater: A 250-mL Erlenmeyer flask containing 100
mL of water heated to boiling on a dedicated one-beaker hotplate (Corning
PC-35 or equivalent). The mixing coil in 4.2.4 is immersed in the boiling
water to speed kinetics of the hydride forming reactions and increase
solubility of interfering reduced metal precipitates.
4.2.6 Gas-Liquid Separator: A glass apparatus for collecting
liquid and gaseous products (P. S. Analytical accessory or equivalent)
which allows the liquid fraction to drain to waste and gaseous products
above the liquid to be swept by a regulated carrier gas (argon) out of the
cell for analysis. To avoid undue carrier gas dilution, the gas volume
above the liquid should not exceed 20 mL. See Figure 1 for an acceptable
separator shape.
4.2.7 Condenser: Moisture picked up by the carrier gas must be
removed before encountering the hot absorbance cell. The moist carrier
gas with the hydrides is dried by passing the gasses through a small (< 25
mL) volume condenser coil (Ace Glass Model 6020-02 or equivalent) that is
cooled to 5°C by a water chiller (Neslab RTE-110 or equivalent). Cool tap-
water in place of a chiller is acceptable.
4.2.8 Flow Meter: A meter capable of regulating up to 1 L/min of
argon carrier gas is recommended.
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4.3 Absorbance Cell: A 17-cm or longer quartz tube T-cell (windowless is
strongly suggested) is recommended, as shown in Figure 1 (Varian Model VGA-76
accessory or equivalent). The cell is held in place by a holder that positions
the cell about 1 cm over a conventional AA air-acetylene burner head. In
operation, the cell is heated to around 900°C by an air-acetylene flame.
4.4 Atomic absorption spectrophotometer: Single- or dual- channel,
single- or double-beam instrument having a grating monochromator, photomultiplier
detector, adjustable slits, a wavelength range of 190 to 800 nm, and provisions
for interfacing with a strip-chart recorder.
4.5 Burner: As recommended by the particular instrument manufacturer for
an air-acetylene flame. An appropriate mounting bracket attached to the burner
that suspends the quartz absorbance cell between 1 and 2 cm above the burner slot
is required.
4.6 Selenium hollow cathode lamp or selenium electrodeless discharge lamp
and power supply. Super-charged hollow-cathode lamps or EDL lamps are
recommended for maximum sensitivity.
4.7 Strip-chart recorder (optional): Connect to output of
spectrophotometer.
5.0 REAGENTS
5.1 Reagent water : Water must be monitored for impurities. Refer to
Chapter 1 for definition of Reagent water.
5.2 Concentrated nitric acid (HN03): Acid must be analyzed to determine
levels of impurities. If a method blank is
-------
QUARTZ CELL
A A OURNCR
TO
CHILLER
•OISCONNECTE
UIIRINO S*XSn
ANALYSIS
20 TURN COIL
(TEFLON)
__—Ą DRAIN
VALVC
(BLANK)
Figure 1. Continuous-flow sodium borohydride/hydride generator apparatus
setup and an AAS sample introduction system
7742-4
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5.7 4% Sodium Borohydride (NaBHJ: A 4 % sodium borohydride solution (20
g reagent-grade NaBH4 plus 2 g sodium hydroxide dissolved in 500 mL of reagent
water) must be prepared for conversion of the selenium to its hydride.
5.8 Selenium solutions:
5.8.1 Selenium standard stock solution (1,000 mg/L): Either
procure certified aqueous standards from a supplier (Spex, Inorganic
Ventures, or equivalent) and verify by comparison with a second standard,
or dissolve 0.3453 g of selenious acid (assay 96.6% of H2Se03) in 200 ml
of reagent water (1 mL = 1 mg Se).
5.8.2 Selenium working stock solution: Pipet 1 ml selenium
standard stock solution into all volumetric flask and bring to volume
with reagent water containing 1.5 ml concentrated HNOj/liter. The
concentration of this solution is 1 mg Se/L (1 ml = 1 \ig Se).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All sample containers must be prewashed with detergents, acids, and
reagent water. Plastic and glass containers are both suitable.
6.3 Special containers (e.g., containers used for volatile organic
analysis) may have to be used if very volatile selenium compounds are suspected
to be present in the samples.
6.4 Aqueous samples must be acidified to a pH of <2 with nitric acid.
6.5 Nonaqueous samples shall be refrigerated, when possible, and analyzed
as soon as possible.
7.0 PROCEDURE
7.1 Place a 100-mL portion of an aqueous sample or extract or 1.000 g of
a dried solid sample in a 250-mL digestion beaker. Digest aqueous samples and
extracts according to Method 3010. Digest solid samples according to Method 3050
(furnace AA option) with the following modifications: add 5 mL of concentrated
hydrochloric acid just prior to the final volume reduction stage to aid in
conversion of selenium to its plus four state; the final volume reduction should
be to less than 5 mL but not to dryness to adequately remove excess hydrogen
peroxide (see note). After dilution to volume, further dilution with diluent may
be necessary if the analyte is known to exceed 750 \ig/l or if interferents are
expected to exceed 1000 mg/L in the digestate.
7742-5 Revision 0
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Note:For solid digestions, the volume reduction stage is critical to obtain
accurate data. Close monitoring of each sample is necessary when this
critical stage in the digestion is reached.
7.2 Prepare samples for hydride analysis by adding 1.00 g urea, and 20 ml
concentrated HC1 to a 5.00 ml aliquot of digested sample in a 50-mL volumetric
flask. Heat in a water bath to dissolve salts and reduce selenium (at least 30
minutes is suggested). Bring flask to volume with reagent water before
analyzing. A ten-fold dilution correction must be made in the final
concentration calculations.
7.3 Prepare working standards from the standard stock selenium solution.
Transfer 0, 0.5, 1.0, 1.5, 2.0, and 2.5 ml of standard to 100-mL volumetric
flasks and bring to volume with diluent. These concentrations will be 0, 5, 10,
15, 20, and 25 jig Se/L.
7.4 If EP extracts (Method 1310) are being analyzed for selenium, the
method of standard additions must be used. Spike appropriate amounts of working
standard selenium solution to three 25 ml aliquots of each unknown. Spiking
volumes should be kept less than 0.250 ml to avoid excessive spiking dilution
errors.
7.5 Set up instrumentation and hydride generation apparatus and fill
reagent containers. The sample and blank flows should be set around 4.2 mL/min,
and the borohydride flow around 2.1 mL/min. The argon carrier gas flow is
adjusted to about 200 mL/min. For the AA, use the 196.0-nm wavelength and 2.0-nm
slit width without background correction. Begin all flows and allow 10 minutes
for warm-up.
7.6 Place sample feed line into a prepared sample solution and start pump
to begin hydride generation. Wait for a maximum steady-state signal on the
strip-chart recorder. Switch to blank sample and watch for signal to decline to
baseline before switching to the next sample and beginning the next analysis.
Run standards first (low to high), then unknowns. Include appropriate QA/QC
solutions, as required. Prepare calibration curves and convert absorbances to
concentration. See following analytical flowchart.
CAUTION: The hydride of selenium 1s very toxic. Precautions must be
taken to avoid Inhaling the gas.
7.7 If the method of standard additions was employed, plot the measured
concentration of the spiked samples and unspiked sample versus the spiked
concentrations. The spiked concentration axis intercept will be the method of
standard additions concentration. If the plot does not result in a straight
line, a nonlinear interference is present. This problem can sometimes be
overcome by dilution or addition of other reagents if there is some knowledge
about the waste. If the method of standard additions was not required, then the
concentration is determined from a standard calibration curve.
7742-6 Revision 0
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8.0 QUALITY CONTROL
8.1 Refer to Section 8.0 of Method 7000A.
9.0 METHOD PERFORMANCE
9.1 The relative standard deviation obtained by a single laboratory for
7 replicates of a contaminated soil was 18% for selenium at 8.2 ug/L in solution.
10.0 REFERENCES
1. Methods for Chemical Analysis of Water and Wastes, EPA-600/4-82-055,
December 1982, Method 206.3.
2. "Evaluation of Hydride Atomic Absorption Methods for Antimony, Arsenic,
Selenium, and Tin", an EMSL-LV internal report under Contract 68-03-3249,
Job Order 70.16, prepared for T. A. Hinners by D. E. Dobb, and J. D.
Lindner of Lockheed Engineering and Sciences Co., and L. V. Beach of the
Varian Corporation.
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METHOD 7742
SELENIUM (ATOMIC ABSORPTION, GASEOUS BOROHYDRIDE)
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METHOD 3510B
SEPARATORY FUNNEL LIQUID-LIQUID EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 This method describes a procedure for isolating organic compounds from
aqueous samples. The method also describes concentration techniques suitable
for preparing the extract for the appropriate determinative methods described
in Section 4.3 of Chapter Four.
1.2 This method is applicable to the isolation and concentration of water-
insoluble and slightly water-soluble organics in preparation for a variety of
chromatographic procedures.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample, usually 1 liter, at a specified pH (see
Table 1), is serially extracted with methylene chloride using a separatory
funnel. The extract is dried, concentrated, and, as necessary, exchanged into
a solvent compatible with the cleanup or determinative method to be used (see
Table 1 for appropriate exchange solvents).
3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 Under basic extraction conditions required to separate analytes for
the packed columns of Method 8250, the decomposition of some analytes has been
demonstrated. Organochlorine pesticides may dechlorinate, phthalate esters may
exchange, and phenols may react to form tannates. These reactions increase with
increasing pH, and are decreased by the shorter reaction times available in
Method 3510. Methods 3520/8270, 3510/8270, and 3510/8250, respectively, are
preferred over Method 3520/8250 for the analysis of these classes of compounds.
4.0 APPARATUS AND MATERIALS
4.1 Separatory funnel - 2 liter, with Teflon stopcock.
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom and a Teflon stopcock.
NOTE; Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits may
be purchased. Use a small pad of Pyrex glass wool to retain the adsorbent.
Prewash the glass wool pad with 50 ml of acetone followed by 50 mL of
elution solvent prior to packing the column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus (Kontes K-570025-0500).
3510B - 1 Revision 2
November 1990
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4.3.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.6 Vials - 2 ml, glass with Teflon lined screw-caps or crimp tops.
4.7 pH indicator paper - pH range including the desired extraction pH.
4.8 Erlenmeyer flask - 250 ml.
4.9 Syringe - 5 ml.
4.10 Graduated cylinder - 1 liter.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination. Reagents should be stored
in glass to prevent the leaching of contaminants from plastic containers.
5.2 Organic-free reagent water - All references to water in this method
refer to.organic-free reagent water, as defined in Chapter One.
5.3 Sodium hydroxide solution (ION), NaOH. Dissolve 40 g NaOH in organic-
free reagent water and dilute to 100 ml.
5.4 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
3510B - 2 Revision 2
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5.5 Sulfuric acid solution (1:1 v/v), H2S04. Slowly add 50 ml of H2S04
(sp. gr. 1.84) to 50 mL of organic-free reagent water.
5.6 Extraction/exchange solvents
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, CH3CH(OH)CH3 - Pesticide quality or equivalent.
5.6.4 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.5 Acetonitrile, CH3CN - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Using a 1 liter graduated cylinder, measure 1 liter (nominal) of
sample and transfer it quantitatively to the separatory funnel. If high
concentrations are anticipated, a smaller volume may be used and then diluted
with organic-free reagent water to 1 liter. Add 1.0 mL of the surrogate
standards to all samples, spikes, and blanks (see Method 3500 and the
determinative method to be used, for details on the surrogate standard solution
and the matrix spike solution). For the sample in each analytical batch selected
for spiking, add 1.0 mL of the matrix spiking standard. For base/neutral-acid
analysis, the amount added of the surrogates and matrix spiking compounds should
result in a final concentration of 100 ng//iL of each base/neutral analyte and
200 ng//uL of each acid analyte in the extract to be analyzed (assuming a 1 ^L
injection). If Method 3640, Gel-Permeation Cleanup, is to be used, add twice
the volume of surrogates and matrix spiking compounds since half the extract is
lost due to loading of the GPC column.
7.2 Check the pH of the sample with wide-range pH paper and, if necessary,
adjust the pH to that indicated in Table 1 for the specific determinative method
that will be used to analyze the extract.
7.3 Add 60 mL of methylene chloride to the separatory funnel.
7.4 Seal and shake the separatory funnel vigorously for 1-2 minutes with
periodic venting to release excess pressure.
NOTE: Methylene chloride creates excessive pressure very rapidly; therefore,
initial venting should be done immediately after the separatory funnel has
been sealed and shaken once. Venting of the separatory funnel should be
into a hood to avoid needless exposure of the analyst to solvent vapors.
3510B - 3
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7.5 Allow the organic layer to separate from the water phase for a minimum
of 10 minutes. If the emulsion interface between layers is more than one-third
the size of the solvent layer, the analyst must employ mechanical techniques to
complete the phase separation. The optimum technique depends upon the sample
and may include stirring, filtration of the emulsion through glass wool,
centrifugation, or other physical methods. Collect the solvent extract in an
Erlenmeyer flask. If the emulsion cannot be broken (recovery of < 80% of the
methylene chloride, corrected for the water solubility of methylene chloride),
transfer the sample, solvent, and emulsion into the extraction chamber of a
continuous extractor and proceed as described in Method 3520, Continuous Liquid-
Liquid Extraction.
7.6 Repeat the extraction two more times using fresh portions of solvent
(Sections 7.3 through 7.5). Combine the three solvent extracts.
•
7.7 If further pH adjustment and extraction is required, adjust the pH
of the aqueous phase to the desired pH indicated in Table 1. Serially extract
three times with 60 mL of methylene chloride, as outlined in Sections 7.3
through 7.5. Collect and combine the extracts and label the combined extract
appropriately.
7.8 If performing GC/MS analysis (Method 8270), the acid/neutral and base
extracts may be combined prior to concentration. However, in some situations,
separate concentration and analysis of the acid/neutral and base extracts may
be preferable (e.g. if for regulatory purposes the presence or absence of
specific acid/neutral or base compounds at low concentrations must be determined,
separate extract analyses may be warranted).
7.9 Perform the concentration using the Kuderna-Danish (K-D) Technique
(Sections 7.10.1 through 7.10.4).
7.10 K-D Technique
7.10.1 Assemble a Kuderna-Danish (K-D) concentrator by attaching
a 10 mL concentrator tube to a 500 mL evaporation flask. Dry the extract
by passing it through a drying column containing about 10 cm of anhydrous
sodium sulfate. Collect the dried extract in a K-D concentrator. Rinse
the Erlenmeyer flask, which contained the solvent extract, with 20-30 mL
of methylene chloride and add it to the column to complete the quantitative
transfer.
7.10.2 Add one or two clean boiling chips to the flask and attach
a three ball Snyder column. Prewet the Snyder column by adding about 1
mL of methylene chloride to the top of the column. Place the K-D apparatus
on a hot water bath (15-20°C above the boiling point of the solvent) so
that the concentrator tube is partially immersed in the hot water and the
entire lower rounded surface of the flask is bathed with hot vapor. Adjust
the vertical position of the apparatus and the water temperature as
required to complete the concentration in 10-20 minutes. At the proper
rate of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid reaches
1 mL, remove the K-D apparatus from the water bath and allow it to drain
and cool for at least 10 minutes.
3510B - 4 Revision 2
November 1990
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7.10.3 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add 50 mL of the exchange solvent,
a new boiling chip, and reattach the Snyder column. Concentrate the
extract, as described in Section 7.11, raising the temperature of the water
bath, if necessary, to maintain proper distillation.
7.10.4 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 mL of methylene chloride or
exchange solvent. If sulfur crystals are a problem, proceed to Method 3660
for cleanup. The extract may be further concentrated by using the
technique outlined in Section 7.11 or adjusted to 10.0 ml with the solvent
last used.
7.11 If further concentration is indicated in Table 1, either the micro-
Snyder column technique (7.11.1) or nitrogen blowdown technique (7.11.2) is used
to adjust the extract to the final volume required.
7.11.1 Micro-Snyder Column Technique
e
7.11.1.1 If further concentration is indicated in Table 1,
add another clean boiling chip to the concentrator tube and attach
a two ball micro-Snyder column. Prewet the column by adding 0.5 ml
of methylene chloride or exchange solvent to the top of the column.
Place the K-D apparatus in a hot water bath so that the concentrator
tube is partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 5-10 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus from the water bath and
allow it to drain and cool for at least 10 minutes. Remove the
Snyder column and rinse the flask and its lower joints into the
concentrator tube with 0.2 ml of extraction solvent. Adjust the
final volume to 1.0-2.0 ml, as indicated in Table 1, with solvent.
7.11.2 Nitrogen Blowdown Technique
7.11.2.1 Place the concentrator tube in a warm bath (35°C) and
evaporate the solvent volume to 0.5 mL using a gentle stream of
clean, dry nitrogen (filtered through a column of activated carbon).
CAUTION: New plastic tubing must not be used between the carbon trap and the
sample, since it may introduce interferences.
7.11.2.2 The internal wall of the tube must be rinsed down
several times with methylene chloride or appropriate solvent during
the operation. During evaporation, the tube solvent level must be
positioned to avoid water condensation. Under normal procedures,
the extract must not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below 1 ml, semi volatile
analytes may be lost.
3510B - 5
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7.12 The extract may now be analyzed for the target analytes using the
appropriate determinative technique(s) (see Section 4.3 of this Chapter). If
analysis of the extract will not be performed immediately, stopper the
concentrator tube and store refrigerated. If the extract will be stored longer
than 2 days it should be transferred to a vial with a Teflon lined screw-cap or
crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3510B - 6 Revision 2
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TABLE 1.
SPECIFIC EXTRACTION CONDITIONS FOR VARIOUS DETERMINATIVE METHODS
Determinative
method
8040
8060
8061
8070
8080
8081
8090
8100
8110
8120
8121
8140
8141
8250bc
8270bd
8310
8321
8410
Initial
extraction
PH
<2
as received
as received
as received
5-9
5-9
5-9
as received
as received
as received
as received
6-8
as received
>11
>11
as received
as received
as received
Secondary
extraction
PH
none
none
none
none
none
none
none
none
none
none
none
none
none
<2
<2
none
none
none
Exchange
solvent
required
for
analysis
2-propanol
hexane
hexane
methanol
hexane
hexane
hexane
none
hexane
hexane
hexane
hexane
hexane
none
none
acetonitrile
methanol
methyl ene chloride
Exchange
solvent
required
for
cleanup
hexane
hexane
hexane
methyl ene chloride
hexane
hexane
hexane
cyclohexane
hexane
hexane
hexane
hexane
hexane
_
-
-
-
methyl ene chloride
Vol ume
of extract
required
for
cleanup (ml)
1.0
2.0
2.0
2.0
10.0
10.0
2.0
2.0
2.0
2.0
2.0
10.0
10.0
-
-
-
-
10.0
Final
extract
vol ume
for
analysis
1.0, 10
10.0
10.0
10.0
10.0
10.0
1.0
1.0
10.0
1.0
1.0
10.0
10.0
1.0
1.0
1.0
1.0
(ml)
.Oa
0.0 (dry)
a Phenols may be analyzed, by Method 8040, using a 1.0 ml 2-propanol extract by GC/FID. Method 8040 also contains an optional
derivatization procedure for phenols which results in a 10 ml hexane extract to be analyzed by GC/ECD.
b The specificity of GC/MS may make cleanup of the extracts unnecessary. Refer to Method 3600 for guidance on the cleanup
procedures available if required.
c Loss of phthalate esters, organochlorine pesticides and phenols can occur under these extraction conditions (see Section 3.2).
d Extraction pH sequence may be reversed to better separate acid and neutral waste components. Excessive pH adjustments may
result in the loss of some analytes (see Section 3.2).
3510B - 7
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METHOD 3510B
SEPARATORY FUNNEL LIQUID-LIQUID EXTRACTION
Yes
7.1 Add
surrogate
slands . to al 1
samples, spikes
and blanks
7.7 Collect
and combine
extracts and
label
7.8
CC/MS
analysis
(Method 8250
8270) being
performed?
7.8 Combine
base/neutral
extracts
prior to
concentration
7.2. Check
and adjust pH
7.3-7.6
Extract 3
times
7.9-7.11
Concentrate
extract
7.12
Ready for
7.7 Further
extractions
required?
3510B - 8
Revision 2
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METHOD 3520B
CONTINUOUS LIQUID-LIQUID EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 This method describes a procedure for isolating organic compounds
from aqueous samples. The method also describes concentration techniques
suitable for preparing the extract for the appropriate determinative steps
described in Section 4.3 of Chapter Four.
1.2 This method is applicable to the isolation and concentration of water-
insoluble and slightly soluble organics in preparation for a variety of
chromatographic procedures.
1.3 Method 3520 is designed for extraction solvents with greater density
than the sample. Continuous extraction devices are available for extraction
solvents that are less dense than the sample. The analyst must demonstrate the
effectiveness of any such automatic extraction device before employing it in
sample extraction.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample, usually 1 liter, is placed into a
continuous liquid-liquid extractor, adjusted, if necessary, to a specific pH
(see Table 1), and extracted with organic solvent for 18-24 hours. The extract
is dried, concentrated, and, as necessary, exchanged into a solvent compatible
with the cleanup or determinative method being employed (see Table 1 for
appropriate exchange solvents).
3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 Under basic extraction conditions required to separate analytes for
the packed columns of Method 8250, the decomposition of some analytes has been
demonstrated. Organochlorine pesticides may dechlorinate, phthalate esters may
exchange, and phenols may react to form tannates. These reactions increase with
increasing pH, and are decreased by the shorter reaction times available in
Method 3510. Methods 3520/8270, 3510/8270, and 3510/8250, respectively, are
preferred over Method 3520/8250 for the analysis of these classes of compounds.
4.0 APPARATUS AND MATERIALS
4.1 Continuous liquid-liquid extractor - Equipped with Teflon or glass
connecting joints and stopcocks requiring no lubrication (Kontes 584200-0000,
584500-0000, 583250-0000, or equivalent).
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom and a Teflon stopcock.
3520B - 1 Revision 2
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NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed
by 50 ml of elution solvent prior to packing the column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.6 Vials - 2 ml, glass with Teflon lined screw-caps or crimp tops.
4.7 pH indicator paper - pH range including the desired extraction pH.
4.8 Heating mantle - Rheostat controlled.
4.9 Syringe - 5 ml.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination. Reagents should be stored
in glass to prevent the leaching of contaminants from plastic containers.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium hydroxide solution (ION), NaOH. Dissolve 40 g NaOH in organic-
free reagent water and dilute to 100 ml.
3520B - 2 Revision 2
November 1990
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5.4 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.5 Sulfuric acid solution (1:1 v/v), H2S04. Slowly add 50 ml of H2S04
(sp. gr. 1.84) to 50 ml of organic-free reagent water.
5.6 Extraction/exchange solvents
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.6.4 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.5 Acetonitrile, CH3CN - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Using a 1 liter graduated cylinder, measure out 1 liter (nominal) of
sample and transfer it quantitatively to the continuous extractor. If high
concentrations are anticipated, a smaller volume may be used and then diluted
with organic-free reagent water to 1 liter. Check the pH of the sample with
wide-range pH paper and adjust the pH, if necessary, to the pH indicated in Table
1 using 1:1 (V/V) sulfuric acid or 10 N sodium hydroxide. Pipet 1.0 mL of the
surrogate standard spiking solution into each sample into the extractor and mix
well. (See Method 3500 and the determinative method to be used, for details on
the surrogate standard solution and the matrix spike solution.) For the sample
in each analytical batch selected for spiking, add 1.0 mL of the matrix spiking
standard. For base/neutral-acid analysis, the amount of the surrogates and
matrix spiking compounds added to the sample should result in a final
concentration of 100 ng//iL of each base/neutral analyte and 200 ng/juL of each
acid analyte in the extract to be analyzed (assuming a 1 pi injection). If
Method 3640, Gel-Permeation Cleanup, is to be used, add twice the volume of
surrogates and matrix spiking compounds since half the extract is lost due to
loading of the GPC column.
7.2 Add 300-500 mL of methylene chloride to the distilling flask. Add
several boiling chips to the flask.
7.3 Add sufficient water to the extractor to ensure proper operation and
extract for 18-24 hours.
3520B - 3 Revision 2
November 1990
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7.4 Allow to cool; then detach the boiling flask. If extraction at a
secondary pH is not required (see Table 1), the extract is dried and concentrated
using one of the techniques referred to in Section 7.7.
7.5 Carefully, while stirring, adjust the pH of the aqueous phase to the
second pH indicated in Table 1. Attach a clean distilling flask containing
500 mL of methylene chloride to the continuous extractor. Extract for 18-24
hours, allow to cool, and detach the distilling flask.
7.6 If performing GC/MS analysis (Method 8270), the acid/neutral and base
extracts may be combined prior to concentration. However, in some situations,
separate concentration and analysis of the acid/neutral and base extracts may
be preferable (e.g. if for regulatory purposes the presence or absence of
specific acid/neutral and base compounds at low concentrations must be
determined, separate extract analyses may be warranted).
7.7 Perform concentration using the Kuderna-Danish (K-D) Technique
(Sections 7.8.1 through 7.8.4).
7.8 K-D Technique
7.8.1 Assemble a Kuderna-Danish (K-D) concentrator by attaching a
10 mL concentrator tube to a 500 mL evaporation flask. Dry the extract
by passing it through a drying column containing about 10 cm of anhydrous
sodium sulfate. Collect the dried extract in a K-D concentrator. Rinse
the flask which contained the solvent extract with 20-30 mL of methylene
chloride and add it to the column to complete the quantitative transfer.
7.8.2 Add one or two clean boiling chips to the flask and attach
a three ball Snyder column. Prewet the Snyder column by adding about 1 mL
of methylene chloride to the top of the column. Place the K-D apparatus
on a hot water bath (15-20°C above the boiling point of the solvent) so
that the concentrator tube is partially immersed in the hot water and the
entire lower rounded surface of the flask is bathed with hot vapor. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete the concentration in 10-20 minutes. At the proper
rate of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid reaches
1 mL, remove the K-D apparatus from the water bath and allow it to drain
and cool for at least 10 minutes. Remove the Snyder column and rinse the
flask and its lower joints into the concentrator tube with 1-2 mL of
extraction solvent.
7.8.3 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add 50 mL of the exchange solvent,
a new boiling chip, and reattach the Snyder column. Concentrate the
extract, as described in Section 7.9, raising the temperature of the water
bath, if necessary, to maintain proper distillation.
7.8.4 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 mL of methylene chloride or
exchange solvent. If sulfur crystals are a problem, proceed to Method
3660 for cleanup. The extract may be further concentrated by using the
3520B - 4 Revision 2
November 1990
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techniques outlined In Section 7.9 or adjusted to 10.0 ml with the solvent
last used.
7.9 If further concentration is indicated in Table 1, either the micro-
Snyder column technique (7.9.1) or nitrogen blowdown technique (7.9.2) is used
to adjust the extract to the final volume required.
7.9.1 Micro-Snyder Column Technique
7.9.1.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro-Snyder column. Prewet
the column by adding 0.5 ml of methylene chloride or exchange solvent
to the top of the column. Place the K-D apparatus in a hot water
bath so that the concentrator tube is partially immersed in the hot
water. Adjust the vertical position of the apparatus and the water
temperature, as required, to complete the concentration in 5-
10 minutes. At the proper rate of distillation the balls of the
column will actively chatter, but the chambers will not flood. When
the apparent volume of liquid reaches 0.5 ml, remove the K-D
apparatus from the water bath and allow it to drain and cool for at
least 10 minutes. Remove the Snyder column, rinse the flask and its
lower joints into the concentrator tube with 0.2 ml of methylene
chloride or exchange solvent, and adjust the final volume to 1.0 to
2.0 ml, as indicated in Table 1, with solvent.
7.9.2 Nitrogen Blowdown Technique
7.9.2.1 Place the concentrator tube in a warm bath (35°C)
and evaporate the solvent volume to 0.5 ml using a gentle stream of
clean, dry nitrogen (filtered through a column of activated carbon).
CAUTION: New plastic tubing must not be used between the carbon trap and the
sample, since it may introduce interferences.
7.9.2.2 The internal wall of the tube must be rinsed down
several times with methylene chloride or appropriate solvent during
the operation. During evaporation, the tube solvent level must be
positioned to avoid water condensation. Under normal procedures,
the extract must not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below 1 ml, semi volatile
analytes may be lost.
7.10 The extract may now be analyzed for the target analytes using the
appropriate determinative technique(s) (see Section 4.3 of this Chapter). If
analysis of the extract will not be performed immediately, stopper the
concentrator tube and store refrigerated. If the extract will be stored longer
than 2 days it should be transferred to a vial with a Teflon lined screw-cap or
crimp top, and labeled appropriately.
3520B - 5
Revision 2
November 1990
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample-preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3520B - 6 Revision 2
November 1990
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TABLE 1.
SPECIFIC EXTRACTION CONDITIONS FOR VARIOUS DETERMINATIVE METHODS
Determinative
method
8040
8060
8061
8070
8080
8081
8090
8100
8110
8120
8121
8140
8141
8250b'c
8270b'd
8310
8321
8410
Initial
extraction
pH
<2
as received
as received
as received
5-9
5-9
5-9
as received
as received
as received
as received
6-8
as received
>11
<2
as received
as received
as received
Secondary
extraction
PH
none
none
none
none
none
none
none
none
none
none
none
none
none
<2
>11
none
none
none
Exchange
solvent
required
for
analysis
2-propanol
hexane
hexane
methanol
hexane
hexane
hexane
none
hexane
hexane
hexane
hexane
hexane
none
none
acetonitrile
methanol
methyl ene chloride
Exchange
solvent
required
for
cleanup
hexane
hexane
hexane
methyl ene chloride
hexane
hexane
hexane
cyclohexane
hexane
hexane
hexane
hexane
hexane
-
-
-
-
methyl ene chloride
Vol ume
of extract
required
for
cleanup (ml)
1.0
2.0
2.0
2.0
10.0
10.0
2.0
2.0
2.0
2.0
2.0
10.0
10.0
-
-
-
-
10.0
Final
extract
vol ume
for
analysis (mL)
1.0,10.0"
10.0
10.0
10.0
10.0
10.0
1.0
1.0
10.0
1.0
1.0
10.0
10.0
1.0
1.0
1.0
1.0
0.0 (dry)
a Phenols may be analyzed, by Method 8040, using a 1.0 mL 2-propanol extract by GC/FID. Method 8040 also contains an optional
derivatization procedure for phenols which results in a 10 mL hexane extract to be analyzed by GC/ECD.
b The specificity of GC/MS may make cleanup of the extracts unnecessary. Refer to Method 3600 for guidance on the cleanup
procedures available if required.
c Loss of phthalate esters, organochlorine pesticides and phenols can occur under these extraction conditions (see Section 3.2).
d If further separation of major acid and neutral components is required, Method 3650, Acid-Base Partition Cleanup, is
recommended. Reversal of the Method 8270 pH sequence is not recommended as analyte losses are more severe under the base
first continuous extraction (see Section 3.2).
3520B - 7
Revision 2
November 1990
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METHOD 3520B
CONTINUOUS LIQUID-LIQUID EXTRACTION
c
Start
7.1 Add appropriate
surrogate and
matrix spiking
solutions
7 . 2 Add methylene
chloride to
distilling flask
7 . 3 Add reagent
Hater to extractor;
extract for 18-24
hours
7.5 Adjust pH of
aqueous phase;
extract for 18-24
hours with clean
flask
Yes
7.6 Combine acid
and base/neutral
extracts prior to
concentration
7.7-7.8 Concentrate
extract
Yes
7.8.3 Add exchange
solvent:
concentrate extract
7 9 Further
concentrate extract
if necessary;
adjust final volume
7.10 Analyze using
organic techniques
8000
Series
Methods
3520B - 8
Revision 2
November 1990
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METHOD 3540B
SOXHLET EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 Method 3540 Is a procedure for extracting nonvolatile and semi-
volatile organic compounds from solids such as soils, sludges, and wastes.
The Soxhlet extraction process ensures intimate contact of the sample matrix
with the extraction solvent.
1.2 This method is applicable to the isolation and concentration of
water insoluble and slightly water soluble organics in preparation for a
variety of chromatographic procedures.
2.0 SUMMARY OF METHOD
2.1 The solid sample is mixed with anhydrous sodium sulfate, placed in
an extraction thimble or between two plugs of glass wool, and extracted using
an appropriate solvent in a Soxhlet extractor. The extract is then dried,
concentrated, and, as necessary, exchanged into a solvent compatible with the
cleanup or determinative step being employed.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
4.0 APPARATUS AND MATERIALS
4.1 Soxhlet extractor - 40 mm ID, with 500 mL round bottom flask.
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom.
NOTE; Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 mL of acetone followed by
50 mL of elution solvent prior to packing the column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus
4.3.1 Concentrator tube - 10 mL, graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation
of extracts.
4.3.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
3540B - 1 Revision 2
November 1990
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4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh
(silicon carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The bath should be used in a hood.
4.6 Vials - Glass, 2 ml capacity, with Teflon lined screw or crimp top.
4.7 Glass or paper thimble or glass wool - Contaminant free.
4.8 Heating mantle - Rheostat controlled.
4.9 Disposable glass pasteur pipet and bulb.
4.10 Apparatus for determining percent dry weight.
4.10.1 Oven - Drying.
4.10.2 Desiccator.
4.10.3 Crucibles - Porcelain or disposable aluminum.
4.11 Apparatus for grinding
4.12 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may
be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na?S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene
chloride, a method blank must be analyzed, demonstrating that there is no
interference from the sodium sulfate.
3540B - 2 Revision 2
November 1990
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5.4 Extraction solvents
5.4.1 Soil/sediment and aqueous sludge samples shall be extracted
using either of the following solvent systems:
5.4.1.1 Acetone/Hexane (1:1) (v/v), CH3COCH3/CH6H14.
Pesticide quality or equivalent.
NOTE: This solvent system has lower disposal cost and lower toxicity.
5.4.1.2 Methylene chloride/Acetone (1:1 v/v),
CH2C12/CH3COCH3. Pesticide quality or equivalent.
5.4.2 Other samples shall be extracted using the following:
5.4.2.1 Methylene chloride, CH2C12. Pesticide quality or
equivalent.
5.4.2.2 Toluene/Methanol (10:1) (v/v), C6H5CH3/CH3OH.
Pesticide quality or equivalent.
5.5 Exchange solvents
5.5.1 Hexane, C6H14. Pesticide quality or equivalent.
5.5.2 2-Propanol, (CH3)2CHOH. Pesticide quality or equivalent.
5.5.3 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.5.4 Acetonitrile, CH3CN. Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analysis,
Section 4.1.
7.0 PROCEDURE
7.1 Sample Handling
7.1.1 Sediment/soil samples - Decant and discard any water layer
on a sediment sample. Mix sample thoroughly, especially composited
samples. Discard any foreign objects such as sticks, leaves, and rocks.
7.1.2 Waste samples - Samples consisting of multiphases must be
prepared by the phase separation method in Chapter Two before
extraction. This procedure is for solids only.
7.1.3 Dry waste samples amenable to grinding - Grind or otherwise
subdivide the waste so that it either passes through a 1 mm sieve or can
be extruded through a 1 mm hole. Introduce sufficient sample into the
grinding apparatus to yield at least 10 g after grinding.
3540B - 3 Revision 2
November 1990
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7.1.4 Gummy, fibrous or oily materials not amenable to grinding
should be cut, shredded, or otherwise broken up to allow mixing, and
maximum exposure of the sample surfaces for extraction. The
professional judgment of the analyst is required for handling these
difficult matrixes.
7.2 Determination of sample % dry weight - In certain cases, sample
results are desired based on dry weight basis. When such data is desired, a
portion of sample for this determination should be weighed out at the same
time as the portion used for analytical determination.
WARNING: The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from a heavily
contaminated hazardous waste sample.
7.2.1 Immediately after weighing the sample for extraction, weigh
5-10 g of the sample into a tared crucible. Determine the % dry weight
of the sample by drying overnight at 105°C. Allow to cool in a
desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.3 Blend 10 g of the solid sample with 10 g of anhydrous sodium
sulfate and place in an extraction thimble. The extraction thimble must drain
freely for the duration of the extraction period. A glass wool plug above and
below the sample in the Soxhlet extractor is an acceptable alternative for the
thimble. Add 1.0 mL of the surrogate standard spiking solution onto the
sample (see Method 3500 for details on the surrogate standard and matrix
spiking solutions). For the sample in each analytical batch selected for
spiking, add 1.0 mL of the matrix spiking standard. For base/neutral-acid
analysis, the amount added of the surrogates and matrix spiking compounds
should result in a final concentration of 100 ng//iL of each base/neutral
analyte and 200 ng//zL of each acid analyte in the extract to be analyzed
(assuming a 1 nl injection). If Method 3640, Gel Permeation Chromatography
Cleanup, is to be used, add twice the volume of surrogates and matrix spiking
compounds since half the extract is lost due to loading of the GPC column.
7.4 Place approximately 300 ml of the extraction solvent (Section 5.4)
into a 500 mL round bottom flask containing one or two clean boiling chips.
Attach the flask to the extractor and extract the sample for 16-24 hours at
4-6 cycles/hr.
7.5 Allow the extract to cool after the extraction is complete.
7.6 Assemble a Kuderna-Danish (K-D) concentrator by attaching a 10 mL
concentrator tube to a 500 mL evaporation flask.
7.7 Dry the extract by passing it through a drying column containing
about 10 cm of anhydrous sodium sulfate. Collect the dried extract in a K-D
concentrator. Wash the extractor flask and sodium sulfate column with 100 to
125 mL of extraction solvent to complete the quantitative transfer.
3540B - 4 Revision 2
November 1990
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7.8 Add one or two clean boiling chips to the flask and attach a three
ball Snyder column. Prewet the Snyder column by adding about 1 ml of
methylene chloride to the top of the column. Place the K-D apparatus on a hot
water bath (15-20°C above the boiling point of the solvent) so that the
concentrator tube is partially immersed in the hot water and the entire lower
rounded surface of the flask is bathed with hot vapor. Adjust the vertical
position of the apparatus and the water temperature, as required, to complete
the concentration in 10-20 minutes. At the proper rate of distillation the
balls of the column will actively chatter, but the chambers will not flood.
When the apparent volume of liquid reaches 1-2 ml, remove the K-D apparatus
from the water bath and allow it to drain and cool for at least 10 minutes.
7.9 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add approximately 50 mL of the exchange
solvent and a new boiling chip, and reattach the Snyder column. Concentrate
the extract as described in Section 7.8, raising the temperature of the water
bath, if necessary, to maintain proper distillation. When the apparent volume
again reaches 1-2 ml, remove the K-D apparatus from the water batch and allow
it to drain and cool for at least 10 minutes.
7.10 Remove the Snyder column and rinse the flask and its lower joints
into the concentrator tube with 1-2 ml of methylene chloride or exchange
solvent. If sulfur crystals are a problem, proceed to Method 3660 for
cleanup. The extract may be further concentrated by using the techniques
described in Section 7.11 or adjusted to 10.0 ml with the solvent last used.
7.11 If further concentration is indicated in Table 1, either micro
Snyder column technique (Section 7.11.1) or nitrogen blowdown technique
(Section 7.11.2) is used to adjust the extract to the final volume required.
7.11.1 Micro Snyder Column Technique
7.11.1.1 Add another one or two clean boiling chips to
the concentrator tube and attach a two ball micro Snyder column.
Prewet the column by adding about 0.5 ml of methylene chloride or
exchange solvent to the top of the column. Place the K-D
apparatus in a hot water bath so that the concentrator tube is
partially immersed in the hot water. Adjust the vertical position
of the apparatus and the water temperature, as required, to
complete the concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus from the water bath and
allow it to drain and cool for at least 10 minutes. Remove the
Snyder column and rinse the flask and its lower joints with about
0.2 ml of solvent and add to the concentrator tube. Adjust the
final volume to 1.0-2.0 ml, as indicated in Table 1, with solvent.
7.11.2 Nitrogen Blowdown Technique
7.11.2.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the
required level using a gentle stream of clean, dry nitrogen
(filtered through a column of activated carbon).
3540B - 5 Revision 2
November 1990
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CAUTION; Do not use plasticized tubing between the carbon trap and the
sample.
7.11.2.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be
positioned to prevent water from condensing into the sample (i.e.,
the solvent level should be below the level of the water bath).
Under normal operating conditions, the extract should not be
allowed to become dry.
CAUTION; When the volume of solvent is reduced below 1 ml, semivolatile
analytes may be lost.
7.12 The extracts obtained may now be analyzed for the target analytes
using the appropriate organic technique(s) (see Section 4.3 of this Chapter).
If analysis of the extract will not be performed immediately, stopper the
concentrator tube and store in a refrigerator. If the extract will be stored
longer than 2 days, it should be transferred to a vial with a Teflon lined
screw cap or crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for
the Analysis of Pollutants Under the Clean Water Act; Final Rule and
Interim Final Rule and Proposed Rule," October 26, 1984.
3540B - 6 Revision 2
November 1990
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TABLE 1.
SPECIFIC EXTRACTION CONDITIONS FOR VARIOUS DETERMINATIVE METHODS
Determinative
method
8040a
8060
8061
8070
8080
8081
8090
8100
8110
8120
8121
8140
8141
8250a'c
8270C
8310
8321
8410
Extraction
PH
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
Exchange
solvent
required
for
analysis
2-propanol
hexane
hexane
methanol
hexane
hexane
hexane
none
hexane
hexane
hexane
hexane
hexane
none
none
acetonitrile
methanol
methyl ene chloride
Exchange Volume
solvent of extract
required required
for for
cleanup cleanup (ml)
hexane
hexane
hexane
methyl ene chloride
hexane
hexane
hexane
cyclohexane
hexane
hexane
hexane
hexane
hexane
--
--
--
--
methylene chloride
1.0
2.0
2.0
2.0
10.0
10.0
2.0
2.0
2.0
2.0
2.0
10.0
10.0
--
--
--
--
10.0
Final
extract
vol ume
for
analysis (ml)
1.0, 10.0"
10.0
10.0
10.0
10.0
10.0
1.0
1.0
10.0
1.0
1.0
10.0
10.0
1.0
1.0
1.0
1.0
0.0 (dry)
a To obtain separate acid and base/neutral extracts, Method 3650 should be performed following
concentration of the extract to 10.0 mL.
b Phenols may be analyzed by Method 8040, using a 1.0 ml 2-propanol extract by GC/FID. Method 8040 also
contains an optical derivatization procedure for phenols which results in a 10 mL hexane extract to be
analyzed by GC/ECD.
The specificity of GC/MS may make cleanup of the extracts unnecessary.
on the cleanup procedures available if required.
3540B - 7
Refer to Method 3600 for guidance
Revision 2
November 1990
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METHOD 3540B
SOXHLET EXTRACTION
c
Start
7.1 Use appropriate
sample handling
technique
7.2 Determine
sample % dry weight
7.3 Add appropriate
surrogate and
matrix spiking
s tandards
7.4 Add extraction
solvent to flask:
extract for 16-24
hours
7.5 Cool extract
7.6 Assemble K-D
concent rator
7.7 Dry and collect
extract in K-D
concentrator
7.8 Concentrate
using Snyder column
and K-D apparatus
7.12 Analyze using
organic techniques
8000
Series
Methods
7.9 Add exchang
solvent,
reconcentrate
extract
3540B - 8
Revision 2
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METHOD 3541
AUTOMATED SOXHLET EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 This method extracts polychlorinated biphenyls (PCBs) from soil,
sediment, sludges, and waste solids. The method uses a commercially available,
unique, three stage extraction system to achieve PCB recovery comparable to
Method 3540, but in a much shorter time. The two differences between this
extraction method and Method 3540 are Sections 7.10 and 7.11. In the initial
extraction stage, the sample-loaded extraction thimble is immersed into the
boiling solvent. This ensures very rapid intimate contact between the specimen
and solvent and rapid recovery of the PCB. In the second stage the thimble is
elevated above the solvent, and is rinse-extracted as in Method 3540. In the
third stage, the solvent is evaporated, as would occur in the Kuderna-Danish
(K-D) concentration step in Method 3540. The concentrated extract is then ready
for measurement of the PCB concentrations using Method 8080 or 8081.
1.2 The method is applicable to the extraction and concentration of water
insoluble or slightly water soluble PCBs in preparation for gas chromatographic
measurement of the PCB concentration of the sample.
2.0 SUMMARY OF METHOD
2.1 After air drying of the samples (EPA Method 600/4-81-055, Interim
Methods for the Sampling and Analysis of Priority Pollutants in Sediments and
Fish Tissue, Section 3.1.3), the sample is ground to 100-200 mesh (150 /im to
75 /itm). The powdered sample is extracted using 1:1 acetonerhexane as the
extraction solvent, as detailed below. The extract is then concentrated and
exchanged into pure hexane prior, to final gas chromatographic PCB measurement.
2.2 This method is applicable to soils, clays, wastes and sediments
containing from 1 to 50 /ug of PCB per gram of sample. It has been statistically
evaluated at 5 and 50 /Ltg/g of Aroclors 1254 and 1260, and found to be equivalent
to Method 3540 (Soxhlet Extraction). Higher concentrations of PCB are measured
following volumetric dilution with hexane.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 If cleanup is necessary, the Florisil and/or sulfur procedures may
be employed. In such cases, proceed with Method 3620, followed by, if necessary,
Method 3660, using the 10 ml hexane extracts obtained from Section 7.14.
3541 - 1 Revision 0
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4.0 APPARATUS AND MATERIALS
4.1 Automated Soxhlet Extraction System - With controlled, heated oil
bath (Soxtec, or equivalent). See Figure 1. Apparatus is used in a hood.
4.2 Cellulose extraction thimbles - Contamination free (Fisher No.
1522-0018, or equivalent).
4.3 Syringe - 5 ml.
4.4 Apparatus for Determining Percent Dry Weight
4.4.1 Drying Oven.
4.4.2 Desiccator.
4.4.3 Crucibles, porcelain.
4.4.4 Balance, analytical.
4.5 Apparatus for Grinding - If the sample will not pass through a 1 mm
standard sieve or cannot be extruded through a 1 mm opening, it should be
processed into a homogeneous sample that meets these requirements. Gummy,
fibrous, or oily materials may be mixed with anhydrous sodium sulfate to improve
grinding efficiency. Disassemble grinder between samples, according to
manufacturer's instructions, and clean with soap and water, followed by acetone
and hexane rinses.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.4 Acetone/hexane (1:1 (v/v)), CH3COCH3/C6H14. Pesticide quality or
equivalent.
5.5 Hexane, C6H14. Pesticide quality or equivalent.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample handling
7.1.1 Sediment/soil samples - Decant and discard any water layer on
a sediment sample. Mix sample thoroughly, especially composited samples.
Discard any foreign objects such as sticks, leaves, and rocks. Air dry
the sample at room temperature for 48 hours in a glass tray or on hexane
cleaned aluminum foil, or dry the sample by mixing with anhydrous sodium
sulfate until a free-flowing powder is obtained.
7.1.2 Dry waste samples amenable to grinding - Grind or otherwise
subdivide the waste so that it either passes through a 1 mm sieve or can
be extruded through a 1 mm hole. Introduce sufficient sample into the
grinding apparatus to yield at least 20 g after grinding.
7.1.2.1 Gummy, fibrous or oily materials not amenable to
grinding should be cut, shredded, or otherwise broken up to allow
mixing, and maximum exposure of the sample surfaces for extraction.
The professional judgment of the analyst is required for handling
these difficult matrices.
7.1.3 Waste samples - Samples consisting of multiple phases must be
prepared by the phase separation in Chapter Two before extraction. This
procedure is for solids gnly.
7.2 Determination of sample percent dry weight - In certain cases, sample
results are desired based on dry weight basis. When such data is desired, a
portion of sample for this determination should be weighed out at the same time
as the portion used for analytical determination.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.
7.2.1 Immediately after weighing the sample for extraction, weigh
5-10 g of the sample into a tared crucible. Determine the % dry weight
of the sample by drying overnight at 105°C. Allow to cool in a desiccator
before weighing:
% dry weight = q of dry sample x 100
g of sample
7.3 Grind sufficient dried sample from Section 7.1.2 or 7.1.3 to yield
20 g of powder. After grinding, samples should pass through a 10 mesh sieve.
7.4 Weigh 10 g of sample into extraction thimbles.
3541 - 3 Revision 0
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7.5 Check the oil level in the automated Soxhlet unit and add oil if
needed. See service manual for details.
7.6 Press the "MAINS" button, observe that the switch lamp is now "ON".
7.7 Open the cold water tap for the reflux condensers. Adjust the flow
to 2 L/min to prevent solvent loss through the condensers.
7.8 Transfer weighed samples into the condensers. Raise the knob to the
"BOILING" position. The magnet will now fasten to the thimble. Lower the knob
to the "RINSING" position. The thimble will now hang just below the condenser
valve.
7.9 Insert the extraction cups containing boiling chips, and load each
with 50 mL of solvent (1:1 (v/v) hexaneracetone). Using the cup holder, lower
the locking handle, ensuring that the safety catch engages. The cups are now
clamped into position.
7.10 Move the extraction knobs to the "BOILING" position. The thimbles
are now immersed in solvent. Set the timer for 60 minutes. The condenser valves
must be in the "OPEN" position. Run for the preset time.
7.11 Move the extraction knobs to the "RINSING" position. The thimbles
will now hang above the solvent surface. Set timer for 60 minutes. Condenser
valves are still open. Run for the preset time.
7.12 After rinse time has elapsed, close the condenser valves by turning
each a quarter-turn, clockwise.
7.13 When all but 1 or 2 mL of solvent have been collected, open the
system and remove the cups. Let the solvent air-evaporate from this point.
7.14 Quantitatively transfer contents of cups to 10 mL collection vials
using hexane. Dilute to volume.
NOTE: The recovery solvent volume can be adjusted by adding solvent at the top
of the condensers. For more details concerning use of the extractor, see
the operating manual for the automated extraction system.
7.15 Shutdown
7.15.1 Turn "OFF" main switch.
7.15.2 Turn "OFF" cold water tap.
7.15.3 Check to ensure that all condensers are free of solvent.
7.16 The extract is now ready for cleanup or analysis, depending on the
extent of interfering co-extractives.
3541 - 4 Revision 0
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8.0 QUALITY CONTROL
8.1 Any reagent blanks or matrix spike samples should be subjected to
exactly the same analytical procedures as those used on actual samples.
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
8.3 The analyst must prepare method blanks to check for cross-
contamination and routinely check the integrity of the instrument seals.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
2. Stewart, J. "EPA Verification Experiment for Validation of the SOXTEC"
PCB Extraction Procedure"; Oak Ridge National Laboratory, Oak Ridge, TN,
37831-6138; October 1988.
3541 - 5 Revision 0
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Figure 1
Automated Soxhlet Extraction System
Condenser
Thimble
Glass Wool Plug
Sample
Aluminum beaker (cup)
Hot plate
3541 - 6
Revision 0
November 1990
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METHOD 3541
AUTOMATED SOXHLET EXTRACTION
7.3 Grind
dried
sample.
7.5 Check
oil level in
Soxhlet unit.
7.8 Transfer
samples into
condensers.
Adjust position
of magnet and
thimble.
Stop
3541 - 7
Revision 0
November 1990
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METHOD 3550B
ULTRASONIC EXTRACTION
See DISCLAIMER-1. See manufacturer's specifications for operational settings.
1.0 SCOPE AND APPLICATION
1.1 Method 3550 is a procedure for extracting nonvolatile and semi-
volatile organic compounds from solids such as soils, sludges, and wastes.
The ultrasonic process ensures intimate contact of the sample matrix with the
extraction solvent.
1.2 The method is divided into two sections, based on the expected
concentration of organics in the sample. The low concentration method
(individual organic components of < 20 mg/Kg) uses a larger sample size and a
more rigorous extraction procedure (lower concentrations are more difficult to
extract). The medium/high concentration method (individual organic components
of > 20 mg/Kg) is much simpler and therefore faster.
1.3 It is highly recommended that the extracts be cleaned up prior to
analysis. See Chapter Four (Cleanup), Section 4.2.2, for applicable methods.
2.0 SUMMARY OF METHOD
2.1 Low concentration method - A 30 g sample is mixed with anhydrous
sodium sulfate to form a free flowing powder. This is solvent extracted three
times using ultrasonic extraction. A portion of the extract is removed for
cleanup and/or analysis.
2.2 Medium/high concentration method - A 2 g sample is mixed with
anhydrous sodium sulfate to form a free flowing powder. This mixture is
solvent extracted three times using ultrasonic extraction. The extract is
separated from the sample by vacuum filtration or centrifugation. The extract
is ready for cleanup and/or analysis following concentration.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
4.0 APPARATUS AND MATERIALS
4.1 Apparatus for grinding dry waste samples.
4.2 Ultrasonic preparation - A horn type device equipped with a
titanium tip, or a device that will give equivalent performance, shall be
used.
3550B - 1 Revision 2
November 1990
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4.2.1 Ultrasonic Disrupter - The disrupter must have a minimum
power wattage of 300 watts, with pulsing capability. A device designed
to reduce the cavitation sound is recommended. Follow the manufacturers
instructions for preparing the disrupter for extraction of samples with
low and medium/high concentration.
Use a 3/4" horn for the low concentration method and a 1/8" tapered
microtip attached to a 1/2" horn for the medium/high concentration
method.
4.3 Sonabox - Recommended with above disrupters for decreasing
cavitation sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.4 Apparatus for determining percent dry weight.
4.4.1 Oven - Drying.
4.4.2 Desiccator.
4.4.3 Crucibles - Porcelain or disposable aluminum.
4.5 Pasteur glass pipets - 1 ml, disposable.
4.6 Beakers - 400 ml.
4.7 Vacuum or pressure filtration apparatus.
4.7.1 Buchner funnel.
4.7.2 Filter paper - Whatman No. 41 or equivalent.
4.8 Kuderna-Danish (K-D) apparatus.
4.8.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation
of extracts.
4.8.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.8.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.8.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.8.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.9 Boiling chips - Solvent extracted, approximately 10/40 mesh
(silicon carbide or equivalent).
4.10 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The batch should be used in a hood.
3550B - 2 Revision 2
November 1990
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4.11 Balance - Top loading, capable of accurately weighing to the
nearest 0.01 g.
4.12 Vials - 2 ml, for GC autosampler, with Teflon lined screw caps or
crimp tops.
4.13 Glass scintillation vials - 20 ml, with Teflon lined screw caps.
4.14 Spatula - Stainless steel or Teflon.
4.15 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed by
50 ml of elution solvent prior to packing the column with adsorbent.
4.16 Syringe - 5 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise specified, it is intended that all inorganic reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other
grades may be used, provided it is first ascertained that the reagent is of
sufficiently high purity to permit its use without lessening the accuracy of
the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na?S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene
chloride, a method blank must be analyzed, demonstrating that there is no
interference from the sodium sulfate.
5.4 Extraction solvents.
5.4.1 Low concentration soil/sediment and aqueous sludge samples
shall be extracted using a solvent system that gives optimum,
reproducible recovery for the matrix/analyte combination to be measured.
Suitable solvent choices are given in Table 1.
5.4.2 Methylene chloriderAcetone, CH2C12:CH3COCH3 (1:1, v:v).
Pesticide quality or equivalent.
5.4.3 Methylene chloride, CH2C12. Pesticide quality or
equivalent.
3550B - 3 Revision 2
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5.4.4 Hexane, C6H14. Pesticide quality or equivalent.
5.5 Exchange solvents.
5.5.1 Hexane, C6H14. Pesticide quality or equivalent.
5.5.2 2-Propanol, (CH3)2CHOH. Pesticide quality or equivalent.
5.5.3 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.5.4 Acetonitrile, CH3CN. Pesticide quality or equivalent.
5.5.5 Methanol, CH3OH. Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample handling
7.1.1 Sediment/soil samples - Decant and discard any water layer
on a sediment sample. Mix sample thoroughly, especially composited
samples. Discard any foreign objects such as sticks, leaves, and rocks.
7.1.1.2 Determine the dry weight of the sample (Section
7.2) remaining after decanting. Measurement of soil pH may be
required.
7.1.2 Waste samples - Samples consisting of multiphases must be
prepared by the phase separation method in Chapter Two before
extraction. This procedure is for solids only.
7.1.3 Dry waste samples amenable to grinding - Grind or otherwise
subdivide the waste so that it either passes through a 1 mm sieve or can
be extruded through a 1 mm hole. Introduce sufficient sample into the
grinder to yield at least 100 g after grinding.
7.1.4 Gummy, fibrous or oily materials not amenable to grinding
should be cut, shredded, or otherwise broken up to allow mixing, and
maximum exposure of the sample surfaces for extraction. The
professional judgment of the analyst is required for handling of these
difficult matrices.
7.2 Determination of percent dry weight - In certain cases, sample
results are desired based on a dry weight basis. When such data is desired,
or required, a portion of sample for this determination should be weighed out
at the same time as the portion used for analytical determination.
3550B - 4 Revision 2
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WARNING; The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from drying a
heavily contaminated hazardous waste sample.
7.2.1 Immediately after weighing the sample for extraction, weigh
5-10 g of the sample into a tared crucible. Determine the % dry weight
of the sample by drying overnight at 105°C. Allow to cool in a
desiccator before weighing:
% dry weight = g of dry sample x 100
g of sample
7.3 Extraction method for samples expected to contain low
concentrations of organics and pesticides (< 20 mg/Kg):
7.3.1 The following step should be performed rapidly to avoid
loss of the more volatile extractables. Weigh approximately 30 g of
sample into a 400 ml beaker. Record the weigh to the nearest 0.1 g.
Nonporous or wet samples (gummy or clay type) that do not have a free-
flowing sandy texture must be mixed with 60 g of anhydrous sodium
sulfate, using a spatula. If required, more sodium sulfate may be
added. After addition of sodium sulfate, the sample should be free
flowing. Add 1 ml of surrogate standards to all samples, spikes,
standards, and blanks (see Method 3500 for details on the surrogate
standard solution and the matrix spike solution). For the sample in
each analytical batch selected for spiking, add 1.0 ml of the matrix
spiking standard. For base/neutral-acid analysis, the amount added of
the surrogates and matrix spiking compounds should result in a final
concentration of 100 ng//iL of each base/neutral analyte and 200 ng//iL
of each acid analyte in the extract to be analyzed (assuming a 1 p.1
injection). If Method 3640, Gel-Permeation Cleanup, is to be used, add
twice the volume of surrogates and matrix spiking compounds since half
of the extract is lost due to loading of the GPC column. Immediately
add 100 ml of 1:1 methylene chloride:acetone.
7.3.2 Place the bottom surface of the tip of the #207 3/4 in.
disrupter horn about 1/2 in. below the surface of the solvent, but above
the sediment layer.
7.3.3 Extract ultrasonically for 3 minutes, with output control
knob set at 10 (full power) and with mode switch on Pulse (pulsing
energy rather than continuous energy) and percent-duty cycle knob set at
50% (energy on 50% of time and off 50% of time). Do not use microtip
probe.
7.3.4 Decant and filter extracts through Whatman No. 41
filter paper using vacuum filtration or centrifuge, and decant
extraction solvent.
7.3.5 Repeat the extraction two or more times with two additional
100 ml portions of solvent. Decant off the solvent after each
ultrasonic extraction. On the final ultrasonic extraction, pour the
entire sample into the Buchner funnel and rinse with extraction solvent.
3550B - 5 Revision 2
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7.3.6 Assemble a Kuderna-Danish (K-D) concentrator by attaching a
10 ml concentrator tube to a 500 ml evaporator flask.
7.3.7 Dry the extract by passing it through a drying column
containing about 10 cm of anhydrous sodium sulfate. Collect the dried
extract in the K-D concentrator. Wash the extractor flask and sodium
sulfate column with 100-125 ml of extraction solvent to complete the
quantitative transfer.
7.3.8 Add one to two clean boiling chips to the evaporation
flask, and attach a three ball Snyder column. Prewet the Snyder column
by adding about 1 ml methylene chloride to the top. Place the K-D
apparatus on a hot water bath (80-90°C) so that the concentrator tube is
partially immersed in the hot water and the entire lower rounded surface
of the flask is bathed with hot vapor. Adjust the vertical position of
the apparatus and the water temperature, as required, to complete the
concentration in 10-15 min. At the proper rate of distillation the
balls of the column will actively chatter, but the chambers will not
flood with condensed solvent. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus and allow it to drain and cool
for at least 10 min.
7.3.9 If a solvent exchange is required (as indicated in
Table 1), momentarily remove the Snyder column, add 50 ml of the
exchange solvent and a new boiling chip, and re-attach the Snyder
column. Concentrate the extract as described in Section 7.3.8, raising
the temperature of the water bath, if necessary, to maintain proper
distillation. When the apparent volume again reaches 1-2 ml, remove the
K-D apparatus and allow it to drain and cool for at least 10 minutes.
7.3.10 Remove the Snyder column and rinse the flask and its
lower joints into the concentrator tube with 1-2 ml of methylene
chloride or exchange solvent. If sulfur crystals are a problem, proceed
to Method 3660 for cleanup. The extract may be further concentrated by
using the technique outlined in Section 7.3.11 or adjusted to 10.0 ml
with the solvent last used.
7.3.11 If further concentration is indicated in Table 1,
either micro Snyder column technique (Section 7.3.11.1) or nitrogen blow
down technique (Section 7.3.11.2) is used to adjust the extract to the
final volume required.
7.3.11.1 Micro Snyder Column Technique
7.3.11.1.1 Add a clean boiling chip and attach a two
ball micro Snyder column to the concentrator tube. Prewet
the column by adding approximately 0.5 ml of methylene
chloride or exchange solvent through the top. Place the
apparatus in the hot water bath. Adjust the vertical
position and the water temperature, as required, to complete
the concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the liquid reaches an
apparent volume of approximately 0.5 ml, remove the
3550B - 6 Revision 2
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apparatus from the water bath and allow to drain and cool
for at least 10 minutes. Remove the micro Snyder column and
rinse its lower joint with approximately 0.2 ml of
appropriate solvent and add to the concentrator tube.
Adjust the final volume to the volume required for cleanup
or for the determinative method (see Table 1).
7.3.11.2 Nitrogen Blowdown Technique
7.3.11.2.1 Place the concentrator tube in a warm
water bath (approximately 35°C) and evaporate the solvent
volume to the required level using a gentle stream of clean,
dry nitrogen (filtered through a column of activated
carbon).
CAUTION; Do not use plasticized tubing between the carbon trap and the
sample.
7.3.11.2.2 The internal wall of the tube must be
rinsed down several times with the appropriate solvent
during the operation. During evaporation, the solvent level
in the tube must be positioned to prevent water from
condensing into the sample (i.e., the solvent level should
be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to
become dry.
CAUTION; When the volume of solvent is reduced below 1 ml, semivolatile
analytes may be lost.
7.4 If analysis of the extract will not be performed immediately,
stopper the concentrator tube and store refrigerated. If the extract will be
stored longer than 2 days, it should be transferred to a vial with a Teflon
lined cap and labeled appropriately.
7.5 Extraction method for samples expected to contain high
concentrations of organics (> 20 mg/Kg):
7.5.1 Transfer approximately 2 g (record weight to the nearest
0.1 g) of sample to a 20 ml vial. Wipe the mouth of the vial with a
tissue to remove any sample material. Record the exact weight of sample
taken. Cap the vial before proceeding with the next sample to avoid any
cross contamination.
7.5.2 Add 2 g of anhydrous sodium sulfate to sample in the 20 ml
vial and mix well.
7.5.3 Surrogate standards are added to all samples, spikes, and
blanks (see Method 3500 for details on the surrogate standard solution
and on the matrix spike solution). Add 2.0 mL of surrogate spiking
solution to sample mixture. For the sample in each analytical batch
selected for spiking, add 2.0 ml of the matrix spiking standard. For
base/neutral-acid analysis, the amount added of the surrogates and
matrix spiking compounds should result in a final concentration of
3550B - 7 Revision 2
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200 ng/juL of each base/neutral analyte and 400 ng//il_ of each acid
analyte in the extract to be analyzed (assuming a 1 /uL injection). If
Method 3640, Gel-Permeation Cleanup, is to be used, add twice the volume
of surrogates and matrix spiking compounds since half the extract is
lost due to loading of the GPC column.
7.5.4 Immediately add whatever volume of solvent is necessary to
bring the final volume to 10.0 ml considering the added volume of
surrogates and matrix spikes. Disrupt the sample with the 1/8 in.
tapered microtip ultrasonic probe for 2 minutes at output control
setting 5 and with mode switch on pulse and percent duty cycle at 50%.
Extraction solvents are:
1. Nonpolar compounds (i.e., organochlorine pesticides and
PCBs), hexane or appropriate solvent.
2. Extractable priority pollutants, methylene chloride.
7.5.5 Loosely pack disposable Pasteur pipets with 2 to 3 cm Pyrex
glass wool plugs. Filter the extract through the glass wool and collect
5.0 ml in a concentrator tube if further concentration is required.
Follow Section 7.3.11 for details on concentration. Normally, the
5.0 ml extract is concentrated to approximately 1.0 ml or less.
7.5.6 The extract is ready for cleanup or analysis, depending on
the extent of interfering co-extractives.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
8.2 Horn tip and tuning criteria are critical elements in achieving
good method performance. Refer to the manufacturer's specifications.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative method for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for
the Analysis of Pollutants Under the Clean Water Act; Final Rule and
Interim Final Rule and Proposed Rule," October 26, 1984.
2. U.S. EPA, Interlaboratory Comparison Study: Methods for Volatile and
Semi-Volatile Compounds, Environmental Monitoring Systems Laboratory,
Office of Research and Development, Las Vegas, NV, EPA 600/4-84-027,
1984.
3550B - 8 Revision 2
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3. Christopher S. Hein, Paul J. Marsden, Arthur S. Shurtleff, "Evaluation of
Methods 3540 (Soxhlet) and 3550 (Sonicatlon) for Evaluation of Appendix
IX Analytes form Solid Samples", S-CUBED, Report for EPA Contract 68-03-
33-75, Work Assignment No. 03, Document No. SSS-R-88-9436, October 1988.
3550B - 9 Revision 2
November 1990
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TABLE 1.
EFFICIENCY OF EXTRACTION SOLVENT SYSTEMS3
Solvent Svstem°
Compound
4-Bromophenyl phenyl ether
4-Chl oro-3-methyl phenol
bi s(2-Chl oroethoxy)methane
bis(2-Chloroethyl) ether
2-Chl oronaphthal ene
4-Chlorophenyl phenyl ether
1 , 2-Di chl oro benzene
1,3-Di chl orobenzene
Diethyl phthalate
4,6-Dinitro-o-cresol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Heptachlor epoxide
Hexachl orobenzene
Hexachl orobutadi ene
Hexachl orocyclopentadi ene
Hexachl oroethane
5-Nitro-o-toluidine
Nitrobenzene
Phenol
1 , 2 , 4-Tr i chl orobenzene
CAS No.b
101-55-3
59-50-7
111-91-1
111-44-4
91-58-7
7005-72-3
95-50-1
541-73-1
84-66-2
534-52-1
121-14-2
606-20-2
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
99-55-8
98-95-3
108-95-2
120-82-1
ABNC
N
A
N
N
N
N
N
N
N
A
N
N
N
N
N
N
N
B
N
A
N
A
%R
64.2
66.7
71.2
42.0
86.4
68.2
33.3
29.3
24.8
66.1
68.9
70.0
65.5
62.1
55.8
26.8
28.4
52.6
59.8
51.6
66.7
B
SD
6.5
6.4
4.5
4.8
8.8
8.1
4.5
4.8
1.6
8.0
1.6
7.6
7.8
8.8
8.3
3.3
3.8
26.7
7.0
2.4
5.5
%R
56.4
74.3
58.3
17.2
78.9
63.0
15.8
12.7
23.3
63.8
65.6
68.3
58.7
56.5
41.0
19.3
15.5
64.6
38.7
52.0
49.9
SD
0.5
2.8
5.4
3.1
3.2
2.5
2.0
1.7
0.3
2.5
4.9
0.7
1.0
1.2
2.7
1.8
1.6
4.7
5.5
3.3
4.0
C
%R
86.7
97.4
69.3
41.2
100.8
96.6
27.8
20.5
121.1
74.2
85.6
88.3
86.7
95.8
63.4
35.5
31.1
74.7
46.9
65.6
73.4
SD
1.9
3.4
2.4
8.4
3.2
2.5
6.5
6.2
3.3
3.5
1.7
4.0
1.0
2.5
4.1
6.5
7.4
4.7
6.3
3.4
3.6
D
%R
84.5
89.4
74.8
61.3
83.0
80.7
53.2
46.8
99.0
55.2
68.4
65.2
84.8
89.3
76.9
46.6
57.9
27.9
60.6
65.5
84.0
SD
0.4
3.8
4.3
11.7
4.6
1.0
10.1
10.5
4.5
5.6
3.0
2.0
2.5
1.2
8.4
4.7
10.4
4.0
6.3
2.1
7.0
E
%R
73.4
84.1
37.5
4.8
57.0
67.8
2.0
0.6
94.8
63.4
64.9
59.8
77.0
78.1
12.5
9.2
1.4
34.0
13.6
50.0
20.0
SD
1.0
1.6
5.8
1.0
2.2
1.0
1.2
0.6
2.9
2.0
2.3
0.8
0.7
4.4
4.6
1.7
1.2
4.0
3.2
8.1
3.2
a Percent recovery of analytes spiked at 200 mg/Kg into NIST sediment SRM 1645
b Chemical Abstracts Service Registry Number
c Compound Type: A = Acid, B = Base, N = neutral
d A = Methylene chloride
B = Methylene chloride/Acetone (1/1)
C = Hexane/Acetone (1/1)
D = Methyl t-butyl ether
E = Methyl t-butyl ether/Methanol (2/1)
3550B - 10
Revision 2
November 1990
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TABLE 2.
SPECIFIC EXTRACTION CONDITIONS FOR VARIOUS DETERMINATIVE METHODS
Determinative
method
8040a
8060
8061
8070
8080
8081
8090
8100
8110
8120
8121
8140
8141
8250"-c
8270C
8310
8321
8410
Extraction
PH
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
as received
Exchange
solvent
required
for
analysis
2-propanol
hexane
hexane
methanol
hexane
hexane
hexane
none
hexane
hexane
hexane
hexane
hexane
none
none
acetonitrile
methanol
methyl ene chloride
Exchange Volume
solvent of extract
required required
for for
cleanup cleanup (ml)
hexane
hexane
hexane
methyl ene chloride
hexane
hexane
hexane
cyclohexane
hexane
hexane
hexane
hexane
hexane
_.
--
--
--
methyl ene chloride
1.0
2.0
2.0
2.0
10.0
10.0
2.0
2.0
2.0
2.0
2.0
10.0
10.0
-.
--
--
--
10.0
Final
extract
vol ume
for
analysis (ml)
1.0, 10. Ob
10.0
10.0
10.0
10.0
10.0
1.0
1.0
10.0
1.0
1.0
10.0
10.0
1.0
1.0
1.0
1.0
0.0 (dry)
a To obtain separate acid and base/neutral extracts, Method 3650 should be performed following
concentration of the extract to 10.0 ml.
b Phenols may be analyzed, by Method 8040, using a 1.0 ml 2-propanol extract by GC/FID. Method 8040 also
contains an optical derivatization procedure for phenols which results in a 10 ml hexane extract to be
analyzed by GC/ECD.
c The specificity of GC/MS may make cleanup of the extracts unnecessary. Refer to Method 3600 for guidance
on the cleanup procedures available if required.
3550B - 11
Revision 2
November 1990
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METHOD 3550B
ULTRASONIC EXTRACTION
START
7 1 Prepare tample*
using appropriate
method for the
wa*te matriit
7.2 Determine the
percent dry weight
of the sample
752 Add anhydrous
sodium sulfate to
•ample
No
7 5.3 Add surrogate
standard* to all
samples. spikes,
and blanks
7.3.1 Add surrogate
standards to al1
samples. spikes,
and blanks
7.3 2 - 7.3.5
Sonicate sample at
least 3 times
754 Adjust
volume; disrupt
sample with tapered
microtip ultrasonic
probe
7.5.5
through
filter
glass wool
7.3.7 Dry and
collect extract in
K-D concentrator
7.3.8 Concentrate
extract and collect
in K-D concentrator
3550B - 12
Revision 2
November 1990
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METHOD 3550B
continued
73.9 Is
a solvent
exchange
required?
73.9 Add exchange
solvent;
concentrate extract
7.3.10 Use Method
3660 for cleanup
7.3.10 Do
sulfur crystals
form?
7.3.11 Further
concentrate and/or
adjust volume
»[ Cleanup or analyze
3550B - 13
Revision 2
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METHOD 3600B
CLEANUP
1.0 SCOPE AND APPLICATION
1.1 General
1.1.1 Injection of sample extracts, without further cleanup or
isolation of analytes, into a gas or liquid chromatograph can cause
extraneous peaks, deterioration of peak resolution and column efficiency,
and loss of detector sensitivity and can greatly shorten the lifetime of
expensive columns. The following techniques have been applied to extract
purification: partitioning between immiscible solvents; adsorption
chromatography; gel permeation chromatography; chemical destruction of
interfering substances with acid, alkali, or oxidizing agents; and
distillation. These techniques may be used individually or in various
combinations, depending on the extent and nature of the co-extractives.
1.1.2 It is an unusual situation (e.g. with some water samples)
when extracts can be directly determined without further treatment. Soil
and waste extracts often require a combination of cleanup methods. For
example, when analyzing for organochlorine pesticides and PCBs, it may be
necessary to use gel permeation chromatography (GPC), to eliminate the high
boiling material and a micro alumina or Florisil column to eliminate
interferences with the analyte peaks on the GC/ECD.
1.2 Specific
1.2.1 Adsorption column chromatography - Alumina (Methods 3610 and
3611), Florisil (Method 3620), and silica gel (Method 3630) are useful for
separating analytes of a relatively narrow polarity range away from
extraneous, interfering peaks of a different polarity.
1.2.2 Acid-base partitioning (Method 3650) - Useful for separating
acidic or basic organics from neutral organics. It has been applied to
analytes such as the chlorophenoxy herbicides and phenols.
1.2.3 Gel permeation chromatography (GPC) (Method 3640) - The most
universal cleanup technique for a broad range of semivolatile organics and
pesticides. It is capable of separating high molecular-weight material
from the sample analytes. It has been used successfully for all the
semivolatile base, neutral, and acid compounds associated with the EPA
Priority Pollutant and the Superfund Target Compound list for GC/MS
analysis for semivolatiles and pesticides. GPC is usually not applicable
for eliminating extraneous peaks on a chromatogram which interfere with
the analytes of interest.
1.2.4 Sulfur cleanup (Method 3660) - Useful in eliminating sulfur
from sample extracts, which may cause chromatographic interference with
analytes of interest.
3600B - 1 Revision 2
November 1990
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1.2.5 Table 1 indicates the recommended cleanup techniques for the
indicated groups of compounds. This information can also be used as
guidance for compounds that are not listed. Compounds that are chemically
similar to these groups of compounds should follow a similar elution
pattern.
2.0 SUMMARY OF METHOD
2.1 Refer to the specific cleanup method for a summary of the procedure.
3.0 INTERFERENCES
3.1 Analytical interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware. All of these
materials must be routinely demonstrated to be free of interferences, under the
conditions of the analysis, by running laboratory reagent blanks.
3.2 More extensive procedures than those outlined in the methods may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Refer to the specific cleanup method for apparatus and materials
needed.
5.0 REAGENTS
5.1 Refer to the specific cleanup method for the reagents needed.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Prior to using the cleanup procedures, samples should undergo solvent
extraction. Chapter Two, Section 2.3.3, may be used as a guide for choosing the
appropriate extraction procedure based on the physical composition of the waste
and on the analytes of interest in the matrix (see also Method 3500 for a general
description of the extraction technique). For some organic liquids, extraction
prior to cleanup may not be necessary.
7.2 In most cases, the extracted sample is then analyzed by one of the
determinative methods available in Section 4.3 of this chapter. If the analytes
of interest are not able to be determined due to interferences, cleanup is
performed.
3600B - 2 Revision 2
November 1990
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7.3 Many of the determinative methods specify cleanup methods that should
be used when determining particular analytes (e.g. Method 8060, gas
chromatography of phthalate esters, recommends using either Method 3610 (Alumina
column cleanup) or Method 3620 (Florisil column cleanup) if interferences prevent
analysis). However, the experience of the analyst may prove invaluable in
determining which cleanup methods are needed. As indicated in Section 1.0 of
this method, many matrices may require a combination of cleanup procedures in
order to ensure proper analytical determinations.
7.4 Guidance for cleanup is specified in each of the methods that follow.
The amount of extract cleanup required prior to the final determination depends
on the selectivity of both the extraction procedure and the determinative method
and the required detection limit.
7.5 Following cleanup, the sample is concentrated to whatever volume is
required in the determinative method. Analysis follows as specified in the
determinative procedure (Section 4.3 of this Chapter).
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered by the cleanup technique before the cleanup is applied
to actual samples. For sample extracts that are cleaned up, the associated
quality control samples (e.g. spikes, blanks, replicates, and duplicates) must
also be processed through the same cleanup procedure.
8.3 The analysis using each determinative method (GC, GC/MS, HPLC)
specifies instrument calibration procedures using stock standards. It is
recommended that cleanup also be performed on a series of the same type of
standards to validate chromatographic elution patterns for the compounds of
interest and to verify the absence of interferences from reagents.
9.0 METHOD PERFORMANCE
9.1 Refer to the specific cleanup method for performance data.
10.0 REFERENCES
10.1 Refer to the specific cleanup method.
3600B - 3 Revision 2
November 1990
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TABLE 1.
RECOMMENDED CLEANUP TECHNIQUES FOR INDICATED GROUPS OF COMPOUNDS
Determinative8 Cleanup Method
Analyte Group Method Option
Phenols 8040 3630", 3640, 3650, 8040C
Phthalate esters 8060, 8061 3610, 3620, 3640
Nitrosamines 8070 3610, 3620, 3640
Organochlorine pesticides & PCBs 8080, 8081 3620, 3640, 3660
Nitroaromatics and cyclic ketones 8090 3620, 3640
Polynuclear aromatic hydrocarbons 8100 3611, 3630, 3640
Chlorinated hydrocarbons 8120, 8121 3620, 3640
Organophosphorus pesticides 8140, 8141 3620
Chlorinated herbicides 8150, 8151 8150d
Priority pollutant semivolatiles 8250, 8270 3640, 3650, 3660
Priority pollutant semivolatiles 8410 3640
Petroleum waste 8250, 8270 3611, 3650
a The GC/MS Methods, 8250 and 8270, are also appropriate determinative methods for
all analyte groups, unless lower detection limits are required.
b Cleanup applicable to derivatized phenols.
c Method 8040 includes a derivatization technique followed by GC/ECD analysis, if
interferences are encountered using GC/FID.
d Methods 8150 and 8151 incorporate an acid-base cleanup step as an integral part
of the method.
3600B - 4 Revision 2
November 1990
-------
METHOD 3600B
CLEANUP
START
I
7.1 Do
solvent
extraction
1
7.2 Analyze
analyte by a
determinative
method from
Sec. 4.3
analyte.-
undeterminabl
7.3 Use cleanup
method
specified for
the determina-
tive method
7.5
Concentrate
sample to
required
volume
3600B - 5
Revision 2
November 1990
-------
METHOD 3630B
SILICA GEL CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Silica gel is a regenerative adsorbent of amorphous silica with weakly
acidic properties. It is produced from sodium silicate and sulfuric acid.
Silica gel can be used for column chromatography and is for separating the
analytes from interfering compounds of a different chemical polarity.
1.2 General applications (Gordon and Ford):
1.2.1 Activated: Heated at 150-160°C for several hours. USES:
Separation of hydrocarbons.
1.2.2 Deactivated: Containing 10-20% water. USES: An adsorbent
for most functionalities with ionic or nonionic characteristics, including
alkaloids, sugar esters, glycosides, dyes, alkali metal cations, lipids,
glycerides, steroids, terpenoids and plasticizers. The disadvantages of
deactivated silica gel are that the solvents methanol and ethanol decrease
adsorbent activity.
1.3 Specific applications: This method includes guidance for cleanup of
sample extracts containing polynuclear aromatic hydrocarbons, derivatized
phenolic compounds, polychlorinated biphenyls (PCBs) and single component
pesticides. When only PCBs are to be measured, this method can be used in
conjunction with sulfuric acid/permanganate cleanup (Method 3665).
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of adsorbent, topped
with a water adsorbent, and then loaded with the sample to be analyzed. Elution
of the analytes is effected with a suitable solvent(s) leaving the interfering
compounds on the column. The eluate is then concentrated.
3.0 INTERFERENCES
3.1 A reagent blank should be analyzed for the compounds of interest
prior to the use of this method. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Chromatographic column - 250 mm long x 10 mm ID; with Pyrex glass wool
at bottom and a Teflon stopcock.
3630B - 1 Revision 2
November 1990
-------
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed
by 50 ml of elution solvent prior to packing the column with adsorbent.
4.2 Beakers - 500 ml.
4.3 Kuderna-Danish (K-D) apparatus
4.3.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Vials - 10, 25 ml, glass with Teflon lined screw-caps or crimp tops.
4.5 Muffle furnace.
4.6 Reagent bottle - 500 ml.
4.7 Water bath - Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.8 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.9 Erlenmeyer flasks - 50 and 250 mL.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
3630B - 2 Revision 2
November 1990
-------
5.3 Silica gel. 100/200 mesh desiccant (Davison Chemical grade 923 or
equivalent). Before use, activate for at least 16 hr. at 130°C in a shallow
glass tray, loosely covered with foil.
5.4 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.5 Eluting solvents
5.5.1 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.5.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.5.3 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.5.4 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.5.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.5.6 Pentane, C5H12 - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Polynuclear aromatic hydrocarbons
7.1.1 Before the silica gel cleanup technique can be utilized, the
extract solvent must be exchanged to cyclohexane. The exchange is performed
as follows:
7.1.1.1 Following K-D concentration of the extract to 1-2 mL
using the macro-Snyder column, allow the apparatus to cool and drain
for at least 10 minutes. Add one or two clean boiling chips to the
K-D flask. Add 4 mL of exchange solvent and attach a two ball micro-
Snyder column. Prewet the Snyder column by adding about 0.5 mL of
methylene chloride to the top of the column. Place the K-D apparatus
on a hot water bath (15-20°C above the boiling point of the solvent)
so that the concentrator tube is partially immersed in the hot water
and the entire lower rounded surface of the flask is bathed with hot
vapor. Adjust the vertical position of the apparatus and the water
temperature, as required, to complete the concentration in 5-10
minutes. At the proper rate of distillation the balls of the column
will actively chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5-1 mL, remove the K-D apparatus
3630B - 3 Revision 2
November 1990
-------
from the water bath and allow it to drain and cool for at least 10
minutes.
Caution: When the volume of solvent is reduced below 1 ml, semivolatile
analytes may be lost.
7.1.1.2 Remove the micro-Snyder column and rinse its lower
joint into the concentrator tube with a minimum amount of exchange
solvent. Adjust the extract volume to about 2 ml.
7.1.2 Prepare a slurry of 10 g of activated silica gel in methylene
chloride and place this into a 10 mm ID chromatographic column. Tap the
column to settle the silica gel and elute the methylene chloride. Add 1
to 2 cm of anhydrous sodium sulfate to the top of the silica gel.
7.1.3 Preelute the column with 40 ml of pentane. The rate for all
elutions should be about 2 mL/min. Discard the eluate and just prior to
exposure of the sodium sulfate layer to the air, transfer the 2 ml
cyclohexane sample extract onto the column using an additional 2 mL
cyclohexane to complete the transfer. Just prior to exposure of the sodium
sulfate layer to the air, add 25 mL of pentane and continue the elution
of the column. Discard this pentane eluate.
7.1.4 Next, elute the column with 25 ml of methylene chloride/
pentane (2:3)(v/v) into a 500 ml K-D flask equipped with a 10 ml
concentrator tube. Concentrate the collected fraction to whatever volume
is required (1-10 ml). Proceed with HPLC or GC analysis. Components that
elute in this fraction are:
Acenaphthene
Acenaphthylene
Anthracene
6enzo(a)anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Benzo(g,h,i)perylene
Benzo(k)fl uoranthene
Chrysene
Dibenzo(a,h)anthracene
Fluoranthene
Fluorene
Indeno(l,2,3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
7.2 Derivatized phenols
7.2.1 This silica gel cleanup procedure is performed on sample
extracts that have undergone pentafluorobenzyl bromide derivatization as
described in Method 8040.
3630B - 4 Revision 2
November 1990
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7.2.2 Place 4.0 g of activated silica gel into a 10 mm ID
chromatographic column. Tap the column to settle the silica gel and add
about 2 g of anhydrous sodium sulfate to the top of the silica gel.
7.2.3 Preelute the column with 6 ml of hexane. The rate for all
elutions should be about 2 mL/min. Discard the eluate and just prior to
exposure of the sodium sulfate layer to the air, pipet onto the column 2 mL
of the hexane solution that contains the derivatized sample or standard.
Elute the column with 10.0 ml of hexane and discard the eluate.
7.2.4 Elute the column, in order, with 10.0 ml of 15% toluene in
hexane (Fraction 1); 10.0 mL of 40% toluene in hexane (Fraction 2); 10.0 ml
of 75% toluene in hexane (Fraction 3); and 10.0 ml of 15% 2-propanol in
toluene (Fraction 4). All elution'mixtures are prepared on a volume:volume
basis. Elution patterns for the phenolic derivatives are shown in Table 1.
Fractions may be combined, as desired, depending upon the specific phenols
of interest or level of interferences. Proceed with GC analysis (Method
8040).
7.3 PCBs and single component pesticides:
7.3.1 Place a portion of activated silica gel (normally 20 g) into
a glass jar and deactivate it with organic-free reagent water to bring the
moisture content to 3.3 percent. Mix the contents of the glass jar
thoroughly and equilibrate for 6 hours. Store the deactivated silica gel
in a sealed glass jar inside a desiccator. Transfer a 3 g portion into
a 10 mm ID glass chromatographic column and top it with 2 to 3 cm of
anhydrous sodium sulfate.
7.3.2 Add 10 ml of hexane to the top of the column to wet and rinse
the sodium sulfate and silica gel. Just prior to exposure of the sodium
sulfate layer to air, stop the hexane eluate flow by closing the stopcock
on the chromatographic column. Discard the eluate.
7.3.3 Transfer the sample extract (2 ml) onto the column. Rinse
the extract vial twice with 1 to 2 ml of hexane and add each rinse to the
column. Elute the column with 80 ml of hexane (Fraction I) at a rate of
about 5 mL/min. Remove the collection flask and set it aside for later
concentration. Elute the column with 50 mL of hexane (Fraction II) and
collect the eluate. Perform a third elution with 15 mL of methylene
chloride (Fraction III). The elution patterns for the organochlorine
pesticides, Aroclor-1016, and Aroclor-1260 are shown in Table 2.
7.3.4 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane (Sections 7.1.1.1 and 7.1.1.2). Proceed with
GC analysis.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
3630B - 5 Revision 2
November 1990
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8.2 The analyst should demonstrate that the compounds of Interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
9.1 Table 1 provides performance information on the fractionation of
phenolic derivatives using this method.
9.2 Table 2 provides performance information on the fractionation of PCBs
and single component pesticides using this method.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3630B - 6 Revision 2
November 1990
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TABLE 1
SILICA GEL FRACTIONATION OF PFBB DERIVATIVES
Percent Recovery by Fraction8
Parameter 123
2-Chlorophenol
2-Nitrophenol
Phenol
2, 4 -Dimethyl phenol
2,4-Dichlorophenol
2 , 4 , 6-Tri chl orophenol
4-Chloro-3-methyl phenol
Pentachl orophenol
4-Nitrophenol
90
90
95
95
50 50
84
75 20
1
9
10
7
1
14
1
90
90
Eluant composition:
Fraction 1 - 15% toluene in hexane.
Fraction 2 - 40% toluene in hexane.
Fraction 3 - 75% toluene in hexane.
Fraction 4 - 15% 2-propanol in toluene.
3630B - 7 Revision 2
November 1990
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TABLE 2
DISTRIBUTION AND PERCENT RECOVERIES OF ORGANOCHLORINE
PESTICIDES AND PCBs AS AROCLORS IN SILICA GEL COLUMN FRACTIONS8-"-0'"'6
Compound
alpha-BHCf
beta-BHC
gamma -BHC
delta-BHC
Heptachlor
Aldrin
Heptachlor
Technical
Endosulfan
4,4'-DDE
Dieldrin
Endrin
Endosulfan
4,4'-DDDf
Fraction I
Cone. Cone.
1 2
epoxide
chlordane
I
II
109(4.
97(5.
14(5.
86(5.
1)
6)
5)
4)
118(8.7)
104(1.6)
22(5.3)
94(2.8)
Endrin aldehyde
Endosulfan
4,4'-DDTf
sulfate
4,4'-Methoxychlor
Toxaphene'
Aroclor-1016
Aroclor-1260
86(4.
91(4.
0)
1)
87(6.1)
95(5.0)
Fraction II Fraction I
Cone. Cone. Cone.
1 2 1
82(1.
107(2.
91(3.
92(3.
95(4.
19(6.8) 39(3.6) 29(5.
95(5.
96(6.
85(10
97(4.
102(4.
81(1.
93(4.
86(13.4) 73(9.1) 15(17
99(9.
15(2.4) 17(1.4) 73(9.
7)
1)
6)
5)
7)
0)
1)
0)
.5)
4)
6)
9)
9)
•7)
9)
4)
II
Cone.
2
74(8.0)
98(12.5)
85(10.7)
83(10.6)
88(10.2)
37(5.1)
87(10.2)
87(10.6)
71(12.3)
86(10.4)
92(10.2)
76(9.5)
82(9.2)
8.7(15.0)
82(10.7)
84(10.7)
Total
Cone.
1
82(1.
107(2.
91(3.
92(3.
109(4.
97(5.
95(4.
62(3.
95(5.
86(5.
96(6.
Recovery
Cone.
2
7)
1)
6)
5)
1)
6)
7)
3)
1)
4)
0)
85(10.5)
97(4.
102(4.
81(1.
93(4.
4)
6)
9)
9)
101(5.3)
99(9.
88(12
86(4.
91(4.
9)
.0)
0)
1)
74(8.
98(12
85(10
83(10
118(8.
104(1.
88(10
98(1.
87(10
94(2.
87(10
0)
.5)
.7)
.6)
7)
6)-
.2)
9)
.2)
8)
.6)
71(12.3)
86(10
92(10
76(9.
82(9.
82(23
82(10
101(10
87(6.
95(5.
•4)
.2)
5)
2)
•7)
.7)
•1)
1)
0)
3630B - 8
Revision 2
November 1990
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TABLE 2
(Continued)
Effluent composition: Fraction I, 80 ml hexane; Fraction II, 50 ml hexane; Fraction III, 15 ml methylene
chloride.
Concentration 1 is 0.5 /Ltg per column for BHCs, heptachlor, aldrin, heptachlor epoxide, and endosulfan
I; 1.0 /ig per column for dieldrin, endosulfan II, 4,4'-DDD, 4,4'-DDE, 4,4'-DDT, endrin, endrin aldehyde,
and endosulfan sulfate; 5 /zg per column for 4,4'-methoxychlor and technical chlordane; 10 ug per column
for toxaphene, Aroclor-1016, and Aroclor-1260.
For Concentration 2, the amounts spiked are 10 times as high as those for Concentration 1.
Values given represent the average recovery of three determinations; numbers in parentheses are the
standard deviation; recovery cutoff point is 5 percent.
Data obtained with standards, as indicated in footnotes b and c, dissolved in 2 ml hexane.
It has been found that because of batch-to-batch variation in the silica gel material, these compounds
cross over in two fractions and the amounts recovered in each fraction are difficult to reproduce.
3630B - 9 Revision 2
November 1990
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METHOD 3630B
ililCA GEL CLEANUP
Start
7.1.1 Exchange
extract «olv«nt to
eyclohexana during
K-D procedure
7.1.2 Prepare
tlurry activated
•ilica gel, prepare
column
Derivatixed
Phenol*
7.3.1 Deactivate
•ilica gel, prepare
column
7.2.1 Do PFBB
derivatixation on
•ample extract
(8040)
7.3.2 Elute the CC
column with hexane
7.1.3 Preelute
column with
pentane, transfer
extract unto column
and elute with
pentane
7.1.4 Elute column
with
CHiCli/pentane;
concentrate
collected fraction;
adjuit volume
7.2.2 Place
activated liliea
gel in
chromatographio
column; add
anhydrou* tU|SO(
7.3.3 Transfer
extract unto column
and elute with
hexane
7.2.3 Preelute
column with hexane;
pipet hexane
•olution on column;
elute
7.3.4 Exchange the
extraction solvent
to hexane (Section*
7.1.1.1 and
7.1.1.2)
7.2.4 Elute column
with hexane
•olution
Aaalyie
by CC
(Method
8040)
3630B - 10
Revision 2
November 1990
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METHOD 3640A
GEL-PERMEATION CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Gel-permeation chromatography (GPC) is a size exclusion cleanup
procedure using organic solvents and hydrophobic gels in the separation of
synthetic macromolecules (1). The packing gel is porous and is characterized
by the range or uniformity (exclusion range) of that pore size. In the choice
of gels, the exclusion range must be larger than the molecular size of the
molecules to be separated (2). A cross linked divinyl benzene-styrene copolymer
(SX-3 Bio Beads or equivalent) is specified for this method.
1.2 General cleanup application - GPC is recommended for the elimination
from the sample of lipids, polymers, copolymers, proteins, natural resins and
polymers, cellular components, viruses, steroids, and dispersed high-molecular-
weight compounds (2). GPC is appropriate for both polar and non-polar analytes,
therefore, it can be effectively used to cleanup extracts containing a broad
range of analytes.
1.3 Specific application - This method includes guidance for cleanup of
sample extracts containing the following analytes from the RCRA Appendix VIII
and Appendix IX lists:
Compound Name CAS No.*
Acenaphthene 83-32-9
Acenaphthylene 208-96-8
Acetophenone 98-86-2
2-Acetylaminofluorene 53-96-3
Aldrin 309-00-2
4-Aminobiphenyl 92-67-1
Aniline 62-53-3
Anthracene 120-12-7
Benomyl 17804-35-2
Benzenethiol 108-98-5
Benzidine 92-87-5
Benz(a)anthracene 56-55-3
Benzo(b)fluoranthene 205-99-2
Benzo(a)pyrene 50-32-8
Benzo(ghi)perylene 191-24-2
Benzo(k)fluoranthene 207-08-9
Benzoic acid 65-85-0
Benzotrichloride 98-07-7
Benzyl alcohol 100-51-6
Benzyl chloride 100-44-7
alpha-BHC 319-84-6
beta-BHC 319-85-7
gamma-BHC 58-89-9
3640A - 1 Revision 1
November 1990
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Compound Name
CAS No.'
delta-BHC
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
2-sec-butyl-4,6-dinitrophenol (Dinoseb)
Carbazole
Carbendazim
alpha-Chlordane
gamma-Chlordane
4-Chloro-3-methylphenol
4-Chloroaniline
Chiorobenzilate
Bi s(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenol
3-Chlorophenol
4-Chlorophenyl phenyl ether
3-Chloropropionitrile
Chrysene
2-Cresol
3-Cresol
4-Cresol
Cyclophosphamide
ODD
DDE
DDT
Di-n-butyl phthalate
Dial late
Dibenzo(a,e)pyrene
Dibenzo(a,i)pyrene
Dibenz(a,j)acridine
Dibenz(a,h)anthracene
Dibenzofuran
Dibenzothiophene
1,2-Di bromo-3-chloropropane
1,2-Dlbromoethane
trans-l,4-Dichloro-2-butene
cis-l,4-Dichloro-2-butene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dlchlorobenzene
3,3'-Dichlorobenzidine
2,6-Dichlorophenol
2,4-Dichlorophenoxyacetic acid (2,4-D)
2,4-Dichlorophenol
2,4-Dichlorotoluene
l,3-Dichloro-2-propanol
319-86-8
101-55-3
85-68-7
88-85-7
86-74-8
10605-21-7
5103-71-9
5566-34-7
59-50-7
106-47-8
510-15-6
111-91-1
111-44-4
108-60-1
91-58-7
95-57-8
106-48-9
108-43-0
7005-72-3
542-76-7
218-01-9
95-48-7
108-39-4
106-44-5
50-18-0
72-54-8
72-55-9
50-29-3
84-74-2
2303-16-4
192-65-4
189-55-9
224-42-0
53-70-3
132-64-9
132-65-0
96-12-8
106-93-4
110-57-6
1476-11-5
95-50-1
106-46-7
541-73-1
91-94-1
87-65-0
94-75-7
120-83-2
95-73-8
96-23-1
3640A - 2
Revision 1
November 1990
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Compound Name
Dieldrin
Di ethyl phthalate
Dimethoate
Dimethyl phthalate
p-Dimethylaminoazobenzene
7,12-Dimethyl-benz(a)anthracene
2,4-Dimethylphenol
3,3-Dimethylbenzidine
4,6-Dinitro-o-cresol
1,3-Dinitrobenzene
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
Diphenyl ether
1 , 2-Diphenyl hydrazi ne
Disulfoton
Endosulfan sulfate
Endosulfan I
Endosulfan II
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methane sulfonate
Ethyl methacrylate
Bis(2-ethylhexyl) phthalate
Famphur
Fluorene
Fluoranthene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi ene
Hexachl oroethane
Hexachl oropropene
Indeno( 1 , 2 , 3-cd) pyrene
Isodrin
Isophorone
cis-Isosafrole
trans-Isosafrole
Kepone
Malononitrile
Merphos
Methoxychlor
3-Methyl chol anthrene
2-Methyl naphthalene
Methyl parathion
4,4'-Methylene-bis(2-chloroaniline)
CAS No.a
60-57-1
84-66-2
60-51-5
131-11-3
60-11-7
57-97-6
105-67-9
119-93-7
534-52-1
99-65-0
51-28-5
121-14-2
606-20-2
122-39-4
101-84-8
122-66-7
298-04-4
1031-07-8
959-98-8
33213-65-9
72-20-8
7421-93-4
53494-70-5
62-50-0
97-63-2
117-81-7
52-85-7
86-73-7
206-44-0
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
1888-71-7
193-39-5
465-73-6
78-59-1
17627-76-8
4043-71-4
143-50-0
109-77-3
150-50-5
72-43-5
56-49-5
91-57-6
298-00-0
101-14-4
3640A - 3
Revision 1
November 1990
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Compound Name
Naphthalene
1,4-Naphthoqulnone
2-Naphthylamine
1-Naphthylamine
5-Nitro-o-toluidine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenol
4-Nitrophenol
N-Nitrosodi-n-butylamine
N-Nitrosodiethanolamine
N-Nitrosodiethylamine
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi-n-propylamine
N-Ni trosomethyl ethyl ami ne
N-Nitrosomorphol ine
N-Nitrosopiperidine
N-Nitrosopyrolidine
Di-n-octyl phthalate
Parathion
Pentachl orobenzene
Pentachl oroethane
Pentachl oronitrobenzene (PCNB)
Pentachl orophenol
Phenacetin
Phenanthrene
Phenol
1,2-Phenylenediamine
Phorate
2-Picoline
Pronamide
Pyrene
Resorcinol
Safrole
1,2,4, 5-Tetrachl orobenzene
2,3,5 , 6-Tetrachl oroni trobenzene
2,3,5,6-Tetrachlorophenol
2,3,4 , 6-Tetrachl orophenol
Tetraethyl dithiopyrophosphate (Sulfotep)
Thiosemicarbazide
2-Toluidine
4-Toluidine
Thiourea, l-(o-chlorophenyl)
Toluene-2,4-diamine
1 , 2 , 3-Tri chl orobenzene
1, 2, 4-Trichl orobenzene
CAS No."
91-20-3
130-15-4
91-59-8
134-32-7
99-55-8
88-74-4
99-09-2
100-01-6
98-95-3
79-46-9
100-02-7
924-16-3
1116-54-7
55-18-5
62-75-9
86-30-6
621-64-7
10595-95-6
59-89-2
100-75-4
930-55-2
117-84-0
56-38-2
608-93-5
76-01-7
82-68-8
87-86-5
62-44-2
85-01-8
108-95-2
95-54-5
298-02-2
109-06-8
23950-58-5
129-00-0
108-46-3
94-59-7
95-94-3
117-18-0
935-95-5
58-90-2
3689-24-5
79-19-6
106-49-0
95-53-4
5344-82-1
95-80-7
87-61-6
120-82-1
3640A - 4
Revision 1
November 1990
-------
Compound Name CAS No."
2,4,6-Trichlorophenol 88-06-2
2,4,5-Trichlorophenol 95-95-4
2,4,5-Trichlorophenoxyacetic acid (2,4,5-T) 93-76-5
2,4,5-Trichlorophenoxypropionic acid (2,4,5-TP) 93-72-1
Warfarin 81-81-2
8 Chemical Abstract Services Registry Number.
Table 1 presents average percent recovery and percent RSD data for these
analytes, as well as the retention volumes of each analyte on a single 6PC
system. Retention volumes vary from column to column. Figure 1 provides
additional information on retention volumes for certain classes of compounds.
The data for the semivolatiles was determined by GC/MS, whereas, the pesticide
data was determined by GC/ECD or GC/FPD. Compounds not amenable to GC were
determined by HPLC. Other analytes may also be appropriate for this cleanup
technique, however, recovery through the GPC should be >70%.
1.4 Normally, this method is most efficient for removing high boiling
materials that condense in the injection port area of a gas chromatograph (GC)
or the front of the GC column. This residue will ultimately reduce the
chromatographic separation efficiency or column capacity because of adsorption
of the target analytes on the active sites. Pentachlorophenol is especially
susceptible to this problem. GPC, operating on the principal of size exclusion,
will not usually remove interference peaks that appear in the chromatogram since
the molecular size of these compounds is relative similar to the target analytes.
Separation cleanup techniques, based on other molecular characteristics (i.e.,
polarity), must be used to eliminate this type of interference.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of preswelled absorbent,
and is flushed with solvent for an extended period. The column is calibrated
and then loaded with the sample extract to be cleaned up. Elution is effected
with a suitable solvent(s) and the product is then concentrated.
3.0 INTERFERENCES
3.1 A reagent blank should be analyzed for the compound of interest prior
to the use of this method. The level of interferences must be below the
estimated quantitation limits (EQLs) of the analytes of interest before this
method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
3640A - 5 Revision 1
November 1990
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4.0 APPARATUS
4.1 Gel-permeation chromatography system - GPC Autoprep Model 1002 A or B,
or equivalent, Analytical Biochemical Laboratories, Inc. Systems that perform
very satisfactorily have also been assembled from the following components - an
HPLC pump, an auto sampler or a valving system with sample loops, and a fraction
collector. All systems, whether automated or manual, must meet the calibration
requirements of Section 7.2.2.
4.1.1 Chromatographic column - 700 mm x 25 mm ID glass column.
Flow is upward. (Optional) To simplify switching from the UV detector
during calibration to the GPC collection device during extract cleanup,
attach a double 3-way valve (Rheodyne Type 50 Teflon Rotary Valve #10-262
or equivalent) so that the column exit flow can be shunted either to the
UV flow-through cell or to the GPC collection device.
4.1.2 Guard column - (Optional) 5 cm, with appropriate fittings
to connect to the inlet side of the analytical column (Supelco 5-8319 or
equivalent).
4.1.3 Bio Beads (S-X3) - 200-400 mesh, 70 gm (Bio-Rad Laboratories,
Richmond, CA, Catalog 152-2750 or equivalent). An additional 5 gm of Bio
Beads is required if the optional guard column is employed. The quality
of Bio Beads may vary from lot to lot because of excessive fines in some
lots. The UV chromatogram of the Calibration solution should be very
similar to that in Figure 2, and the backpressure should be within 6-10
psi. Also, the gel swell ratio in methylene chloride should be in the
range of 4.4 - 4.8 mL/gm. In addition to fines having a detrimental effect
on chromatography, they can also pass through the column screens and damage
the valve.
4.1.4 Ultraviolet detector - Fixed wavelength (254 nm) with a semi-
prep flow-through cell.
4.1.5 Strip chart recorder, recording integrator or laboratory
data system.
4.1.6 Syringe - 10 mL with Luerlok fitting.
4.1.7 Syringe filter assembly, disposable - Bio-Rad "Prep Disc"
sample filter assembly #343-0005, 25 mm, and 5 micron filter discs or
equivalent. Check each batch for contaminants. Rinse each filter assembly
(prior to use) with methylene chloride if necessary.
4.2 Analytical balance - 0.0001 g.
4.3 Volumetric flasks, Class A - 10 mL to 1000 mL
4.4 Graduated cylinders
5.0 REAGENTS
5.1 Methylene chloride, CH2C12. Pesticide quality or equivalent.
3640A - 6 Revision 1
November 1990
-------
5.1.1 Some brands of methyl ene chloride may contain unacceptably
high levels of acid (HC1). Check the pH by shaking equal portions of
methylene chloride and water, then check the pH of the water layer.
5.1.1.1 If the pH of the water layer is < 5, filter the
entire supply of solvent through a 2 in. x 15 in. glass column
containing activated basic alumina. This column should be
sufficient for processing approximately 20-30 liters of solvent.
Alternatively, a different supply of methylene chloride should be
found.
5.2 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.3 n-Butyl chloride, CH3CH2CH2CH2C1 . Pesticide quality or equivalent.
5.4 GPC Calibration Solution. Prepare a calibration solution in methylene
chloride containing the following analytes (in elution order):
Compound mq/L
corn oil 25,000
bis(2-ethylhexyl) phthalate 1000
methoxychlor 200
perylene 20
sulfur 80
Note: Sulfur is not very soluble in methylene chloride, however, it is soluble
in warm corn oil. Therefore, one approach is to weigh out the corn oil,
warm it and transfer the weighed amount of sulfur into the warm corn oil.
Mix it and then transfer into a volumetric flask with methylene chloride,
along with the other calibration compounds.
Store the calibration solution in an amber glass bottle with a Teflon
lined screw-cap at 4°C, and protect from light. (Refrigeration may cause the
corn oil to precipitate. Before use, allow the calibration solution to stand
at room temperature until the corn oil dissolves.) Replace the calibration
standard solution every 6 months, or more frequently if necessary.
5.5 Corn Oil Spike for Gravimetric Screen. Prepare a solution of corn
oil in methylene chloride (5 mg/100
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 It is very important to have consistent laboratory temperatures during
an entire GPC run, which could be 24 hours or more. If temperatures are not
consistent, retention times will shift, and the dump and collect times determined
3640A - 7 Revision 1
November 1990
-------
by the calibration standard will no longer be appropriate. The ideal laboratory
temperature to prevent outgassing of the methylene chloride is 72°F.
7.2 GPC Setup and Calibration
7.2.1 Column Preparation
7.2.1.1 Weigh out 70 gm of Bio Beads (SX-3). Transfer them
to a quart bottle with a Teflon lined cap or a 500 ml separatory
funnel with a large bore stopcock, and add approximately 300 ml of
methylene chloride. Swirl the container to ensure the wetting of
all beads. Allow the beads to swell for a minimum of 2 hours.
Maintain enough solvent to sufficiently cover the beads at all
times. If a guard column is to be used, repeat the above with 5 gm
of Bio Beads in a 125 ml bottle or a beaker, using 25 ml of
methylene chloride.
7.2.1.2 Turn the column upside down from its normal position,
and remove the inlet bed support plunger (the inlet plunger is
longer than the outlet plunger). Position and tighten the outlet
bed support plunger as near the end as possible, but no closer than
5 cm (measured from the gel packing to the collar).
7.2.1.3 Raise the end of the outlet tube to keep the solvent
in the GPC column, or close the column outlet stopcock if one is
attached. Place a small amount of solvent in the column to minimize
the formation of air bubbles at the base of poured column packing.
7.2.1.4 Swirl the bead/solvent slurry to get a homogeneous
mixture and, if the wetting was done in a quart bottle, quickly
transfer it to a 500 ml separatory funnel with a large bore
stopcock. Drain the excess methylene chloride directly into the
waste beaker, and then start draining the slurry into the column
by placing the separatory funnel tip against the column wall. This
will help to minimize bubble formation. Swirl occasionally to keep
the slurry homogeneous. Drain enough to fill the column. Place the
tubing from the column outlet into a waste beaker below the column,
open the stopcock (if attached) and allow the excess solvent to
drain. Raise the tube to stop the flow and close the stopcock when
the top of the gel begins to look dry. Add additional methylene
chloride to just rewet the gel.
7.2.1.5 Wipe any remaining beads and solvent from the inner
walls of the top of the column with a laboratory tissue. Loosen the
seal slightly on the other plunger assembly (long plunger) and
insert it into the column. Make the seal just tight enough so that
any beads on the glass surface will be pushed forward, but loose
enough so that the plunger can be pushed forward.
CAUTION: Do not tighten the seal if beads are between the seal and the glass
surface because this can damage the seal and cause leakage.
3640A - 8 Revision 1
November 1990
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7.2.1.6 Compress the column as much as possible without
applying excessive force. Loosen the seal and gradually pull out
the plunger. Rinse and wipe off the plunger. Slurry any remaining
beads and transfer them into the column. Repeat Section 7.2.1.5 and
reinsert the plunger. If the plunger cannot be inserted and pushed
in without allowing beads to escape around the seal, continue
compression of the beads without tightening the seal, and loosen and
remove the plunger as described. Repeat this procedure until the
plunger is successfully inserted.
7.2.1.7 Push the plunger until it meets the gel, then compress
the column bed about four centimeters.
7.2.1.8 Pack the optional 5 cm column with approximately 5 gm
of preswelled beads (different guard columns may require different
amounts). Connect the guard column to the inlet of the analytical
column.
7.2.1.9 Connect the column inlet to the solvent reservoir
(reservoir should be placed higher than the top of the column) and
place the column outlet tube in a waste container. Placing a
restrictor in the outlet tube will force air out of the column more
quickly. A restrictor can be made from a piece of capillary
stainless steel tubing of 1/16" OD x 10/1000" ID x 2". Pump
methylene chloride through the column at a rate of 5 mL/min for one
hour.
7.2.1.10 After washing the column for at least one hour,
connect the column outlet tube, without the restrictor, to the inlet
side of the UV detector. Connect the system outlet to the outlet
side of the UV detector. A restrictor (same size as in Section
7.2.1.9) in the outlet tube from the UV detector will prevent bubble
formation which causes a noisy UV baseline. The restrictor will not
effect flow rate. After pumping methylene chloride through the
column for an additional 1-2 hours, adjust the inlet bed support
plunger until approximately 6-10 psi backpressure is achieved. Push
the plunger in to increase pressure or slowly pull outward to reduce
pressure.
7.2.1.11 When the GPC column is not to be used for several
days, connect the column outlet line to the column inlet to prevent
column drying and/or channeling. If channeling occurs, the gel must
be removed from the column, reswelled, and repoured as described
above. If drying occurs, methylene chloride should be pumped through
the column until the observed column pressure is constant and the
column appears wet. Always recalibrate after column drying has
occurred to verify retention volumes have not changed.
7.2.2 Calibration of the GPC Column
7.2.2.1 Using a 10 ml syringe, load sample loop #1 with
calibration solution (Section 5.6). With the ABC automated system,
the 5 ml sample loop requires a minimum of 8 ml of the calibration
solution. Use a firm, continuous pressure to push the sample onto
3640A - 9 Revision 1
November 1990
-------
the loop. Switch the valve so that GPC flow is through the UV flow-
through cell.
7.2.2.2 Inject the calibration solution and obtain a UV trace
showing a discrete peak for each component. Adjust the detector
and/or recorder sensitivity to produce a UV trace similar to Figure
2 that meets the following requirements. Differences between
manufacturers' cell volumes and detector sensitivities may require
a dilution of the calibration solution to achieve similar results.
An analytical flow-through detector cell will require a much less
concentrated solution than the semi-prep cell, and therefore the
analytical cell is not acceptable for use.
7.2.2.3 Following are criteria for evaluating the UV
chromatogram for column condition.
7.2.2.3.1 Peaks must be observed, and should be
symmetrical, for all compounds in the calibration solution.
7.2.2.3.2 Corn oil and phthalate peaks must exhibit
>85% resolution.
7.2.2.3.3 Phthalate and methoxychlor peaks must exhibit
>85% resolution.
7.2.2.3.4 Methoxychlor and perylene peaks must exhibit
>85% resolution.
7.2.2.3.5 Perylene and sulfur peaks must not be
saturated and must exhibit >90% baseline resolution.
7.2.2.4 Calibration for Semivolatiles - Using the information
from the UV trace, establish appropriate collect and dump time
periods to ensure collection of all target analytes. Initiate column
eluate collection just before elution of bis(2-ethylhexyl) phthalate
and after the elution of the corn oil. Stop eluate collection
shortly after the elution of perylene. Collection should be stopped
before sulfur elutes. Use a "wash" time of 10 minutes after the
elution of sulfur. Each laboratory is required to establish its
specific time sequences. See Figure 2 for general guidance on
retention time. Figure 1 illustrates retention volumes for different
classes of compounds.
7.2.2.5 Calibration for Organochlorine Pesticides/PCBs -
Determine the elution times for the phthalate, methoxychlor,
perylene, and sulfur. Choose a dump time which removes >85% of the
phthalate, but collects >95% of the methoxychlor. Stop collection
after the elution of perylene, but before sulfur elutes.
7.2.2.6 Verify the flow rate by collecting column eluate for
10 minutes in a graduated cylinder and measure the volume, which
should be 45-55 ml (4.5-5.5 mL/min). If the flow rate is outside
of this range, corrective action must be taken, as described above.
Once the flow rate is within the range of 4.5-5.5 mL/min, record the
3640A - 10 Revision 1
November 1990
-------
column pressure (should be 6-10 psi) and room temperature. Changes
in pressure, solvent flow rate, and temperature conditions can affect
analyte retention times, and must be monitored. If the flow rate
and/or column pressure do not fall within the above ranges, a new
column should be prepared. A UV trace that does not meet the
criteria in Section 7.2.2.3 would also indicate that a new column
should be prepared. It may be necessary to obtain a new lot of Bio
Beads if the column fails all the criteria.
7.2.2.7 Reinject the calibration solution after appropriate
collect and dump cycles have been set, and the solvent flow and
column pressure have been established.
7.2.2.7.1 Measure and record the volume of collected
GPC eluate in a graduated cylinder. The volume of GPC eluate
collected for each sample extract processed may be used to
indicate problems with the system during sample processing.
7.2.2.7.2 The retention times for bis(2-ethylhexyl)
phthalate and perylene must not vary more than ±5% between
calibrations. If the retention time shift is >5%, take
corrective action. Excessive retention time shifts are caused
by:
7.2.2.7.2.1 Poor laboratory temperature control
or system leaks.
7.2.2.7.2.2 An unstabilized column that requires
pumping methylene chloride through it for several more
hours or overnight.
7.2.2.7.2.3 Excessive laboratory temperatures,
causing outgassing of the methylene chloride.
7.2.2.8 Analyze a GPC blank by loading 5 ml of methylene
chloride into the GPC. Concentrate the methylene chloride that
passes through the system during the collect cycle using a Kuderna-
Danish (KD) evaporator. Analyze the concentrate by whatever
detectors will be used for the analysis of future samples. Exchange
the solvent if necessary. If the blank exceeds the estimated
quantitation limit of the analytes, pump additional methylene
chloride through the system for 1-2 hours. Analyze another GPC blank
to ensure the system is sufficiently clean. Repeat the methylene
chloride pumping if necessary.
7.3 Extract Preparation
7.3.1 Adjust the extract volume to 10.0 ml. The solvent extract
must be primarily methylene chloride. All other solvents, e.g. 1:1
methylene chloride/acetone, must be concentrated to 1 ml (or as low as
possible if a precipitate forms) and diluted to 10.0 mL with methylene
chloride. Thoroughly mix the sample before proceeding.
3640A - 11 Revision 1
November 1990
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7.3.2 Filter the extract through a 5 micron filter disc by attaching
a syringe filter assembly containing the filter disc to a 10 ml syringe.
Draw the sample extract through the filter assembly and into the 10 mL
syringe. Disconnect the filter assembly before transferring the sample
extract into a small glass container, e.g. a 15 ml culture tube with a
Teflon lined screw cap. Alternatively, draw the extract into the syringe
without the filter assembly. Attach the filter assembly and force the
extract through the filter and into the glass container. The latter is
the preferred technique for viscous extracts or extracts with a lot of
solids. Particulate larger than 5 microns may scratch the valve, which
may result in a system leak and cross-contamination of sample extracts in
the sample loops. Repair of the damaged valve is quite expensive.
NOTE; Viscosity of a sample extract should not exceed the viscosity of 1:1
water/glycerol. Dilute samples that exceed this viscosity.
7.4 Screening the Extract
7.4.1 Screen the extract to determine the concentration of dissolved
residue by evaporating a 100 /iL aliquot to dryness and weighing the
residue. The concentration of dissolved residue loaded on the GPC column
cannot exceed 0.500 g. Concentrations exceeding 0.500 g will very likely
result in incomplete extract cleanup and contamination of the GPC switching
valve (which results in cross-contamination of sample extracts).
7.4.1.1 Transfer 100 /nL of the filtered extract from Section
7.3.2 to a tared aluminum weighing dish.
7.4.1.2 A suggested evaporation technique is to use a heat
lamp. Set up a 250 watt heat lamp in a hood so that it is 8 + 0.5 cm
from a surface covered with a clean sheet of aluminum foil. Surface
temperature should be 80-100°C (check temperature by placing a
thermometer on the foil and under the lamp). Place the weighing dish
under the lamp using tongs. Allow it to stay under the lamp for 1
min. Transfer the weighing dish to an analytical balance or a micro
balance and weigh to the nearest 0.1 mg. If the residue weight is
less than 10 mg/100 /*L, then further weighings are not necessary.
If the residue weight is greater than 10 mg/100 pi, then determine
if constant weight has been achieved by placing the weighing dish
and residue back under the heat lamp for 2 or more additional 0.5
min. intervals. Reweigh after each interval. Constant weight is
achieved when three weights agree within ±10%.
7.4.1.3 Repeat the above residue analysis on a blank and a
spike. Add 100 n\. of the same methylene chloride used for the
sample extraction, to a weighing dish and determine residue as above.
Add 100 jil_ of a corn oil spike (5 mg/100 pi) to another weighing
dish and repeat the residue determination.
7.4.2 A residue weight of 10 mg/100 pi of extract represents 500 mg
in 5 mL of extract. Any sample extracts that exceed the 10 mg/100 /xL
residue weight must be diluted so that the 5 mL loaded on the GPC column
does not exceed 0.500 g. When making the dilution, keep in mind that a
3640A - 12 Revision 1
November 1990
-------
minimum volume of 8 ml is required when loading the ABC GPC unit.
Following is a calculation that may be used to determine what dilution is
necessary if the residue exceeds 10 mg.
Y ml taken = 10 ml final x 10 ma maximum
for dilution volume X mg of residue
Example:
Y mL taken = 10 ml final x 10 ma maximum
for dilution volume 15 mg of residue
Y mL taken for dilution = 6.7 ml
Therefore, taking 6.7 mL of sample extract from Section 7.3.2, and
diluting to 10 mL with methylene chloride, will result in 5 mL of diluted
extract loaded on the GPC column that contains 0.500 g of residue.
NOTE; This dilution factor must be included in the final calculation of analyte
concentrations. In the above example, the dilution factor is 1.5.
7.5 GPC Cleanup
7.5.1 Calibrate the GPC at least once per week following the
procedure outlined in Sections 7.2.2 through 7.2.2.6. Ensure that UV
trace requirements, flow rate and column pressure criteria are acceptable.
Also, the retention time shift must be <5% when compared to retention times
in the last calibration UV trace.
7.5.1.1 If these criteria are not met, try cleaning the column
by loading one or more 5 mL portions of butyl chloride and running
it through the column. Butyl chloride removes the discoloration and
particulate that may have precipitated out of the methylene chloride
extracts. If a guard column is being used, replace it with a new
one. This may correct the problem. If column maintenance does not
restore acceptable performance, the column must be repacked with new
Bio Beads and calibrated.
7.5.2 Draw a minimum of 8 mL of extract (diluted, if necessary,
and filtered) into a 10 mL syringe.
7.5.3 Attach the syringe to the turn lock on the injection port.
Use firm, continuous pressure to push the sample onto the 5-mL sample
loop. If the sample is difficult to load, some part of the system may be
blocked. Take appropriate corrective action. If the back pressure is
normal (6-10 psi), the blockage is probably in the valve. Blockage may
be flushed out of the valve by reversing the inlet and outlet tubes and
pumping solvent through the tubes. (This should be done before sample
loading.)
NOTE; Approximately 2 mL of the extract remains in the lines between the
injection port and the sample loop; excess sample also passes through the
sample loop to waste.
3640A - 13 Revision 1
November 1990
-------
7.5.4 After loading a loop, and before removing the syringe from
the injection port, index the 6PC to the next loop. This will prevent
loss of sample caused by unequal pressure in the loops.
7.5.5 After loading each sample loop, wash the loading port with
methylene chloride in a PTFE wash bottle to minimize cross-contamination.
Inject approximately 10 ml of methylene chloride to rinse the common tubes.
7.5.6 After loading all the sample loops, index the GPC to the 00
position, switch to the "RUN" mode and start the automated sequence.
Process each sample using the collect and dump cycle times established in
Section 7.2.2.
7.5.7 Collect each sample in a 250 ml Erlenmeyer flask, covered
with aluminum foil to reduce solvent evaporation, or directly into a
Kuderna-Danish evaporator. Monitor sample volumes collected. Changes in
sample volumes collected may indicate one or more of the following
problems:
7.5.7.1 Change in solvent flow rate, caused by channeling
in the column or changes in column pressure.
7.5.7.2 Increase in column operating pressure due to the
absorption of particles or gel fines onto either the guard column
or the analytical column gel, if a guard column is not used.
7.5.7.3 Leaks in the system or significant variances in
room temperature.
7.6 Concentrate the extract by the standard K-D technique (see any of
the extraction methods, Section 4.2 of this chapter). See the determinative
methods (Chapter Four, Section 4.3) for the final volume.
7.7 It should be remembered that only half of the sample extract is
processed by the GPC (5 ml of the 10 ml extract is loaded onto the GPC column),
and thus, a dilution factor of 2 (or 2 multiplied by any dilution factor in
Section 7.4.2) must be used for quantitation of the sample in the determinative
method.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 3600 for specific quality control
procedures.
8.2 The analyst should demonstrate that the compound(s) of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
3640A - 14 Revision 1
November 1990
-------
9.0 METHOD PERFORMANCE
9.1 Refer to Table 1 for single laboratory performance data.
10.0 REFERENCES
1. Wise, R.H.; Bishop, D.F.; Williams, R.T.; Austern, B.M. "Gel Permeation
Chromatography in the GC/MS Analysis of Organics in Sludges"; U.S. EPA
Municipal Environmental Research Laboratory: Cincinnati, Ohio 45268.
2. Czuczwa, J.; Alford-Stevens, A. "Optimized Gel Permeation Chromatographic
Cleanup for Soil, Sediment, Waste and Waste Oil Sample Extracts for GC/MS
Determination of Semivolatile Organic Pollutants, JAOAC, submitted April
1989.
3. Marsden, P.J.; Taylor, V.; Kennedy, M.R. "Evaluation of Method 3640 Gel
Permeation Cleanup"; Contract No. 68-03-3375, U.S. Environmental Protection
Agency, Cincinnati, Ohio, pp. 100, 1987.
3640A - 15 Revision 1
November 1990
-------
TABLE 1
GPC RECOVERY AND RETENTION VOLUMES FOR RCRA
APPENDIX VIII ANALYTES
Compound
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetylaminofluorene
Aldrin
4-Aminobiphenyl
Aniline
Anthracene
Benomyl
Benzenethiol
Benzidine
Benz(a)anthracene
Benzo(b)fl uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzo(k)fl uoranthene
Benzoic acid
Benzotrichloride
Benzyl alcohol
Benzyl chloride
alpha-BHC
beta-BHC
gamma- BHC
delta-BHC
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
2-sec-butyl-4,6-dinitrophenol (Dinoseb)
Carbazole
Carbendazim
alpha-Chlordane
gamma-Chlordane
4-Chl oro-3-methyl phenol
4-Chloroaniline
Chi orobenzi late
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
2 -Chi oronaphthal ene
2-Chlorophenol
3-Chlorophenol
4-Chlorophenol
4-Chlorophenyl phenyl ether
3-Chloropropionitrile
Chrysene
2-Cresol
3-Cresol
% Rec1
97
72
94
97
99
96
93
89
131
92
95
100
93
93
90
91
66
93
95
99
84
94
93
102
93
104
103
99
131
97
93
87
88
92
89
76
83
89
90
86
87
98
80
102
91
70
% RSD2
2
10
7
2
9
7
4
2
8
11
5
3
5
3
6
4
7
7
17
4
13
9
4
7
1
3
18
5
8
2
2
1
3
5
1
2
2
1
1
3
2
2
5
1
1
3
Ret. Vol.3 (ml
196-235
196-235
176-215
156-195
196-215
176-215
196-235
196-235
146-195
196-235
176-215
196-235
196-235
196-235
196-235
196-235
176-195
176-215
176-215
176-215
196-215
196-215
196-215
216-255
176-215
136-175
176-195
196-255
146-195
196-235
196-215
196-255
196-235
176-235
156-195
156-215
156-195
196-235
196-215
196-215
196-215
176-215
176-215
196-235
196-215
196-215
3640A - 16
Revision 1
November 1990
-------
TABLE 1 (continued)
Compound
4-Cresol
Cyclophosphamide
ODD
DDE
DDT
Di-n-butyl phthalate
Dial late
Dibenzo(a,e)pyrene
Dibenzo(a,i)pyrene
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Dibenzothiophene
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
trans- l,4-Dichloro-2-butene
cis-l,4-Dichloro-2-butene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,6-Dichlorophenol
2,4-Dichlorophenoxyacetic acid (2,4-D)
2,4-Dichlorophenol
2,4-Dichlorotoluene
l,3-Dichloro-2-propanol
Dieldrin
Di ethyl phthalate
Dimethoate
3,3'-Dimethoxybenzidinea
Dimethyl phthalate
p-Dimethylaminoazobenzene
7,12-Dimethyl-benz(a)anthracene
2,4-Dimethylphenol
3, 3' -Dimethyl benzi dine
4,6-Dinitro-o-cresol
1,3-Dinitrobenzene
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
Diphenyl ether
1 , 2-Di phenyl hydrazi ne
Disulfoton
Endosulfan sulfate
Endosulfan I
Endosulfan II
Endrin
% Rec1
88
114
94
94
96
104
97
94
99
117
92
94
94
83
121
107
106
81
81
81
98
86
80
87
70
73
100
103
79
15
100
96
77
93
93
100
99
118
93
101
95
67
92
81
94
99
92
95
%RSD2
2
10
4
2
6
3
6
10
8
9
5
1
3
2
8
6
6
1
1
1
3
3
NA
2
9
13
5
3
15
11
1
1
1
2
2
1
2
7
4
2
6
12
1
15
2
8
6
6
Ret. Vol.3 (ml
196-215
146-185
196-235
196-235
176-215
136-175
156-175
216-235
216-235
176-195
196-235
176-235
196-235
176-215
196-215
176-195
176-215
196-235
196-235
196-235
176-215
196-215
76-215
96-215
196-235
176-215
196-215
136-195
146-185
156-195
156-195
176-215
176-215
176-215
156-215
156-195
156-195
176-195
156-195
156-175
176-235
196-215
176-215
146-165
176-195
176-215
196-215
196-215
3640A - 17
Revision 1
November 1990
-------
TABLE 1 (continued)
Compound
Endrin aldehyde
Endrin ketone
Ethyl methane sulfonate
Ethyl methacrylate
Bis(2-ethylhexyl) phthalate
Famphur
Fluorene
Fluoranthene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutadl ene
Hexachl orocycl opentadi ene
Hexachl oroethane
Hexachl oropropene
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
cis-Isosafrole
trans- Isosaf role
Kepone
Malononitrile
Merphos
Methoxychlor
3-Methyl chol anthrene
2 -Methyl naphthal ene
Methyl parathion
4,4'-Methylene-bis(2-chloroaniline)
Naphthalene
1,4-Naphthoqulnone
2-Naphthylamine
1-Naphthylamine
5-Nitro-o-toluidine
2-Nitroanillne
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenol
4-Nitrophenol
N-Nitroso-di -n-butyl amine
N-Nitrosodiethanolamine
N-Nitrosodiethyl amine
N-Ni trosodimethyl ami ne
N-Nitrosodiphenyl amine
N-Nitrosodi-n-propyl amine
N-Ni trosomethyl ethyl amine
N-Nitrosomorpholine
N-Nitrosopiperidine
% Rec1
97
94
62
126
101
99
95
94
85
91
108
86
89
85
91
79
98
68
90
88
102
111
93
94
74
67
84
96
95
73
94
96
77
96
96
103
86
95
77
89
104
94
86
99
85
83
86
84
%RSD2
1
4
7
7
1
NA
1
1
2
11
2
2
3
1
2
13
5
7
4
16
NA
9
12
6
12
6
13
1
7
7
8
6
2
8
2
8
2
3
3
4
3
2
13
2
4
7
4
4
Ret. Vol.3 (ml
176-215
176-215
176-235
176-195
120-145
126-165
176-235
196-235
195-215
156-195
196-235
176-215
176-215
196-235
196-235
216-255
196-235
156-195
176-215
156-195
196-235
156-195
126-165
156-195
176-195
196-215
146-185
176-215
196-215
176-215
196-235
196-235
176-195
176-215
176-215
176-215
176-195
176-195
196-215
156-175
146-185
156-175
156-195
156-195
156-175
156-175
156-195
156-195
3640A - 18
Revision 1
November 1990
-------
TABLE 1 (continued)
Compound % Rec1 %RSD2 Ret. Vol.3 (ml)
N-Nitrosopyrolidine
Di-n-octyl phthalate
Parathion
Pentachlorobenzene
Pentachloroethane
Pentachloronitrobenzene (PCNB)
Pentachlorophenol
Phenacetin
Phenanthrene
Phenol
1 , 2-Phenyl enedi ami ne
Phorate
2-Picoline
Pronamide
Pyrene
Resorcinol
Safrole
Streptozotocin"
1,2,4,5-Tetrachlorobenzene
2,3,5 , 6-Tetrachl oro-ni trobenzene
2,3,4,6-Tetrachlorophenol
2,3,5 , 6-Tetrachl orophenol
Tetraethyl dithiopyrophosphate (Sulfotep)
Thiosemicarbazide
2-Toluidine
4-Toluidine
Thiourea, l-(o-chlorophenyl)
Toluene- 2, 4-di ami ne
1 , 2 , 3-Tri chl orobenzene
1,2,4-Trichlorobenzene
2, 4, 5-Tri chl orophenol
2, 4, 6-Tri chl orophenol
2,4,5-Trichlorophenoxyacetic acid (2,4,5-T)
2,4,5-Trichlorophenoxypropionic acid
Warfarin
92
83
109
95
74
91
102
100
94
83
91
74
99
105
98
70
93
6
96
85
95
96
89
74
92
87
75
69
87
89
77
95
71
67
94
1
4
14
2
1
8
1
3
2
2
1
NA
14
15
2
6
1
48
2
9
1
7
14
3
3
8
11
7
1
1
1
1
23
NA
2
156-175
120-156
146-170
196-235
196-235
156-195
196-215
156-195
196-235
156-195
196-215
116-135
156-215
156-195
215-235
196-215
176-215
225-245
196-235
176-215
196-215
196-215
116-135
146-185
176-235
176-235
166-185
176-215
196-235
196-235
216-235
216-235
156-235
216-215
166-185
NA = Not applicable, recovery presented as the average of two determinations.
a Not an appropriate analyte for this method.
1 The percent recovery is based on an average of three recovery values.
2 The % relative standard deviation is determined from three recovery values.
3 These Retention Volumes are for guidance only as they will differ from column to
column and from system to system.
3640A - 19 Revision 1
November 1990
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Figure 1
GPC RETENTION VOLUME OF CLASSES OF ANALYTES
W//////////////,
PAH1!
CHLOROBENZENES
PHTHALAT8
OROANOPHOSPHATE
PESTICIDES
CORN OIL —
NITROSAMINE3, NITROAROMATICS
AROMATIC AMINES
NITROPHENOLS
CHLOROPHENOLS
ORQANOCHIORINE
PESTICIOES/PCB't
W///////W////A HERBICIDES (6 160)
— PCP
C-Collect
10
20
30 40
TIME (minutes)
50
60
70
3640A - 20
Revision 1
November 1990
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Figure 2
UV CHROMATOGRAM OF THE CALIBRATION SOLUTION
Injection
5 fflLs
on column
— 0 minutes
Corn oil
25 mg/nL
Bis(2-ethylheayl) .pbthtiate
1.0 mg/nL
Methoxychlor
0.2 ng/mL
Perylene
0.02 mg/mL .
Sulfur
0.08 rng/oL :—
IS minuces
.""' 30 minutes
~,_.^,; ; 45 minutes
700 mm X25 am
70 g Bio-Beads SX
Bed length - 490
CH,C12 at 5.0 uL
254 nu
col iiw^0i._."0^—o«;
1?I_..T._!"_::.':—ILT-— ," ' '..m.-J.i: ~1_ "im_"..'....."JlJ 60 minutes
3640A - 21
Revision 1
November 1990
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METHOD 3640A
GEL-PERMEATION CLEANUP
(START)
7.1 Ensure ambient temp, consistent
throughout GPC run.
i
i
7.2 GPC Setup and Calibration
7.2.1 Column Preparation
7.2.1.1 Place Bio Beads and MeCI
in a container. Swirl and
allow beads to swell.
7.2.1.2 Remove column inlet bed
support plunger. Position
and tighten outlet bed
support plunger to column
end.
7.2.1.3 Ensure GPC column outlet
contains solvent. Place
small amt. solvent in
column to minimize bubble
formation.
7.2.1.4 Transfer bead mixture into
sep. funnel. Drain excess
solvent; drain beads Into
column. Keep beads wet
throughout.
7.2.1.5 Loosen seal on opposite
plunger assembly, insert
into column.
7.2.1.6 Compress column. Slurry
remaining beads and repeat
Section 7.2.1.5 and column
compression.
7.2.1.7 Compress column bed
approximately four cm.
7.2.1.8 Pack option 5 cm. guard
column w/roughly 5 gm.
preswellea beads.
7.2.1.9 Connect column inlet to
solvent reservoir. Pump
MeCI at 5 ml./min. for
1 hr.
7.2.1.10 Connect column outlet to
UV-Vis detector. Place
restrictor at detector outlet.
Run MeCI for additional
1-2 hrs. Compress column
bed to provide 6-10 psi
backpressure.
7.2.1.11 Connect outlet line to column
inlet when column not in
use. Repack column when
channeling is observed.
Assure consistent
backpressure when beads are
rewefted after drying.
3640A - 22
Revision 1
November 1990
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METHOD 3640A
continued
7.2.2 Calibration of tht GPC Column
7.2.2.1
i
i
Load sample loop with
calibration solution.
1
7.2.2.2
!
Inject calibration soln.; adjust
recorder or detector sensitivity
to produce similar UV trace as
Fig. 2 .
1
7.2.2.3
1
Evaluation criteria for UV
chromatogrom.
I
7.2.2.4
r
Calibration for Semivolatiles
Use information from UV
trace to obtain collect and
dump times. Initiate collection
before bis(2-ethylhexyl) phthalate.
stop after perylene. Stop run
before sulfur elutes.
1
7.2.2.5
r
Calibration for Organochlorine
Pesticides/PCBs
Choose dump time which removes
> 85% phthalate. but collects at
times > 95% methoxychlor. Stop
collection between perylene and
sulfur elution.
i
7.2.2.6
t
Verify column flow rate and
backpressure. Correct
inconsistencies when criteria
are not met.
7.2.2.7 Reinject calibration soln. when
collect and dump cycles are set.
and column criteria are met.
7.2.2.7.1 Measure and record volume
of GPC eluate.
7.2.2.7.2 Correct for retention time
shifts of > +/- 5% for
bis(2-e»hylhexyl) phthalate
and perylene.
7.2.2.8 Inject and analyze GPC blank
for column cleanliness. Pump
through MeCI as column wash.
3640A - 23
Revision 1
November 1990
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METHOD 3640A
continued
7.3 Extract Preparation
7.3.1 Adjust extract volume to 10 ml.
Primary solvent should be MeCI.
7.3.2 Filter extract through 5 micron
filter disc/syringe assembly into
small gloss container.
7.4 Screening the Extract
7.4.1 Screen extract by determining
residue wt. of 100 ul aliquot.
7.4.1.1 Transfer 100 ul of filtered
extract from Section 7.3.2
to tared aluminum weighing
dish.
1
7.4.1.2 Evaporate extract solvent
under heating lamp. Weigh
residue to nearest 0.1 mg.
I
7.4.1.3 Repeat residue analysis of
Section 7.4.1.2 w/blonk
and spike sample.
I
7.4.2 Use dilution example to
determine neccessory dilution
when residue wts. > 10 mg.
7.5 GPC Cleanup
7.5.1
i
P
Calibrate GPC weekly. Assure
column criteria, UV trace, retention
time shift criteria are met.
7.5.1.1
Clean column w/butyl chloride
loadings, or replacement of guard
column.
1
i
| 7.5.2 Draw 8 ml. extract into syringe. |
| 7.5.3 Load sample Into injection loop
7.5.4 Index GPC to next loop to
prevent sample loss.
7.5.5 Wash sample port w/MeCI
between sample loadings.
7.5.6 At end of loadings, index GPC to
00, switch to "RUN" mode, start
automated sequence.
7.5.7 Collect sample into aluminum foil
covered Erlenmeyer flask or into
Kuderno-Donish evaporator.
7.6 Concentrate extract by std.
Kuderno-Donish technique.
7.7 Note dilution factor of GPC method
Into final determinations.
STOP
3640A - 24
Revision 1
November 1990
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METHOD 3665
SULFURIC ACID/PERMANGANATE CLEANUP
1.0 SCOPE AND APPLICATION
1.1 This method is suitable for the rigorous cleanup of sample extracts
prior to analysis for polychlorinated biphenyls. This method should be used
whenever elevated baselines or overly complex chromatograms prevent accurate
quantitation of PCBs. This method cannot be used to cleanup extracts for other
target analytes, as it will destroy most organic chemicals including the
pesticides aldrin, dieldrin, endrin, endosulfan (I and II), and endosulfan
sulfate.
2.0 SUMMARY OF METHOD
2.1 An extract is solvent exchanged to hexane, then the hexane is
sequentially treated with (1) concentrated sulfuric acid and (2) 5% aqueous
potassium permanganate. Appropriate caution must be taken with these corrosive
reagents.
2.2 Blanks and replicate analysis samples must be subjected to the same
cleanup as the samples associated with them.
2.3 It is important that all the extracts be exchanged to hexane before
initiating the following treatments.
3.0 INTERFERENCES
3.1 This technique will not destroy chlorinated benzenes, chlorinated
naphthalenes (Halowaxes), and a number of chlorinated pesticides.
4.0 APPARATUS
4.1 Syringe or Class A volumetric pipet, glass; 1.0, 2.0 and 5.0 mL.
4.2 Vials - 1, 2 and 10 mL, glass with Teflon lined screw caps or crimp
tops.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 mL graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
3665 - 1 Revision 0
November 1990
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4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Vortex mixer.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sulfuric acid/Water, H2SOyH20, (1:1, v/v).
5.4 Hexane, C6H14 - Pesticide grade or equivalent.
5.5 Potassium permanganate, KMn04, 5 percent aqueous solution (w/v).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sulfuric acid cleanup
7.1.1 Using a syringe or a volumetric pipet, transfer 1.0 or 2.0 mL
of the hexane extract to a 10 mL vial and, in a fume hood, carefully add
5 mL of the 1:1 sulfuric acid/water solution.
7.1.2 The volume of hexane extract used depends on the requirements
of the GC autosampler used by the laboratory. If the autosampler functions
reliably with 1 mL of sample volume, 1.0 mL of extract should be used.
If the autosampler requires more than 1 mL of sample volume, 2.0 mL of
extract should be used.
CAUTION: Make sure that there is no exothermic reaction nor evolution of gas
prior to proceeding.
3665 - 2 Revision 0
November 1990
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7.1.3 Cap the vial tightly and vortex for one minute. A vortex
must be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks, AVOID SKIN CONTACT,
SULFURIC ACID BURNS.
7.1.4 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer; it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.1.5 If a clean phase separation is achieved, proceed to
Section 7.1.8.
7.1.6 If the hexane layer is colored or the emulsion persists for
several minutes, remove the sulfuric acid layer from the vial and dispose
of it properly. Add another 5 ml of the clean 1:1 sulfuric acid/water.
NOTE: Do not remove any hexane at this stage of the procedure.
7.1.7 Vortex the sample for one minute and allow the phases to
separate.
7.1.8 Transfer the hexane layer to a clean 10 ml vial.
7.1.9 Add an additional 1 ml of hexane to the sulfuric acid layer,
cap and shake. This second extraction is done to ensure quantitative
transfer of the PCBs and Toxaphene.
7.1.10 Remove the second hexane layer and combine with the hexane
from Section 7.1.8.
7.2 Permanganate cleanup
7.2.1 Add 5 ml of the 5 percent aqueous potassium permanganate
solution to the combined hexane fractions from 7.1.10.
CAUTION: Make sure that there is no exothermic reaction nor evolution of gas
prior to proceeding.
7.2.2 Cap the vial tightly and vortex for 1 minute. A vortex must
be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks. AVOID SKIN CONTACT,
POTASSIUM PERMANGANATE BURNS.
7.2.3 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer, it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.2.4 If a clean phase separation is achieved, proceed to
Section 7.2.7.
7.2.5 If the hexane layer is colored or the emulsion persists for
several minutes, remove the permanganate solution from the vial via a
3665 - 3 Revision 0
November 1990
-------
glass pipette and dispose of it properly. Add another 5 ml of the clean
aqueous permanganate solution.
NOTE: Do not remove any hexane at this stage of the procedure.
7.2.6 Vortex the sample and allow the phases to separate.
7.2.7 Transfer the hexane layer to a clean 10 ml vial.
7.2.8 Add an additional 1 ml of hexane to the permanganate layer,
cap the vial securely and shake. This second extraction is done to ensure
quantitative transfer of the PCBs and Toxaphene.
7.2.9 Remove the second hexane layer and combine with the hexane
from Section 7.2.7.
7.3 Final preparation
7.3.1 Reduce the volume of the combined hexane layers to the
original volume (1 or 2 ml) using the Kuderna-Danish Technique
(Section 7.3.1.1).
7.3.1.1 Add one or two clean boiling chips to the flask and
attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 ml of hexane to the top of the column. Place the K-D
apparatus on a hot water bath (15-20°C above the boiling point of
the solvent) so that the concentrator tube is partially immersed in
the hot water and the entire lower rounded surface of the flask is
bathed with hot vapor. Adjust the vertical position of the apparatus
and the water temperature, as required, to complete the concentration
in 10-20 minutes. At the proper rate of distillation the balls of
the column will actively chatter, but the chambers will not flood.
When the apparent volume of liquid reaches 1-2 ml, remove the K-D
apparatus from the water bath and allow it to drain and cool for at
least 10 minutes.
7.3.1.2 Remove the Snyder column and rinse the flask and its
lower joints into the concentrator tube with 1-2 ml of hexane. The
extract may be further concentrated by using either the micro Snyder
column technique (Section 7.3.2) or nitrogen blowdown technique
(Section 7.3.3).
7.3.2 Micro Snyder Column Technique
7.3.2.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top of the column.
Place the K-D apparatus in a hot water bath so that the concentrator
tube is partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 5-10 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus from the water bath and
3665 - 4 Revision 0 „
November 1990
-------
allow It to drain and cool for at least 10 minutes. Remove the
Snyder column and rinse the flask and Its lower joints with about
0.2 ml of hexane and add to the concentrator tube. Adjust the final
volume to 1.0-2.0 ml, as required, with hexane.
7.3.3 Nitrogen Slowdown Technique
7.3.3.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the required
level using a gentle stream of clean, dry nitrogen (filtered through
a column of activated carbon).
CAUTION; Do not use plasticized tubing between the carbon trap and the sample.
7.3.3.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be positioned
to prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become
dry.
7.3.4 Remove any remaining organochlorine pesticides from the
extracts using Florisil Column Cleanup (Method 3620) or Silica Gel Cleanup
(Method 3630).
7.3.5 The extracts obtained may now be analyzed for the target
analytes using the appropriate organic technique(s) (see Section 4.3 of
this Chapter). If analysis of the extract will not be performed
immediately, stopper the concentrator tube and store in a refrigerator.
If the extract will be stored longer than 2 days, it should be transferred
to a vial with a Teflon lined screw cap or crimp top, and labeled
appropriately.
8.0 QUALITY CONTROL
8.2 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 No performance data are currently available.
10.0 REFERENCES
None required.
3665 - 5 Revision 0
November 1990
-------
METHOD 3665
SULFURIC ACID/PERMANGANATF CLEANUP
7 1 1 Carefully
combine hexane
with 1: 1
H,SO,/H,0
solution
7.1.2
Transfer the
appropriate
volume to
vial
7.1.3-714
Cap , vortex.
and allow
phase
separation
/I . 1 . <
f phi
separi
^ clea
.
7.]
Tram
hexane
to clei
,
> Is N.
ise
ition
in? .
Yes
8
fer
layer
in vial
7.1.9 Add
hexane to
H.SO, layer,
cap and shake
7.1.10
Combine two
hexane layer*
716 Remove
\ No and dispose
J > HiSO. solution.
/ add clean H.SO.
solution
7.1.7 Cap.
vortex, and
separation
^^i
<
7.2.1 Add
KMnO,
solution
1
72.2-723
Cap, vortex,
and allow
phase
separation
/T1H l*^
' phase N
. clean? .
Yes
72.7
Transfer
hexane layer
to clean vial
728 Add
hexane to
KMnO. layer,
cap and shake
1
72.9 Combine
two hexane
layers
\ No
• >
7.2.5 Remove
and dispose
KMnO. solution,
add clean KMnO.
solution
7.2.6 Cap.
vortex , and
allow phase
separation
7.3.1-7.3.3
Reduce volume
using K-D
and/or nitrogen
blowdown tech.
I
7.3.4 Use
Method 3620 or
Method 3630 to
further remove
contaminants
1
7 . 3 . 5 Stopper
and
refrigerate
for further
analysis
*
( Stop J
3665 - 6
Revision 0
November 1990
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METHOD 5040A
ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC SAMPLING TRAIN fVOSTh
GAS CHROMATOGRAPHY/MASS SPECTROMETRY TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 5040 was formerly Method 3720 In the Second Edition of this
manual.
1.2 This method covers the determination of volatile principal organic
hazardous constituents (POHCs), collected on Tenax and Tenax/charcoal sorbent
cartridges using a volatile organic sampling train, VOST (1). Much of the
description for purge-and-trap GC/MS analysis is described in Method 8240 of
this chapter. Because the majority of gas streams sampled using VOST will
contain a high concentration of water, the analytical method is based on the
quantitative thermal desorption of volatile POHCs from the Tenax and
Tenax/charcoal traps and analysis by purge-and-trap GC/MS. For the purposes of
definition, volatile POHCs are those POHCs with boiling points less than 100°C.
1.3 This method is applicable to the analysis of Tenax and Tenax/ charcoal
cartridges used to collect volatile POHCs from wet stack gas effluents from
hazardous waste incinerators.
1.4 The sensitivity of the analytical method for a particular volatile
POHC depends on the level of interferences and the presence of detectable levels
of volatile POHCs 1n blanks. The desired target detection limit of the
analytical method is 0.1 ng/L (20 ng on a single pair of traps) for a particular
volatile POHC desorbed from either a single pair of Tenax and Tenax/charcoal
cartridges or by thermal desorption of up to six pairs of traps onto a single
pair of Tenax and Tenax/charcoal traps. The resulting single pair of traps is
then thermally desorbed and analyzed by purge-and-trap GC/MS.
1.5 This method is recommended for use only by experienced mass
spectroscopists or under the close supervision of such qualified persons.
2.0 SUMMARY OF METHOD
2.1 A schematic diagram of the analytical system is shown in Figure 1.
The contents of the sorbent cartridges are spiked with an internal standard and
thermally desorbed for 10 min at 180°C with organic-free nitrogen or helium gas
(at a flow rate of 40 mL/min), bubbled through 5 ml of organic-free reagent
water, and trapped on an analytical adsorbent trap. After the 10 min.
desorption, the analytical adsorbent trap is rapidly heated to 180°C, with the
carrier gas flow reversed so that the effluent flow from the analytical trap is
directed into the GC/MS. The volatile POHCs are separated by temperature
programmed gas chromatography and detected by low-resolution mass spectrometry.
The concentrations of volatile POHCs are calculated using the internal standard
technique.
5040A - 1 Revision 1
November 1990
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3.0 INTERFERENCES
3.1 Refer to Methods 3500 and 8240.
4.0 APPARATUS AND MATERIALS
4.1 Thermal desorption unit:
4.1.1 The thermal desorption unit (for Inside/Inside VOST
cartridges, use Supelco "clamshell" heater; for Inside/Outside VOST
cartridges, user-fabricated unit Is required) should be capable of
thermally desorblng the sorbent resin tubes. It should also be capable
of heating the tubes to 180 ± 10°C with flow of organic-free nitrogen or
helium through the tubes.
4.2 Purge-and-trap unit:
4.2.1 The purge-and-trap unit consists of three separate pieces of
equipment: the sample purger, trap, and the desorber. It should be
capable of meeting all requirements of Method 5030 for analysis of
purgeable organic compounds from water.
4.3 GC/MS system: As described In Method 8240.
5.0 REAGENTS
5.1 Organic-free reagent water. All references to water 1n this method
refer to organic-free reagent water, as defined 1n Chapter One.
5.2 Methanol, CH3OH - Pesticide grade, or equivalent.
5.3 Analytical trap reagents:
5.3.1 2,6-Diphenylene oxide polymer: Tenax (60/80 mesh), chromato-
graphic grade or equivalent.
5.3.2 Methyl silicone packing: 3% OV-1 on Chromosorb W (60/80
mesh) or equivalent.
5.3.3 Silica gel: Davison Chemical (35/00 mesh), Grade 15, or
equivalent.
5.3.4 Charcoal: Petroleum-based (SKC Lot 104 or equivalent).
5.4 Stock standard solution:
5.4.1 Stock standard solutions will be prepared from pure standard
materials or purchased as certified solutions. The stock standards should
be prepared in methanol using assayed liquids or gases, as appropriate.
Because of the toxicity of some of the organohalides, primary dilutions
of these materials should be prepared in a hood. A NIOSH/MESA approved
5040A - 2 Revision 1
November 1990
-------
toxic gas respirator should be used when the analyst handles high
concentrations of such materials.
5.4.2 Fresh stock standards should be prepared weekly for volatile
POHCs with boiling points of <35°C. All other standards must be replaced
monthly, or sooner if comparison with check standards indicates a problem.
5.5 Secondary dilution standards:
5.5.1 Using stock standard solutions, prepare, in methanol,
secondary dilution standards that contain the compounds of interest, either
singly or mixed together. The secondary dilution standards should be
prepared at concentrations such that the desorbed calibration standards
will bracket the working range of the analytical system.
5.6 4-Bromofluorobenzene (BFB) standard:
5.6.1 Prepare a 25 ng//xL solution of BFB in methanol.
5.7 Deuterated benzene:
5.7.1 Prepare a 25 ng//iL solution of benzene-d6 in methanol.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Refer to Method 0030, Chapter Ten.
6.2 Sample trains obtained from the VOST should be analyzed within 2-6
weeks of sample collection.
7.0 PROCEDURE
7.1 Assembly of PTD device:
7.1.1 Assemble a purge-and-trap desorption device (PTD) that meets
all the requirements of Method 5030 (refer to Figure 1).
7.1.2 Connect the thermal desorption device to the PTD device.
Calibrate the PTD-GC/MS system using the internal standard technique.
7.2 Internal standard calibration procedure:
7.2.1 This approach requires the use of deuterated benzene as the
internal standard for these analyses. Other internal standards may be
proposed for use in certain situations. The important criteria for
choosing a particular compound as an internal standard are that it be
similar in analytical behavior to the compounds of interest and that it
can be demonstrated that the measurement of ,the internal standard be
unaffected by method or matrix interferences. Other internal standards
that have been used are d10-ethylbenzene and d4-l,2-dichloroethane. One
adds 50 ng of BFB to all sorbent cartridges (in addition to one or more
5040A - 3 Revision 1
November 1990
-------
internal standards) to provide continuous monitoring of the GC/MS
performance relative to BFB.
7.2.2 Prepare calibration standards at a minimum of three
concentration levels for each analyte of interest.
7.2.3 The calibration standards are prepared by spiking a blank
Tenax or Tenax/charcoal trap with a methanolic solution of the calibration
standards (including 50 ng of the internal standard, such as deuterated
benzene), using the flash evaporation technique. The flash evaporation
technique requires filling the needle of a 5.0 juL syringe with clean
methanol and drawing air into the syringe to the 1.0 /uL mark. This is
followed by drawing a methanolic solution of the calibration standards
(containing 25 /xg/ML of the internal standard) to the 2.0 ML mark. The
glass traps should be attached to the injection port of a gas chromatograph
while maintaining the injector temperature at 160°C. The carrier gas flow
through the traps should be maintained at about 50 mL/min.
7.2.4 After directing the gas flow through the trap, the contents
of the syringe should be slowly expelled through the gas chromatograph
injection port over about 15 sec. After 25 sec have elapsed, the gas flow
through the trap should be shut off, the syringe removed, and the trap
analyzed by the PTD-GC/MS procedure outlined in Method 8240. The total
flow of gas through the traps during addition of calibration standards to
blank cartridges, or internal standards to sample cartridges, should be
25 mL or less.
7.2.5 Analyze each calibration standard for both Tenax and Tenax/
charcoal cartridges according to Section 7.3. Tabulate the area response
of the characteristic ions of each analyte against the concentration of
the internal standard and calculate the response factor (RF) for each
compound, using Equation 1.
RF - ASC|S/A|SCS (1)
where:
As =Area of the characteristic ion for the analyte to be measured.
A,s =Area of the characteristic ion for the internal standard.
Cls =Amount (ng) of the internal standard.
Cs =Amount (ng) of the volatile POHC in calibration standard.
If the RF value over the working range is a constant (<10% RSD),
the RF can be assumed to be invariant, and the average RF can be used for
calculations. Alternatively, the results can be used to plot a calibration
curve of response ratios, As/A,,, versus RF.
7.2.6 The working calibration curve or RF must be verified on each
working day by the measurement of one or more of the calibration standards.
5040A - 4 Revision 1
November 1990
-------
If the response varies by more than ±25% for any analyte, a new calibration
standard must be prepared and analyzed for that analyte.
7.3 The schematic of the PTD-GC/MS system is shown in Figure 1. The
sample cartridge is placed in the thermal desorption apparatus (for Inside/
Inside VOST cartridges, use Supelco "clamshell" heater; for Inside/Outside VOST
cartridges, user fabricated unit is required) and desorbed in the purge-and-trap
system by heating to 180°C for 10 min at a flow rate of 40 mL/min. The desorbed
components pass into the bottom of the water column, are purged from the water,
and collected on the analytical adsorbent trap. After the 10 min desorption
period, the compounds are desorbed from the analytical adsorbent trap into the
GC/MS system according to the procedures described in Method 8240.
7.4 Qualitative analysis
7.4.1 The qualitative identification of compounds determined by this
method is based on retention time, and on comparison of the sample mass
spectrum, after background correction, with characteristic ions in a
reference mass spectrum. The reference mass spectrum must be generated
by the laboratory using the conditions of this method. The characteristic
ions from the reference mass spectrum are defined to be the three ions of
greatest relative intensity, or any ions over 30% relative intensity if
less than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.4.1.1 The intensities of the characteristic ions of a
compound maximize in the same scan or within one scan of each other.
Selection of a peak by a data system target compound search routine,
where the search is based on the presence of a target chromatographic
peak containing ions specific for the target compound at a compound-
specific retention time, will be accepted as meeting this criterion.
7.4.1.2 The RRT of the sample component is within ± 0.06 RRT
units of the RRT of the standard component.
7.4.1.3 The relative intensities of the characteristic ions
agree within 30% of the relative intensities of these ions in the
reference spectrum. (Example: For an ion with an abundance of 50%
in the reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.4.1.4 Structural isomers that produce very similar mass
spectra should be identified as individual isomers if they have
sufficiently different GC retention times. Sufficient GC resolution
is achieved if the height of the valley between two isomer peaks is
less than 25% of the sum of the two peak heights. Otherwise,
structural isomers are identified as isomeric pairs.
7.4.1.5 Identification is hampered when sample components
are not resolved chromatographically and produce mass spectra
containing ions contributed by more than one analyte. When gas
chromatographic peaks obviously represent more than one sample
component (i.e., a broadened peak with shoulder(s) or a valley
between two or more maxima), appropriate selection of analyte spectra
5040A - 5 Revision 1
November 1990
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and background spectra is important. Examination of extracted ion
current profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is apparent),
the identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting compound.
7.4.2 For samples containing components not associated with the
calibration standards, a library search may be made for the purpose of
tentative identification. The necessity to perform this type of
identification will be determined by the type of analyses being conducted.
Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference spectrum
(ions > 10% of the most abundant ion) should be present in the sample
spectrum.
(2) The relative intensities of the major ions should agree within
+ 20%. (Example: For an ion with an abundance of 50% in the standard
spectrum, the corresponding sample ion abundance must be between 30 and
70%).
(3) Molecular ions present in the reference spectrum should be
present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination or
presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the sample
spectrum because of background contamination or coeluting peaks. Data
system library reduction programs can sometimes create these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison of the
sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
7.5 Quantitative analysis
7.5.1 When an analyte has been qualitatively identified,
quantitation should be based on the integrated abundance from the EICP of
the primary characteristic ion chosen for that analyte. If the sample
produces an interference for the primary characteristic ion, a secondary
characteristic ion should be used.
7.5.1.1 Using the internal standard calibration procedure,
the amount of analyte in the sample cartridge is calculated using
the response factor (RF) determined in Section 7.2.5 and Equation 2.
Amount of POHC = ASC|S/A|SRF (2)
5040A - 6 Revision 1
November 1990
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where:
Ag =Area of the characteristic ion for the analyte to be measured.
A,s = Area for the characteristic ion of the internal standard.
C,8 = Amount (ng) of internal standard.
7.5.1.2 The choice of methods for evaluating data collected
using VOST for incinerator trial burns is a regulatory decision.
The procedures used extensively by one user are outlined below.
7.5.1.3 The total amount of the POHCs of interest collected
on a pair of traps should be summed.
7.5.1.4 The observation of high concentrations of POHCs of
interest in blank cartridges indicates possible residual
contamination of the sorbent cartridges prior to shipment to and
use at the site. Data that fall in this category (especially data
indicating high concentrations of POHCs in blank sorbent cartridges)
should be qualified with regard to validity, and blank data should
be reported separately. The applicability of data of this type to
the determination of ORE is a regulatory decision. Continued
observation of high concentrations of POHCs in blank sorbent
cartridges indicates that procedures for cleanup, monitoring,
shipment, and storage of sorbent cartridges by a particular user be
investigated to eliminate this problem.
7.5.1.5 If any internal standard recoveries fall outside the
control limits established in Section 8.4, data for all analytes
determined for that cartridge(s) must be qualified with the
observation.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 0030 for sample preparation procedures.
8.2 Before performing any analyses, the analyst must demonstrate the
ability to generate acceptable precision and accuracy with this method. This
ability is established as described in Section 7.2.
8.3 The laboratory must spike all Tenax and Tenax/charcoal cartridges
with the internal standard(s) to monitor continuing laboratory performance.
This procedure is described in Section 7.2.
8.4 To establish the ability to generate acceptable accuracy and
precision, the analyst must spike blank Tenax and Tenax/charcoal cartridges with
the analytes of interest at two concentrations in the working range.
8.4.1 The average response factor (R) and the standard deviation
(S) for each must be calculated.
5040A - 7 Revision 1
November 1990
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8.4.2 The average recovery and standard deviation must fall within
the expected range for determination of volatile POHCs using this method.
The expected range for recovery of volatile POHCs using this method is 50-
150%.
8.5 Each day, the analyst must demonstrate through analysis of blank
Tenax and Tenax/charcoal cartridges and organic-free reagent water that
interferences from the analytical system are under control.
8.6 The daily GC/MS performance tests required for this method are
described in Method 8240.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. Protocol for Collection and Analysis of Volatile POHC's Using VOST.
EPA/600/8-84-007, March 1984.
2. Validation of the Volatile Organic Sampling Train (VOST) Protocol. Volumes
I and II. EPA/600/4-86-014a, January 1986.
5040A - 8 Revision 1
November 1990
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METHOD 5040A
ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC SAMPLING TRAIN (VOST1
GAS CHROMATOGRAPHY/MASS SPECTROMETRY TECHNIQUE
Start
7 1.1 Assemble
purge and trap
desorption
device
7 1.2 Connect
thermal
desorption
device;
calib. system
7.2.1 Select
internal
standa rd
7.2.3 Prep
calibrati
standards u
flash evapo
techniqu
ing
at.
7.2.4 Direct
gas flow
through traps
7 2.4 Expel
contents of
syringe through
CC injection
port
7.2.4 Analyze
trap by P-T-D
CC/MS
procedure
(Method 8240)
725 Analyze
each ca1ib.
standard for
both cartridges
(see 7.3)
7.2.S Tabulate
area response
and calculate
response factor
7.2.6 Verify
response
factor each
day
7 3 Place
sample
cartridge in
desorp . appar . ;
desorb in P-T
7 . 3 Desorb
into CC/MS
system
(Method 8240)
74.1
Quantatively
identify
volatile POHCs
(Method 8240)
7.5.1 Use
primary
characteristic
ion for
quantification
7.5.1.1
Calculate
amount of
analyte in
sample
(
7 5 1 . 3 Sum
amount of POHCs
of interest for
eacK pair of
traps
7 5-1.4 Analyze
blanks for
signs of
residual
contamina ti on .
7 . 5 . 1 . 5 Compare
int. std.
recoveries to
section 8 . 4
control 1 imi ts
;"~~*x
Stop
_^"
5040A - 9
Revision 1
November 1990
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METHOD 5050
BOMB COMBUSTION METHOD FOR SOLID WASTE
1.0 SCOPE AND APPLICATION
1.1 This method describes the sample preparation steps necessary to
determine total chlorine in solid waste and virgin and used oils, fuels and
related materials, including: crankcase, hydraulic, diesel, lubricating and fuel
oils, and kerosene by bomb oxidation and titration or ion chromatography.
Depending on the analytical finish chosen, other halogens (bromine and fluorine)
and other elements (sulfur and nitrogen) may also be determined.
1.2 The applicable range of this method varies depending on the
analytical finish chosen. In general, levels as low as 500 /zg/g chlorine in the
original oil sample can be determined. The upper range can be extended to
percentage levels by dilution of the combustate.
1.3 This standard may involve hazardous materials, operations, and
equipment. This standard does not purport to address all of the safety problems
associated with its use. It is the responsibility of the user of this standard
to establish appropriate safety and health practices and determine the
applicability of regulatory limitations prior to use. Specific safety statements
are given in Section 3.0.
2.0 SUMMARY OF METHOD
2.1 The sample is oxidized by combustion in a bomb containing oxygen
under pressure. The liberated halogen compounds are absorbed in a sodium
carbonate/sodium bicarbonate solution. Approximately 30 to 40 minutes are
required to prepare a sample by this method. Samples with a high water content
(> 25%) may not combust efficiently and may require the addition of a mineral oil
to facilitate combustion. Complete combustion is still not guaranteed for such
samples.
2.2 The bomb combustate solution can then be analyzed for the following
elements as their anion species by one or more of the following methods:
Method Title
9252 Chloride (Titrimetric, Mercuric Nitrate)
9253 Chloride (Titrimetric, Silver Nitrate)
9056 Anion Chromatography Method (Chloride, Sulfate, Nitrate,
Phosphate, Fluoride, Bromide)
5050 - 1 Revision 0
November 1990
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NOTE: Strict adherence to all of the provisions prescribed hereinafter
ensures against explosive rupture of the bomb, or a blowout,
provided the bomb is of proper design and construction and in good
mechanical condition. It is desirable, however, that the bomb be
enclosed in a shield of steel plate at least 1/2 in. (12.7 mm)
thick, or equivalent protection be provided against unforeseeable
contingencies.
3.0 INTERFERENCES
3.1 Samples with very high water content (> 25%) may not combust
efficiently and may require the addition of a mineral oil to facilitate
combustion.
3.2 To determine total nitrogen in samples, the bombs must first be
purged of ambient air. Otherwise, nitrogen results will be biased high.
4.0 APPARATUS AND MATERIALS
4.1 Bomb, having a capacity of not less than 300 ml, so constructed
that it will not leak during the test, and that quantitative recovery of the
liquids from the bomb may be readily achieved. The inner surface of the bomb may
be made of stainless steel or any other material that will not be affected by the
combustion process or products. Materials used in the bomb assembly, such as the
head gasket and lead-wire insulation, shall be resistant to heat and chemical
action and shall not undergo any reaction that will affect the chlorine content
of the sample in the bomb.
4.2 Sample cup, platinum or stainless steel, 24 mm in outside diameter
at the bottom, 27 mm in outside diameter at the top, 12 mm in height outside, and
weighing 10 to 11 g.
4.3 Firing wire, platinum or stainless steel, approximately No. 26 B
& S gage.
4.4 Ignition circuit, capable of supplying sufficient current to ignite
the nylon thread or cotton wicking without melting the wire.
NOTE: The switch in the ignition circuit shall be of the type that
remains open, except when held in closed position by the operator.
4.5 Nylon sewing thread, or Cotton Wicking, white.
4.6 Funnel, to fit a 100-mL volumetric flask.
4.7 Class A volumetric flasks, 100-mL, one per sample.
4.8 Syringe, 5- or 10-mL disposable plastic.
4.9 Apparatus for specific analysis methods are given in the methods.
4.10 Analytical balance: capable of weighing to 0.0001 g.
5050 - 2 Revision 0
November 1990
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5.0 REAGENTS
5.1 Purity of reagents. Reagent-grade chemicals shall be used in all
tests. Unless otherwise indicated, it is intended that all reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Oxygen. Free of combustible material and halogen compounds,
available at a pressure of 40 atm.
WARNING:
Oxygen vigorously accelerates combustion (see Appendix Al.l)
5.4 Sodium bicarbonate/sodium carbonate solution. Dissolve 2.5200 g
NaHC03 and 2.5440 g Na2C03 in reagent water and dilute to 1 L.
5.5 White oil. Refined.
5.6 Reagents and materials for specific analysis methods are given in
the methods.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Ensure that the portion of the sample used for the test is repre-
sentative of the sample.
6.3 To minimize losses of volatile halogenated solvents that may be
present in the sample, keep the field and laboratory samples as free of headspace
as possible.
6.4 Because used oils may contain toxic and/or carcinogenic substances
appropriate field and laboratory safety procedures should be followed.
7.0 PROCEDURE
7.1
Sample Preparation
7.1.1 Preparation of bomb and sample. Cut a piece of firing wire
approximately 100 mm in length and attach the free ends to the terminals.
Arrange the wire so that it will be just above and not touching the sample
cup. Loop a cotton thread around the wire so that the ends will extend
into the sampling cup. Pipet 10 mL of the NaHCOj/NajCOj solution into the
bomb, wetting the sides. Take an aliquot of the oil sample of approxi-
mately 0.5 g using a 5- or 10-mL disposable plastic syringe, and place in
the sample cup. The actual sample weight is determined by the difference
5050 - 3
Revision 0
November 1990
-------
between the weight of the empty and filled syringe. Do not use more than
1 g of sample.
NOTE: After repeated use of the bomb for chlorine determination, a film
may be noticed on the inner surface. This dullness should be
removed by periodic polishing of the bomb. A satisfactory method
for doing this is to rotate the bomb in a lathe at about 300 rpm
and polish the inside surface with Grit No. 2/0 or equivalent
paper1 coated with a light machine oil to prevent cutting, and
then with a paste of grit-free chromic oxide and water. This
procedure will remove all but very deep pits and put a high polish
on the surface. Before using the bomb, it should be washed with
soap and water to remove oil or paste left from the polishing
operation. Bombs with porous or pitted surfaces should never be
used because of the tendency to retain chlorine from sample to
sample.
NOTE: If the sample is not readily combustible, other nonvolatile,
chlorine-free combustible diluents such as white oil may be
employed. However, the combined weight of sample and nonvolatile
diluent shall not exceed 1 g. Some solid additives are relatively
insoluble but may be satisfactorily burned when covered with a
layer of white oil.
NOTE: The practice of alternately running samples high and low in
chlorine content should be avoided whenever possible. It is
difficult to rinse the last traces of chlorine from the walls of
the bomb, and the tendency for residual chlorine to carry over
from sample to sample has been observed in a number of
laboratories. When a sample high in chlorine has preceded one low
in chlorine content, the test on the low-chlorine sample should
be repeated, and one or both of the low values thus obtained
should be considered suspect if they do not agree within the
limits of repeatability of this method.
NOTE: Do not use more than 1 g total of sample and white oil or other
chlorine-free combustible material. Use of excess amounts of
these materials could cause a buildup of dangerously high pressure
and possible rupture of the bomb.
7.1.2 Addition of oxygen. Place the sample cup in position
and arrange the thread so that the end dips into the sample. Assemble the
bomb and tighten the cover securely. Admit oxygen slowly (to avoid
blowing the oil from the cup) until a pressure is reached as indicated in
Table 1.
NOTE: Do not add oxygen or ignite the sample if the bomb has been
jarred, dropped, or tiled.
1Emery Polishing Paper grit No. 2/0 may be purchased from the Behr-Manning
Co., Troy, NY.
2Chromic oxide may be purchased from J.T. Baker & Co., Phillipsburg, NJ.
5050 - 4 Revision 0
November 1990
-------
7.1.3 Combustion. Immerse the bomb in a cold water bath.
Connect the terminals to the open electrical circuit. Close the circuit
to ignite the sample. Remove the bomb from the bath after immersion for
at least 10 minutes. Release the pressure at a slow, uniform rate such
that the operation requires at least 1 min. Open the bomb and examine the
contents. If traces of unburned oil or sooty deposits are found, discard
the determination, and thoroughly clean the bomb before using it again.
7.1.4 Collection of halogen solution. Using reagent water and
a funnel, thoroughly rinse the interior of the bomb, the sample cup, the
terminals, and the inner surface of the bomb cover into a 100-mL
volumetric flask. Dilute to the mark with reagent water.
7.1.5 Cleaning procedure for bomb and sample cup. Remove any
residual fuse wire from the terminals and the cup. Using hot water, rinse
the interior of the bomb, the sample cup, the terminals, and the inner
surface of the bomb cover. (If any residue remains, first scrub the bomb
with Alconox solution). Copiously rinse the bomb, cover, and cup with
reagent water.
7.2 Sample Analysis. Analyze the combustate for chlorine or other
halogens using the methods listed in Step 2.2. It may be necessary to dilute the
samples so that the concentration will fall within the range of standards.
7.3 Calculations. Calculate the concentrations of each element
detected in the sample according to the following equation:
<=«. * Ve- X DF (1)
' " 5
where:
C0 = concentration of element in the sample, /*g/g
^com = concentration of element in the combustate, ng/ml
V = total volume of combustate, ml
Dh = dilution factor
W0 = weight of sample combusted, g.
Report the concentration of each element detected in the sample in
micrograms per gram.
Example: A 0.5-g oil sample was combusted, yielding 10 ml of combustate.
The combustate was diluted to 100 ml total volume and analyzed for chloride,
which was measured to be 5 /xg/mL. The concentration of chlorine in the original
sample is then calculated as shown below:
5 ug x (10 ml) x (10)
C0 = _mL (2)
0.5 g
C0 = 1,000 MS (3)
g
5050 - 5 Revision 0
November 1990
-------
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 One sample in ten should be bombed twice. The results should agree
to within 10%, expressed as the relative percent difference of the results.
8.3 Analyze matrix spike and matrix spike duplicates - spike samples with
the elements of interest at a level commensurate with the levels being
determined. The spiked compounds should be similar to those expected in the
sample. Any sample suspected of containing > 25% water should also be spiked
with organic chlorine.
8.4 For higher levels (e.g., percent levels), spiking may be
inappropriate. For these cases, samples of known composition should be
combusted. The results should agree to within 10% of the expected result.
8.5 Quality control for the analytical method(s) of choice should be
followed.
9.0 PERFORMANCE
See analytical methods referenced in Step 2.2.
10.0 REFERENCES
1. ASTM Method D 808-81, Standard Test Method for Chlorine in New and Used
Petroleum Products (Bomb Method). 1988 Annual Book of ASTM Standards. Volume
05.01 Petroleum Products and Lubricants.
2. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract No.
68-01-7075, WA 80. July 1988.
5050 - 6 Revision 0
November 1990
-------
TABLE 1.
GAGE PRESSURES
Capacity of bomb, ml
Minimum
gage
pressure , atm
Maximum
gage
pressure , atm
300 to 350
350 to 400
400 to 450
450 to 500
38
35
30
27
40
37
32
29
aThe minimum pressures are specified to provide sufficient oxygen for complete
combustion, and the maximum pressures represent a safety requirement. Refer to
manufacturers' specifications for appropriate gage pressure, which may be lower
than those listed here.
APPENDIX
Al. PRECAUTIONARY STATEMENTS
Al.1 Oxygen
vigorously
Warning—Oxygen
accelerates combustion.
Keep oil and grease away. Do
not use oil or grease on regulators,
gages, or control equipment.
Use only with equipment
conditioned for oxygen service by
careful cleaning to remove oil,
grease, and other combustibles.
Keep combustibles away from
oxygen and eliminate ignition
sources.
Keep surfaces clean to prevent
ignition or explosion, or both, on
contact with oxygen.
Always use a pressure
regulator. Release regulator tension
before opening cylinder valve.
All equipment and containers
used must be suitable and recommended
for oxygen service.
Never attempt to transfer
oxygen from cylinder in which it is
received to any other cylinder. Do
not mix gases in cylinders.
Do not drop cylinder. Make
sure cylinder is secured at all
times.
Keep cylinder valve closed when
not in use.
Stand away from outlet when
opening cylinder valve.
For technical use only. Do not
use for inhalation purposes.
Keep cylinder out of sun and
away from heat.
Keep cylinders from corrosive
environment.
Do not use cylinder without
label.
Do not use dented or damaged
cylinders.
See Compressed Gas Association
booklets G-4 and G4.1 for details of
safe practice in the use of oxygen.
5050 - 7
Revision 0
November 1990
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METHOD 5050
BOMB COMBUSTION METHOD FOR SOLID WASTE
START
7.1.1 Prepare bomb
and sample
I
7.1.2 Slowly add
oxygen to sample
cup
1
7.1.3 Immerse bomb
in co 1 d wa ter ;
igni te sample ;
remove bomb from
water ; release
pressure; open bomb
1
7.1.4 Rinse bomb,
sample cup.
terminals, and bomb
cover with water
p4
715 Rinse bomb,
sample cup,
terminals, and bomb
cover with hot
water
1
7.2 Analyze
combustate
1
7 . 3 Calculate
concentration of
each element
detected
/^ ^\
/ \
I STOP
\^ J
5050 - 8
Revision 0
November 1990
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METHOD 5041
Protocol for Analysis of Sorbent Cartridges from Volatile Organic
Sampling Train; Wide-bore Capillary Column Technique
1.0 SCOPE AND APPLICATION
1.1 This method describes the analysis of volatile principal organic
hazardous constituents (POHCs) collected from the stack gas effluents of
hazardous waste incinerators using the VOST methodology (1). For a comprehensive
description of the VOST sampling methodology see Method 0030. The following
compounds may be determined by this method:
Compound Name
CAS No.'
Acetone
Acrylonitrile
Benzene
Bromodi chloromethane
Bromoform"
Bromomethane0
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chlorodi bromomethane
Chloroethane0
Chloroform
Chloromethane0
Dibromomethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans-1,2-Dichloroethene
1,2-Di chloropropane
cis-l,3-Dichloropropene
trans-1,3-Dichloropropene
Ethyl benzene"
lodomethane
Methylene chloride
Styrene"
1,1,2,2-Tetrachloroethane"
Tetrachloroethene
Toluene
1,1,1-Tri chloroethane
1,1,2-Tri chloroethane
Trichloroethene
Tri chlorof1uoromethane
l,2,3-Trichloropropaneb
Vinyl chloride6
Xylenes"
67-64-1
107-13-1
71-43-2
75-27-4
75-25-2
74-83-9
75-15-0
56-23-5
108-90-7
124-48-1
75-00-3
67-66-3
74-87-3
74-95-3
75-35-3
107-06-2
75-35-4
156-60-5
78-87-5
10061-01-5
10061-02-6
100-41-4
74-88-4
75-09-2
100-42-5
79-34-5
127-18-4
108-88-3
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
75-01-4
5041 - 1
Revision 0
November 1990
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a Chemical Abstract Services Registry Number.
b Boiling point of this compound is above 132°C. Method 0030 is not
appropriate for quantitative sampling of this analyte.
c Boiling point of this compound is below 30°C. Special precautions must
be taken when sampling for this analyte by Method 0030. Refer to Section 1.3 for
discussion.
1.2 This method is most successfully applied to the analysis of non-polar
organic compounds with boiling points between 30°C and 100°C. Data are applied
to the calculation of destruction and removal efficiency (ORE), with limitations
discussed below.
1.3 This method may be applied to analysis of many compounds which boil
above 100°C, but Method 0030 is always inappropriate for collection of compounds
with boiling points above 132°C. All target analytes with boiling points greater
than 132°C are so noted in the target analyte list presented in Section 1.1. Use
of Method 0030 for collection of compounds boiling between 100°C and 132°C is
often possible, and must be decided based on case by case inspection of
information such as sampling method collection efficiency, tube desorption
efficiency, and analytical method precision and bias. An organic compound with
a boiling point below 30°C may break through the sorbent under the conditions
used for sample collection. Quantitative values obtained for compounds with
boiling points below 30°C must be qualified, since the value obtained represents
a minimum value for the compound if breakthrough has occurred. In certain cases,
additional QC measures may have been taken during sampling very low boilers with
Method 0030. This information should be considered during the data
interpretation stage.
When Method 5041 is used for survey analyses, values for compounds boiling
above 132°C may be reported and qualified since the quantity obtained represents
a minimum value for the compound. These minimum values should not be used for
trial burn ORE calculations or to prove insignificant risk.
1.4 The VOST analytical methodology can be used to quantitate volatile
organic compounds that are insoluble or slightly soluble in water. When
volatile, water soluble compounds are included in the VOST organic compound
analyte list, quantitation limits can be expected to be approximately ten times
higher than quantitation limits for water insoluble compounds (if the compounds
can be recovered at all) because the purging efficiency from water (and possibly
from Tenax-GC*) is poor.
1.5 Overall sensitivity of the method is dependent upon the level of
interferences encountered in the sample and the presence of detectable
concentrations of volatile POHCs in blanks. The target detection limit of this
method is 0.1 /*g/m3 (ng/L) of flue gas, to permit calculation of a ORE equal to
or greater than 99.99% for volatile POHCs which may be present in the waste
stream at 100 ppm. The upper end of the range of applicability of this method
is limited by the dynamic range of the analytical instrumentation, the overall
loading of organic compounds on the exposed tubes, and breakthrough of the
volatile POHCs on the sorbent traps used to collect the sample. Table 1 presents
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retention times and characteristic ions for volatile compounds which can be
determined by this method. Table 2 presents method detection limits for a range
of volatile compounds analyzed by the wide-bore VOST methodology.
1.6 The wide-bore VOST analytical methodology is restricted to use by,
or under the supervision of, analysts experienced in the use of sorbent media,
purge-and-trap systems, and gas chromatograph/mass spectrometers, and skilled
in the interpretation of mass spectra and their use as a quantitative tool.
2.0 SUMMARY OF METHOD
2.1 The sorbent tubes are thermally desorbed by heating and purging with
organic-free helium. The gaseous effluent from the tubes is bubbled through
pre-purged organic-free reagent water and trapped on an analytical sorbent trap
in a purge-and-trap unit (Figure 2). After desorption, the analytical sorbent
trap is heated rapidly and the gas flow from the analytical trap is directed to
the head of a wide-bore column under subambient conditions. The volatile organic
compounds desorbed from the analytical trap are separated by temperature
programmed high resolution gas chromatography and detected by continuously
scanning low resolution mass spectrometry (Figure 3). Concentrations of volatile
organic compounds are calculated from a multi-point calibration curve, using the
method of response factors.
3.0 INTERFERENCES
3.1 Sorbent tubes which are to be analyzed for volatile organic compounds
can be contaminated by diffusion of volatile organic compounds (particularly
Freon* refrigerants and common organic solvents) through the external container
(even through a Teflon* lined screw cap on a glass container) and the Swagelok®
sorbent tube caps during shipment and storage. The sorbent tubes can also be
contaminated if organic solvents are present in the analytical laboratory. The
use of blanks is essential to assess the extent of any contamination. Field
blanks must be prepared and taken to the field. The end caps of the tubes are
removed for the period of time required to exchange two pairs of traps on the
VOST sampling apparatus. The tubes are recapped and shipped and handled exactly
as the actual field samples are shipped and handled. At least one pair of field
blanks is included with each six pairs of sample cartridges collected.
3.2 At least one pair of blank cartridges (one Tenax-GC®, one
Tenax-6C*/charcoal) shall be included with shipment of cartridges to a hazardous
waste incinerator site as trip blanks. These trip blanks will be treated like
field blanks except that the end caps will not be removed during storage at the
site. This pair of traps will be analyzed to monitor potential contamination
which may occur during storage and shipment.
3.3 Analytical system blanks are required to demonstrate that
contamination of the purge-and-trap unit and the gas chromatograph/mass
spectrometer has not occurred or that, in the event of analysis of sorbent tubes
with very high concentrations of organic compounds, no compound carryover is
occurring. Tenax® from the same preparation batch as the Tenax* used for field
sampling should be used in the preparation of the method (laboratory) blanks.
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A sufficient number of cleaned Tenax® tubes from the same batch as the field
samples should be reserved in the laboratory for use as blanks.
3.4 Cross contamination can occur whenever low-concentration samples are
analyzed after high-concentration samples, or when several high-concentration
samples are analyzed sequentially. When an unusually concentrated sample is
analyzed, this analysis should be followed by a method blank to establish that
the analytical system is free of contamination. If analysis of a blank
demonstrates that the system is contaminated, an additional bake cycle should
be used. If the analytical system is still contaminated after additional baking,
routine system maintenance should be performed: the analytical trap should be
changed and conditioned, routine column maintenance should be performed (or
replacement of the column and conditioning of the new column, if necessary), and
bakeout of the ion source (or cleaning of the ion source and rods, if required).
After system maintenance has been performed, analysis of a blank is required to
demonstrate that the cleanliness of the system is acceptable.
3.5 Impurities in the purge gas and from organic compounds out-gassing
in tubing account for the majority of contamination problems. The analytical
system must be demonstrated to be free from contamination under the conditions
of the analysis by analyzing two sets of clean, blank sorbent tubes with organic-
free reagent purge water as system blanks. The analytical system is acceptably
clean when these two sets of blank tubes show values for the analytes which are
within one standard deviation of the normal system blank. Use of plastic
coatings, non-Teflon* thread sealants, or flow controllers with rubber
components should be avoided.
3.6 VOST tubes are handled in the laboratory to spike standards and to
position the tubes within the desorption apparatus. When sorbent media are
handled in the laboratory atmosphere, contamination is possible if there are
organic solvents in use anywhere in the laboratory. It is therefore necessary
to make daily use of system blanks to monitor the cleanliness of the sorbents
and the absence of contamination from the analytical system. A single set of
system blank tubes shall be exposed to normal laboratory handling procedures
and analyzed as a sample. This sample should be within one standard deviation
of normal VOST tube blanks to demonstrate lack of contamination of the sorbent
media.
3.7 If the emission source has a high concentration of non-target organic
compounds (for example, hydrocarbons at concentrations of hundreds of ppm), the
presence of these non-target compounds will interfere with the performance of
the VOST analytical methodology. If one or more of the compounds of interest
saturates the chromatographic and mass spectrometric instrumentation, no
quantitative calculations can be made and the tubes which have been sampled under
the same conditions will yield no valid data for any of the saturated compounds.
In the presence of a very high organic loading, even if the compounds of interest
are not saturated, the instrumentation is so saturated that the linear range has
been surpassed. When instrument saturation occurs, it is possible that compounds
of interest cannot even be identified correctly because a saturated mass
spectrometer may mis-assign masses. Even if compounds of interest can be
identified, accurate quantitative calculations are impossible at detector
saturation. No determination can be made at detector saturation, even if the
target compound itself is not saturated. At detector saturation, a negative bias
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will be encountered In analytical measurements and no accurate calculation can
be made for the Destruction and Removal Efficiency if analytical values may be
biased negatively.
3.8 The recoveries of the surrogate compounds, which are spiked on the
VOST tubes immediately before analysis, should be monitored carefully as an
overall indicator of the performance of the methodology. Since the matrix of
stack emissions is so variable, only a general guideline for recovery of 50-150%
can be used for surrogates. The analyst cannot use the surrogate recoveries as
a guide for correction of compound recoveries. The surrogates are valuable only
as a general indicator of correct operation of the methodology. If surrogates
are not observed or if recovery of one or more of the surrogates is outside the
50-150% range, the VOST methodology is not operating correctly. The cause of
the failure in the methodology is not obvious. The matrix of stack emissions
contains large amounts of water, may be highly acidic, and may contain large
amounts of target and non-target organic compounds. Chemical and surface
interactions may be occurring on the tubes. If recoveries of surrogate compounds
are extremely low or surrogate compounds cannot even be identified in the
analytical process, then failure to observe an analyte may or may not imply that
the compound of interest has been removed from the emissions with a high degree
of efficiency (that is, the Destruction and Removal Efficiency for that analyte
is high).
4.0 APPARATUS AND MATERIALS
4.1 Tube desorption apparatus: Acceptable performance of the methodology
requires: 1) temperature regulation to ensure that tube temperature during
desorption is regulated to 180°C ± 10°; 2) good contact between tubes and the
heating apparatus to ensure that the sorbent bed is thoroughly and uniformly
heated to facilitate desorption of organic compounds; and 3) gas-tight
connections to the ends of the tubes to ensure flow of desorption gas through
the tubes without leakage during the heating/desorption process. A simple
clamshell heater which will hold tubes which are 3/4" in outer diameter will
perform acceptably as a desorption apparatus.
4.2 Purge-and-trap device: The purge-and-trap device consists of three
separate pieces of equipment: a sample purge vessel, an analytical trap, and
a desorber. Complete devices are commercially available from a variety of
sources, or the separate components may be assembled. The cartridge thermal
desorption apparatus is connected to the sample purge vessel by 1/8" Teflon*
tubing (unheated transfer line). The tubing which connects the desorption
chamber to the sample purge vessel should be as short as is practical.
4.2.1 The sample purge vessel is required to hold 5 ml of organic-
free reagent water, through which the gaseous effluent from the VOST tubes
is routed. The water column should be at least 3 cm deep. The gaseous
headspace between the water column and the analytical trap must have a
total volume of less than 15 ml. The purge gas must pass through the water
column as finely divided bubbles with a diameter of less than 3 mm at the
origin. The sample purger shown in Figure 4 meets these requirements.
Alternate sample purging vessels may be used if equivalent performance is
demonstrated.
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4.2.2 The analytical trap must be at least 25 cm and have an
internal diameter of at least 0.105 in. The analytical trap must contain
the following components:
2,6-diphenylene oxide polymer: 60/80 mesh, chromatograph grade
(Tenax-GC*, or equivalent)
methyl silicone packing: OV-1 (3%) on Chromosorb-W 60/80
mesh, or equivalent
silica gel: 35/60 mesh, Davison grade 15 or
equivalent
coconut charcoal: prepare from Barneby Cheney,
CA-580-26, or equivalent, by
crushing through 26 mesh screen.
The proportions are: 1/3 Tenax-GC*, 1/3 silica gel, and 1/3
charcoal, with approximately 1.0 cm of methyl silicone packing. The
analytical trap should be conditioned for four hours at 180°C with gas
flow (10 mL/min) prior to use in sample analysis. During conditioning,
the effluent of the trap should not be vented to the analytical column.
The thermal desorption apparatus is connected to the injection system of
the mass spectrometer by a transfer line which is heated to 100°C.
4.2.3 The desorber must be capable of rapidly heating the analytical
trap to 180°C for desorption. The polymer section of the trap should not
exceed 180°C, and the remaining sections should not exceed 220°C, during
bake-out mode.
4.3 Gas chromatograph/mass spectrometer/data system:
4.3.1 Gas chromatograph: An analytical system complete with a
temperature programmable oven with sub-ambient temperature capabilities
and all required accessories, including syringes, analytical columns, and
gases.
4.3.2 Chromatographic column: 30 m x 0.53 mm ID wide-bore fused
silica capillary column, 3 2m film thickness, DB-624 or equivalent.
4.3.3 Mass spectrometer: capable of scanning from 35-260 amu every
second or less, using 70 eV (nominal) electron energy in the electron
ionization mode and producing a mass spectrum that meets all of the
criteria in Table 3 when 50 ng of 4-bromofluorobenzene (BFB) is injected
into the water in the purge vessel.
4.3.4 Gas chromatograph/mass spectrometer interface: Any gas
chromatograph to mass spectrometer interface that gives acceptable
calibration points at 50 ng or less per injection of each of the analytes,
and achieves the performance criteria for 4-bromofluorobenzene shown in
Table 3, may be used. If a glass jet separator is used with the wide-bore
column, a helium make-up flow of approximately 15 ml, introduced after the
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end of the column and prior to the entrance of the effluent to the
separator, will be required for optimum performance.
4.3.5 Data system: A computer system that allows the continuous
acquisition and storage on machine readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows searching any gas chromatographic/mass spectrometric data file for
ions of a specified mass and plotting such ion abundances versus time or
scan number. This type of plot is defined as an Extracted Ion Current
Profile (EICP). Software must also be available that allows the
integration of the ion abundances in any EICP between specified time or
scan number limits. The most recent version of the EPA/NIST Mass Spectral
Library should also be available.
4.4 Wrenches: 9/16", 1/2", 7/16", and 5/16".
4.5 Teflon* tubing: 1/8" diameter.
4.6 Syringes: 25 /il_ syringes (2), 10 /xL syringes (2).
4.7 Fittings: 1/4" nuts, 1/8" nuts, 1/16" nuts, 1/4" to 1/8" union, 1/4"
to 1/4" union, 1/4" to 1/16" union.
4.8 Adjustable stand to raise the level of the desorption unit, if
required.
4.9 Volumetric flasks: 5 ml, class A with ground glass stopper.
4.10 Injector port or equivalent, heated to 180°C for loading standards
onto VOST tubes prior to analysis.
4.11 Vials: 2 ml, with Teflon* lined screw caps or crimp tops.
4.12 Syringe: 5 ml, gas-tight with shutoff valve.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2.1 It is advisable to maintain the stock of organic-free reagent
water generated for use in the purge-and-trap apparatus with a continuous
stream of inert gas bubbled through the water. Continuous bubbling of the
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Inert gas maintains a positive pressure of Inert gas above the water as
a safeguard against contamination.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. To avoid
contamination with other laboratory solvents, it is advisable to maintain a
separate stock of methanol for the preparation of standards for VOST analysis
and to regulate the use of this methanol very carefully.
5.4 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially prepared
stock standards can be used if they are verified against EPA standards. If EPA
standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable. Stock standard solutions must be prepared in high purity methanol.
All preparation of standards should take place in a hood, both to avoid
contamination and to ensure safety of the analyst preparing the standards.
5.4.1 Place about 4 ml of high purity methanol in a 5 ml volumetric
flask. Allow the flask to stand, unstoppered, for about 10 min, or until
all alcohol wetted surfaces have dried.
5.4.1.1 Add appropriate volumes of neat liquid chemicals or
certified solutions, using a syringe of the appropriate volume.
Liquid which is added to the volumetric flask must fall directly
into the alcohol without contacting the neck of the flask. Gaseous
standards can be purchased as methanol solutions from several
commercial vendors.
5.4.1.2 Dilute to volume with high purity methanol, stopper,
and then mix by inverting the flask several times. Calculate
concentration by the dilution of certified solutions or neat
chemicals.
5.4.2 Transfer the stock standard solution into a Teflon* sealed
screw cap bottle. An amber bottle may be used. Store, with minimal
headspace, at -10°C to -20°C, and protect from light.
5.4.3 Prepare fresh standards every two months for gases. Reactive
compounds such as styrene may need to be prepared more frequently. All
other standards must be replaced after six months, or sooner if comparison
with check standards indicates a problem.
5.5 Secondary dilution standards: Using stock standard solutions,
prepare, in high purity methanol, secondary dilution standards containing the
compounds of interest, either singly or mixed together. Secondary dilution
standards must be stored with minimal headspace and should be checked frequently
for signs of degradation or evaporation, especially just prior to preparing
calibration standards from them.
5.6 Surrogate standards: The recommended surrogates are toluene-d6,
4-bromofluorobenzene, and l,2-dichloroethane-d4. Other compounds may be used as
surrogate compounds, depending upon the requirements of the analysis. Surrogate
compounds are selected to span the elution range of the compounds of interest.
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Isotopically labeled compounds are selected to preclude the observation of the
same compounds In the stack emissions. More than one surrogate is used so that
surrogate measurements can still be made even if analytical interferences with
one or more of the surrogate compounds are encountered. However, at least three
surrogate compounds should be used to monitor the performance of the methodology.
A stock surrogate compound solution in high purity methanol should be prepared
as described in Section 5.4, and a surrogate standard spiking solution should
be prepared from the stock at a concentration of 250 /ig/10 mL in high purity
methanol. Each pair of VOST tubes (or each individual VOST tube, if the tubes
are analyzed separately) must be spiked with 10 /ul_ of the surrogate spiking
solution prior to GC/MS analysis.
5.7 Internal standards: The recommended internal standards are
bromochloromethane, 1,4-difluorobenzene, and chlorobenzene-d5. Other compounds
may be used as internal standards as long as they have retention times similar
to the compounds being analyzed by GC/MS. The internal standards should be
distributed through the chromatographic elution range. Prepare internal standard
stock and secondary dilution standards in high purity methanol using the
procedures described in Sections 5.2 and 5.3. The secondary dilution standard
should be prepared at a concentration of 25 mg/L of each of the internal standard
compounds. Addition of 10 pi of this internal standard solution to each pair
of VOST tubes (or to each VOST tube, if the tubes are analyzed individually) is
the equivalent of 250 ng total.
5.8 4-Bromofluorobenzene (BFB) standard: A standard solution containing
25 ng//il_ of BFB in high purity methanol should be prepared for use as a tuning
standard.
5.9 Calibration standards: Calibration standards at a minimum of five
concentrations will be required from the secondary dilution of stock standards
(see Sections 5.2 and 5.3). A range of concentrations for calibration can be
obtained by use of different volumes of a 50 mg/L methanol solution of the
calibration standards. One of the concentrations used should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in field samples but should not exceed the linear range of the GC/MS analytical
system (a typical range for a calibration would be 10, 50, 100, 350, and 500 ng,
for example). Each calibration standard should contain each analyte for
detection by this method. Store calibration standards for one week only in a
vial with no headspace.
5.10 Great care must be taken to maintain the integrity of all standard
solutions. All standards of volatile compounds in methanol must be stored at
-10° to -20°C in amber bottles with Teflon* lined screw caps or crimp tops. In
addition, careful attention must be paid to the use of syringes designated for
a specific purpose or for use with only a single standard solution since cross
contamination of volatile organic standards can occurs very readily.
/
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See Method 0030 for the VOST Sampling Methodology.
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6.2 VOST samples are collected on paired cartridges. The first of the
pair of sorbent cartridges is packed with approximately 1.6 g of Tenax-GC®
resin. The second cartridge of the pair is packed with Tenax-GC* and petroleum
based charcoal (3:1 by volume; approximately 1 g of each). In sampling, the
emissions gas stream passes through the Tenax-GC* layer first and then through
the charcoal layer. The Tenax-GC* is cleaned and reused; charcoal is not reused
when tubes are prepared. Sorbent is cleaned and the tubes are packed. The tubes
are desorbed and subjected to a blank check prior to being sent to the field.
When the tubes are used for sampling (see Figure 5 for a schematic diagram of
the Volatile Organic Sampling Train (VOST)), cooling water is circulated to the
condensers and the temperature of the cooling water is maintained near 0°C. The
end caps of the sorbent cartridges are placed in a clean, screw capped glass
container during sample collection.
6.3 After the apparatus is leak checked, sample collection is
accomplished by opening the valve to the first condenser, turning on the pump,
and sampling at a rate of 1 liter/min for 20 minutes. The volume of sample for
any pair of traps should not exceed 20 liters. An alternative set of conditions
for sample collection requires sampling at a reduced flow rate, where the overall
volume of sample collected is 5 liters at a rate of 0.25 L/min for 20 minutes.
The 20 minute period is required for collecting an integrated sample.
6.4 Following collection of 20 liters of sample, the train is leak checked
a second time at the highest pressure drop encountered during the run to minimize
the chance of vacuum desorption of organics from the Tenax*.
6.5 The train is returned to atmospheric pressure and the two sorbent
cartridges are removed. The end caps are replaced and the cartridges are placed
in a suitable environment for storage and transport until analysis. The sample
is considered invalid if the leak test does not meet specifications.
6.6 A new pair of cartridges is placed in the VOST, the VOST is leak
checked, and the sample collection process is repeated until six pairs of traps
have been exposed.
6.7 All sample cartridges are kept in coolers on cold packs after exposure
and during shipment. Upon receipt at the laboratory, the cartridges are stored
in a refrigerator at 4°C until analysis.
7.0 PROCEDURE
7.1 Recommended operating conditions for cartridge desorber,
purge-and-trap unit, and gas chromatograph/mass spectrometer using the wide-bore
column are:
Cartridge Desorption Oven
Desorb Temperature 180°C
Desorb Time 11 minutes
Desorption Gas Flow 40 mL/min
Desorption/Carrier Gas Helium, Grade 5.0
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Purqe-and-Trap Concentrator
Analytical Trap Desorption Flow
Purge Temperature
Purge Time
Analytical Trap Desorb Temperature
Analytical Trap Desorb Time
Gas Chromatoaraph
Column
Carrier Gas Flow
Makeup Gas Flow
Injector Temperature
Transfer Oven Temperature
Initial Temperature
Initial Hold Time
Program Rate
Final Temperature
Final Hold Time
Mass Spectrometer
Manifold Temperature
Scan Rate
Mass Range
Electron Energy
Source Temperature
2.5 mL/min helium
Ambient
11 minutes
180°C
5 minutes
DB-624, 0.53 mm ID x 30 m thick film
(3 Mm) fused silica capillary, or
equivalent
15 mL/min
15 mL/min
200°C
240°C
5°C
2 minutes
6°C/min
240°C
1 minute, or until elution ceases
105°C
1 sec/cycle
35-260 amu
70 eV (nominal)
According to
specifications
manufacturer's
7.2 Each GC/MS system must be hardware tuned to meet the criteria in
Table 3 for a 50 ng injection of 4-bromofluorobenzene (2 ML injection of the
BFB standard solution into the water of the purge vessel). No analyses may be
initiated until the criteria presented in Table 3 are met.
7.3 Assemble a purge-and-trap device that meets the specifications in
Method 5030. Condition the analytical trap overnight at 180°C in the purge
mode, with an inert gas flow of at least 20 mL/min. Prior to use each day,
condition the trap for 10 minutes by backflushing at 180°C, with the column at
220°C.
7.4 Connect the purge-and-trap device to a gas chromatograph.
7.5 Assemble a VOST tube desorption apparatus which meets the requirements
of Section 4.1.
unit.
7.6 Connect the VOST tube desorption apparatus to the purge-and-trap
7.7 Calibrate the instrument using the internal standard procedure, with
standards and calibration compounds spiked onto cleaned VOST tubes for
calibration.
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7.7.1 Compounds in methanolic solution are spiked onto VOST tubes
using the flash evaporation technique. To perform flash evaporation, the
injector of a gas chromatograph or an equivalent piece of equipment is
required.
7.7.1.1 Prepare a syringe with the appropriate volume of
methanolic standard solution (either surrogates, internal standards,
or calibration compounds).
7.7.1.2 With the injector port heated to 180°C, and with an
inert gas flow of 10 mL/min through the injector port, connect the
paired VOST tubes (connected as in Figure 1, with gas flow in the
same direction as the sampling gas flow) to the injector port;
tighten with a wrench so that there is no leakage of gas. If
separate tubes are being analyzed, an individual Tenax® or
Tenax*/charcoal tube is connected to the injector.
7.7.1.3 After directing the gas flow through the VOST tubes,
slowly inject the first standard solution over a period of 25
seconds. Wait for 5 sec before withdrawing the syringe from the
injector port.
7.7.1.4 Inject a second standard (if required) over a period
of 25 seconds and wait for 5 sec before withdrawing the syringe from
the injector port.
7.7.1.5 Repeat the sequence above as required until all of
the necessary compounds are spiked onto the VOST tubes.
7.7.1.6 Wait for 30 seconds, with gas flow, after the last
spike before disconnecting the tubes. The total time the tubes are
connected to the injector port with gas flow should not exceed 2.5
minutes. Total gas flow through the tubes during the spiking process
should not exceed 25 ml to prevent break through of adsorbed
compounds during the spiking process. To allow more time for
connecting and disconnecting tubes, an on/off valve may be installed
in the gas line to the injector port so that gas is not flowing
through the tubes during the connection/disconnection process.
7.8 Prepare the purge-and-trap unit with 5 ml of organic-free reagent
water in the purge vessel.
7.9 Connect the paired VOST tubes to the gas lines in the tube desorption
unit. The tubes must be connected so that the gas flow during desorption will
be opposite to the flow of gas during sampling: i.e., the tube desorption gas
passes through the charcoal portion of the tube first. An on/off valve may be
installed in the gas line leading to the tube desorption unit in order to prevent
flow of gas through the tubes during the connection process.
7.10 Initiate tube desorption/purge and heating of the VOST tubes in the
desorption apparatus.
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7.11 Set the oven of the gas chromatograph to subambient temperatures by
cooling with liquid nitrogen.
7.12 Prepare the GC/MS system for data acquisition.
7.13 At the conclusion of the tube/water purge time, attach the analytical
trap to the gas chromatograph, adjust the purge-and-trap device to the desorb
mode, and Initiate the gas chromatograph1c program and the GC/MS data
acquisition. Concurrently, Introduce the trapped materials to the gas
chromatographlc column by rapidly heating the analytical trap to 180°C while
backflushing the trap with Inert gas at 2.5 mL/min for 5 mln. Initiate the
program for the gas chromatograph and simultaneously Initiate data acquisition
on the GC/MS system.
7.14 While the analytical trap 1s being desorbed Into the gas
chromatograph, empty the purging vessel. Wash the purging vessel with a minimum
of two 5 ml flushes of organic-free reagent water (or methanol followed by
organic-free reagent water) to avoid carryover of analytes into subsequent
analyses.
7.15 After the sample has been desorbed, recondition the analytical trap
by employing a bake cycle on the purge-and-trap unit. The analytical trap may
be baked at temperatures up to 220°C. However, extensive use of high
temperatures to recondition the trap will shorten the useful life of the
analytical trap. After approximately 11 minutes, terminate the trap bake and
cool the trap to ambient temperatures in preparation for the next sample. This
procedure is a convention for reasonable samples and should be adequate if the
concentration of contamination does not saturate the analytical system. If the
organic compound concentration is so high that the analytical system is saturated
beyond the point where even extended system bakeout is not sufficient to clean
the system, a more extensive system maintenance must be performed. To perform
extensive system maintenance, the analytical trap is replaced and the new trap
1s conditioned. Maintenance is performed on the GC column by removing at least
one foot from the front end of the column. If the chromatography does not
recover after column maintenance, the chromatographic column must be replaced.
The ion source should be baked out and, if the bakeout 1s not sufficient to
restore mass spectrometric peak shape and sensitivity, the ion source and the
quadrupole rods must be cleaned.
7.16 Initial calibration for the analysis of VOST tubes: It is essential
that calibration be performed in the mode in which analysis will be performed.
If tubes are being analyzed as pairs, calibration standards should be prepared
on paired tubes. If tubes are being analyzed Individually, a calibration should
be performed on individual Tenax® only tubes and Tenax'/charcoal tubes.
7.16.1 Prepare the calibration standards by spiking VOST tubes
using the procedure described in Section 7.7.1. Spike each pair of VOST
tubes (or each of the individual tubes) immediately before analysis.
Perform the calibration analyses in order from low concentration to high
to minimize the compound carryover. Add 5.0 ml of organic-free reagent
water to the purging vessel. Initiate tube desorb/purge according to the
procedure described above.
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7.16.2 Tabulate the area response of the characteristic primary
ions (Table 1) against concentration for each target compound, each
surrogate compound, and each internal standard. The first criterion for
quantitative analysis is correct identification of compounds. The
compounds must elute within + 0.06 retention time units of the elution
time of the standard analyzed on the same analytical system on the day of
the analysis. The analytes should be quantitated relative to the closest
eluting internal standard, according to the scheme shown in Table 4.
Calculate response factors (RF) for each compound relative to the internal
standard shown in Table 4. The internal standard selected for the
calculation of RF is the internal standard that has a retention time
closest to the compound being measured. The RF is calculated as follows:
RF = (VCJ/(VCJ
where: A,, = area of the characteristic ion for the compound
being measured.
Als = area of the characteristic ion for the specific
internal standard.
C,s = concentration of the specific internal standard.
Cx * concentration of the compound being measured.
7.16.3 The average RF must be calculated for each compound. A system
performance check should be made before the calibration curve is used.
Five compounds (the System Performance Check Compounds, or SPCCs) are
checked for a minimum average response factor. These compounds are
chloromethane, 1,1-dichloroethane, bromoform, 1,1,2,2-tetrachloroethane,
and chlorobenzene. The minimum acceptable average RF for these compounds
should be 0.300 (0.250 for bromoform). These compounds typically have RFs
of 0.4 - 0.6, and are used to check compound instability and check for
degradation caused by contaminated lines or active sites in the system.
Examples of these occurrences are:
7.16.3.1 Chloromethane: This compound is the most likely
compound to be lost if the purge flow is too fast.
7.16.3.2 Bromoform: This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in transfer lines may adversely affect
response. Response of the primary quantitation ion (m/z 173) is
directly affected by the tuning for 4-bromofluorobenzene at the ions
of masses 174 and 176. Increasing the ratio of ions 174 and 176 to
mass 95 (the base peak of the mass spectrum of bromofluorobenzene)
may improve bromoform response.
7.16.3.3 1,1,2,2-Tetrachloroethane and 1,1-dichloroethane:
These compounds are degraded by contaminated transfer lines in
purge-and-trap systems and/or active sites in trapping materials.
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7.16.4 Using the response factors from the Initial calibration,
calculate the percent relative standard deviation (%RSD) for the
Calibration Check Compounds (CCCs).
%RSD = (SD/X) x 100
where: %RSD = percent relative standard deviation
RF, = individual RF measurement
RF = mean of 5 initial RFs for a compound (the 5 points
over the calibration range)
SD = standard deviation of average RFs for a compound,
where SD is calculated:
SD
- RF)5
The %RSD for each individual CCC should be less than 30 percent. This
criterion must be met in order for the individual calibration to be valid.
The CCCs are: 1,1-dichloroethene, chloroform, 1,2-dichloropropane, toluene,
ethylbenzene, and vinyl chloride.
7.17 Daily GC/MS Calibration
7.17.1 Prior to the analysis of samples, purge 50 ng of the
4-bromofluorobenzene standard. The resultant mass spectrum for the BFB
must meet all of the criteria given in Table 3 before sample analysis
begins. These criteria must be demonstrated every twelve hours of
operation.
7.17.2 The initial calibration curve (Section 7.16) for each
compound of interest must be checked and verified once every twelve hours
of analysis time. This verification is accomplished by analyzing a
calibration standard that is at a concentration near the midpoint
concentration for the working range of the GC/MS and checking the SPCC
(Section 7.16.3) and CCC (Section 7.16.4).
7.17.3 System Performance Check Compounds (SPCCs): A system
performance check must be made each twelve hours of analysis. If the SPCC
criteria are met, a comparison of response factors is made for all
compounds. This is the same check that is applied during the initial
calibration. If the minimum response factors are not achieved, the system
must be evaluated, and corrective action must be taken before analysis is
allowed to begin. The minimum response factor for volatile SPCCs is 0.300
(0.250 for bromoform). If these minimum response factors are not achieved,
some possible problems may be degradation of the standard mixture,
contamination of the injector port, contamination at the front end of the
analytical column, and active sites in the column or chromatographic
system. If the problem is active sites at the front end of the analytical
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column, column maintenance (removal of approximately 1 foot from the front
end of the column) may remedy the problem.
7.17.4 Calibration Check Compounds: After the system performance
check has been met, CCCs listed in Section 7.16.4 are used to check the
validity of the initial calibration. Calculate the percent difference
using the following equation:
(RF, - RFC) x 100
% Difference =
RF,
where: RF, = average response factor from initial calibration
RFC = response factor from current calibration check
standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. Benzene, toluene, and
styrene will have problems with response factors if Tenax® decomposition
occurs (either as a result of sampling exposure or temperature
degradation), since these compounds are decomposition products of Tenax9.
If the percent difference for each CCC is less than 25%, the initial
calibration is assumed to be valid. If the criterion of percent difference
less than 25% is not met for any one CCC, corrective action MUST be taken.
Problems similar to those listed under SPCCs could affect this criterion.
If a source of the problem cannot be determined after corrective action
is taken, a new five-point calibration curve MUST be generated. The
criteria for the CCCs MUST be met before quantitative analysis can begin.
7.17.5 Internal standard responses and retention times in the check
calibration standard must be evaluated immediately after or during data
acquisition. If the retention time for any internal standard changes by
more than 30 seconds from the last check calibration (12 hr), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. A factor which may influence the retention
times of the internal standards on sample tubes is the level of overall
organic compound loading on the VOST tubes. If the VOST tubes are very
highly loaded with either a single compound or with multiple organic
compounds, retention times for standards and compounds of interest will
be affected. If the area for the primary ion of any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration check, the gas chromatograph and mass spectrometer should be
inspected for malfunctions and corrections must be made, as appropriate.
If the level of organic loading of samples is high, areas for the primary
ions of both compounds of interest and standards will be adversely
affected. Calibration check standards should not be subject to variation,
since the concentrations of organic compounds on these samples are set to
be within the linear range of the instrumentation. If instrument
malfunction has occurred, analyses of samples performed under conditions
of malfunction may be invalidated.
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7.18 GC/MS Analysis of Samples
7.18.1 Set up the cartridge desorption unit, purge-and-trap unit,
and GC/MS as described above.
7.18.2 BFB tuning criteria and daily GC/MS calibration check
criteria must be met before analyzing samples.
7.18.3 Adjust the helium purge gas flow rate (through the cartridges
and purge vessel) to approximately 40 mL/min. Optimize the flow rate to
provide the best response for chloromethane and bromoform, if these
compounds are analytes. A flow rate which is too high reduces the recovery
of chloromethane, and an insufficient gas flow rate reduces the recovery
of bromoform.
7.18.4 The first analysis performed after the tuning check and the
calibration or daily calibration check is a method blank. The method blank
consists of clean VOST tubes (both Tenax* and Tenax*/charcoal) which are
spiked with surrogate compounds and internal standards according to the
procedure described in Section 7.7.1. The tubes which are used for the
method blanks should be from the same batch of sorbent as the sorbent used
for the field samples. After the tubes are cleaned and prepared for
shipment to the field, sufficient pairs of tubes should be retained from
the same batch in the laboratory to provide method blanks during the
analysis.
7.18.5 The organic-free reagent water for the purge vessel for the
analysis of each of the VOST samples should be supplied from the labor-
atory inventory which is maintained with constant bubbling of inert gas
to avoid contamination.
7.18.6 If the analysis of a pair of VOST tubes has a concentration
of analytes that exceeds the initial calibration range, no reanalysis of
desorbed VOST tubes is possible. An additional calibration point can be
added to bracket the higher concentration encountered in the samples so
that the calibration database encompasses six or more points.
Alternatively, the data may be flagged in the report as "extrapolated
beyond the upper range of the calibration." The use of the secondary ions
shown in Table 1 is permissible only in the case of interference with the
primary quantitation ion. Use of secondary ions to calculate compound
concentration in the case of saturation of the primary ion is not an
acceptable procedure, since a negative bias of an unpredictable magnitude
is introduced into the quantitative data when saturation of the mass
spectrum of a compound is encountered. If high organic loadings, either
of a single compound or of multiple compounds, are encountered, it is vital
that a method blank be analyzed prior to the analysis of another sample
to demonstrate that no compound carryover is occurring. If concentrations
of organic compounds are sufficiently high that carryover problems are
profound, extensive bakeout of the purge-and-trap unit will be required.
Complete replacement of the contaminated analytical trap, with the
associated requirement for conditioning the new trap, may also be required
for VOST samples which show excessive concentrations of organic compounds.
Other measures which might be required for decontamination of the
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analytical system Include bakeout of the mass spectrometer, replacement
of the filament of the mass spectrometer, cleaning of the ion source of
the mass spectrometer, and/or (depending on the nature of the
contamination) maintenance of the chromatographic column or replacement
of the chromatographic column, with the associated requirement for
conditioning the new chromatographic column.
7.19 Data Interpretation
7.19.1 Qualitative analysis:
7.19.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The reference
mass spectrum must be generated by the laboratory using the
conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.19.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound specific
retention time will be accepted as meeting this criterion.
7.19.1.1.2 The RRT of the sample component is within
+0.06 RRT units of the RRT of the standard component.
7.19.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum. (Example:
For an ion with an abundance of 50% in the reference spectrum,
the corresponding abundance in a sample spectrum can range
between 20% and 80%.)
7.19.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
7.19.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
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appropriate selection of analyte spectra and background spectra
is important. Examination of extracted ion current profiles
of appropriate ions can aid in the selection of spectra, and
in qualitative identification of compounds. When analytes
coelute (i.e., only one chromatographic peak is apparent), the
identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting
compound.
7.19.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for
the purpose of tentative identification. The necessity to perform
this type of identification will be determined by the type of
analyses being conducted. Guidelines for making tentative
identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in
the standard spectrum, the corresponding sample ion abundance must
be between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from
the sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison
of sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
7.19.2 Quantitative analysis:
7.19.2.1 When a compound has been identified, the quantitative
analysis of that compound will be based on the integrated abundance
from the extracted ion current profile of the primary characteristic
ion for that compound (Table 1). In the event that there is
interference with the primary ion so that quantitative measurements
cannot be made, a secondary ion may be used. Note: Use of a
secondary ion to perform quantitative calculations in the event of
the saturation of the primary ion is not an acceptable procedure
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because of the unpredictable extent of the negative bias which is
introduced. Quantitative calculations are performed using the
internal standard technique. The internal standard used to perform
quantitative calculations shall be the internal standard nearest the
retention time of a given analyte (see Table 4).
7.19.2.2 Calculate the amount of each identified analyte from
the VOST tubes as follows:
Amount (ng) = (ASC|S)/(A(SRF)
where: As = area of the characteristic ion for the
analyte to be measured.
Als = area of the characteristic ion of the
internal standard.
Cls = amount (ng) of the internal standard.
7.19.2.3 The choice of methods for evaluating data collected
using the VOST methodology for incinerator trial burns is a
regulatory decision. Various procedures are used to decide whether
blank correction should be performed and how blank correction should
be performed. Regulatory agencies to which VOST data are submitted
also vary in their preferences for data which are or which are not
blank corrected.
7.19.2.4 The total amount of the POHCs of interest collected
on a pair of traps should be summed.
7.19.2.5 The occurrence of high concentrations of analytes
on method blank cartridges indicates possible residual contamination
of sorbent cartridges prior to shipment and use at the sampling site.
Data with high associated blank values must be qualified with respect
to validity, and all blank data should be reported separately. No
blank corrections should be made in this case. Whether or not data
of this type can be applied to the determination of Destruction and
Removal Efficiency is a regulatory decision. Continued observation
of high concentrations of analytes on blank sorbent cartridges
indicates that procedures for cleanup and quality control for the
sampling tubes are inadequate. Corrective action MUST be applied
to tube preparation and monitoring procedures to maintain method
blank concentrations below detection limits for analytes.
7.19.2.6 Where applicable, an estimate of concentration for
noncalibrated components in the sample may be made. The formulae
for quantitative calculations presented above should be used with
the following modifications: The areas A,, and Als should be from the
total ion chromatograms, and the Response Factor for the
noncalibrated compound should be assumed to be 1. The nearest
eluting internal standard free from interferences in the total ion
chromatogram should be used to determine the concentration. The
concentration which is obtained should be reported indicating: (1)
that the value is an estimate; and (2) which internal standard was
used.
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7.19.2.7 If any internal standard recoveries fall outside
the control limits established in Section 8.4, data for all analytes
determined for that cartridge(s) must be qualified with the
observation. Report results without correction for surrogate
compound recovery data. When duplicates are analyzed, report the
data obtained with the sample results.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum Quality Control requirements of
are specified in Chapter One. In addition, this program should consist of an
initial demonstration of laboratory capability and an ongoing analysis of check
samples to evaluate and document data quality. The laboratory must maintain
records to document the quality of the data generated. Ongoing data quality
checks are compared with established performance criteria to determine if the
results of analyses meet the performance characteristics of the method. When
sample analyses indicate atypical method performance, a quality control check
standard (spiked method blank) must be analyzed to confirm that the measurements
were performed in an in-control mode of instrument operation.
8.2 Before processing any samples, the analyst should demonstrate, through
the analysis of a method blank (laboratory blank sorbent tubes, reagent water
purge) that interferences from the analytical system, glassware, sorbent tube
preparation, and reagents are under control. Each time a new batch of sorbent
tubes is analyzed, a method blank should be processed as a safeguard against
chronic laboratory contamination. Blank tubes which have been carried through
all the stages of sorbent preparation and handling should be used in the
analysis.
8.3 The experience of the analyst performing the 6C/MS analyses is
invaluable to the success of the analytical methods. Each day that the analysis
is performed, the daily calibration check standard should be evaluated to
determine if the chromatographic and tube desorption systems are operating
properly. Questions that should be asked are: Do the peaks look normal? Is
the system response obtained comparable to the response from previous
calibrations? Careful examination of the chromatogram of the calibration
standard can indicate whether column maintenance is required or whether the
column is still usable, whether there are leaks in the system, whether the
injector septum requires replacing, etc. If changes are made to the system (such
as change of a column), a calibration check must be carried out and a new
multipoint calibration must be generated.
8.4 Required instrument quality control is found in the following
sections:
8.4.1 The mass spectrometer must be tuned to meet the specifications
for 4-bromofluorobenzene in Section 7.2 (Table 3).
8.4.2 An initial calibration of the tube
desorption/purge-and-trap/GC/MS must be performed as specified in Section
7.7.
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8.4.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.16.3 and the CCC criteria in Section 7.16.4 each twelve hours
of instrument operation.
8.5 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.5.1 A quality control (QC) check sample concentrate is required
containing each analyte at a concentration of 10 mg/L in high purity
methanol. The QC check sample concentrate may be prepared from pure
standard materials or purchased as certified solutions. If the QC check
sample concentrate is prepared by the laboratory, the QC check sample
concentrate must be prepared using stock standards prepared independently
from the stock standards used for calibration.
8.5.2 Spike four pairs of cleaned, prepared VOST tubes with 10 /xL
of the QC check sample concentrate and analyze these spiked VOST tubes
according to the method beginning in Section 7.0.
8.5.3 Calculate the average recovery (X) in ng and the standard
deviation of the recovery (s) in ng for each analyte using the results of
the four analyses.
8.5.4 The average recovery and standard deviation must fall within
the expected range for determination of volatile organic compounds using
the VOST analytical methodology. The expected range for recovery of
volatile organic compounds using this method is 50-150%. Standard
deviation will be compound dependent, but should, in general, range from
15 to 30 ng. More detailed method performance criteria must be generated
from historical records in the laboratory or from interlaboratory studies
coordinated by the Environmental Protection Agency. Since the additional
steps of sorbent tube spiking and desorption are superimposed upon the
methodology of Method 8240, direct transposition of Method 8240 criteria
is questionable. If the recovery and standard deviation for all analytes
meet the acceptance criteria, the system performance is acceptable and the
analysis of field samples may begin. If any individual standard deviation
exceeds the precision limit or any individual recovery falls outside the
range for accuracy, then the system performance is unacceptable for that
analyte. NOTE: The large number of analytes listed in Table 1 presents
a substantial probability that one or more will fail at least one of the
acceptance criteria when all analytes for this method are determined.
8.5.5 When one or more of the analytes tested fails at least one
of the acceptance criteria, the analyst must proceed according to one of
the alternatives below.
8.5.5.1 Locate and correct the source of any problem with the
methodology and repeat the test for all the analytes beginning with
Section 8.5.2.
8.5.5.2 Beginning with Section 8.5.2, repeat the test only
for those analytes that have failed to meet acceptance criteria.
Repeated failure, however, will confirm a general problem either
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with the measurement system or with the applicability of the
methodology to the particular analyte (especially if the analyte in
question is not listed in Table 1). If the problem is identified
as originating in the measurement system, locate and correct the
source of the problem and repeat the test for all compounds of
interest beginning with Section 8.5.2.
8.6 To determine acceptable accuracy and precision limits for surrogate
standards, the following procedure should be performed.
8.6.1 For each sample analyzed, calculate the percent recovery of
each surrogate compound in the sample.
8.6.2 Once a minimum of thirty samples has been analyzed, calculate
the average percent recovery (p) and the standard deviation of the percent
recovery (s) for each of the surrogate compounds.
8.6.3 Calculate the upper and lower control limits for method
performance for each surrogate standard. This calculation is performed
as follows:
Upper Control Limit (UCL) = p + 3s
Lower Control Limit (LCL) = p - 3s
For reference, the comparable control limits for recovery of the
surrogate compounds from water and soil in Method 8240 are:
4-Bromofluorobenzene Water: 86-115% Soil: 74-121%
l,2-Dichloroethane-d4 Water: 76-114% Soil: 70-121%
Toluene-d8 Water: 88-110% Soil: 81-117%
The control limits for the VOST methodology would be expected to be
similar, but exact data are not presently available. Individual laboratory
control limits can be established by the analysis of replicate samples.
8.6.4 If surrogate recovery is not within the limits established
by the laboratory, the following procedures are required: (1) Verify that
there are no errors in calculations, preparation of surrogate spiking
solutions, and preparation of internal standard spiking solutions. Also,
verify that instrument performance criteria have been met. (2) Recalculate
the data and/or analyze a replicate sample, if replicates are available.
(3) If all instrument performance criteria are met and recovery of
surrogates from spiked blank VOST tubes (analysis of a method blank) is
acceptable, the problem is due to the matrix. Emissions samples may be
highly acidic and may be highly loaded with target and non target organic
compounds. Both of these conditions will affect the ability to recover
surrogate compounds which are spiked on the field samples. The surrogate
compound recovery is thus a valuable indicator of the interactions of
sampled compounds with the matrix. If surrogates spiked immediately before
analysis cannot be observed with acceptable recovery, the implications for
target organic analytes which have been sampled in the field must be
assessed very carefully. If chemical or other interactions are occurring
on the exposed tubes, the failure to observe an analyte may not necessarily
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imply that the Destruction and Removal Efficiency for that analyte is
high.
8.7 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples analyzed. Field duplicates may be analyzed to assess the precision of
the environmental measurements. When doubt exists over the identification of
a peak on the chromatogram, confirmatory techniques such as gas chromatography
with a dissimilar column or a different ionization mode using a mass spectrometer
may be used, if replicate samples showing the same compound are available.
Whenever possible, the laboratory should analyze standard reference materials
and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined in Chapter One. The MDL
concentrations listed in Table 2 were obtained using cleaned blanked VOST tubes
and reagent water. Similar results have been achieved with field samples. The
MDL actually achieved in a given analysis will vary depending upon instrument
sensitivity and the effects of the matrix. Preliminary spiking studies indicate
that under these conditions, the method detection limit for spiked compounds in
extremely complex matrices may be larger by a factor of 500-1000.
10.0 REFERENCES
1. Protocol for Collection and Analysis of Volatile POHCs Using VOST.
EPA/600/8-84-007, March, 1984.
2. Validation of the Volatile Organic Sampling Train (VOST) Protocol. Volumes
I and II. EPA/600/4-86-014A, January, 1986.
3. U. S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for
Analysis of Pollutants Under the Clean Water Act, Method 624," October 26,
1984.
4. Bellar, T. A., and J. J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
739-744, 1974.
5. Bellar, T. A., and J. J. Lichtenberg, "Semi-Automated Headspace Analysis
of Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds," in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp 108-129, 1979.
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TABLE 1.
RETENTION TIMES AND CHARACTERISTIC IONS FOR VOLATILE COMPOUNDS
WHICH CAN BE ANALYZED BY METHOD 5041
Retention
Compound Time (min)
Acetone
Acrylonitrile
Benzene
Bromochl oromethane
Bromodichloromethane
4-Bromof 1 uorobenzene
Bromoform
Bromomethane
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chi orodi bromomethane
Chloroethane
Chloroform
Chi oromethane
Di bromomethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1,2-Dichloroethene
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans- 1,3-Dichloropropene
1 ,4-Di f 1 uorobenzene
Ethyl benzene
lodomethane
Methyl ene chloride
Styrene
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,1,1 -Tri chl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1 , 2 , 3-Tri chl oropropane
Vinyl chloride
Xylenes*
7.1
8.6
13.3
12.0
16.0
23.4
22.5
4.1
7.1
12.6
20.5
19.3
4.2
12.2
3.0
15.4
10.0
13.3
6.4
8.6
15.2
17.0
18.2
14.2
21.1
7.0
8.1
22.3
24.0
18.6
17.4
12.4
18.4
14.5
5.1
24.0
3.2
22.2
Primary Ion
Mass
43
53
78
128
83
95
173
94
76
117
112
129
64
83
50
93
63
62
96
96
63
75
75
114
106
142
84
104
83
164
92
97
97
130
101
75
62
106
Secondary Ion(s)
Mass(es)
58
52, 51
52, 77
49, 130, 51
85, 129
174, 176
171, 175, 252
96, 79
78
119, 121
114, 77
208, 206
66, 49
85, 47
52, 49
174, 95
65, 83
64, 98
61, 98
61, 98
62, 41
77, 39
77, 39
63, 88
91
127, 141
49, 51, 86
78, 103
85, 131, 133
129, 131, 166
91, 65
99, 117
83, 85, 99
95, 97, 132
103, 66
110, 77, 61
64, 61
91
* The retention time given is for m- and p-xylene, which coelute on the Megabore
column. o-Xylene elutes approximately 50 seconds later.
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TABLE 2.
PRELIMINARY METHOD DETECTION LIMITS AND BOILING POINTS
FOR VOLATILE ORGANICS ANALYZED BY METHOD 5041*
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methyl ene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1 -Di chloroethane
trans-l,2-Dichloroethene
Chloroform
1,2-Dichl oroethane
1,1,1-Trichloroethane
Carbon tetrachloride
Bromodi chloromethane
1,1,2 , 2-Tetrachl oroethane"
1 , 2 -Di chl oropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1 , 1 , 2-Tri chl oroethane
Benzene
cis-l,3-Djk:hloropropene
Bromoform"
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene"
Styrene"
Tr i chl orof 1 uoromethane
lodomethane
Acrylonitrile
Dibromomethane
1 , 2 , 3 -Tr i chl oropropane"
total Xylenes"
CAS Number
74-87-3
74-83-9
75-01-4
75-00-3
75-09-2
67-64-1
75-15-0
75-35-4
75-35-3
156-60-5
67-66-3
107-06-2
71-55-6
56-23-5
75-27-4
79-34-5
78-87-5
10061-02-6
79-01-6
124-48-1
79-00-5
71-43-2
10061-01-5
75-25-2
127-18-4
108-88-3
108-90-7
100-41-4
100-42-5
75-69-4
74-88-4
107-13-1
74-95-3
96-18-4
Detection
Limit, ng
58
26
14
21
9
35
11
14
12
11
11
13
8
8
11
23
12
17
11
21
26
26
27
26
11
15
15
21
46
17
9
13
14
37
22
Boiling
Point, °C
-24
4
-13
13
40
56
46
32
57
48
62
83
74
77
88
146
95
112
87
122
114
80
112
150
121
111
132
136
145
24
43
78
97
157
138-144
* The method detection limit (MDL) is defined as the minimum concentration of a
substance that can be measured and reported with 99% confidence that the analyte
concentration is greater than zero and is determined from analysis of a sample in
a given matrix containing the analyte. The detection limits cited above were
determined according to Title 40 CFR, Part 136, Appendix B, using standards spiked
onto clean VOST tubes. Since clean VOST tubes were used, the values cited above
represent the best that the methodology can achieve. The presence of an emissions
matrix will affect the ability of the methodology to perform at its optimum level.
** Not appropriate for quantitative sampling by Method 0030.
5041 - 26
Revision 0
November 1990
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TABLE 3.
KEY ION ABUNDANCE CRITERIA FOR 4-BROMOFLUOROBENZENE
Mass Ion Abundance Criteria
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95%, but less than 101% of mass 174
177 5 to 9% of mass 176
5041 - 27 Revision 0
November 1990
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TABLE 4.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
Bromochloromethane
Acetone
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chloromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
Trichloroethene
trans-1,2-Dichloroethene
lodomethane
Methylene chloride
Tri chlorof1uoromethane
Vinyl chloride
1.4-Di f1uorobenzene
Benzene
Bromodichloromethane
Bromoform
Carbon tetrachloride
Chlorodi bromomethane
Dibromomethane
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-1,3-Dichl oropropene
1,1,1-Trichloroethane
1,1,2-Tri chloroethane
Ch1orobenzene-d5
4-Bromofluorobenzene (surrogate)
Chlorobenzene
Ethyl benzene
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
Toluene-d8 (surrogate)
1,2,3-Tri chloropropane
Xylenes
5041 - 28
Revision 0
November 1990
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i 1
V 00
0 ©
Figure 1. Cartridge Desbrptlon Flow
5041 - 29
Revision 0
November 1990
-------
cn
O
t»
O
(D
O
•o
i— O
I
U> 3
Curtridge Oesorplion Unit
1/8" Teflon Tubing
Stand to Raise
Clam Shell Oven
Z30
15
-------
«Q
C
(D
CO
O
(71
O
At
.O
O
A>
o
-h
O
o>
Tube
Oesorplion
Unit
^
Purge and Trap
Apparatus
Gas
Chromatograph
d • i \
^Interlace 1 *•
Mass
Spectrometer
i
I Data System |
1
l
Storage Media
for Archive
CO
ve
CO
If
-------
o
Water Fill Line
Sintered Glass Frit
Gas Flow
Figure 4. Sample Purge Vessel
5041 - 32
Revision 0
November 1990
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cn
o
CO
CO
10
c
(D
cn
co
o
3-
(D
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C*
O
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Class Wool
Particalal.
Slack
(or lesl system)
2
a»
CO
Bl
Exhaust
Condensata
Trap
Impinger
Silica Gel
o
to
-------
METHOD 5041
Protocol for Analysis of Sorbent Cartridges from Volatile Organic
Sampling Train; Wide-bore Capillary Column Technique
C su
7 1 Conditions for
Cartridge
Desorption Oven.
Purge & Trap
Concentrator. CC.
and MS
7 2 Daily, tune
the CC/MS with
BFB and cheek
calib curve
See sect 7 17
73 -76
Assemble the
sys tern.
771 Calibrate the
instrument system us-
ing the internal std
procedure. Stds and
calibration cmpds are
spiked into cleaned
UOST tubes using the
flash evaporation
technique.
7.8 Prep the
purge & trap
unit Kith 5ml
organic-free
reagent Hater.
7 9 Connect
paired VOST
tubes to the
gas lines for
desorption.
7 10 Initiate
tube
desorption/
purge and
heating
7 11 Set the CC
oven to subam-
bient tempera-
ture with
liquid nitrogen
7 12 Prep the
CC/MS system
for data
aquisition.
7 13 After the
tube/water purge
time, attach the
analytical trap to
the CC/MS for
desorption.
7 .14 Hash purg-
ing vessel with
two Sml flushes
of organic-free
reagent water.
7.15 Recondition the
analytical trap by
baking it out at
tamps up to 220 C for
11 min. Trap replace-
ment may be necessary
if the analytical
trap is saturated
beyond cleanup.
7 16.1 Prep
calib stds as
in 7.7.1. Add
*ater to vessel
and desorb.
7 16 2
Tabulate the
area response
of all cmpds
of interest
7.16 3
Calculate the
average RF for
each compound
of interest
7.16 4 Calcu-
late the XRSD
for the CCCs.
The XRSD must
be <30V
7 18 CC/MS
analysis of
samples.
7 19 1 Qualita-
tive analysis
of data and
ident. guide-
1ines of cmpds.
7 19 2 Quanti-
tative analysis
of data for the
compounds of
interest.
5041 - 34
Revision 0
November 1990
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METHOD 5100
DETERMINATION OF THE VOLATILE ORGANIC CONCENTRATION OF WASTE SAMPLES
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the determination of the volatile organic
concentration of hazardous wastes.
1.2 Performance of this method should not be attempted by persons
unfamiliar with the operation of a flame ionization detector (FID) or a Hall
electrolytic conductivity detector (HECD), because knowledge beyond the scope
of this presentation is required.
2.0 SUMMARY OF METHOD
2.1 A sample of waste is collected from a source as close to the point
of generation as practical. The sample is then heated and purged with nitrogen
to separate the volatile organic compounds. Part of the sample is analyzed for
carbon concentration, as methane, with an FID, and part of the sample is analyzed
for chlorine concentration, as chloride, with an HECD. The volatile organic
concentration is the sum of the measured carbon and chlorine concentrations of
the sample.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A field blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
4.0 APPARATUS AND MATERIALS
4.1 Sampling. The following equipment is required:
4.1.1 Static Mixer. Installed in-line or as a by-pass loop, sized
so that the drop size of the dispersed phase is no greater than 1,000 urn.
If the installation of the mixer is in a by-pass loop, then the entire
waste stream must be diverted through the mixer.
4.1.2 Tap. Installed no further than two pipe diameters downstream
of the static mixer outlet.
4.1.3 Sampling Tube. Flexible Teflon, 0.25 in. ID.
5100 - 1 Revision 0
November 1990
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4.1.4 Sample Container. Borosilicate glass or Teflon, 15 to 50 ml,
and a Teflon lined screw cap capable of forming an air tight seal.
4.1.5 Cooling Coil. Fabricated from 0.25 in. ID 304 stainless steel
tubing with a thermocouple at the coil outlet.
4.2 Analysis. The following equipment is required:
4.2.1 Purging Apparatus. For separating the volatile organics from
the waste sample. A schematic of the system is shown in Figure 1. The
purging apparatus consists of the following major components:
4.2.1.1 Purging Chamber. A glass container to hold the sample
while it is heated and purged with dry nitrogen. Exact dimensions
are shown in Figure 3.
The cap of the purging chamber is equipped with three fittings:
one for a mechanical stirrer (fitted with the #11 Ace thread), one
for a thermometer (top fitting), and one for the Teflon exit tubing
(side fitting) as shown in Figure 3.
The base of the purging chamber is a 50 mm inside diameter (ID)
cylindrical glass tube. One end of the tube is fitted with a 50 mm
Ace-thread fitting, while the other end is sealed. Near the sealed
end in the side wall is a fitting for a glass purging lance.
4.2.1.2 Purging Lance. Glass tube, 6 mm ID by 15.25 cm long,
bent into an "L" shape. The "L" end of the tube is sealed, and then
pierced with fifteen holes, each 1 mm in diameter.
4.2.1.3 Mechanical Stirrer. Stainless steel or Teflon
stirring rod driven by an electric motor.
4.2.1.4 Coalescing Filter. Porous fritted disc incorporated
into a container with the same dimensions as the purging chamber.
The details of the design are shown in Figure 3.
4.2.1.5 Constant Temperature Bath. Capable of maintaining
a temperature around the purging chamber and coalescing filter of
75 ± 5°C.
4.2.1.6 Three-way Valves. Two, manually operated, stainless
steel.
4.2.1.7 Flow Controller. Capable of maintaining a purge gas
flow rate of 6 ± 0.006 L/min.
4.2.1.8 Rotameters. Two for monitoring the air flow through
the purging system (0-20 L/min).
4.2.1.9 Sample Splitters. Two heated flow restrictors. At
a purge rate of up to 6 L/min, one will supply a constant flow of
70 to 100 mL/min to the analyzers. The second will split the
analytical flow between the FID and the HECD. The approximate flow
5100 - 2 Revision 0
November 1990
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to the FID will be 40 mL/min and to the HECD will be 15 mL/nrin, but
the exact flow must be adjusted to be compatible with the Individual
detector and to meet Its linearity requirement.
4.2.1.10 Adsorbent Tube. To hold 10 g of activated charcoal.
Excess purge gas Is vented through the adsorbent tube to prevent any
potentially hazardous materials from entering the laboratory.
4.2.2 Volatile Organic Measurement System. Consisting of an FID
to measure the carbon concentration of the sample, and an HECD to measure
the chlorine concentration (as chloride).
4.2.2.1 FID. An FID meeting the following specifications Is
required:
4.2.2.1.1 Linearity. A linear response (+ 5 percent)
over the operating range as demonstrated by the procedures
established in Section 8.1.1.
4.2.2.1.2 Range. A full scale range of 50 pg carbon/sec
to 50 jug carbon/sec. Signal attenuators shall be available
to produce a minimum signal response of 10 percent of full
scale.
4.2.2.1.3 Data Recording System. Analog strip chart
recorder or digital integration system compatible with the FID
for permanently recording the output of the detector.
4.2.2.2 HECD. An HECD meeting the following specifications
is required:
4.2.2.2.1 Linearity. A linear response (+ 10 percent)
over the response range as demonstrated by the procedures in
Section 8.1.2.
4.2.2.2.2 Range. A full scale range of 5.0 pg/sec to
500 ng/sec chloride. Signal attenuators shall be available
to produce a minimum signal response of 10 percent of full
scale.
4.2.2.2.3 Data Recording System. Analog strip chart
recorder or digital integration system compatible with the
output voltage range of HECD.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
Indicated, it 1s Intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without adversely impacting the accuracy of the determination.
5100 - 3 Revision 0
November 1990
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5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water as defined in Chapter One.
5.3 Sampling.
5.3.1 Polyethylene glycol (PEG), 98 percent pure with an average
molecular weight of 400. Remove any organic compounds that may be detected
as volatile organics already present in the polyethylene glycol before it
is used, by heating it to 250°C and purging it with nitrogen at a flow rate
of 1 to 2 L/min for 2 hours. Waste PEG must be disposed of properly
(consult local, State and Federal guidelines and regulations).
5.4 Analysis.
5.4.1 Sample Separation. The following are required for the sample
purging step:
5.4.1.1 Polyethylene glycol. Same as Section 5.3.1.
5.4.1.2 Silicone, Mineral, or Peanut Oil. For use as the heat
dispersing medium in the constant temperature bath.
5.4.1.3 Purging Gas. Zero grade nitrogen (N2), containing
less than 1 ppm carbon.
5.4.2 Volatile Organics Measurement. The following are required
for measuring the volatile organic concentrations:
5.4.2.1 Hydrogen (H2). Zero grade H2, 99.999 percent pure.
5.4.2.2 Combustion Gas. Zero grade air or oxygen, as required
by the FID.
5.4.2.3 FID Calibration Gases.
5.4.2.3.1 Low-level Calibration Gas. Gas mixture
standard with a nominal concentration of 35 ppm (v/v) propane
in N2.
5.4.2.3.2 Mid-level Calibration Gas. Gas mixture
standard with a nominal concentration of 175 ppm (v/v) propane
in N2.
5.4.2.3.3 High-level Calibration Gas. Gas mixture
standard with a nominal concentration of 350 ppm (v/v) propane
in N2.
5.4.2.4 HECD Calibration Gases.
5.4.2.4.1 Low-level Calibration Gas. Gas mixture
standard with a nominal concentration of 20 ppm (v/v) 1,1-
dichlproethene in N2.
5100 - 4 Revision 0
November 1990
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5.4.2.4.2 Mid-level Calibration Gas. Gas mixture
standard with a nominal concentration of 100 ppm (v/v) 1,1-
dichloroethene in N2.
5.4.2.4.3 High-level Calibration Gas. Gas mixture
standard with a nominal concentration of 200 ppm (v/v) 1,1-
dichloroethene in N2.
5.4.2.5 n-Propanol, CH3CH2CH2OH. ACS grade or better.
5.4.2.6 Electrolyte Solution. For use in the conductivity
detector. Mix together 500 ml of water and 500 ml of n-propanol and
store in a glass container.
5.4.2.7 Charcoal. Activated coconut, 12 to 30 mesh.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See Volume One, Section B, Chapter Four, "Organic Analytes,"
Section 4.1.
6.2 Sampling Plan Design and Development. Use the procedures given in
Volume Two, Part III, Chapter Nine, "Sampling Plan."
6.3 Waste in Enclosed Pipes. Sample as close as practical to the point
of waste generation in order to minimize the loss of organics. Assemble the
sampling apparatus as shown in Figure 4. Install the static mixer in the process
line or in a by-pass line. Locate the tap within two pipe diameters of the
static mixer outlet.
6.4 Prepare the sampling containers as follows: Pour into the container
an amount of PEG equal to the total volume of the sample container, less 10 mL.
PEG will reduce, but not eliminate, the loss of volatile organic compounds during
sample collection. Weigh the sample container with the screw cap, the PEG and
any labels to the nearest 0.01 g, and record the weight (mst). Before sampling,
store the containers in an ice bath until the temperature of the PEG is less than
4°C.
6.5 Begin sampling by purging the sample lines and cooling coil with at
least four volumes of waste. Collect the purged material in a separate container
and dispose of it properly.
6.6 After purging, stop the sample flow and direct the sampling tube to
a preweighed sample container, prepared as described in Section 6.4. Keep the
tip of the tube below the surface of the PEG during sampling to minimize contact
with the atmosphere. Sample at a flow rate such that the temperature of the
waste is less than 10°C. Fill the sample container and immediately cap it
(within 5 seconds) so that a minimum headspace exists in the container. Store
immediately in a cooler and cover with ice.
6.7 Alternative sampling techniques may be used upon the approval of the
Administrator.
5100 - 5 Revision 0
November 1990
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7.0 PROCEDURE
7.1 Sample Recovery. Remove the sample container from the cooler, and
wipe the exterior of the container to remove any extraneous Ice, water, or other
debris. Reweigh the sample container and sample to the nearest 0.01 g, and
record the weight (msf). Pour the contents of the sample container into the
purging flask. Rinse the sample container three times with PEG, transferring
the rinsings to the purging flask after each rinse. The total volume of PEG in
the purging flask shall be approximately 50 ml. Add approximately 50 ml of
water.
7.2 Apparatus Assembly. Assemble the purging apparatus as shown in Figure
2, leaving the purging chamber out of the constant temperature bath. Adjust the
stirring rod so that it nearly reaches the bottom of the chamber. Position the
sparger so that it is within 1 cm of the bottom, but does not interfere with the
stirring rod. Lower the thermometer so that it extends into the liquid.
7.3 Sample Analysis. Turn on the constant temperature bath and allow the
temperature to equilibrate at 75 ± 5°C. Turn the bypass valve so that the purge
gas bypasses the purging chamber. Turn on the purge gas. Allow both the FID
and the HECD to warm up until a stable baseline is achieved on each detector.
Pack the adsorbent tube with 10 g of charcoal. Replace the charcoal after each
run and dispose of the spent charcoal properly. Place the assembled chamber in
the constant temperature bath. When the temperature of the PEG reaches 75 ± 5°C,
turn the bypass valve so that the purge gas flows through the purging chamber.
Begin recording the response of the FID and the HECD. Compare the readings
between the two rotameters in the system. If the readings differ by more than
five percent, stop the purging and determine the source of the discrepancy before
resuming.
As purging continues, monitor the output of the FID to make certain that
the separation is proceeding correctly, and that the results are being properly
recorded. Every 10 minutes, read and record the purge flow rate and the liquid
temperature. Continue purging for 30 minutes.
7.4 Initial Performance Check of Purging System. Before placing the
system in operation, after a shutdown of greater than six months, and after any
major modification, conduct the linearity checks described in Sections 7.4.1 and
7.4.2. Install all calibration gases at the three-way calibration gas valve.
See Figure 1.
7.4.1 FID Linearity Check and Calibration. With the purging system
operating as in Section 7.3, allow the FID to establish a stable baseline.
Set the secondary pressure regulator of the calibration gas cylinder to
the same pressure as the purge gas cylinder, and inject the calibration
gas by turning the calibration gas valve to switch flow from the purge gas
to the calibration gas. Continue the calibration gas flow for
approximately two minutes before switching to the purge gas. Make
triplicate injections of each calibration gas (Section 5.4.2.3), and then
calculate the average response factor for each concentration (R,), as well
as the overall mean of the response factor values, R0. The instrument
linearity is acceptable if each R, is within 5 percent of R0 and if the
relative standard deviation (Section 7.7.10) for each set of triplicate
5100 - 6 Revision 0
November 1990
-------
injections is less than 5 percent. Record the overall mean value of the
propane response factor values as the FID calibration response factor, R0.
7.4.2 HECD Linearity Check and Calibration. With the purging system
operating as in Section 7.3, allow the HECD to establish a stable baseline.
Set the secondary pressure regulator of the calibration gas cylinder to
the same pressure as the purge gas cylinder, and inject the calibration
gas by turning the calibration gas valve to switch flow from the purge gas
to the calibration gas. Continue the calibration gas flow for about two
minutes before switching to the purge gas. Make triplicate injections of
each calibration gas (Section 5.4.2.4), and then calculate the average
response factor for each concentration, R,h, as well as the overall mean of
the response factors, Roh. The instrument linearity is acceptable if each
R,h (Section 7.7.5) is within 10 percent of Roh and if the relative standard
deviation (Section 7.7.10) for each set of triplicate injections is less
than 10 percent. Record the overall mean value of the chlorine response
factors as the HECD response factor, Roh.
7.5 Daily Calibrations.
7.5.1 FID Daily Calibration. Inject duplicate samples from the mid-
level FID calibration gas (Section 5.4.2.3.2) as described in Section
7.4.1, and calculate the average daily response factor (DR,). System
operation is adequate if the DR, is within 5 percent of the R0 calculated
during the initial performance test (Section 7.4.1). Use the DR, for
calculation of carbon content in the samples.
7.5.2 HECD Daily Calibration. Inject duplicate samples from the
mid-level HECD calibration gas (Section 5.4.2.4.2) as described in Section
7.4.2, and calculate the average daily response factor DR,h. The system
operation is adequate if the DR,h is within 10 percent of the Roh calculated
during the initial performance test (Section 7.4.2). Use the DR,h for
calculation of chlorine in the samples.
7.6 Water Blank. Transfer about 60 ml of organic-free reagent water into
the purging chamber. Add 50 mL of PEG to the purging chamber. Treat the blank
as described in Sections 7.2 and 7.3.
7.7 Calculations
7.7.1 Nomenclature.
A,, = Area under the water blank response curve, counts.
A8 = Area under the sample response curve, counts.
C = Concentration of volatile organic in the sample, ppm(w/w).
Cc = Concentration of FID calibration gas, ppm(v/v).
Ch = Concentration of HECD calibration gas, ppm(v/v).
DR, = Average daily response factor of the FID, ng C/counts.
5100 - 7 Revision 0
November 1990
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DR,h =Average daily response factor of the HECD detector, ng CV/counts.
mco = Mass of carbon, as methane, in the FID calibration standard, ng.
mch = Mass of chloride in the HECD calibration standard, jug.
ms = Mass of the waste sample, g.
msc = Mass of carbon, as methane, in the sample, jug.
msf = Mass of sample container and waste sample, g.
msh = Mass of chloride in the sample, ng.
mst = Mass of sample container prior to sampling, g.
m,,,, = Mass of volatile organic in the sample, ng.
Pa = Ambient barometric pressure in the laboratory, Torr.
Qc = Flowrate of calibration gas, L/min.
tc = Length of time standard gas is delivered to the analyzer, min.
Ta = Ambient temperature in the laboratory, °K.
7.7.2 Mass of Carbon, as Methane in the FID Calibration Gas.
mco = k2 Cc tc Qc (PyiJ Eq. 1
where k2 = 0.5773 /ig C-°K/ /zl -Torr
7.7.3 Mass of Chloride in the HECD Detector Calibration Gas.
mch - k3 Ch tc Qc (PyTJ Eq. 2
where k3 = 1.1371 09 Cl -°K/Ml -Torr
7.7.4 FID Response Factor.
R, = mco/A Eq. 3
7.7.5 HECD Response Factor.
Rth = mch/A Eq. 4
5100 - 8 Revision 0
November 1990
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7.7.6 Mass of Carbon in the Sample.
msc = DR, (A. - Ab) Eq. 5
7.7.7 Mass of Chloride in the Sample.
msh = DR,h (A. - A,) Eq. 6
7.7.8 Mass of Volatile Organic in the Sample.
m = msc + msh Eq. 7
7.7.9 Standard Deviation.
SD = lOOx [ z(x,-x)2/(n-l)]1/2 Eq. 8
7.7.10 Relative Standard Deviation.
RSD = SD/x Eq. 9
7.7.11 Mass of Sample.
ms = msf - mst Eq. 10
7.7.12 Concentration of Volatile Organic in Waste.
C = m^/m., Eq. 11
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific Quality Control procedures.
8.2 Maintain a record of performance of all system checks and
calibrations.
8.2 Calibrate analytical balance against standard weights.
5100 - 9 Revision 0
November 1990
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9.0 METHOD PERFORMANCE
9.1 Performance data are not currently available.
10.0 REFERENCES
1. "Determination of the Volatile Organic Content of Waste Samples" Method
25D; Proposed Amendment to 40 CFR Part 60, Appendix A, January 1989.
5100 - 10 Revision 0
November 1990
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CALIBRATION GAS
VALVE FLOW
METER
Ul
i—•
o
o
FLOW
COALESCING METER
PURGING
CHAMBER
X
Vent
FID
SPLITTER
HECD
«o
-------
FIGURE 2
ROTAMETER
DATHIIOATCR/
CONlROLLER
STIRRING
MOTOR
DETECTORS
OIL 0 ATI I
PURGING CHAMBER
COALESCING FILTER
5100 - 12
Revision 0
November 1990
-------
FIGURE 3
Purging Chamber
TRUBORE
STIRRER
#7ACETHRED
#SOACETHRED
BUNA-N
0-RING
u
p
o
a
#7ACETHRED
5100 - 13
Revision 0
November 1990
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FIGURE 4
WASTE UNE
FROM SOURCE
STATIC MIXER
00
t
VALVES
OPTIONAL PUMP
REDUCER (1/4 " TUBE FITTING)
TEFLON OR STAINLESS STEEL COIL
ICED ATI!
SAMPLE CONTAINER
5100 - 14
Revision 0
November 1990
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METHOD 5100
DETERMINATION OF THE VOLATILE ORGANIC CONCENTRATION OF WASTE SAMPLES
7.1 Pour sample
contents in purgt
flask
7.1 Rinse sampe
container 3 times
with OOP
7.1 Assemble
purging apparatus
as shown in
Figure 1
7.2 Equlibrat*
the system
7.2 Oinct purgt
gas through
purgt chombir
7.2 Record
response of HO
and HECO
7.2 Stop
operations end
readjust
5100 - 15
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METHOD 5110
DETERMINATION OF ORGANIC PHASE VAPOR PRESSURE IN WASTE SAMPLES
1.0 SCOPE AND APPLICATION
1.1 This method is applicable for determining the organic phase vapor
pressure of waste samples from treatment, storage, and disposal facilities
(TSDF).
1.2 Performance of this method should not be attempted by persons
unfamiliar with the operation of a Flame lonization Detector (FID) nor by those
who are unfamiliar with source sampling, because knowledge beyond the scope of
this presentation is required.
2.0 SUMMARY OF METHOD
2.1 A waste sample is collected from a source as close to the point of
generation as practical. The headspace vapor of the sample is analyzed for
carbon content by a headspace analyzer, which uses an FID.
3.0 INTERFERENCES
3.1 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A field sample blank prepared
from organic-free reagent water and carried through sampling and subsequent
storage and handling can serve as a check on such contamination.
3.2 Contamination by carryover can occur whenever a low-concentration
sample is analyzed after a high-concentration sample. To reduce carryover, the
sample syringe must be rinsed out between samples with organic-free reagent
water. Whenever an unusually concentrated sample is encountered, it should be
followed by an analysis of organic-free reagent water. It may be necessary to
wash out the syringe with detergent, rinse with distilled water, and dry in a
150°C oven between analyses.
3.3 Before processing daily samples, the analyst should demonstrate that
the entire analytical system is free from interference by the analysis of an
organic-free reagent water or solvent blank.
4.0 APPARATUS AND MATERIALS
4.1 Sampling. The following equipment is required:
4.1.1 Sample Containers. Vials, glass, with butyl rubber septa,
Perkin-Elmer Corporation Part Numbers 0105-0129 (glass vials), 6001^0728
(gray butyl rubber septa, plug style), 0105-0131 (butyl rubber septa), or
equivalent. The seal must be made from butyl rubber. Silicone rubber
seals are not acceptable.
5110 - 1 Revision 0
January 1990
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4.1.2 Vial Sealer. Perkin-Elmer Number 105-0106, or equivalent.
4.1.3 Gas-Tight Syringe. Perkin-Elmer Number 00230117, or
equivalent.
4.2 The following equipment is required if sampling from an enclosed
pipe:
4.2.1 Static Mixer. Installed in-line or as a by-pass loop, sized
so that the drop size of the dispersed phase is not greater that 1,000 Aim.
If the installation of the mixer is in a by-pass loop, then the entire
waste stream must be diverted through the mixer.
4.2.2 Tap.
4.2.3 Tubing, Teflon, 0.25 in. ID.
4.2.4 Cooling Coil. Stainless steel (304), 0.25 in. ID, equipped
with a thermocouple at the coil outlet.
4.3 Analysis. The following equipment is required:
4.3.1 Balanced Pressure Headspace Sampler. Perkin-Elmer HS-6, HS-
100, or equivalent, equipped with a glass bead column instead of a
chromatographic column.
4.3.2 Flame lonization Detector. An FID meeting the following
specifications is required:
4.3.2.1 Linearity. A linear response (±5 percent) over the
operating range, as demonstrated by the procedures established in
Sections 7.2.2 and 8.1.1.
4.3.2.2 Range. A full scale range of 1 to 10,000 ppm CH4.
Signal attenuators should be available to produce a minimum signal
response of 10 percent of full scale.
4.3.3 Data Recording System. Analog strip chart recorder or digital
integration system compatible with the FID for permanently recording the
output of the detector.
4.3.4 Thermometer. Capable of reading temperatures in the range
of 30° to 60°C with an accuracy of ±0.1°C.
5.0 REAGENTS
5.1 Analysis. The following reagents are required for analysis:
5.1.1 Hydrogen (H2). Zero grade.
5.1.2 Carrier Gas. Zero grade nitrogen, containing less than 1 ppm
carbon and less than 1 ppm carbon dioxide.
5110 - 2 Revision 0
January 1990
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5.1.3 Combustion Gas. Zero grade air or oxygen, as required by the
FID.
5.2 Calibration and Linearity Check.
5.2.1 Stock Cylinder Gas Standard. 100 percent propane. The
manufacturer shall (a) certify the gas composition to be accurate to ±3
percent or better (see Section 5.2.1.1); (b) recommend a maximum shelf
life over which the gas concentration does not change by greater than ±5
percent from the certified value; and (c) affix the date of gas cylinder
preparation, certified propane concentration, and recommended maximum
shelf life to the cylinder before shipment to the buyer.
5.2.1.1 Cylinder Standards Certification. The manufacturer
shall certify the concentration of the calibration gas in the
cylinder by (a) directly analyzing the cylinder and (b) calibrating
his analytical procedure on the day of cylinder analysis. To
calibrate his analytical procedure, the manufacturer shall use, as
a minimum, a three-point calibration curve.
5.2.1.2 Verification of Manufacturer's Calibration Standards.
Before using, the manufacturer shall verify the concentration of each
calibration standard by (a) comparing it to gas mixtures prepared
in accordance with the procedure described in Section 7.1 of Method
106 of 40 CFR Part 61, Appendix B, or by (b) calibrating it against
Standard Reference Materials (SRMs), prepared by the National
Institute of Science and Technology, if such SRMs are available.
The agreement between the initially determined concentration value
and the verification concentration value must be within ±5 percent.
The manufacturer must reverify all calibration standards on a time
interval that is consistent with the shelf life of the cylinder
standards sold.
5.3 Blanks
5.3.1 Organic-free reagent water. All references to water in this
method refer to organic-free reagent water as defined in Chapter One.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
6.2 Sampling Plan Design and Development. Use the procedures given in
Chapter Nine, "Sampling Plan."
6.3 Collect samples according to the procedures in Chapter 9, or, if it
is necessary to sample from an enclosed pipe, sample according to the procedures
described below.
6.3.1 The apparatus designed to sample from an enclosed pipe is
shown in Figure 1. The apparatus consists of an in-line static mixer, a
tap, a cooling coil immersed in an ice bath, a flexible Teflon tube
5110 - 3 Revision 0
January 1990
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connected to the outlet of the cooling coil, and sample container. Locate
the tap within two pipe diameters of the static mixer outlet. Install the
static mixer in the process line or in a by-pass line.
6.3.2 Begin sample collection by purging the sample lines and
cooling coil with at least four volumes of waste. Collect the purged
material in a separate container.
6.3.3 After purging, stop the sample flow and transfer the Teflon
sampling tube to a sample container. Sample at a flow rate such that the
temperature of the waste is <10°C. Fill the sample container halfway (±5
percent) and cap immediately (within 5 seconds).
6.3.4 Store the collected samples on ice or in a refrigerator until
analysis.
6.3.5 Alternative sampling techniques may be used upon the approval
of the Administrator.
7.0 PROCEDURE
7.1 Calibration
7.1.1 Maintain a record of each item.
7.1.2 Use the procedures in Section 7.1.3 to calibrate the headspace
analyzer and FID, and to check for linearity before the system is first
placed in operation, after any shutdown that is longer than 6 months, and
after any modification of the system.
7.1.3 Calibration and Linearity. Use the procedures in Section
6.2.1 of Method 18 of 40 CFR Part 60, Appendix A, to prepare the standards
and calibrate the flowmeters, using propane as the standard gas. Fill the
calibration standard vials halfway (±5 percent) with organic-free reagent
water. Prepare a minimum of three concentrations that will bracket the
applicable cutoff. For a cutoff of 5.2 kPa (0.75 psi), prepare nominal
concentrations of 30,000, 50,000, and 70,000 ppm as propane. For a cutoff
of 27.6 kPa (4.0 psi), prepare nominal concentrations of 200,000, 300,000,
and 400,000 ppm as propane.
7.1.3.1 Use the procedures in Section 7.2.3 to measure the
FID response of each standard. Use a linear regression analysis to
calculate the values for the slope (k) and the y-intercept (b). Use
the procedures in Section 7.2 and 7.3 to test the calibration and
the linearity.
7.1.4 Daily FID Calibration Check. Check the calibration at the
beginning and at the end of the daily runs by using the following
procedures. Prepare two calibration standards at the nominal cutoff
concentrations using the procedures in Section 7.1.3 Place one at the
beginning and end of the daily run. Measure the FID response of the daily
calibration standard. Use the values for k and b obtained from the most
recent calibration and use Equation 4 to calculate the concentration of
5110 - 4 Revision 0
January 1990
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the daily standard. Use an equation similar to Equation 2 to calculate
the percent difference between the daily standard and Cs. If the percent
difference is within five, then the previous values for k and b can be
used. Otherwise, use the procedures in Section 7.1.3 to recalibrate the
FID.
7.2 Analysis.
7.2.1 Allow one hour for the headspace vials to equilibrate at the
temperature specified in the regulation. Allow the FID to warm until a
stable baseline is achieved on the detector.
7.2.2 Check the calibration of the FID daily, using the procedures
in Section 7.1.4.
7.2.3 Follow the manufacturer's recommended procedures for the
normal operation of the headspace sampler and FID.
7.2.4 Use the procedures in Sections 7.3.4 and 7.3.5 to calculate
the organic vapor pressure in the samples.
7.2.5 Monitor the output of the detector to make certain that the
results are being properly recorded.
7.3 Calculations
7.3.1 Nomenclature
A = Measurement of the area under the response curve, counts.
b = y-intercept of the linear regression line.
Ca = Measured vapor phase organic concentration of sample, ppm as
propane.
Cma = Average measured vapor phase organic concentration of standard, ppm
as propane.
Cm = Measured vapor phase organic concentration of standard, ppm as
propane.
Cs = Calculated standard concentration, ppm as propane.
k = Slope of the linear regression line.
Pbar = Atmosphere pressure at analysis conditions, mm Hg (in. Hg).
p* = Organic vapor pressure in the sample, kPa (psi).
B = 1.333 x 10"6 kPa/[(mm Hg)(ppm)], 4.91 x 10'7 psi/ [(in.Hg)(ppm)])
7.3.2 Linearity. Use Equation 1 to calculate the measured standard
concentration for each standard vial.
5110 - 5 Revision 0
January 1990
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cm = k A + b Eq. 1
7.3.2.1 Calculate the average measured standard concentration
(Cma) for each set of triplicate standards, and use Equation 2 to
calculate the percent difference between Cma and Cs
f - f
Ls ^ma
Percent Difference = x 100 Eq. 2
Cs
The instrument linearity is acceptable if the percent
difference is less than or equal to five for each standard.
7.3.3 Relative standard Deviation (RSD). Use Equation 3 to
calculate the RSD for each triplicate set of standards.
100
%RSD =
n (Cm - Cma)2
Z Eq. 3
'"1 (n - 1)
The calibration is acceptable if the RSD is within five percent for
each standard concentration.
7.3.4 Concentration of Organics in the Headspace. Use Equation 4
to calculate the concentration of vapor phase organics in each sample.
Ca = k A + b Eq. 4
7.3.5 Vapor Pressure of Organics in the Headspace. Use Equation
5 to calculate the vapor pressure of organics in the sample.
P" - B Pbar Ca Eq. 5
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific Quality Control procedures.
8.'2 Maintain a record of performance of all system checks and
calibrations.
9.0 METHOD PERFORMANCE
9.1 No performance data are currently available.
10.0 REFERENCES
1. "Determination of Vapor Phase Organic Concentrations in Waste Samples,"
Method 25E; Proposed Amendment to 40 CFR Part 60, Appendix A, January 1989.
5110 - 6 Revision 0
January 1990
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FIGURE 1
WASTE LINE
FROM SOURCE
STATIC MIXER
t
VALVES
OPTIONAL PUMP
REDUCER (1/4 " TUBE FITTING)
TEFLON OR STAINLESS STEEL COIL f ft " )
ICEDATII
SAMPLE CONTAINER
5110 - 7
Revision 0
January 1990
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METHOD 5110
DETERMINATION OF ORGANIC PHASE VAPOR PRESSURE IN WASTE SAMPLES
Start
7.2.1 Allow
system to
equilibrate
for 1 hour.
7.2.2 Do daily
FID calibration
check using
procedures from
section 7.1.4.
7.2.3 Operate
headspace
sampler and FID
according to
manufacturer.
7.2.4 Monitor
detector
output to
assure proper
recording.
7.3.2 Calculate
linearity
according to
equations
given.
7.3.3 Calculate
relative
standard
deviation of
standards.
7.3.4 Calculate
concentration
of organics in
the headspace.
7.3.5 Calculate
the vapor
pressure of the
organics in the
headspace.
Stop
5110 - 8
Revision 0
January 1990
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METHOD 801OB
HALOGENATED VOLATILE ORGANICS
1.0 SCOPE AND APPLICATION
1.1 Method 8010 is used to determine the concentration of various volatile
halogenated organic compounds. The following compounds can be determined by this
method:
ADorooriate Techniaue
Compound Name
Allyl chloride
Benzyl Chloride
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Bromoacetone
Bromobenzene
Bromod i chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chloroacetaldehyde
Chlorobenzene
Chloroethane
2-Chloroethanol
2-Chloroethyl vinyl ether
Chloroform
1-Chlorohexane
Chl oromethane
Chloromethyl methyl ether
Chloroprene
4-Chlorotoluene
Di bromochl oromethane
1 , 2-Di bromo-3-chl oropropane
Dibromomethane
1,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 , 2-Di chl oroethene
Di chl oromethane
1 , 2 -Di chl oropropane
1, 3-Di chl oro-2-propanol
cis-1, 3-Di chl oropropene
trans-l,3-Dichloropropene
CAS No.a
107-05-1
100-44-7
111-91-1
39638-32-9
598-31-2
108-86-1
75-27-4
75-25-2
74-83-9
56-23-5
107-20-0
108-90-7
75-00-3
107-07-03
110-75-8
67-66-3
544-10-5
74-87-3
107-30-2
126-99-8
106-43-4
124-48-1
96-12-8
74-95-3
95-50-1
541-73-1
106-46-7
764-41-0
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
75-09-2
78-87-5
96-23-1
10061-01-5
10061-02-6
Purge-and-Trap
b
PP
PP
b
PP
b
b
b
b
b
b
b
b
PP
b
b
pc
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
PP
b
b
Direct
Injection
b
b
pc
b
b
b
b
b
b
b
b
b
b
b
b
b
pc
b
pc
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
8010B - 1
Revision 2
November 1990
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Appropriate Technique
Direct
Compound Name CAS No.a Purge-and-Trap Injection
Epichlorhydrin
Ethyl ene di bromide
Methyl iodide
1,1,2, 2 -Tetrachl oroethane
1,1,1 , 2-Tetrachl oroethane
Tetrachl oroethene
1,1,1-Trichloroethane
1 , 1 , 2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1 , 2 , 3 -Tri chl oropropane
Vinyl Chloride
106-89-8
106-93-4
74-88-4
79-34-5
630-20-6
127-18-4
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
75-01-4
PP
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
a Chemical Abstract Services Registry Number
b Adequate response using this technique
pp Poor purging efficiency, resulting in high EQLs
pc Poor chromatographic performance.
1.2 Table 1 indicates compounds that may be analyzed by this method and
lists the method detection limit for each compound in organic-free reagent water.
Table 2 lists the estimated quantitation limit for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8010 provides gas chromatographic conditions for the detection
of halogenated volatile organic compounds. Samples can be introduced into the
GC using direct injection or purge-and-trap (Method 5030). Ground water samples
must be analyzed using Method 5030. A temperature program is used in the gas
chromatograph to separate the organic compounds. Detection is achieved by a
electrolytic conductivity detector (HECD).
2.2 The method provides an optional gas chromatographic column that may
be helpful in resolving the analytes from co-eluting non-target compounds and
for analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
8010B - 2 Revision 2
November 1990
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detector, analytical
columns, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 8 ft x 0.1 in. ID stainless steel or glass
column packed with 1% SP-1000 on Carbopack-B 60/80 mesh or
equivalent.
4.1.2.2 Column 2 - 6 ft x 0.1 in. ID stainless steel or glass
column packed with chemically bonded n-octane on Porasil-C 100/120
mesh (Durapak) or equivalent.
4.1.3 Detector - Electrolytic conductivity (HECD).
4.2 Sample introduction apparatus, refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes, 5 ml Luerlok glass hypodermic and a 5 ml, gas-tight with
shutoff valve.
4.4 Volumetric flask, Class A, 10, 50, 100, 500, and 1,000 ml with a
ground glass stopper.
4.5 Microsyringe, 10 and 25 /itL with a 0.006 in. ID needle (Hamilton 702N
or equivalent) and a 100 pi.
4.6 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. Store away from
other solvents.
8010B - 3 Revision 2
November 1990
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5.4 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
methanol using assayed liquids or gases, as appropriate. Because of the toxicity
of some of the organohalides, primary dilutions of these materials should be
prepared in a hood.
5.4.1 Place about 9.8 ml of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes until all alcohol-wetted surfaces have dried. Weigh the
flask to the nearest 0.0001 g.
5.4.2 Add the assayed reference material, as described below.
5.4.2.1 Liquids. Using a 100 pi syringe, immediately add two
or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.4.2.2 Gases. To prepare standards for any compounds that
boil below 30°C (e.g. bromomethane, chloroethane, chloromethane,
dichlorodifluoromethane, trichlorofluoromethane, vinyl chloride),
fill a 5 ml valved gas-tight syringe with the reference standard to
the 5.0 ml mark. Lower the needle to 5 mm above the methanol
meniscus. Slowly introduce the reference standard above the surface
of the liquid. The heavy gas rapidly dissolves in the methanol.
This may also be accomplished by using a lecture bottle equipped with
a Hamilton Lecture Bottle Septum (#86600). Attach Teflon tubing to
the side-arm relief valve and direct a gentle stream of gas into the
methanol meniscus.
5.4.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.4.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.4.5 Prepare fresh standards every 2 months, for gases or for
reactive compounds such as 2-chloroethylvinyl ether. All other standards
must be replaced after 6 months, or sooner if comparison with check
standards indicates a problem.
5.5 Secondary dilution standards. Using stock standard solutions, prepare
secondary dilution standards in methanol, as needed, containing the compounds
of interest, either singly or mixed together. The secondary dilution standards
should be prepared at concentrations such that the aqueous calibration standards
prepared in Section 5.6 will bracket the working range of the analytical system.
Secondary dilution standards should be stored with minimal headspace for
8010B - 4 Revision 2
November 1990
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volatiles and should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from them.
5.6 Calibration standards. Prepare calibration standards in organic-free
reagent water from the secondary dilution of the stock standards, at a minimum
of five concentrations. One of the concentrations should be at a concentration
near, but above, the method detection limit. The remaining concentrations should
correspond to the expected range of the concentrations found in real samples or
should define the working range of the GC. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Table 1 may be included). In order to prepare accurate aqueous standard
solutions, the following precautions must be observed.
5.6.1 Do not inject more than 20 /uL of alcoholic standards into
100 ml of water.
5.6.2 Use a 25 p.i Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to deliver
reproducible volumes of methanolic standards into water).
5.6.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.6.4 Mix aqueous standards by inverting the flask three times only.
5.6.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.6.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.6.7 Aqueous standards are not stable and should be discarded after
one hour, unless properly sealed and stored. The aqueous standards can
be stored up to 24 hours, if held in sealed vials with zero headspace.
5.7 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples. The
compounds recommended for use as surrogate spikes (Section 5.8) have been used
successfully as internal standards, because of their generally unique retention
times.
5.7.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Section 5.5.
5.7.2 Prepare a spiking solution containing each of the internal
standards using the procedures described in Sections 5.4 and 5.5. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 ng//*L of each internal standard compound. The
8010B - 5 : Revision 2
November 1990
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addition of 10 juL of this standard to 5.0 ml of sample or calibration
standard would be equivalent to 30 M9/L-
5.7.3 Analyze each calibration standard according to Section 7.0,
adding 10 p.1 of internal standard spiking solution directly to the
syringe.
5.8 Surrogate standards - The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and organic-free reagent water
blank with surrogate halocarbons. A combination of bromochloromethane,
bromochlorobenzene and bromofluorobenzene is recommended to encompass the range
of temperature program used in this method. From stock standard solutions
prepared as in Section 5.4, add a volume to give 750 /ig of each surrogate to
45 mL of organic-free reagent water contained in a 50 ml volumetric flask, mix,
and dilute to volume for a concentration of 15 ng//iL. Add 10 /il_ of this
surrogate spiking solution directly into the 5 ml syringe with every sample and
reference standard analyzed. If the internal standard calibration procedure is
used, the surrogate compounds may be added directly to the internal standard
spiking solution (Section 5.7.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph using
either direct injection or purge-and-trap (Method 5030). Method 5030 may be
used directly on ground water samples or low-concentration contaminated soils
and sediments. For medium-concentration soils or sediments, methanolic
extraction, as described in Method 5030, may be necessary prior to purge-and-
trap analysis.
7.2 Gas chromatographic conditions (Recommended)
7.2.1 Column 1:
Helium flow rate = 40 mL/min
Temperature program:
Initial temperature = 45°C, hold for 3 minutes
Program = 45°C to 220°C at 8°C/min
Final temperature = 220°C, hold for 15 minutes.
7.2.2 Column 2:
Helium flow rate = 40 mL/min
Temperature program:
Initial temperature = 50°C, hold for 3 minutes
Program = 50°C to 170°C at 6°C/min
Final temperature = 170°C, hold for 4 minutes.
8010B - 6 Revision 2
November 1990
-------
7.3 Calibration. The procedure for internal or external calibration may
be used. Refer to Method 8000 for a description of each of these procedures.
Use Table 1 and Table 2 for guidance on selecting the lowest point on the
calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Section 7.4.1).
7.4 Gas chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap) or the direct injection method (see
Section 7.4.1.1). If the internal standard calibration technique is used,
add 10 /iL of internal standard to the sample prior to purging.
7.4.1.1 In very limited applications (e.g. aqueous process
wastes) direct injection of the sample onto the GC column with a
10 juL syringe may be appropriate. The detection limit is very high
(approximately 10,000 /ig/L) therefore, it is only permitted where
concentrations in excess of 10,000 /xg/L are expected or for water-
soluble compounds that do not purge. The system must be calibrated
by direct injection (bypassing the purge-and-trap device).
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times on the two
columns for a number of organic compounds analyzable using this method.
An example of the separation achieved by Column 1 is shown in Figure 1.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Refer to Method 8000 for guidance on calculation of
concentration.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using the second GC column is recommended.
7.4.7 If the response for a peak is off-scale, prepare a dilution
of the sample with organic-free reagent water. The dilution must be
performed on a second aliquot of the sample which has been properly sealed
and stored prior to use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8010B - 7 Revision 2
November 1990
-------
8.2 Mandatory quality control to validate the GC system operation is found
in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each analyte of interest at a concentration of 10 mg/L in
methanol.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both Tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following are required:
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
9.0 METHOD PERFORMANCE
9.1 This method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked
at six concentrations over the range 8.0-500 M9/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the analyte, and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and by the calibration procedure used.
10.0 REFERENCES
1. Bellar, T.A.; Lichtenberg, J.J. «L. Amer. Water Works Assoc. 1974, 66(12),
pp. 739-744.
8010B - 8 Revision 2
November 1990
-------
2. Bellar, T.A.; Lichtenberg, J.J., Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds, Measurement of Organic Pollutants in Water and Wastewater; Van
Hall, Ed.; ASTM STP 686, pp 108-129, 1979.
3. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters: Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane"; report for EPA
Contract 68-03-2635 (in preparation).
4. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act: Final Rule and Interim
Final Rule and Proposed Rule", October 26, 1984.
5. "EPA Method Validation Study 23, Method 601 (Purgeable Halocarbons)"; Report
for EPA Contract 68-03-2856 (in preparation).
6. Gebhart, J.E., S.V. Lucas, S.J. Naber, A.M. Berry, T.H. Danison and H.M.
Burkholder, "Validation of SW-846 Methods 8010, 8015, and 8020"; Report for
EPA Contract 68-03-1760, Work Assignment 2-15; US EPA, EMSL-Cincinnati,
1987.
8010B - 9 Revision 2
November 1990
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
FOR HALOGENATED VOLATILE ORGANICS
Compound
Ally chloride
Benzyl chloride*'0
Bis(2-chloroethoxy)methane"
Bis(2-chloroisopropyl) ether"
Bromobenzene
Bromodi chl oromethane
Bromoform*
Bromomethane*
Carbon tetrachloride*
Chl oroacetal dehyde*
Chlorobenzene"
Chl oroethane
Chloroform*
1-Chlorohexane
2-Chloroethyl^ vinyl ether*
Chl oromethane*
Chloromethyl methyl ether*
4-Chlorotoluene
Di bromochl oromethane
1 , 2-Di bromo-3-chl oropropane*
Dibromomethane*
1,2-Dichlorobenzene] ,
1,3-Dichlorobenzene] '
1,4-Dichlorobenzene*
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane*'"
1,1-Dichloroethane"
1,2-Dichloroethane]
1,1-Dichloroethene*
trans-l,2-Dichlproethene*
Di chl oromethane*
1, 2-Di chl oropropane*
trans-l,3-Dichloropropene*
Ethyl ene di bromide
1,1,2, 2 -Tetrachl oroethane]
1,1,1 , 2-Tetrachl oroethane*
Tetrachl oroethene*
1,1,1 -Tri chl oroethane
1 , 1 , 2-Trichl oroethane"
Tri chl oroethene*
Tri chl orof 1 uoromethane]
1,2, 3 -Tri chl oropropane*
Vinyl Chloride*
CAS
Registry
Number
107-05-1
100-44-7
111-91-1
39638-32-9
108-86-1
75-27-4
75-25-2
74-83-9
56-23-5
107-20-0
108-90-7
75-00-3
67-66-3
544-10-5
110-75-8
74-87-3
107-30-2
106-43-4
124-48-1
96-12-8
74-95-3
95-50-1
541-73-1
106-46-7
764-41-0
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
75-09-2
78-87-5
10061-02-5
106-93-4
79-34-5
630-20-6
127-18-4
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
75-01-4
Retention Time
(minutes)
Column 1 Column 2
10.17
30.29
38.60
34.79
29.05
15.44
21.12
2.90
14.58
(b)
25.49
5.18
12.62
26.26
19.23
1.40
8.88
34.46
18.22
28.09
13.83
37.96
36.88
38.64
23.45
3.68
11.21
13.14
10.04
11.97
7.56
16.69
16.976
19.59
23.12
21.10
23.05
14.48
18.27
17.40
9.26
22.95
3.25
(b)
(b)
(b)
(b)
(b)
14.62
19.17
7.05
11.07
(b)
18.83
8.68
12.08
(b)
(b)
5.28
(b)
(b)
16.62
(b)
14.92
23.52
22.43
22.33
(b)
(b)
12.57
15.35
7.72
9.38
10.12
16.62
16.60
(b)
(b)
21.70
14.97
13.10
18.07
13.12
(b)
(b)
5.28
Method
Detection
Limit"
(M9/L)
(b)
(b)
(b)
(b)
(b)
0.002
0.020
0.030
0.003
(b)
0.001
0.008
0.002
(b)
0.130
0.010
(b)
(b)
(b)
0.030
(b)
(b)
(b)
(b)
(b)
(b)
0.002
0.002
0.003
0.002
(b)
(b)
0.340
(b)
0.010
(b)
0.001
0.003
0.007
0.001
(b)
(b)
0.006
8010B - 10
Revision 2
November 1990
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TABLE 1.
Continued
a = Using purge-and-trap method (Method 5030)
b = Not determined
* = Appendix VIII compounds
c = Demonstrated very erratic results when tested by purge-and-trap
d = See Section 4.10.2 of Method 5030 for guidance on selection of trapping
material
e = Estimated retention time
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix Factor"
Ground water ', 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
Sample EQLs are highly matrix-dependent. The EQLs listed herein
are provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)].
For non-aqueous samples, the factor is on a wet-weight basis.
8010B - 11 Revision 2
November 1990
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TABLE 3.
CALIBRATION AND QC ACCEPTANCE CRITERIA8
Analyte
Bromod i chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethylvinyl ether
Chloroform
Chl oromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 , 2-Dichl oroethene
Dichl oromethane
1,2-Dichloropropane
ci s-1 ,3-Dichl oropropene
trans- 1,3-Di chl oropropene
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
1,1,1 -Tri chl oroethane
1 , 1 , 2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
Vinyl chloride
Range
for Q
(M9/L)
15.2-24.8
14.7-25.3
11.7-28.3
13.7-26.3
14.4-25.6
15.4-24.6
12.0-28.0
15.0-25.0
11.9-28.1
13.1-26.9
14.0-26.0
9.9-30.1
13.9-26.1
16.8-23.2
14.3-25.7
12.6-27.4
12.8-27.2
15.5-24.5
14.8-25.2
12.8-27.2
12.8-27.2
9.8-30.2
14.0-26.0
14.2-25.8
15.7-24.3
15.4-24.6
13.3-26.7
13.7-26.3
Limit
for S
(M9/U
4.3
4.7
7.6
5.6
5.0
4.4
8.3
4.5
7.4
6.3
5.5
9.1
5.5
3.2
5.2
6.6
6.4
4.0
5.2
7.3
7.3
9.2
5.4
4.9
3.9
4.2
6.0
5.7
Range
for x
(M9/L)
10.7-32.0
5.0-29.3
3.4-24.5
11.8-25.3
10.2-27.4
11.3-25.2
4.5-35.5
12.4-24.0
D-34.9
7.9-35.1
1.7-38.9
6.2-32.6
11.5-25.5
11.2-24.6
13.0-26.5
10.2-27.3
11.4-27.1
7.0-27.6
10.1-29.9
6.2-33.8
6.2-33.8
6.6-31.8
8.1-29.6
10.8-24.8
9.6-25.4
9.2-26.6
7.4-28.1
8.2-29.9
Range
P> PS
(%)
42-172
13-159
D-144
43-143
38-150
46-137
14-186
49-133
D-193
24-191
D-208
7-187
42-143
47-132
51-147
28-167
38-155
25-162
44-156
22-178
22-178
8-184
26-162
41-138
39-136
35-146
21-156
28-163
Q = Concentration measured in QC check sample, in M9/L.
s = Standard deviation of four recovery measurements, in M9/L-
X = Average recovery for four recovery measurements, in Aig/L.
P, Ps = Percent recovery measured.
D = Detected; result must be greater than zero.
a Criteria from 40 CFR Part 136 for Method 601 and were calculated assuming
a QC check sample concentration of 20 M9/L-
8010B - 12
Revision 2
November 1990
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION3
Analyte
Bromodichloromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethyl vinyl ether"
Chloroform
Chloromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 , 2 -Di chl oroethene
Di chloromethane
l,2-Dichloropropaneb
cis-l,3-Dichloropropeneb
trans-l,3-Dichloropropeneb
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
1,1,1 -Trichl oroethane
1, 1, 2 -Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
Vinyl chloride
Accuracy, as
recovery, x'
(M9/L)
1.12C-1.02
0.96C-2.05
0.76C-1.27
0.98C-1.04
l.OOC-1.23
0.99C-1.53
l.OOC
0.93C-0.39
0.77C+0.18
0.94C+2.72
0.93C+1.70
0.95C+0.43
0.93C-0.09
0.95C-1.08
1.04C-1.06
0.98C-0.87
0.97C-0.16
0.91C-0.93
l.OOC
l.OOC
l.OOC
0.95C+0.19
0.94C+0.06
0.90C-0.16
0.86C+0.30
0.87C+0.48
0.89C-0.07
0.97C-0.36
Single analyst
precision, s/
(M9/L)
0.11X+0.04
0.12X+0.58
0.28X+0.27
0.15X+0.38
0.15X-0.02
0.14X-0.13
0.20X
0.13X+0.15
0.28X-0.31
0.11X+1.10
0.20X+0.97
0.14X+2.33
0.15X+0.29
0.08X+0.17
0.11X+0.70
0.21X-0.23
0.11X+1.46
0.11X+0.33
0.13X
0.18X
0.18X
0.14X+2.41
0.14X+0.38
0.15X+0.04
0.13X-0.14
0.13X-0.03
0.15X+0.67
0.13X+0.65
Overall
precision,
S' (M9/L)
0.20X+1.00
0.21X+2.41
0.36X+0.94
0.20X+0.39
0.18X+1.21
0.17X+0.63
0.35X
0.19X-0.02
0.52X+1.31
0.24X+1.68
0.13X+6.13
0.26X+2.34
0.20X+0.41
0.14X+0.94
0.15X+0.94
0.29X-0.04
0.17X+1.46
0.21X+1.43
0.23X
0.32X
0.32X
0.23X+2.79
0.18X+2.21
0.20X+0.37
0.19X+0.67
0.23X+0.30
0.26X+0.91
0.27X+0.40
x' = Expected recovery for one or more measurements of a sample containing
s/ =
C
X
a concentration of C, in M9/L.
Expected single analyst standard deviation of measurements at an average
concentration of x, in M9/L.
= Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in M9/L.
= True value for the concentration, in p.g/1.
= Average recovery found for measurements of samples containing a
concentration of C, in jug/L.
a From 40 CFR Part 136 for Method 601.
b Estimates based upon the performance in a single laboratory.
8010B - 13
Revision 2
November 1990
-------
FIGURE 1.
GAS CHROMATOGRAM OF HALOGENATED VOLATILE ORGANICS
8010B - 14
Revision 2
November 1990
-------
METHOD 8010B
HALOGENATED VOLATILE ORGANICS
7 3 Calibrate
(refer to
Method 8000)
7.4.1 Introduce
•ample into CC
by direct
injection or
purge-and-trap.
742 folio.
Method 8000
for analyiia
sequence.
ete.
744 Record
volume purged
or
injected,and
peak liiet.
7.4.S Calculate
concentration*
(refer to
Method 8000)
7 4 6 Are
interference*
•u*peeted?
7 4.7 !• peak
reaponte off
icala?
746 Analyie
•ample uoing
teeond CC
eoluam.
747 Dilute
second
aliquot of
•anple
8010B - 15
Revision 2
November 1990
-------
METHOD 8020B
AROMATIC VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8020 is used to determine the concentration of various aromatic
volatile organic compounds. The following compounds can be determined by this
method:
Compound Name
CAS No.'
Appropriate Technique
Direct
Purge-and-Trap Injection
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Ethyl benzene
2-Picoline
Pyridine
Styrene
Toluene
Thiophenol (Benzenethiol)
o-Xylene
m-Xylene
p-Xylene
71-43-2
108-90-7
95-50-1
541-73-1
106-46-7
100-41-4
109-06-8
110-86-1
100-42-5
108-88-3
108-98-5
95-47-6
108-38-3
106-42-3
b
b
b
b
b
b
PP
pc
b
b
pc
b
b
b
b
b
b
b
b
b
b
pc
b
b
pc
b
b
b
a Chemical Abstract Services Registry Number.
b adequate response by this technique.
pp Poor purging efficiency, resulting in high EQLs
pc Poor chromatographic performance.
1.2 Table 1 lists the method detection limit for each target analyte in
organic-free reagent water. Table 2 lists the estimated quantitation limit (EQL)
for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8020 provides chromatographic conditions for the detection of
aromatic volatile compounds. Samples can be introduced into the GC using direct
injection or purge-and-trap (Method 5030). Ground water samples must be
determined using Method 5030. A temperature program is used in the gas
chromatograph to separate the organic compounds. Detection is achieved by a
photo-ionization detector (PID).
8020B -1
Revision 2
November 1990
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2.2 If interferences are encountered, the method provides an optional gas
chromatographic column that may be helpful in resolving the analytes from the
interferences and for analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Method 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A field sample blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detectors, column
supplies, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Column 1: 6 ft x 0.082 in. ID #304 stainless steel
or glass column packed with 5% SP-1200 and 1.75% Bentone-34 on
100/120 mesh Supelcoport, or equivalent.
4.1.2.2 Column 2: 8 ft x 0.1 in. ID stainless steel or glass
column packed with 5% l,2,3-Tris(2-cyanoethoxy)propane on 60/80 mesh
Chromosorb W-AW, or equivalent.
4.1.3 Detector - Photoionization (PID) (h-Nu Systems, Inc. Model
PI-51-02 or equivalent).
4.2 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes - A 5 ml Luerlok glass hypodermic and a 5 ml, gas-tight with
shutoff valve.
4.4 Volumetric flask, Class A - 10, 50, 100, 500, and 1,000 ml with a
ground glass stopper.
4.5 Microsyringe - 10 and 25 /xL with a 0.006 in. ID needle (Hamilton 702N
or equivalent) and a 100 fj,i.
4.6 Analytical balance - 0.0001 g.
8020B -2 Revision 2
November 1990
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5.0 REAGENTS
5.1 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Methanol (CH3OH) - pesticide quality or equivalent. Store away from
other solvents.
5.3 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
methanol using assayed liquids. Because of the toxicity of benzene and
1,4-dichlorobenzene, primary dilutions of these materials should be prepared in
a hood.
5.3.1 Place about 9.8 ml of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 min or until all alcohol wetted surfaces have dried. Weigh the
flask to the nearest 0.0001 g.
5.3.2 Using a 10 pi syringe, immediately add two or more drops of
assayed reference material to the flask; then reweigh. The liquid must
fall directly into the alcohol without contacting the neck of the flask.
5.3.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.3.4 Transfer the stock standard solution into a Teflon-sealed
screw-cap bottle. Store, with minimal headspace, at 4°C and protect from
light.
5.3.5 All standards must be replaced after 6 months, or sooner if
comparison with check standards indicates a problem.
5.4 Secondary dilution standards: Using stock standard solutions, prepare
in methanol secondary dilution standards, as needed, that contain the compounds
of interest, either singly or mixed together. The secondary dilution standards
should be prepared at concentrations such that the aqueous calibration standards
prepared in Section 5.4 will bracket the working range of the analytical system.
Secondary dilution standards should be stored with minimal headspace for
volatiles and should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from them.
5.5 Calibration standards: Calibration standards at a minimum of five
concentrations are prepared in organic-free reagent water from the secondary
dilution of the stock standards. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
8020B -3 Revision 2
November 1990
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in real samples or should define the working range of the GC. Each standard
should contain each analyte for detection by this method (e.g., some or all of
the compounds listed in the target analyte list may be included). In order to
prepare accurate aqueous standard solutions, the following precautions must be
observed.
5.5.1 Do not inject more than 20 /il_ of alcoholic standards into
100 ml of organic-free reagent water.
5.5.2 Use a 25 /iL Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to deliver
reproducible volumes of methanolic standards into water).
5.5.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.5.4 Mix aqueous standards by inverting the flask three times only.
5.5.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.5.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.5.7 Aqueous standards are not stable and should be discarded after
1 hr, unless properly sealed and stored. The aqueous standards can be
stored up to 24 hr, if held in sealed vials with zero headspace.
5.6 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
Alpha,alpha,alpha-trifluorotoluene has been used successfully as an internal
standard.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest as described in Section 5.5.
5.6.2 Prepare a spiking solution containing each of the internal
standards using the procedures described in Sections 5.3 and 5.4. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 mg/L of each internal standard compound, the addition
of 10 jitL of this standard to 5.0 ml of sample or calibration standard
would be equivalent to 30 /xg/L.
5.6.3 Analyze each calibration standard according to Section 7.0,
adding 10 nl of internal standard spiking solution directly to the
syringe.
8020B -4 Revision 2
November 1990
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5.7 Surrogate standards: The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and organic-free reagent water
blank with surrogate compounds (bromochlorobenzene, bromofluorobenzene, 1,1,1-
trifluorotoluene, fluorobenzene, and difluorobenzene are recommended) which
encompass the range of the temperature program used in this method. From stock
standard solutions prepared as in Section 5.3, add a volume to give 750 jig of
each surrogate to 45 ml of organic-free reagent water contained in a 50 ml
volumetric flask, mix, and dilute to volume for a concentration of 15 ng//uL.
Add 10 /nL of this surrogate spiking solution directly into the 5 ml syringe with
every sample and reference standard analyzed. If the internal standard
calibration procedure is used, the surrogate compounds may be added directly to
the internal standard spiking solution (Section 5.6.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For medium-concentration soils or sediments, methanolic extraction,
as described in Method 5030, may be necessary prior to purge-and-trap analysis.
7.2 Gas chromatography conditions (Recommended):
7.2.1 Column 1:
Carrier gas (He) flow rate:
For lower boiling compounds:
Initial temperature:
Temperature program:
For higher boiling compounds:
Initial temperature:
Temperature program:
36 mL/min
50°C, hold for 2 min;
50°C to 90°C at 6°C/nrin, hold until
compounds have eluted.
all
50°C, hold for 2 min;
50°C to 110°C at 3°C/min, hold until
all compounds have eluted.
Column 1 provides outstanding separations for a wide variety of
aromatic hydrocarbons. Column 1 should be used as the primary analytical
column because of its unique ability to resolve para-, meta-, and ortho-
aromatic isomers.
7.2.2 Column 2:
Carrier gas (He) flow rate:
Initial temperature:
Temperature program:
30 mL/min
40°C, hold for 2 min;
40°C to 100°C at 2°C/min, hold until all
compounds have eluted.
8020B -5
Revision 2
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Column 2, an extremely high polarity column, has been used for a
number of years to resolve aromatic hydrocarbons from alkanes in complex
samples. However, because resolution between some of the aromatics is not
as efficient as with Column 1, Column 2 should be used as a confirmatory
column.
7.3 Calibration: Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Section 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis:
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap method) or the direct injection method.
If the internal standard calibration technique is used, add 10 ML of
internal standard to the sample prior to purging.
7.4.1.1 Direct injection: In very limited applications (e.g.,
aqueous process wastes), direct injection of the sample into the GC
system with a 10 /xL syringe may be appropriate. The detection limit
is very high (approximately 10,000 /ig/L); therefore, it is only
permitted when concentrations in excess of 10,000 ng/L are expected
or for water soluble compounds that do not purge. The system must
be calibrated by direct injection (bypassing the purge-and-trap
device).
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention~time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times and detection
limits for a number of organic compounds analyzable using this method.
An example of the separation achieved by Column 1 is shown in Figure 1.
Figure 2 shows an example of the separation achieved using Column 2.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using the second GC column is recommended.
7.4.7 If the response for a peak is off scale, prepare a dilution
of the sample with organic-free reagent water. The dilution must be
8020B -6 Revision 2
November 1990
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performed on a second aliquot of the sample which has been properly sealed
and stored prior to use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Mandatory quality control to validate the GC system operation is found
in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each parameter of interest at a concentration of 10 ng//uL
in methanol.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both Tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following is required.
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze
the sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
9.0 METHOD PERFORMANCE
9.1 This method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked
at six concentrations over the range 2.1 - 500 M9/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the parameter and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 4.
8020B -7 Revision 2
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9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and by the calibration procedure used.
10.0 REFERENCES
1. Bellar, T.A., and J.J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
pp. 739-744, 1974.
2. Bellar, T.A., and J.J. Lichtenberg, "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds", in Van Hall (ed.), Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
3. Dowty, B.J., S.R. Antoine, and J.L. Laseter, "Quantitative and Qualitative
Analysis of Purgeable Organics by High Resolution Gas Chromatography and
Flame lonization Detection", in Van Hall, ed., Measurement of Organic
Pollutants in Water and Wastewater. ASTM STP 686, pp. 24-35, 1979.
4. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane. Report for EPA
Contract 68-03-2635 (in preparation).
5. "EPA Method Validation Study 24, Method 602 (Purgeable Aromatics)", Report
for EPA Contract 68-03-2856 (in preparation).
6. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule", October 26, 1984.
7. Gebhart, J.E., S.V. Lucas, S.J. Naber, A.M. Berry, T.H. Danison and H.M.
Burkholder, "Validation of SW-846 Methods 8010, 8015, and 8020"; Report for
EPA Contract 68-03-1760, Work Assignment 2-15; US EPA, EMSL-Cincinnati,
1987.
8020B -8
Revision 2
November 1990
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
FOR AROMATIC VOLATILE ORGANICS
Compound
Benzene
Chlorobenzene
1,4-Dichlorobenzene
1,3-Dichlorobenzene
1,2-Di chlorobenzene
Ethyl Benzene
Styrene
Toluene
o-Xylene
m-Xylene
p-Xylene
Retention
(min)
Col. 1
2.59
9.38
16.42
17.54
20.60
8.12
11.00
5.14
10.54
9.77
9.18
time
Col. 2
2.75
8.02
16.2
15.0
19.4
6.25
(b)
4.25
(b)
(b)
(b)
Method
detection
limit8
(M9/L)
0.06
0.13
0.11
0.4
0.12
0.01
0.12
0.01
0.03
0.13
0.08
a Using purge-and-trap method (Method 5030).
b Not determined.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQLs)
FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
a Sample EQLs are highly matrix dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
b EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For
non-aqueous samples, the factor is on a wet-weight basis.
8020B -9 Revision 2
November 1990
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Parameter
Benzene
Chl orobenzene
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Ethyl benzene
Toluene
Range
for Q
(M9/L)
15.4-24.6
16.1-23.9
13.6-26.4
14.5-25.5
13.9-26.1
12.6-27.4
15.5-24.5
Limit
for s
(M9/L)
4.1
3.5
5.8
5.0
5.5
6.7
4.0
Range
for x
(M9/L)
10.0-27.9
12.7-25.4
10.6-27.6
12.8-25.5
11.6-25.5
10.0-28.2
11.2-27.7
Range
p p
r» rs
39-150
55-135
37-154
50-141
42-143
32-160
46-148
Q - Concentration measured in QC check sample, in M9/L.
s = Standard deviation of four recovery measurements, in M9/L-
x = Average recovery for four recovery measurements, in M9/L.
P, Ps =Percent recovery measured.
D = Detected; result must be greater than zero.
a Criteria from 40 CFR Part 136 for Method 602, and were calculated assuming
as check sample concentration of 20 /ig/L. These criteria are based
directly upon the method performance data in Table 4. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 1.
8020B -10
Revision 2
November 1990
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION
Parameter
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Ethyl benzene
Toluene
Accuracy, as
recovery, x'
0.92C+0.57
0.95C+0.02
0.93C+0.52
0.96C-0.04
0.93C-0.09
0.94C+0.31
0.94C+0.65
Single analyst
precision, s/
(M9/L)
0.09X+0.59
0.09X+0.23
0.17X-0.04
0.15X-0.10
O.lBx+0.28
0.17X+0.46
0.09X+0.48
Overall
precision,
S' (M9/L)
0.21X+0.56
0.17X+0.10
0.22X+0.53
0.19X+0.09
0.20X+0.41
0.26x+0.23
0.18x-0.71
x'
s/
S'
Expected recovery for one or more measurements of a sample containing
concentration C, in M9/L.
Expected single analyst standard deviation of measurements at an average
concentration of x, in
Expected interlaboratory_ standard deviation of measurements at an average
concentration found of x, in /ig/L.
True value for the concentration, in M9/L-
Average recovery found for measurements of samples containing a
concentration of C, in M9/L-
8020B -11
Revision 2
November 1990
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Figure 1
I
Column: S%SM200/1.7S%B«nloiw34
Profram: 6QOC-2 IMftutM. 6°C/Min. 10 IW°C
Ovtwtor: Phototonintion
Samptt: 0.401«/1 Standard Mi«tur«
8 10 12 14
RETENTION TIME (MINUTES)
16
18
20 22
An example of the separation achieved using Column 1.
8020B -12
Revision 2
November 1990
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Figure 2
(2«Cy»no«ho«v)
40»C-2 Minum
*hetoioniation
••"pit: 2.0 MI/I Standard Mixture
to 100«C
An example of the separation achieved using Column 2.
8020B -13
Revision 2
November 1990
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METHOD 8020B
AROMATIC VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
Start
7.1 Introduce
compound! into gas
chromatograph by
direct injection or
purge-and-trap
(Method 5030)
7 44 Record volume
purged or injected
and peak sizes
7.2 Set gat
chromatograph
condition
7.3 Calibrate
(rafer to Method
8000)
74.1 Introduce
volatile compound!
into gas
chromatograph by
purge-and•trap or
direct injection
74.2 folio* Method
8000 for analysis
sequence, etc
7.45 Calculate
concentration
(refer to Method
8000)
7.4.6 Ar
analytical
interferences
suspected?
74.7 Is
response for
a peak
off-scale?
7.4.6 Analyze using
second CC column
7.4.7 Dilute second
aliquot of sample
8020B -14
Revision 2
November 1990
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METHOD 8021A
HALOGENATED AND AROMATIC VOLATILES BY GAS CHROMATOGRAPHY USING
ELECTROLYTIC CONDUCTIVITY AND PHOTOIONIZATION DETECTORS
IN SERIES; CAPILLARY TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8021 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Analyte
CAS No/
Appropriate Technique
Direct
Purge-and-Trap Injection
Benzene
Bromobenzene
Bromochl oromethane
Bromodichloromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chi orodi bromomethane
Chloroethane
Chloroform
Chi oromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Di bromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans- 1, 2-Di chl oroethene
71-43-2
108-86-1
74-97-5
75-27-4
75-25-2
74-83-9
104-51-8
135-98-8
98-06-6
56-23-5
108-90-7
124-48-1
75-00-3
67-66-3
74-87-3
95-49-8
106-43-4
96-12-8
106-93-4
74-95-3
95-50-1
541-73-1
106-46-7
75-71-8
75-34-3
107-06-2
75-35-4
156-59-4
156-60-5
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
8021A - 1
Revision 1
November 1990
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Analyte
CAS No.Ł
Appropriate Technique
Direct
Purge-and-Trap Injection
1,2-Dichloropropane
1 , 3 -Di chl oropropane
2 , 2 -Di chl oropropane
1,1-Dichloropropene
cis-l,3-dichloropropene
trans- 1,3-di chl oropropene
Ethyl benzene
Hexachl orobutad i ene
I sopropyl benzene
p-Isopropyl toluene
Methyl ene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2, 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1 , 2 , 3-Tr i chl orobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1,1,2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1 , 2 , 3-Tri chl oropropane
1,2, 4-Trimethyl benzene
1 ,3 , 5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
78-87-5
142-28-9
590-20-7
563-58-6
10061-01-5
10061-02-6
100-41-4
87-68-3
98-82-8
99-87-6
75-09-2
91-20-3
103-65-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
87-61-6
120-82-1
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
95-63-6
108-67-8
75-01-4
95-47-6
108-38-3
106-42-3
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
a Chemical Abstract Services Registry Number.
b Adequate response by thi
pp Poor purging efficiency
i Inappropriate technique
s technique.
resulting in high EQLs.
for this analyte.
pc Poor chromatographic behavior.
1.2 Method detection limits (MDLs) are compound dependent and vary with
purging efficiency and concentration. The MDLs for selected analytes are
presented in Table 1. The applicable concentration range of this method is
compound and instrument dependent but is approximately 0.1 to 200 /xg/L-
Analytes that are inefficiently purged from water will not be detected when
present at low concentrations, but they can be measured with acceptable accuracy
8021A - 2
Revision 1
November 1990
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and precision when present in sufficient amounts. Determination of some
structural isomers (i.e. xylenes) may be hampered by coelution.
1.3 The estimated quantitation limit (EQL) of Method 8021 for an
individual compound is approximately 1 /xg/Kg (wet weight) for soil/sediment
samples, 0.1 mg/Kg (wet weight) for wastes, and 1 p.g/1 for ground water (see
Table 3). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 This method is recommended for use only by analysts experienced in
the measurement of purgeable organics at the low /xg/L level or by experienced
technicians under the close supervision of a qualified analyst.
1.5 The toxicity or carcinogenicity of chemicals used in this method
has not been precisely defined. Each chemical should be treated as a potential
health hazard, and exposure to these chemicals should be minimized. Each
laboratory is responsible for maintaining awareness of OSHA regulations regarding
safe handling of chemicals used in this method. Additional references to
laboratory safety are available for the information of the analyst (references
4 and 6).
1.6 The following method analytes have been tentatively classified as
known or suspected human or mammalian carcinogens: benzene, carbon tetrachloride,
1,4-dichlorobenzene, 1,2-dichloroethane, hexachloro-butadiene, 1,1,2,2-
tetrachloroethane, 1,1,2-trichloroethane, chloroform, 1,2-dibromoethane,
tetrachloroethene, trichloroethene, and vinyl chloride. Pure standard materials
and stock standard solutions of these compounds should be handled in a hood.
A NIOSH/MESA approved toxic gas respirator should be worn when the analyst
handles high concentrations of these toxic compounds.
2.0 SUMMARY OF METHOD
2.1 Method 8021 provides gas chromatographic conditions for the detection
of halogenated and aromatic volatile organic compounds. Samples can be analyzed
using direct injection or purge-and-trap (Method 5030). Ground water samples
must be analyzed using Method 5030 (where applicable). A temperature program
is used in the gas chromatograph to separate the organic compounds. Detection
is achieved by an electrolytic conductivity detector (HECD) and a photoionization
detector (PID) in series.
2.2 Tentative identifications are obtained by analyzing standards under
the same conditions used for samples and comparing resultant GC retention times.
Confirmatory information can be gained by comparing the relative response from
the two detectors. Concentrations of the identified components are measured by
relating the response produced for that compound to the response produced by a
compound that is used as an internal standard.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
8021A - 3 Revision 1
November 1990
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3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
4.0 APPARATUS AND MATERIALS
4.1 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.2 Gas Chromatograph - capable of temperature programming; equipped
with variable-constant differential flow controllers, subambient oven controller,
photoionization and electrolytic conductivity detectors connected with a short
piece of uncoated capillary tubing, 0.32-0.5 mm ID, and data system.
4.2.1 Column - 60 m x 0.75 mm ID VOCOL wide-bore capillary column
wi-th 1.5 fj.m film thickness (Supelco Inc., or equivalent).
4.2.2 Photoionization detector (PID) (Tracer Model 703, or
equivalent).
4.2.3 Electrolytic conductivity detector (HECD) (Tracer Hall Model
700-A, or equivalent).
4.3 Syringes - 5 ml glass hypodermic with Luer-Lok tips.
4.4 Syringe valves - 2-way with Luer ends (Teflon or Kel-F).
4.5 Microsyringe - 25 /xL with a 2 in. x 0.006 in. ID, 22° bevel needle
(Hamilton #702N or equivalent).
4.6 Microsyringes - 10, 100 juL.
4.7 Syringes - 0.5, 1.0, and 5 ml, gas tight with shut-off valve.
4.8 Bottles - 15 ml, Teflon lined with screw-cap or crimp top.
4.9 Analytical balance - 0.0001 g.
4.10 Refrigerator.
4.11 Volumetric flasks, Class A - 10 to 1000 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform
to the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
8021A - 4 Revision 1
November 1990
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used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to
be free of analytes. Store away from other solvents.
5.4 Vinyl chloride, (99.9% pure), CH2=CHC1. Vinyl chloride is available
from Ideal Gas Products, Inc., Edison, New Jersey and from Matheson, East
Rutherford, New Jersey, as well as from other sources. Certified mixtures of
vinyl chloride in nitrogen at 1.0 and 10.0 ppm (v/v) are available from several
sources.
5.5 Stock standards - Stock solutions may either be prepared from pure
standard materials or purchased as certified solutions. Prepare stock standards
in methanol using assayed liquids or gases, as appropriate. Because of the
toxicity of some of the organohalides, primary dilutions of these materials of
the toxicity should be prepared in a hood.
NOTE: If direct injection is used, the solvent system of standards must match
that of the sample. It is not necessary to prepare high concentration
aqueous mixed standards when using direct injection.
5.5.1 Place about 9.8 ml of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes until all alcohol-wetted surfaces have dried. Weigh the
flask to the nearest 0.1 mg.
5.5.2 Add the assayed reference material, as described below.
5.5.2.1 Liquids: Using a 100 pi syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.5.2.2 Gases: To prepare standards for any compounds that
boil below 30°C (e.g. bromomethane, chloroethane, chloromethane,
dichlorodifluoromethane, trichlorofluoromethane, vinyl chloride),
fill a 5 ml valved gas-tight syringe with the reference standard to
the 5.0 ml mark. Lower the needle to 5 mm above the methanol
meniscus. Slowly introduce the reference standard above the surface
of the liquid. The heavy gas rapidly dissolves in the methanol.
This may also be accomplished by using a lecture bottle equipped with
a Hamilton Lecture Bottle Septum (#86600). Attach Teflon tubing to
the side-arm relief valve and direct a gentle stream of gas into the
methanol meniscus.
5.5.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
8021A - 5 Revision 1
November 1990
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calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.5.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap or crimp top. Store, with minimal headspace, at
-10°C to -20°C and protect from light.
5.5.5 Prepare fresh stock standards every two months for gases.
Reactive compounds such as 2-chloroethylvinyl ether and styrene may need
to be prepared more frequently. All other standards must be replaced
after six months. Both gas and liquid standards must be monitored closely
by comparison to the initial calibration curve and by comparison to QC
reference samples. It may be necessary to replace the standards more
frequently if either check exceeds a 25% difference.
5.6 Prepare secondary dilution standards, using stock standard solutions,
in methanol, as needed, that contain the compounds of interest, either singly
or mixed together. The secondary dilution standards should be prepared at
concentrations such that the aqueous calibration standards prepared in Section
5.7 will bracket the working range of the analytical system. Secondary dilution
standards should be stored with minimal headspace for volatiles and should be
checked frequently for signs of degradation or evaporation, especially just prior
to preparing calibration standards from them.
5.7 Calibration standards, at a minimum of five concentration levels
are prepared in organic-free reagent water from the secondary dilution of the
stock standards. One of the concentration levels should be at a concentration
near, but above, the method detection limit. The remaining concentration levels
should correspond to the expected range of the concentrations found in real
samples or should define the working range of the GC. Standards (one or more)
should contain each analyte for detection by this method (e.g. some or all of
the target analytes may be included). In order to prepare accurate aqueous
standard solutions, the following precautions must be observed.
NOTE: Prepare calibration solutions for use with direct injection analyses in
water at the concentrations required.
5.7.1 Do not inject more than 20 jiL of alcoholic standards into
100 ml of water.
5.7.2 Use a 25 p.1 Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to deliver
reproducible volumes of methanolic standards into water).
5.7.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.7.4 Mix aqueous standards by inverting the flask three times.
5.7.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
8021A - 6 Revision 1
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5.7.6 Never use pi pets to dilute or transfer samples or aqueous
standards.
5.7.7 Aqueous standards are not stable and should be discarded
after one hour, unless properly sealed and stored. The aqueous standards
can be stored up to 12 hours, if held in sealed vials with zero headspace.
5.8 Internal standards - Prepare a spiking solution containing
fluorobenzene and 2-bromo-l-chloropropane in methanol, using the procedures
described in Sections 5.5 and 5.6. It is recommended that the secondary dilution
standard be prepared at a concentration of 5 mg/L of each internal standard
compound. The addition of 10 p.1 of such a standard to 5.0 ml of sample or
calibration standard would be equivalent to 10 /xg/L.
5.9 Surrogate standards - The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and reagent blank with two or
more surrogate compounds. A combination of bromochloromethane, 2-bromo-l-
chloropropane, 1,4-dichlorobutane and bromochlorobenzene is recommended to
encompass the range of the temperature program used in this method. From stock
standard solutions prepared as in Section 5.5, add a volume to give 750 /ug of
each surrogate to 45 ml of organic-free reagent water contained in a 50 mL
volumetric flask, mix, and dilute to volume for a concentration of 15 ng//iL.
Add 10 juL of this surrogate spiking solution directly into the 5 ml syringe with
every sample and reference standard analyzed. If the internal standard
calibration procedure is used, the surrogate compounds may be added directly to
the internal standard spiking solution (Section 5.8).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For medium-concentration soils or sediments, methanolic extraction,
as described in Method 5030, may be necessary prior to purge-and-trap analysis.
7.2 Gas chromatography conditions (Recommended)
7.2.1 Oven settings:
Carrier gas (Helium) Flow rate: 6mL/min.
Temperature program
Initial temperature: 10°C, hold for 8 minutes at
Program: 10°C to 180°C at 4°C/nrin
Final temperature: 180°C, hold until all expected
compounds have eluted.
8021A - 7 Revision 1
November 1990
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7.2.2 The carrier gas flow is augmented with an additional 24 mL
of helium flow before entering the photoionization detector. This make-
up gas is necessary to ensure optimal response from both detectors.
7.2.3 These halogen-specific systems eliminate misidentifications
due to non-organohalides which are coextracted during the purge step. A
Tracer Hall Model 700-A detector was used to gather the single laboratory
accuracy and precision data presented in Table 2. The operating conditions
used to collect these data are:
Reactor tube: Nickel, 1/16 in OD
Reactor temperature: 810°C
Reactor base temperature: 250°C
Electrolyte: 100% n-Propyl alcohol
Electrolyte flow rate: 0.8 mL/min
Reaction gas: Hydrogen at 40 mL/min
Carrier gas plus make-up gas: Helium at 30 mL/min
7.2.4 A sample chromatogram obtained with this column is presented
in Figure 5. This column was used to develop the method performance
statements in Section 9.0. Estimated retention times and MDLs that can
be achieved under these conditions are given in Table 1. Other columns
or element specific detectors may be used if the requirements of Section
8.0 are met.
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Section 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap method) or the direct injection method
(see Section 7.4.1.1). If the internal standard calibration technique is
used, add 10 /iL of internal standard to the sample prior to purging.
7.4.1.1 Direct injection - In very limited applications (e.g.
aqueous process wastes) direct injection of the sample into the GC
system with a 10 /iL syringe may be appropriate. The detection limit
is very high (approximately 10,000 M9/L), therefore, it is only
permitted where concentrations in excess of 10,000 /ug/L are expected
or for water-soluble compounds that do not purge. The system must
be calibrated by direct injection (bypassing the purge-and-trap
device).
7.4.2 Follow Section 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
8021A - 8 Revision 1
November 1990
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time windows, and identification criteria. Include a mid-concentration
standard after each group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times on the two
detectors for a number of organic compounds analyzable using this method.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using a second GC column is recommended.
7.4.7 If the response for a peak is off-scale, prepare a dilution
of the sample with organic-free reagent water. The dilution must be
performed on a second aliquot of the sample which has been properly sealed
and stored prior to use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures, and
to Method 8000 for gas chromatographic procedures. Quality control to ensure
the proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Mandatory quality control to validate the GC system operation is
found in Method 8000.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following are required.
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8021A - 9 Revision 1
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9.0 METHOD PERFORMANCE
9.1 Method detection limits for these analytes have been calculated
from data collected by spiking organic-free reagent water at 0.1 p.g/1. These
data are presented in Table 1.
9.2 This method was tested in a single laboratory using organic-free
reagent water spiked at 10 jug/L. Single laboratory precision and accuracy data
for each detector are presented for the method analytes in Table 2.
10.0 REFERENCES
1. Volatile Organic Compounds in Water by Purqe-and-Trap Capillary Column Gas
Chromatoqraphv with Photoionization and Electrolytic Conductivity Detectors
in Series. Method 502.2; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory: Cincinnati, OH, September, 1986.
2. The Determination of Haloqenated Chemicals in Water bv the Purge and Trap
Method. Method 502.1; Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, Ohio 45268, September, 1986.
3. Volatile Aromatic and Unsaturated Organic Compounds in Water by Purge and
Trap Gas Chromatoqraphv. Method 503.1; Environmental Protection Agency,
Environmental Monitoring and Support Laboratory: Cincinnati, Ohio,
September, 1986.
4. Glaser, J.A.; Forest, D.L.; McKee, G.D.; Quave, S.A.; Budde, W.L. "Trace
Analyses for Wastewaters"; Environ. Sci. Technol. 1981, 15, 1426.
5. Bellar, T.A.; Lichtenberg, J.J. The Determination of Synthetic Organic
Compounds in Water by Purge and Sequential Trapping Capillary Column Gas
Chromatographv; U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, Ohio, 45268.
8021A - 10 Revision 1
November 1990
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TABLE 1.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL) FOR
VOLATILE ORGANIC COMPOUNDS ON PHOTOIONIZATION DETECTION (PID) AND
HALL ELECTROLYTIC CONDUCTIVITY DETECTOR (HECD) DETECTORS
Analyte
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene Chloride
trans- 1,2-Di chl oroethene
1,1-Dichloroethane
2 , 2-Di chl oropropane
cis- 1,2-Di chloroethane
Chloroform
Bromochl oromethane
1,1,1 -Tri chl oroethane
1,1-Dichloropropene
Carbon Tetrachloride
Benzene
1,2-Dichloroethane
Tri chl oroethene
1 , 2-Di chl oropropane
Bromodichl oromethane
Dibromomethane
Toluene
1 , 1 , 2-Tri chl oroethane
Tetrachl oroethene
1,3-Dichloropropane
Di bromochl oromethane
1,2-Dibromoethane
Chlorobenzene
Ethyl benzene
1,1,1 , 2-Tetrachl oroethane
m-Xylene
p-Xylene
o-Xylene
Styrene
Isopropyl benzene
Bromoform
1,1,2 , 2-Tetrachl oroethane
1, 2, 3-Tri chl oropropane
PID
Ret. Time8
minute
_b
-
9.88
-
_
-
16.14
-
19.30
-
-
23.11
-
-
-
25.21
-
26.10
-
27.99
-
-
-
31.95
-
33.88
-
-
-
36.56
36.72
-
36.98
36.98
38.39
38.57
39.58
-
-
-
HECD
Ret. Time
minute
8.47
9.47
9.93
11.95
12.37
13.49
16.18
18.39
19.33
20.99
22.88
23.14
23.64
24.16
24.77
25.24
25.47
-
26.27
28.02
28.66
29.43
29.59
-
33.21
33.90
34.00
34.73
35.34
36.59
-
36.80
-
-
-
-
-
39.75
40.35
40.81
PID
MDL
M9/L
0.02
NDC
0.05
0.02
0.02
0.009
0.02
0.01
0.05
0.003
0.005
0.01
0.01
0.02
0.01
0.05
HECD
MDL
M9/L
0.05
0.03
0.04
1.1
0.1
0.03
0.07
0.02
0.06
0.07
0.05
0.01
0.02
0.01
0.03
0.02
0.01
0.03
0.01
0.006
0.02
2.2
ND
0.04
0.03
0.03
0.8
0.01
0.005
1.6
0.01
0.4
8021A - 11
Revision 1
November 1990
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TABLE 1.
(Continued)
Analyte
PID
Ret. Time8
minute
HECD
Ret. Time
minute
PID
MDL
M9/L
HECD
MDL
n-Propylbenzene 40.87
Bromobenzene 40.99
1,3,5-Trimethylbenzene 41.41
2-Chlorotoluene 41.41
4-Chlorotoluene 41.60
tert-Butylbenzene 42.92
1,2,4-Trimethylbenzene 42.71
sec-Butyl benzene 43.31
p-Isopropyltoluene 43.81
1,3-Dichlorobenzene 44.08
1,4-Dichlorobenzene 44.43
n-Butylbenzene 45.20
1,2-Dichlorobenzene 45.71
1,2-Di bromo-3-Chloropropane
1,2,4-Trichlorobenzene 51.43
Hexachlorobutadiene 51.92
Naphthalene 52.38
1,2,3-Trichlorobenzene 53.34
Internal Standards
Fluorobenzene 26.84
2-Bromo-1-chloropropane
41.03
41.45
41.63
44.11
44.47
45.74
48.57
51.46
51.96
53.37
33.08
0.004
0.006
0.004
ND
0.02
0.06
0.05
0.02
0.01
0.02
0.007
0.02
0.05
0.02
0.06
0.06
ND
0.03
0.01
0.01
0.02
0.01
0,
3.
0.
02
0
03
0.02
0.03
Retention times determined on 60 m x 0.75 mm ID VOCOL capillary column.
Program: Hold at 10°C for 8 minutes, then program at 4°C/min to 180°C, and
hold until all expected compounds have eluted.
Dash (-) indicates detector does not respond.
ND = Not determined.
8021A - 12
Revision 1
November 1990
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TABLE 2.
SINGLE LABORATORY ACCURACY AND PRECISION DATA
FOR VOLATILE ORGANIC COMPOUNDS IN WATER"
Photoionization
Detector
Analyte
Benzene
Bromobenzene
Bromochloromethane
Bromodichloromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chloromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1,2 Dichloroethene
trans - 1 , 2 -Di chl oroethene
1 , 2-Di chl oropropane
1,3-Di chl oropropane
2 , 2-Di chl oropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene .
I sopropyl benzene
p-Isopropyl toluene
Recovery,8
%
99
99
-
-
-
-
100
97
98
-
100
-
-
-
NDC
101
-
-
-
-
102
104
103
-
-
-
100
ND
93
-
-
-
103
101
99
98
98
Standard
Deviation
of Recovery
1.2
1.7
-
-
-
-
4.4
2.6
2.3
-
1.0
-
-
-
ND
1.0
-
-
-
-
2.1
1.7
2.2
-
-
-
2.4
ND
3.7
-
-
-
3.6
1.4
9.5
0.9
2.4
Hall Electrolytic
Conductivity Detector
Standard
Recovery,8 Deviation
% of Recovery
_b
97
96
97
106
97
-
-
-
92
103
96
98
96
97
97
86
102
97
109
100
106
98
89
100
100
103
105
99
103
100
105
103
-
98
-
-
.
2.7
3.0
2.9
5.5
3.7
-
-
-
3.3
3.7
3.8
2.5
8.9
2.6
3.1
9.9
3.3
2.7
7.4
1.5
4.3
2.3
5.9
5.7
3.8
2.9
3.5
3.7
3.8
3.4
3.6
3.4
-
8.3
-
-
8021A - 13
Revision 1
November 1990
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TABLE 2.
(Continued)
Analyte
Photoionization
Detector
Recovery,
Standard
Deviation
of Recovery
Hall Electrolytic
Conductivity Detector
Standard
Recovery,8 Deviation
% of Recovery
Methyl ene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2, 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1 -Tri chl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1 , 2 , 3-Tri chl oropropane
1 , 2 , 4-Trimethyl benzene
1,3, 5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
-
102
103
104
-
-
101
99
106
104
-
-
100
-
-
99
101
109
99
100
99
.
6.3
2.0
1.4
-
-
1.8
0.8
1.9
2.2
-
-
0.78
-
-
1.2
1.4
5.4
0.8
1.4
0.9
97
-
-
-
99
99
97
-
98
102
104
109
96
96
99
-
-
95
-
-
~
2.8
-
-
-
2.3
6.8
2.4
-
3.1
2.1
3.4
6.2
3.5
3.4
2.3
-
-
5.6
-
~
Recoveries and standard deviations were determined from seven samples and spiked at
10 /ig/L of each analyte. Recoveries were determined by internal standard method. Internal
standards were: Fluorobenzene for PID, 2-Bromo-l-chloropropane for HECD.
Detector does not respond.
ND = Not determined.
This method was tested in a single laboratory using water spiked at 10
reference 8).
(see
8021A - 14
Revision 1
November 1990
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TABLE 3.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
Sample EQLs are highly matrix dependent. The EQLs listed herein
are provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)].
For non-aqueous samples, the factor is on a wet-weight basis.
8021A - 15 Revision 1
November 1990
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FIGURE 1.
PURGING DEVICE
3MWUMJT
HVAT STftNGC VM.VI
t? CM * GAUQC STWNGC NCIOU
• MM 00 MU
MUT (MM OO
00
8021A - 16
Revision 1
November 1990
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FIGURE 2.
TRAP PACKINGS AND CONSTRUCTION TO INCLUDE DESORB CAPABILITY
PACKING OTTAJL
CONSTRUCTION OCT/ML
8021A - 17
Revision 1
November 1990
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FIGURE 3.
PURGE-AND-TRAP SYSTEM - PURGE MODE
CAAMCftOAS
FLOW CONTROL
uouo
r-COLUMN OVCN
IVQULATO*
COLUMN
OPTIONAL 4*OKT COLUMN
8CLECDON VALVf
TfU^MLffT
AMOK GAS
FLOWCONTKX
1»MOLŁCULAM
8ICVC FH.TW
ANALYTICAL COLUMN
ocvcc
NOTt
AU UNfS •CTWCCN
AND OC SHOULD M HCA7IO
towc
8021A - 18
Revision 1
November 1990
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FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE
COLUMN OVfN
CONFWMATOOT COLUMN
TOOCTfCTOA
PLOW CONTROL
iVMOLfCULAA
MVf RLTIft
8021A - 19
Revision 1
November 1990
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FIGURE 5.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
-------
METHOD 8021A
VOLATILE ORGANIC COMPOUNDS IN WATER BY PURGE AND TRAP CAPILLARY
COLUMN GAS CHRQMATOGRAPHY WITH PHOTO I ON I.Z AT I ON AND ELECTROLYTIC
CONDUCTIVITY DETECTORS IN SERIES
Start
72 Set
chromatographic
condilions.
7.3 Refer to
Method 8000
for
calibration
techniques.
7 4.1 Introduce
sample into CC
using direct
injection or
purge-and-trap.
7.4.4 Record
sample volume
introduced
into CC and
peak sizes.
7.4.5 Refer
to Method
8000 for
calculations.
7.4.6 Are
analytical
interferences
suspected?
7.4.7 Is peak
response off
scale?
Reanalyze
sample uing
second CC
column.
Dilute and
reanalyze
second
aliquot of
sample.
8021A - 21
Revision 1
November 1990
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METHOD 8031
ACRYLONITRILE BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8031 is used to determine the concentration of acrylonitrile
in water. This method may also be applicable to other matrices. The following
compounds can be determined by this method:
Compound Name CAS No.'
Acrylonitrile 107-13-1
a Chemical Abstract Services Registry Number.
1.2 The estimated quantitation limit of Method 8031 for determining the
concentration of acrylonitrile in water is approximately 10
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A measured sample volume is micro-extracted with methyl tert-butyl
ether. The extract is separated by gas chromatography and measured with a
Nitrogen/Phosphorus detector.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that leads to discrete
artifacts and/or elevated baselines in gas chromatograms. All of these materials
must be routinely demonstrated to be free from interferences under the conditions
of the analysis by running laboratory reagent blanks.
3.2 Samples can be contaminated by diffusion of volatile organics around
the septum seal into the sample during handling and storage. A field blank
should be prepared from organic-free reagent water and carried through the
sampling and sample handling protocol to serve as a check on such contamination.
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3.3 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph system
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detector, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Column: Porapak Q - 6 ft., 80/10 Mesh, glass column, or
equivalent.
4.1.3 Nitrogen/Phosphorus detector.
4.2 Materials
4.2.1 Grab sample bottles - 40 ml VOA bottles.
4.2.2 Mixing bottles - 90 ml bottle with a Teflon lined cap.
4.2.3 Syringes - 10 /uL and 50 /uL.
4.2.4 Volumetric flask (Class A) - 100 ml.
4.2.5 Graduated cylinder - 50 ml.
4.2.6 Pipet (Class A) - 5, 15, and 50 ml.
4.2.7 Vials - 10 ml.
4.3 Preparation
4.3.1 Prepare all materials to be used as described in Chapter 4
for volatile organics.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
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5.2 General
5.2.1 Methanol, CH3OH - Pesticide quality, or equivalent.
5.2.2 Organic-free reagent water. All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.3 Methyl tert-butyl ether, CH3Ot-C4H9 - Pesticide quality, or
equivalent.
5.2.4 Acrylonitrile, H2C:CHCN, 98%.
5.3 Stock standard solution
5.3.1 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially
prepared stock standards can be used if they are verified against EPA
standards. If EPA standards are not available for verification, then
standards certified by the manufacturer and verified against a standard
made from pure material is acceptable.
5.3.2 The stock standard solution may be prepared by volume or by
weight. Stock solutions must be replaced after one year, or sooner if
comparison with the check standards indicates a problem.
CAUTION; Acrylonitrile is toxic. Standard preparation should be performed in
a laboratory fume hood.
5.3.2.1 To prepare the stock standard solution by volume:
inject 10 n\. of acrylonitrile (98%) into a 100 ml volumetric flask
with a syringe. Make up to volume with methanol.
5.3.2.2 To prepare the stock standard solution by weight:
Place about 9.8 ml of organic-free reagent water into a 10 ml
volumetric flask before weighing the flask and stopper. Weigh the
flask and record the weight to the nearest 0.0001 g. Add two drops
of pure acrylonitrile, using a 50 pi syringe, to the flask. The
liquid must fall directly into the water, without contacting the
inside wall of the flask. Stopper the flask and then reweigh.
Dilute to volume with organic-free reagent water. Calculate the
concentration from the net gain in weight.
5.4 Working standard solutions
5.4.1 Prepare a minimum of 5 working standard solutions that cover
the range of analyte concentrations expected in the samples. Working
standards of 20, 40, 60, 80, and 100 M9/L may be prepared by injecting 10,
20, 30, 40, and 50 /xL of the stock standard solution prepared in Section
5.3.2.1 into 5 separate 90 ml mixing bottles containing 40 ml of organic-
free reagent water.
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5.4.2 Inject 15 ml of methyl tert-butyl ether into each mixing
bottle, shake vigorously, and let stand 5 minutes, or until layers have
separated.
5.4.3 Remove 5 ml of top layer by pipet, and place in a 10 ml vial.
5.4.4 Keep all standard solutions below 4°C until used.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample Extraction
7.1.1 Pour 40 mL of the sample into a 90 mL mixing bottle. Pipet
15 mL of Methyl tert-butyl ether into the mixing bottle. Shake vigorously
for about 2 min. and let stand for about 5 min. Remove about 5 mL of the
top layer and store in a 10 mL vial.
7.2 Chromatographic Conditions (Recommended)
Carrier Gas (He) flow rate: 35 mL/min.
Column Temperature: 180° C, Isothermal
Injection port temperature: 250° C
Detector temperature: 250° C
Detector Current (DC): 18 volts
Gases: Hydrogen, 3 mL/min; Air, 290 mL/min.
7.3 Calibration of GC
7.3.1 On a daily basis, inject 3 pi of methyl tert-butyl ether
directly into the GC to flush the system. Also purge the system with
methyl tert-butyl ether injections between injections of standards and
samples.
7.3.2 Inject 3 juL of a sample blank (organic-free reagent water
carried through the sample storage procedures and extracted with methyl
tert-butyl ether).
7.3.3 Inject 3 pi of at least five standard solutions: one should
be near the detection limit; one should be near, but below, the expected
concentrations of the analyte; one should be near, but above, the expected
concentrations of the analyte. The range of standard solution
concentrations used should not exceed the working range of the GC system.
7.3.4 Prepare a calibration curve using the peak areas of the
standards (retention time of acrylonitrile under the conditions of Section
7.2 is approximately 2.3 minutes). If the calibration curve deviates
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significantly from a straight line, prepare a new calibration curve with
the existing standards, or, prepare new standards and a new calibration
curve. See Method 8000, Section 7.4.2, for additional guidance on
calibration by the external standard method.
7.4 Sample Analysis
7.4.1 Inject 3 /iL of the sample extract, using the same
chromatographic conditions used to prepare the standard curve. Calculate
the concentration of acrylonitrile in the extract, using the area of the
peak, against the calibration curve prepared in Section 7.3.4.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Prior to preparation of stock solutions, methanol and methyl
tert-butyl ether reagents should be analyzed gas chromatographically under the
conditions described in Section 7.2, to determine possible interferences with
the acrylonitrile peak. If the solvent blanks show contamination, a different
batch of solvents should be used.
9.0 METHOD PERFORMANCE
9.1 Method 8031 was tested in a single laboratory over a period of days.
Duplicate samples and one spiked sample were run for each calculation. The GC
was calibrated daily. Results are presented in Table 1.
10.0 REFERENCES
1. K.L. Anderson, "The Determination of Trace Amounts of Acrylonitrile in
Water by Specific Nitrogen Detector Gas Chromatograph", American Cynamid
Report No. WI-88-13, 1988.
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TABLE 1
SINGLE LABORATORY METHOD PERFORMANCE
CONCENTRATION
SAMPLE SPIKE (jiig/L) % RECOVERY
A 60 100
B 60 105
C 40 86
D 40 100
E 40 88
F 60 94
Average 96
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METHOD 8031
ACRYLONITRILE BY GAS CHRQMATQGRAPHY
Start
7.1.1 Extract
40ml of sample
with methyl
t-butyl ether
in 90ml bottle.
72 Set
chroma tographic
conditions
7.3.1 Flush CC
system with 3ul
methyl t-buty.l
ether.
7.3.2 Analyze
3ul of sample
blank
7.3.3-7.3.4
Establish
calibration
curve with at
least S stds.
7 .4 Sample
Analysis
Stop
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METHOD 8032
ACRYLAMIDE BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8032 is used to determine trace amounts of acrylamide monomer
in aqueous matrices. This method may be applicable to other matrices. The
following compounds can be determined by this method:
Compound Name CAS No.a
Acrylamide 79-06-01
a Chemical Abstract Services Registry Number.
1.2 The method detection limit (MDL) in clean water is 0.032 M9/L.
1.3 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8032 is based on bromination of the acrylamide double bond.
The reaction product (2,3-dibromopropionamide) is extracted from the reaction
mixture with ethyl acetate, after salting out with sodium sulfate. The extract
is cleaned up using a Florisil column, and analyzed by gas chromatography with
electron capture detection (GC/ECD).
2.2 Compound identification should be supported by at least one additional
qualitative technique. Analysis using a second gas chromatographic column or
gas chromatography/mass spectrometry may be used for compound confirmation.
3.0 INTERFERENCES
3.1 No interference is observed from sea water or in the presence of 8.0%
of ammonium ions derived from ammonium bromide. Impurities from potassium
bromide are removed by the Florisil clean up procedure.
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatographic System
4.1.1 Gas chromatograph suitable for on-column injections with all
required accessories, including detector, analytical columns, recorder,
gases, and syringes. A data system for measuring peak heights and/or peak
areas is recommended.
4.1.2 Column: 2 m x 3 mm glass column, 5% FFAP (free fatty acid
polyester) on 60-80 mesh acid washed Chromosorb W, or equivalent.
4.1.3 Detector: electron capture detector.
4.2 Kuderna-Danish (K-D) apparatus.
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Separatory funnel - 150 ml.
4.4 Volumetric flask (Class A) - 100 ml, with ground glass stopper; 25 ml,
amber, with ground glass stopper.
4.5 Syringe - 5 mL.
4.6 Microsyringes - 5 /iL, 100 ML-
4.7 Pipets (Class A).
4.8 Glass column (30 cm x 2 cm).
4.9 Mechanical shaker.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
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such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Ethyl acetate, C2H5C02C2H5. Pesticide quality, or equivalent.
5.3.2 Diethyl ether, C2H5OC2H5. Pesticide quality, or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.3.3 Methanol, CH3OH. Pesticide quality, or equivalent.
5.3.4 Benzene, C6H6. Pesticide quality, or equivalent.
5.3.5 Acetone, CH3COCH3. Pesticide quality, or equivalent.
5.4 Saturated bromine water. Prepare by shaking organic-free reagent
water with bromine and allowing to stand for 1 hour, in the dark, at 4°C. Use
the aqueous phase.
5.5 Sodium sulfate (anhydrous, granular), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.6 Sodium thiosulfate, Na2S203, 1 M aqueous solution.
5.7 Potassium bromide, KBr, prepared for infrared analysis.
5.8 Concentrated hydrobromic acid, HBr, specific gravity 1.48.
5.9 Acrylamide monomer, H2C:CHCONH2, electrophoresis reagent grade,
minimum 95% purity.
5.10 Dimethyl phthalate, C6H4(COOCH3)2, 99.0% purity.
5.11 Florisil (60/100 mesh): Prepare Florisil by activating at 130°C for
at least 16 hours. Alternatively, store Florisil in an oven at 130°C. Before
use, cool the Florisil in a desiccator. Pack 5 g of the Florisil, suspended in
benzene, in a glass column (Section 4.8).
5.12 Stock standard solutions
5.12.1 Prepare a stock standard solution of acrylamide monomer as
specified in Section 5.12.1.1. When compound purity is assayed to be 96%
or greater, the weight can be used without correction to calculate the
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concentration of the stock standard. Commercially prepared standards can
be used at any concentration if they are certified by the manufacturer or
by an independent source.
5.12.1.1 Dissolve 105.3 mg of acrylamide monomer in
organic-free reagent water in a 100 ml volumetric flask, and dilute
to the mark with organic-free reagent water. Dilute the solution of
acrylamide monomer so as to obtain standard solutions containing
0.1 - 10 ng/ml of acrylamide monomer.
5.13 Calibration standards
5.13.1 Dilute the acrylamide stock solution with organic-free
reagent water to produce standard solutions containing 0.1-5 M9/roL of
acrylamide. Prior to injection the calibration standards are reacted and
extracted in the same manner as environmental samples (Section 7).
5.14 Internal standards
5.14.1 The suggested internal standard is dimethyl phthalate.
Prepare a solution containing 100 iig/ml of dimethyl phthalate in ethyl
acetate. The concentration of dimethyl phthalate in the sample extracts
and calibration standards should be 4 M9/mL.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Bromination
7.1.1 Pipet 50 mL of sample into a 100 mL glass stoppered flask.
Dissolve 7.5 g of potassium bromide into the sample, with stirring.
7.1.2 Adjust the pH of the solution with concentrated hydrobromic
acid until the pH is between 1 and 3.
7.1.3 Wrap the flask with aluminum foil in order to exclude light.
Add 2.5 mL of saturated bromine water, with stirring. Store the flask and
contents in the dark, at 0°C, for at least 1 hour.
7.1.4 After reacting the solution for at least an hour, decompose
the excess of bromine by adding 1 M sodium thiosulfate solution, dropwise,
until the color of the solution is discharged.
7.1.5 Add 15 g of sodium sulfate, using a magnetic stirrer to effect
vigorous stirring.
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7.2 Extraction
7.2.1 Transfer the solution into a 150 ml separatory funnel. Rinse
the reaction flask three times with 1 ml aliquots of organic-free reagent
water. Transfer the rinsings into the separatory funnel.
7.2.2 Extract the aqueous solution with two 10 mL portions of ethyl
acetate for 2 min each, using a mechanical shaker (240 strokes per min).
Dry the organic phase with 1 g of sodium sulfate.
7.2.3 Transfer the organic phase into a 25 ml amber volumetric
flask. Rinse the sodium sulfate with three 1.5 ml portions of ethyl
acetate and combine the rinsings with the organic phase.
7.2.4 Add exactly 100 /xg of dimethyl phthalate to the flask and
make the solution up to the 25 ml mark with ethyl acetate. Inject 5 /xL
portions of this solution into the gas chromatograph.
7.3 Florisil cleanup: Whenever interferences are observed, the samples
should be cleaned up as follows.
7.3.1 Transfer the dried extract into a Kuderna-Danish evaporator
with 15 ml of benzene. Evaporate the solvent at 70°C under reduced
pressure, and concentrate the solution to about 3 ml.
7.3.2 Add 50 ml of benzene and subject the solution to Florisil
column chromatography at a flow rate of 3 mL/min. Elute the column first
with 50 ml of diethyl ether/benzene (1:4) at a flow rate of 5 mL/min, and
then with 25 ml of acetone/benzene (2:1) at a flow rate of 2 mL/min.
Discard all of the first eluate and the initial 9 mL portion of the second
eluate, and use the remainder for the determination, using dimethyl
phthalate (4 ng/ml) as an internal standard.
Note: Benzene is toxic, and should be only be used under a ventilated laboratory
hood.
7.4 Gas chromatographic conditions:
Nitrogen carrier gas flow rate: 40 mL/min
Column temperature: 165°C.
Injector temperature: 180°C
Detector temperature: 185°C.
Injection volume: 5 /xL
7.5 Calibration:
7.5.1 Inject 5 /nL of a sample blank (organic-free reagent water
carried through all sample storage, handling, bromination and extraction
procedures).
7.5.2 Prepare standard solutions of acrylamide as described in
Section 5.13.1. Brominate and extract each standard solution as described
in Sections 7.1 and 7.2.
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7.5.2.1 Inject 5 p.1 of each of a minimum of five standard
solutions: one should be near the detection limit; one should be
near, but below, the expected concentrations of the analyte; one
should be near, but above, the expected concentrations of the
analyte.
7.5.2.2 Prepare a calibration curve using the peak areas of
the standards. If the calibration curve deviates significantly from
a straight line, prepare a new calibration curve with the existing
standards, or, prepare new standards and a new calibration curve.
See Method 8000, Section 7.4.3, for additional guidance on
calibration by the internal standard method.
7.5.2.3 Calculate the response factor for each standard
according to Equation 1.
(Ps) (Mis)
RF = Equation 1
(P,s) (MA)
RF = Response factor
Ps = Peak height of acrylamide
Mis = Amount of internal standard injected (ng)
Pls = Peak height of internal standard
MA = Amount of acrylamide injected (ng)
7.5.3 Calculate the mean response factor according to Equation 2,
n
2 RF
i=l
RF = Equation 2
RF = Mean response factor
RF = Response factors from standard analyses (calculated in Equation 1)
n = Number of analyses
7.6 Gas chromatographic analysis:
7.6.1 Inject 5 ^l portions of each sample (containing 4 ng/ml
internal standard) into the gas chromatograph. An example GC/ECD
chromatogram is shown in Figure 1.
7.6.2 The concentration of acrylamide monomer in the sample is given
by Equation 3.
(PA) (Mis)
[A] = — Equation 3
(V,) (V.)
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[A] = Concentration of acryl amide monomer in sample (jug/mL)
PA = Peak height of acryl amide monomer
Mls = Amount of internal standard injected (ng)
Vs = Total volume of sample (ml)
P^ = Peak height of internal standard
RF = Mean response factor from Equation 2
V, = Injection volume (/iL)
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
9.0 METHOD PERFORMANCE
9.1 The following performance data have been generated under the
conditions described in this method:
9.1.1 The calibration curve for Method 8032 is linear over the range
0-5 /xg/L of acryl amide monomer.
9.1.2 The limit of detection for an aqueous solution is 0.032 M9/L.
9.1.3 The yields of the brominated compound are 85.2 + 3.3% and 83.3
+ 0.9%, at fortification concentrations of 1.0 and 5.0 Mg/L, respectively.
9.2 Table 1 provides the recoveries of acryl amide monomer from river
water, sewage effluent, and sea water.
9.3 The recovery of the bromi nation product as a function of the amount
of potassium bromide and hydrobromic acid added to the sample is shown in
Figure 2.
9.4 The effect of the reaction time on the recovery of the bromination
product is shown in Figure 3. The yield was constant when the reaction time
was more than 1 hour.
9.5 Figure 4 shows the recovery of the bromination product as a function
of the initial pH from 1 to 7.35. The yield was constant within this pH range.
The use of conventional buffer solutions, such as sodium acetate - acetic acid
solution or phosphate solution, caused a significant decrease in yield.
10.0 REFERENCES
1. Hashimoto, A., "Improved Method for the Determination of Acrylamide Monomer
in Water by Means of Gas-Liquid Chromatography with an Electron-capture
Detector," Analyst, 101:932-938, 1976.
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TABLE 1
RECOVERY OF ACRYLAMIDE FROM WATER SAMPLES AS
2,3-DIBROMOPROPIONAMIDE
Sampl e
Matrix
Standard
River Water
Sewage
Effluent
Sea Water
Acryl amide
Monomer
Spiked//Ltg
0.05
0.20
0.25
0.20
0.20
0.20
Amount of 2,
Calculated
0.162
0.649
0.812
0.649
0.649
0.649
3-DBPA7jug
Found"
0.138
0.535
0.677
0.531
0.542
0.524
Overall
Bromi nation
Recovery
%b
85.2
82.4
83.3
81.8
83.5
80.7
Recovery of
Acryl amide
Monomer, %b
...
—
—
99.4
101.3
98.8
Coefficient
of
Variation
3.3
1.0
0.9
2.5
3.0
3.5
a 2,3-Dibromopropionamide
b Mean of five replicate determinations
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Figure 1
Typical gas chromatograms of the bromination product obtained from aqueous
acrylamide monomer solution:
A. Untreated
B. With Florisil cleanup
BL. Chromatogram of blank, concentrated five-fold before gas chromatographic
analysis.
Peaks:
1. 2,3-Dibromopropionamide
2. Dimethyl phthalate
4-7. Impurities from potassium bromide
Sample size = 100 mL; acrylamide monomer = 0.1 M9
8032 - 9
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Figure 2
8
o
*»
-------
Figure 3
24
Effect of reaction time on the bromination. Reaction conditions:
50 ml of sample;
0.25 /ig of acrylamide monomer;
7.5 g of potassium bromide;
2.5 ml of saturated bromine water
Extraction conditions:
15 g of sodium sulfate;
extraction at pH 2;
solvent = 10 ml of ethyl acetate (X2)
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Figure 4
1 »
t t 1
012345971
PM
Effect of initial pH on the bromination. Reaction and extraction conditions as
in Figure 3. The pH was adjusted to below 3 with concentrated hydrobromic acid,
and to 4-5 with dilute hydrobromic acid. Reaction at pH 6 was in distilled
water. pH 7.35 was achieved by careful addition of dilute sodium hydroxide
solution. The broken line shows the result obtained by the use of sodium acetate
- acetic acid buffer solution.
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METHOD 8032
ACRYLAMIDE BY GAS CHRQMATOGRAPHY
7.1 Sromination
7.1.1 Dissolve 7.5 gr. KBr into
50 ml. sample in flask.
7.1.2 Adjust soln. pH with
concentrated HBr to between
1 and 3.
7.1.3 Wrap soln. flask w/aluminum.
Add 2.5 ml. satd. bromine
water, stir, store at 0 C for
1 hr.
7.1.4 Add 1 M sodium thiosulfate
dropwise to flask to
decompose excess bromine.
7.1.5 Add 15 gr. sodium sulfate,
and stir.
7.2 Extraction
7.2.1 Transfer flask soln. to
sep. funnel along with
rinses.
7.2.2 Extract soln. twice w/ethyl
acetate. Dry organic phase
using sodium sulfate.
7.2.3 Transfer organic phase and
rinses into amber glass flask.
7.2.4 Add 100 ug. dimethyl
phthalate to flask, dilute to
mark. Inject 5 ul. into GC.
7.3 Florisil Cleanup
7.3.1 Transfer dried extract to K-D
assembly w/benzene.
Concentrate to 3 ml. at 70 C
under reduced pressure.
7.3.2 Add 50 ml. benzene to
solution. Pass soln. through
Florisil column. Elute with
diethyl ether/benzene, then
acetone/benzene. Collect
the second elution train (less
initial 9 ml.) for analysis.
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METHOD 8032
continued
7.4 GC Conditions
7.5 Calibration
7.5.1 Inject 5 ul. sample blank.
7.5.2 Brominqte and extract std.
solns. similar to the samples.
.1 Inject 5 ul. of each of the
minimum 5 stds.
.2 Plot peak area vs. f ].
.3 Calculate response factor
(RF) for each [ ].
7.5.3 Calculate mean RF from
eqn. 2.
7.6 GC Analysis
7.6.1 Inject 5 ul. sample containing
internal std. into GC.
7.6.2 Calculate acrylamide monomer
concentration in sample using
eqn. 3.
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METHOD 8061
PHTHALATE ESTERS BY CAPILLARY GAS CHROMATOGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
1.0 SCOPE AND APPLICATION
1.1 Method 8061 is used to determine the identities and concentrations
of various phthalate esters in liquid, solid and sludge matrices. The following
compounds can be determined by this method:
Compound Name CAS No.'
Benzyl benzoate (I.S.) 120-51-4
Bis(2-n-butoxyethyl) phthalate (BBEP) 117-83-9
Bis(2-ethoxyethyl) phthalate (BEEP) 605-54-9
Bis(2-ethylhexyl) phthalate (DEHP) 117-81-7
Bis(2-methoxyethyl) phthalate (BMEP) 117-82-8
Bis(4-methyl-2-pentyl) phthalate (BMPP) 146-50-9
Butyl benzyl phthalate (BBP) 85-68-7
Diamyl phthalate (DAP) 131-18-0
Di-n-butyl phthalate (DBP) 84-74-2
Dicyclohexyl phthalate (DCP) 84-61-7
Diethyl phthalate (DEP) 84-66-2
Dihexyl phthalate (DHP) 84-75-3
Diisobutyl phthalate (DIBP) 84-69-5
Dimethyl phthalate (DMP) 131-11-3
Dinonyl phthalate 84-76-4
Di-n-octyl phthalate (OOP) 117-84-0
Hexyl 2-ethylhexyl phthalate (HEHP) 75673-16-4
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limits (MDL) for the target
analytes in a water matrix. The MDLs for the components of a specific sample
may differ from those listed in Table 1 because MDLs depend on the nature of
interferences in the sample matrix. Table 2 lists the estimated quantitation
limits (EQL) for other matrices.
1.3 When this method is used to analyze for any or all of the target
analytes, compound identification should be supported by at least one additional
qualitative technique. This method describes conditions for parallel column,
dual electron capture detector analysis which fulfills the above requirement.
Retention time information obtained on two megabore fused-silica open tubular
columns is given in Table 1. Alternatively, gas chromatography/mass spectrometry
could be used for compound confirmation.
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1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 liter for
liquids, 10 to 30 grams for solids and sludges) is extracted by using the
appropriate sample extraction technique specified in Methods 3510, 3540, and
3550. Method 3520 is not recommended for the extraction of aqueous samples
because the longer chain esters (dihexyl phthalate, bis(2-ethylhexyl) phthalate,
di-n-octyl phthalate, and dinonyl phthalate) tend to adsorb to the glassware and
consequently, their extraction recoveries are <40 percent. Aqueous samples are
extracted at a pH of 5 to 7, with methylene chloride, in a separatory funnel
(Method 3510). Alternatively, particulate-free aqueous samples could be filtered
through membrane disks that contain C18-bonded silica. The phthalate esters are
retained by the silica and, later eluted with acetonitrile. Solid samples are
extracted with hexane/acetone (1:1) or methylene chloride/acetone (1:1) in a
Soxhlet extractor (Method 3540) or with an ultrasonic extractor (Method 3550).
After cleanup, the extract is analyzed by gas chromatography with electron
capture detection (GC/ECD).
2.2 The sensitivity of Method 8061 usually depends on the level of
interferences rather than on instrumental limitations. If interferences prevent
detection of the analytes, cleanup of the sample extracts is necessary. Either
Method 3610 or 3620 alone or followed by Method 3660, Sulfur Cleanup, may be used
to eliminate interferences in the analysis. Method 3640, Gel Permeation Cleanup,
is applicable for samples that contain high amounts of lipids and waxes.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Interferences coextracted from the samples will vary considerably
from waste to waste. While general cleanup techniques are referenced or provided
as part of this method, unique samples may require additional cleanup approaches
to achieve desired sensitivities for the target analytes.
3.3 Glassware must be scrupulously clean. All glassware require treatment
in a muffle furnace at 400°C for 2 to 4 hrs, or thorough rinsing with
pesticide-grade solvent, prior to use. Refer to Chapter 4, Section 4.1.4, for
further details regarding the cleaning of glassware. Volumetric glassware should
not be heated in a muffle furnace.
If Soxhlet extractors are baked in the muffle furnace, care must be taken
to ensure that they are dry (breakage may result if any water is left in the
side-arm). Thorough rinsing with hot tap water, followed by deionized water and
acetone is not an adequate decontamination procedure. Even after a Soxhlet
extractor was refluxed with acetone for three days, with daily solvent changes,
the concentrations of bis(2-ethylhexyl) phthalate were as high as 500 ng per
washing. Storage of glassware in the laboratory introduces contamination, even
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if the glassware is wrapped in aluminum foil. Therefore, any glassware used in
Method 8061 should be cleaned immediately prior to use.
3.4 Florisil and alumina may be contaminated with phthalate esters and,
therefore, use of these materials in sample cleanup should be employed
cautiously. If these materials are used, they must be obtained packaged in
glass (plastic packaging will contribute to contamination with phthalate esters).
Washing of these materials prior to use with the solvent(s) used for elution
during extract cleanup was found helpful, however, heating at 3208C for Florisil
and 210"C for alumina is recommended. Phthalate esters were detected in Florisil
cartridge method blanks at concentrations ranging from 10 to 460 ng, with
5 phthalate esters in the 105 to 460 ng range. Complete removal of the phthalate
esters from Florisil cartridges does not seem possible, and it is therefore
desirable to keep the steps involved in sample preparation to a minimum.
3.5 Paper thimbles and filter paper must be exhaustively washed with the
solvent that will be used in the sample extraction. Soxhlet extraction of paper
thimbles.and filter paper for 12 hrs with fresh solvent should be repeated for
a minimum of three times. Method blanks should be obtained before any of the
precleaned thimbles or filter papers are used. Storage of precleaned thimbles
and filter paper in precleaned glass jars covered with aluminum foil is
recommended.
3.6 Glass wool used in any step of sample preparation should be a
specially treated pyrex wool, pesticide grade, and must be baked at 400°C for
4 hrs. immediately prior to use.
3.7 Sodium sulfate must be obtained packaged in glass (plastic packaging
will contribute to contamination with phthalate esters), and must be purified
by heating at 400°C for 4 hrs. in a shallow tray, or by precleaning with
methylene chloride (Section 5.3). To avoid recontamination, the precleaned
material must be stored in glass-stoppered glass bottles, or glass bottles
covered with precleaned aluminum foil. The storage period should not exceed two
weeks. To minimize contamination, extracts should be dried directly in the
glassware in which they are collected by adding small amounts of precleaned
sodium sulfate until an excess of free flowing material is noted.
3.8 The presence of elemental sulfur will result in large peaks which
often mask the region of the compounds eluting before dicyclohexyl phthalate
(Compound No. 14) in the gas chromatograms shown in Figure 1. Method 3660 is
suggested for removal of sulfur.
3.9 Waxes and lipids can be removed by Gel Permeation Chromatography
(Method 3640). Extracts containing high concentrations of lipids are viscous,
and may even solidify at room temperature.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatography
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column and split/splitless injections and
all required accessories, including detector, analytical columns, recorder,
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gases, and syringes. A data system for measuring peak heights and/or peak
areas is recommended.
4.1.1.1 Eight inch injection tee (Supelco, Inc., Catalog
No. 2-3665, or equivalent) or glass Y splitter for megabore columns
(J&W Scientific, "press-fit", Catalog No. 705-0733, or equivalent).
4.1.2 Columns
4.1.2.1 Column 1, 30 m x 0.53 mm ID, 5% phenyl/95% methyl
silicone fused-silica open tubular column (DB-5, J&W Scientific, or
equivalent), 1.5 jum film thickness.
4.1.2.2 Column 2, 30 m x 0.53 mm ID, 14% cyanopropyl phenyl
silicone fused-silica open tubular column (DB-1701, J&W Scientific,
or equivalent), 1.0 p,m film thickness.
4.1.3 Detector - Dual electron capture detector (ECD)
4.2 Glassware, see Methods 3510, 3540, 3550, 3610, 3620, 3640, and 3660
for specifications.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips, approximately 10/40 mesh. Heat to 400°C for 30 min,
or Soxhlet-extract with methylene chloride prior to use.
4.5 Water bath, heated, with concentric ring cover, capable of temperature
control (± 2°C).
4.6 Vacuum system for eluting disposable solid-phase extraction
cartridges.
4.6.1 Vacuum manifold consisting of individually adjustable, easily
accessible flow-control valves for up to 24 cartridges, sample rack,
chemically resistant cover and seals, heavy-duty glass basin, removable
stainless steel solvent guides, built-in vacuum gauge and valve.
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4.6.2 Vacuum trap made of 500 ml side arm flask fitted with a
one-hole stopper and glass tubing.
4.6.3 6 mL, 1 g solid-phase extraction cartridges, LC-Florisil or
equivalent, prepackaged, ready to use.
4.7 Vials - 2 ml, 10 ml, glass with Teflon lined screw-caps or crimp
tops.
4.8 Apparatus for filtration of aqueous samples through extraction disks
(optional).
4.8.1 Vacuum apparatus: (Vac Elut SPS24, Analytichem International,
or equivalent).
4.8.1.1 1 liter suction flask.
4.8.1.2 Disk base.
4.8.1.3 Graduated funnel.
4.8.1.4 Clamp.
4.8.1.5 Vacuum gauge.
4.8.1.6 Pinch clamp.
4.8.1.7 25 x 200 mm test tube.
4.8.2 47 mm C1B-extraction disks (3M-Empore, Analytichem
International, Catalog No. 1214-5004, or equivalent).
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.4 Florisil - 60/80 mesh, activated at 400"C for 16 hrs, then deactivated
with water (3 percent by weight).
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5.5 Alumina - Alumina Woelm N Super I, activated/deactivated as described
for Florisil, or equivalent.
5.6 Solvents:
5.6.1 Hexane, C6H14 - Pesticide quality, or equivalent.
5.6.2 Methylene chloride, CH2C12 - Pesticide quality, or equivalent.
5.6.3 Acetone, CH3COCH3 - Pesticide quality, or equivalent.
5.6.4 Acetonitrile, CH3CN - HPLC grade.
5.6.5 Methanol, CH3OH - HPLC grade.
5.6.6 Diethyl Ether, C2H5OC2H5 - Pesticide quality, or equivalent.
Must be free of peroxides, as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.7 Stock standard solutions:
5.7.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in hexane,
and diluting to volume in a 10 ml volumetric flask. When compound purity
is assayed to be 96 percent or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standard solutions can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.7.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp tops. Store at 4eC and protect from
light. Stock standard solutions should be checked periodically by gas
chromatography for signs of degradation or evaporation, especially just
prior to preparation of calibration standards.
5.7.3 Stock standard solutions must be replaced after 6 months, or
sooner if comparison with check standards indicates a problem.
5.8 Calibration standards: Calibration standards are prepared at a minimum
of five concentrations for each parameter of interest through dilution of the
stock standard solutions with hexane. One of the concentrations should be at
a concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples, or should define the working range of the GC. Calibration
solutions must be replaced after 1 to 2 months, or sooner if comparison with
calibration verification standards indicates a problem.
5.9 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
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affected by method or matrix interferences. Benzyl benzoate has been tested
and found appropriate for Method 8061.
5.9.1 Prepare a spiking solution of benzyl benzoate in hexane at
5000 mg/L. Addition of 10 /iL of this solution to 1 ml of sample extract
is recommended. The spiking concentration of the internal standard should
be kept constant for all samples and calibration standards. Store the
internal standard spiking solution at 4eC in glass vials with Teflon lined
screw-caps or crimp tops. Standard solutions should be replaced when
ongoing QC (Section 8) indicates a problem.
5.10 Surrogate standards: The analyst should monitor the performance of
the extraction, cleanup (when used), analytical system, and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and blank with surrogate compounds. Three surrogates are suggested
for Method 8061: diphenyl phthalate, diphenyl isophthalate, and dibenzyl
phthalate.
5.10.1 Prepare a surrogate standard spiking solution, in acetone,
which contains 50 ng//ul_ of each compound. Addition of 500 juL of this
solution to 1 L of water or 30 g solid sample is equivalent to 25 /tg/L of
water or 830 jug/kg of solid sample. The spiking concentration of the
surrogate standards may be adjusted accordingly, if the final volume of
extract is reduced below 2 ml for water samples or 10 ml for solid samples.
Store the surrogate spiking solution at 4°C in glass vials with Teflon
lined screw-caps or crimp tops. The solution must be replaced after
6 months, or sooner if ongoing QC (Section 8) indicates problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a pH of
5 to 7 with methylene chloride in a separatory funnel (Method 3510).
Method 3520 is not recommended for the extraction of aqueous samples
because the longer chain esters (dihexyl phthalate bis(2-ethylhexyl)
phthalate, di-n-octyl phthalate, and dinonyl phthalate) tend to adsorb to
the glassware and consequently, their extraction recoveries are
<40 percent. Solid samples are extracted with hexane/acetone (1:1) or
methylene chloride/acetone (1:1) in a Soxhlet extractor (Method 3540) or
with an ultrasonic extractor (Method 3550). Immediately prior to
extraction, spike 500 /xL of the surrogate standard spiking solution
(concentration = 50 ng//uL) into 1 L aqueous sample or 30 g solid sample.
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7.1.2 Extraction of particulate-free aqueous samples using
C16-extraction disks (optional):
7.1.2.1 Disk preconditioning: Place the C18-extraction disk
into the filtration apparatus and prewash the disk with 10 to 20 ml
of acetonitrile. Apply vacuum to pull the solvent through the disk.
Maintain vacuum to pull air through for 5 min. Follow with 10 ml
of methanol. Apply vacuum and pull most of the methanol through the
disk. Release vacuum before the disk gets dry. Follow with 10 mL
organic-free reagent water. Apply vacuum and pull most of the water
through the disk. Release the vacuum before the disk gets dry.
7.1.2.2 Sample preconcentration: Add 2.5 mL of methanol to
the 500 ml aqueous sample in order to get reproducible results.
Pour the sample into the filtration apparatus. Adjust vacuum so
that it takes approximately 20 min to process 502.5 ml of sample.
After all of the sample has passed through the membrane disk, pull
air through the disk for 5 to 10 min. to remove any residual water.
7.1.2.3 Sample elution: Break the vacuum and place the tip
of the filter base into the test tube that is contained inside the
suction flask. Add 10 ml of acetonitrile to the graduated funnel,
making sure to rinse the walls of the graduated funnel with the
solvent. Apply vacuum to pass the acetonitrile through the membrane
disk.
7.1.2.4 Extract concentration: Concentrate the extract to
2 mL or less, using either the micro Snyder column technique (Section
7.1.2.4.1) or nitrogen blowdown technique (Section 7.1.2.4.2).
7.1.2.4.1 Micro Snyder Column Technique
7.1.2.4.1.1 Add one or two clean boiling chips
to the concentrator tube and attach a two ball micro
Snyder column. Prewet the column by adding about 0.5 mL
of acetonitrile to the top of the column. Place the K-D
apparatus in a hot water bath (15-20°C above the boiling
point of the solvent) so that the concentrator tube is
partially immersed in the hot water and the entire lower
rounded surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus and the
water temperature, as required, to complete the
concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively
chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5 mL, remove the K-D
apparatus from the water bath and allow it to drain and
cool for at least 10 minutes. Remove the Snyder column
and rinse the flask and its lower joints with about
0.2 mL of solvent and add to the concentrator tube.
Adjust the final volume to 1.0-2.0 mL with solvent.
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7.1.2.4.2 Nitrogen Slowdown Technique
7.1.2.4.2.1 PI ace the concentrator tube i n a warm
water bath (approximately 35°C) and evaporate the solvent
volume to the required level using a gentle stream of
clean, dry nitrogen (filtered through a column of
activated carbon).
CAUTION; Do not use plasticized tubing between the carbon trap and the sample.
7.1.2.4.2.2 The internal wall of the tube must
be rinsed down several times with acetonitrile during
the operation. During evaporation, the solvent level
in the tube must be positioned to prevent water from
condensing into the sample (i.e., the solvent level
should be below the level of the water bath). Under
normal operating conditions, the extract should not be
allowed to become dry.
7.2 Solvent Exchange: Prior to Florisil cleanup or gas chromatographic
analysis, the methylene chloride and methylene chloride/acetone extracts obtained
in Section 7.1.1 must be exchanged to hexane, as described in Sections 7.2.1
through 7.2.3. Exchange is not required for the acetonitrile extracts obtained
in Section 7.1.2.4.
7.2.1 Add one or two clean boiling chips to the flask and attach
a three ball Snyder column. Concentrate the extract as described in
Section 7.1.2.4.1, using 1 ml of methylene chloride to prewet the column,
and completing the concentration in 10-20 minutes. When the apparent
volume of liquid reaches 1-2 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes.
7.2.2 Momentarily remove the Snyder column, add 50 ml of hexane,
a new boiling chip, and attach the macro Snyder column. Concentrate the
extract as described in Section 7.1.2.4.1, using 1 ml of hexane to prewet
the Snyder column, raising the temperature of the water bath, if necessary,
to maintain proper distillation, and completing the concentration in 10-20
minutes. When the apparent volume of liquid reaches 1-2 ml, remove the
K-D apparatus and allow it to drain and cool for at least 10 min.
7.2.3 Remove the Snyder column and rinse the flask and its lower
joint into the concentrator tube with 1 to 2 ml hexane. A 5 ml syringe
is recommended for this operation. Adjust the extract volume to 2 ml for
water samples, using either the micro Snyder column technique (Section
7.1.2.4.1) or nitrogen blowdown technique (Section 7.1.2.4.2), or 10 ml
for solid samples. Stopper the concentrator tube and store at 4°C if
further processing will be performed immediately. If the extract will be
stored for two days or longer, it should be transferred to a glass vial
with a Teflon lined screw-cap or crimp top. Proceed with the gas
chromatographic analysis.
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7.3 Cleanup/Fractionation:
7.3.1 Cleanup may not be necessary for extracts from a relatively
clean sample matrix. If polychlorinated biphenyls (PCBs) and
organochlorine pesticides are known to be present in the sample, proceed
with the procedure outlined in Methods 3610 or 3620. Collect Fraction 1
by eluting with 140 ml (Method 3610) or 100 ml (Method 3620) of 20-percent
diethyl ether in hexane. Note that, under these conditions, bis(2-
methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and bis(2-n-
butoxyethyl) phthalate are not recovered from the Florisil column. The
elution patterns and compound recoveries are given in Table 3.
7.3.2 As an alternative to Method 3620, Florisil Cartridge Cleanup
may be used be used for extract cleanup. With this method,
bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and
bis(2-n-butoxyethyl) phthalate are recovered quantitatively.
7.3.2.1 If PCBs and organochlorine pesticides are known to
be present in the sample, and if Florisil Cartridge Cleanup is
considered, then two fractions are collected: Fraction 1 is eluted
with 5 ml of 20 percent methylene chloride in hexane and Fraction 2
is eluted with 5 ml of 10-percent acetone in hexane. The elution
patterns and compound recoveries are given in Table 4. Fraction 1
contains the organochlorine pesticides and PCBs, and can be
discarded. Fraction 2 contains the phthalate esters and is analyzed
by GC/ECD.
7.4 Gas chromatographic conditions (recommended):
7.4.1 Column 1 and Column 2 (Section 4.1.2):
Carrier gas (He) = 6 mL/min.
Injector temperature = 250°C.
Detector temperature = 320°C.
Column temperature:
Initial temperature = 150°C, hold for 0.5 min.
Temperature program = 150"C to 2208C at 5'C/min., followed by
220°C to 275°C at 3°C/min.
Final temperature = 275°C hold for 13 min.
7.4.2 Table 1 gives the retention times and MDLs that can be
achieved by this method for the 16 phthalate esters. An example of the
separations achieved with the DB-5 and DB-1701 fused-silica open tubular
columns is shown in Figure 1.
7.5 Calibration:
7.5.1 Refer to Method 8000 for proper calibration techniques. Use
Tables 1 and 2 for guidance on selecting the lowest point on the
calibration curve.
7.5.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for the description of each of these
procedures.
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7.6 Gas chromatographic analysis:
7.6.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 juL of internal standard solution at 5000 mg/L
to the sample prior to injection.
7.6.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.6.3 Record the sample volume injected and the resulting peak
areas.
7.6.4 Using either the internal or the external calibration
procedure (Method 8000), determine the identity and the quantity of each
component peak in the sample chromatogram which corresponds to the
compounds used for calibration purposes.
7.6.5 If the response of a peak exceeds the working range of the
system, dilute the extract and reanalyze.
7.6.6 Identify compounds in the sample by comparing the retention
times of the peaks in the sample chromatogram with those of the peaks in
standard chromatograms. The retention time window used to make identifica-
tions is based upon measurements of actual retention time variations over
the course of 10 consecutive injections. Three times the standard
deviation of the retention time can be used to calculate a suggested window
size.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC
specified in Method 3600 and in the specific cleanup method.
8.2 Mandatory quality control to evaluate the GC system operation is
found in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain the test compounds at 5 to 10 ng//iL.
8.3 Calculate the recoveries of the surrogate compounds for all samples,
method blanks, and method spikes. Determine if the recoveries are within limits
established by performing QC procedures outlined in Method 8000.
8.3.1 If the recoveries are not within limits, the following are
required:
8.3.1.1 Make sure there are no errors in calculations,
surrogate solutions and internal standards. Also check instrument
performance.
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8.3.1.2 Recalculate the data and/or reanalyze the extract
if any of the above checks reveal a problem.
8.3.1.3 Reextract and reanalyze the sample if none of the
above are a problem, or flag the data as "estimated concentration."
8.4 An internal standard peak area check must be performed on all samples.
The internal standard must be evaluated for acceptance by determining whether
the measured area for the internal standard deviates by more than 30 percent from
the average area for the internal standard in the calibration standards. When
the internal standard peak area is outside that limit, all samples that fall
outside the QC criteria must be reanalyzed.
8.5 GC/MS confirmation: Any compounds confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by GC/MS
as determined by the laboratory-generated detection limits.
8.5.1 The GC/MS would normally require a minimum concentration of
10 ng/jiL in the final extract for each single-component compound.
8.5.2 The sample extract and associated blank should be analyzed
by GC/MS as per Section 7.0 of Method 8270. Normally, analysis of a blank
is not required for confirmation analysis, however, analysis for phthalates
is a special case because of the possibility for sample contamination
through septum punctures, etc.
8.5.3 A reference standard of the compound must also be analyzed
by GC/MS. The concentration of the reference standard must be at a
concentration that would demonstrate the ability to confirm the phthalate
esters identified by GC/ECD.
8.6 Include a mid-concentration calibration standard after each group of
20 samples in the analysis sequence. The response factors for the
mid-concentration calibration must be within ± 15 percent of the average values
for the multiconcentration calibration.
8.7 Demonstrate through the analyses of standards that the Florisil
fractionation scheme is reproducible. When using the fractionation schemes
given in Methods 3610 or 3620, batch-to-batch variations in the composition of
the alumina or Florisil material may cause variations in the recoveries of the
phthalate esters.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDL concentrations listed in
Table 1 were obtained using organic-free reagent water. Details on how to
determine MDLs are given in Chapter One. The MDL actually achieved in a given
analysis will vary, as it is dependent on instrument sensitivity and matrix
effects.
9.2 This method has been tested in a single laboratory by using different
types of aqueous samples and solid samples which were fortified with the test
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compounds at two concentrations. Single-operator precision, overall precision,
and method accuracy were found to be related to the concentration of the
compounds and the type of matrix. Results of the single-laboratory method
evaluation are presented in Tables 5, 6, and 7.
9.3 The accuracy and
matrix, sample preparation
procedures used.
precision obtained is determined by the sample
technique, cleanup techniques, and calibration
10.0 REFERENCES
1. Glazer, J.A.; Foerst, G.D.; McKee, G.D.; Quave, S.A., and Budde, W.L.,
"Trace Analyses for Wastewaters," Environ. Sci. and Technol. 15: 1426,
1981. v
2. Lopez-Avila, V., Baldin, E., Benedicto, J., Milanes, J., and Beckert, W.F.,
"Application of Open-Tubular Columns to SW-846 GC Methods", EMSL-Las Vegas,
1990.
3. Beckert, W.F. and Lopez-Avila, V., "Evaluation of SW-846 Method 8060 for
Phthalate Esters", Proceedings of Fifth Annual Testing and Quality Assurance
Symposium, USEPA, 1989.
8061 - 13
Revision 0
November 1990
-------
TABLE 1.
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS FOR THE PHTHALATE ESTERS3
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
IS
SU-1
SU-2
SU-3
Compound name
Dimethyl phthalate (DMP)
Diethyl phthalate (DEP)
Diisobutyl phthalate (DIBP)
Di-n-butyl phthalate (DBP)
Bis(4-methyl-2-pentyl) phthalate (BMPP)
Bis(2-methoxyethyl) phthalate (BMEP)
Diamyl phthalate (DAP)
Bis(2-ethoxyethyl) phthalate (BEEP)
Hexyl 2-ethylhexyl phtfralate (HEHP)
Dihexyl phthalate (DHP)
Butyl benzyl phthalate (BBP)
Bis(2-n-butoxyethyl) phthalate (BBEP)
Bis(2-ethylhexyl) phthalate (DEHP)
Dicyclohexyl phthalate (DCP)
Di-n-octyl phthalate (OOP)
Dinonyl phthalate
Benzyl benzoate
Diphenyl phthalate (DPP)
Diphenyl isophthalate (DPIP)
Di benzyl phthalate (DBZP)
Chemical
Abstract
Registry
No.
131-11-3
84-66-2
84-69-5
84-74-2
146-50-9
117-82-8
131-18-0
605-54-9
75673-16-4
84-75-3
85-68-7
117-83-9
117-81-7
84-61-7
117-84-0
84-76-4
120-51-4
84-62-8
744-45-6
523-31-9
Retention time
(min)
Column 1
7.06
9.30
14.44
16.26
18.77
17.02
20.25
19.43
21.07
24.57
24.86
27.56
29.23
28.88
33.33
38.80
12.71
29.46
32.99
34.40
Column 2
6.37
8.45
12.91
14.66
16.27
16.41
18.08
18.21
18.97
21.85
23.08
25.24
25.67
26.35
29.83
33.84
11.07
28.32
31.37
32.65
MDLb
Liquid
(ng/L)
640
250
120
330
370
510
110
270
130
68
42
84
270
22
49
22
c
c
c
c
8061 - 14
Revision 0
November 1990
-------
Table 1. (continued)
Column 1 is a 30 m x 0.53 mm ID DB-5 fused-silica open tubular column (1.5 urn film thickness).
Column 2 is a 30 m 0.53 mm ID DB-1701 fused-silica open tubular column (1.0 /xm film thickness).
Temperature program is 150°C (0.5 min hold) to 220'C at 5°C/min, then to 275°C (13 min hold) at
3eC/min. An 8-in Supelco injection tee or a J&W Scientific press fit glass inlet splitter is used
to connect the two columns to the injection port of a gas chromatograph. Carrier gas helium at
6 mL/min; makeup gas nitrogen at 20 mL/min; injector temperature 250°C; detector temperature
320°C.
MDL is the method detection limit. The MDL was determined from the analysis of seven replicate
aliquots of organic-free reagent water processed through the entire analytical method (extraction,
Florisil cartridge cleanup, and GC/ECD analysis using the single column approach: DB-5 fused-
silica capillary column). MDL = t(n.1-0_gg) x SD where t(rKli099) is the student's t value appropriate for
a 99 percent confidence interval and' a standard deviation with n-1 degrees of freedom, and SD is
the standard deviation of the seven replicate measurements. Values measured were not corrected
for method blanks.
Not applicable.
8061 - 15 Revision 0
November 1990
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQL) FOR VARIOUS MATRICES8
Matrix Factor"
Groundwater 10
Low-concentration soil by ultrasonic extraction 670
with GPC cleanup
High-concentration soil and sludges by ultrasonic 10,000
extraction
Non-water miscible waste 100,000
Sample EQLs are highly matrix dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For
nonaqueous samples, the factor is on a wet weight basis.
8061 - 16 Revision 0
November 1990
-------
TABLE 3.
AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS USING METHODS 3610, 3620, AND
THE ALUMINA AND FLORISIL DISPOSABLE CARTRIDGE PROCEDURE
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Method
3610
alumina8
64.5
62.5
77.0
76.5
89.5
70.5
75.0
67.0
90.5
73.0
87.0
62.5
91.0
84.5
108
71.0
Method
3620
Florisil*
40.0
57.0
80.0
85.0
84.5
0
81.5
0
105
74.5
90.0
0
82.0
83.5
115
72.5
Alumina
cartridge"
101
103
104
108
103
64. lc
103
111
101
108
103
108
97.6
97.5
112
97.3
Florisil
cartridge"
89.4
97.3
91.8
102
105
78. 3e
94.5
93.6
96.0
96.8
98.6
91.5
97.5
90.5
97.1
105
a 2 determinations; alumina and Florisil chromatography performed according
to Methods 3610 and 3620, respectively.
b 2 determinations, using 1 g alumina cartridges; Fraction 1 was eluted with
5 mL of 20-percent acetone in hexane. 40 /ug of each component was spiked
per cartridge.
0 36.8 percent was recovered by elution with an additional 5 mL of
20-percent acetone in hexane.
d 2 determinations, using 1 g Florisil cartridges; Fraction 1 was eluted
with 5 mL of 10-percent acetone in hexane. 40 /ug of each component was
spiked per cartridge.
e 14.4 percent was recovered by elution with an additional 5 mL of
10-percent acetone in hexane.
8061 - 17
Revision 0
November 1990
-------
TABLE 4.
ELUTION AND AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS USING
THE FLORISIL DISPOSABLE CARTRIDGES
Compound
Percent recovery8
Fraction 1 Fraction 2
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis (2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
0
0
0
12
0
0
3.3
0
0
0
0
0
0
0
0
0
130
88
118
121
123
32
94
82
126
62
98
135
110
106
123
102
(52)
(2.8)
(16)
(13)
(5.7)
(31)
(8.3)
(19)
(6.4)
(15)
(6.5)
(34)
(2.7)
(3.3)
(7.0)
(8.7)
The number of determinations was 3. The values given in parentheses are
the percent relative standard deviations of the average recoveries.
8061 - 18
Revision 0
November 1990
-------
TABLE 5.
ACCURACY AND PRECISION DATA FOR EXTRACTION USING
THE 3M-EMPORE DISKS AND METHOD 8061
HPLC-qrade water
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Dlamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dlhexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis (2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Average
recovery
(%)
88.6
92.3
87.6
90.3
87.2
107
93.6
108
93.9
98.4
97.3
94.8
91.3
106
84.9
96.9
Precision
(% RSD)
17.7
10.3
16.2
13.2
9.5
13.6
21.0
8.9
22.4
5.0
2.6
6.3
7.4
19.9
3.8
11.1
Groundwater
Average
recovery
i°/\
(A)
86.6
92.6
89.3
95.0
86.7
113
78.9
102
83.4
97.7
66.0
98.7
96.3
108
90.1
95.2
Precision
(% RSD)
14.3
7.2
1.6
1.5
4.9
2.8
5.8
4.0
8.8
14.8
39.3
6.0
7.9
13.3
6.1
12.7
The number of determinations was 4.
100 MgA per component.
The spiking concentration was
8061 - 19
Revision 0
November 1990
-------
TABLE 6.
ACCURACY AND PRECISION DATA FOR METHOD 3510 AND METHOD 8061a
Spike Concentration
(20 ua/L)
Estuarine
Compound
Dimethyl phthalate
Di ethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Djhexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis (2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Surrogates:
Diphenyl phthalate
Diphenyl isophthalate
Di benzyl phthalate
water
84.0
71.2
76.0
83.2
78.6
73.8
78.2
75.6
84.7
79.8
84.1
78.5
81.4
77.4
74.9
59.5
98.5
95.8
93.9
(4.1)
(3.8)
(6.5)
(6.5)
(2.6)
(1.0)
(7.3)
(3.3)
(5.3)
(7.2)
(6.4)
(3.5)
(4.1)
(6.5)
(4.9)
(6.1)
(2.6)
(1.9)
(4.4)
Leachate
98
82
95
97
87
87
92
90
91
102
105
92
93
88
87
77
113
112
112
.9 (19.6)
.8 (19.3)
.3 (16.9)
.5 (22.3)
.3 (18.2)
.2 (21.7)
.1 (21.5)
.8 (22.4)
.1 (27.5)
(21.5)
(20.5)
.3 (16.1)
.0 (15.0)
.2 (13.2)
.5 (18.7)
.3 (4.2)
(14.9)
(11.7)
(14.0)
Estuarine
Groundwater
87.1
88.5
92.7
91.0
92.6
82.4
88.8
86.4
81.4
90.9
89.6
89.3
90.5
91.7
87.2
67.2
110
109
106
(8.1)
(15.3)
(17.1)
(10.7)
(13.7)
(4.4)
(7.5)
(5.8)
(17.6)
(7.6)
(6.1)
(3.6)
(4.9)
(15.2)
(3.7)
(8.0)
(3.3)
(3.3)
(3.8)
Spike Concentration
(60 ua/U
water
87.1
71.0
99.1
87.0
97.4
82.5
89.2
88.7
107
90.1
92.7
86.1
86.5
87.7
85.1
97.2
110 (
104
111
(7.5)
(7.7)
(19.0)
(8.0)
(15.0)
(5.5)
(2.8)
(4.9)
(16.8)
(2.4)
(5.6)
(6.2)
(6.9)
(9.6)
(8.3)
(7.0)
12.4)
(5.9)
(5.9)
Leachate
112
88.5
100
106
107
99.0
112
109
117
109
117
107
108
102
105
108
95.1
97.1
93.3
(17.5)
(17.9)
(9.6)
(17.4)
(13.3)
(13.7)
(14.2)
(14.6)
(11.4)
(20.7)
(24.7)
(15.3)
(15.1)
(14.3)
(17.7)
(17.9)
(7.2)
(7.1)
(9.5)
Groundwater
90.9 (4.5)
75.3 (3.5)
83.2 (3.3)
87.7 (2.7)
87.6 (2.9)
76.9 (6.6)
92.5 (1.8)
84.8 (5.9)
80.1 (4.1)
88.9 (2.4)
93.0 (2.0)
92.4 (0.6)
91.1 (3.0)
71.9 (2.4)
90.4 (2.0)
90.1 (1.1)
107 (2.4)
106 (2.8)
105 (2.4)
The number of determinations was 3.
the average recoveries.
The values given in parentheses are the percent relative standard deviations of
8061 - 20
Revision 0
November 1990
-------
TABLE 7.
ACCURACY AND PRECISION DATA FOR METHOD 3550 AND METHOD 8061a
Spike Concentration
(1 mq/Ka)
Compound
Dimethyl phthalate
Di ethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Estuarine
sediment
77.9
68.4
103
121
108
26.6
95.0
c
c
103
113
114
c
36.6
c
c
(42.8)
(1.7)
(3.1)
(25.8)
(57.4)
(26.8)
(10.2)
(3.6)
(12.8)
(21.1)
(48.8)
Municipal
sludge
52.1
68.6
106
86.3
97.3
72.7
81.9
66.6
114
96.4
82.8
74.0
76.6
65.8
93.3
80.0
(35.5)
(9.1)
(5.3)
(17.7)
(7.4)
(8.3)
(7.1)
(4.9)
(10.5)
(10.7)
(7.8)
(15.6)
(10.6)
(15.7)
(14.6)
(41.1)
Sandy loam
soil
c
54.7
70.3
72.6
c
0
81.9
c
57.7
77.9
56.5
c
99.2
92.8
84.7
64.2
(6.2)
(3.7)
(3.7)
(15.9)
(2.8)
(2.4)
(5.1)
(25.3)
(35.9)
(9.3)
(17.2)
Spike Concentration
(3 U.Q/Q)
Estuarine
sediment
136
60.2
74.8
74.6
104
19.5
77.3
21.7
72.7
75.5
72.9
38.3
59.5
33.9
36.8
c
(9.6)
(12.5)
(6.0)
(3.9)
(1.5)
(14.8)
(4.0)
(22.8)
(11.3)
(6.8)
(3.4)
(25.1)
(18.3)
(66.1)
(16.4)
Municipal
sludge
64.8 (11.5)
72.8 (10.0)
84.0 (4.6)
113 (5.8)
150 (6.1)
59.9 (5.4)
116 (3.7)
57.5 (9.2)
26.6 (47.6)
80.3 (4.7)
76.8 (10.3)
98.0 (6.4)
85.8 (6.4)
68.5 (9.6)
88.4 (7.4)
156 (8.6)
Sandy loam
soil
70.2
67.0
79.2
70.9
83.9
0
82.1
84.7
28.4
79.5
67.3
62.0
65.4
62.2
115
115
(2.0)
(15.1)
(0.1)
(5.5)
(11.8)
(15.5)
(8.5)
(4.3)
(2.7)
(3.8)
(3.4)
(2.8)
(19.1)
(29.2)
(13.2)
a The number of determinations was 3. The values given in parentheses are the percent relative standard deviations of the
average recoveries. All samples were subjected to Florisil cartridge cleanup.
b The estuarine sediment extract (Florisil, Fraction 1) was subjected to sulfur cleanup (Method 3660 with
tetrabutylammonium sulfite reagent).
c Not able to determine because of matrix interferant.
8061 - 21
Revision 0
November 1990
-------
Figure 1
OB-S
30 m x 0.53 mm ID
1.5-p.m Film
IS
11 12 SU-1 SU-2 SU-3
6 6
5
14
13
16
I-
o
L1J
1 IS
I
SU-2 SU-3
12 SU-1 15 I i 16
113
DB-1701
30 mx 0.53 mm ID
1.0-p.mFilm
10
11
14
V
u
u
u
JL
JL
MxJU^1
i i
10
20
TIME (min)
30
40
GC/ECD chromatograms of a composite phthalate esters standard (concentration 10 ng//iL per
compound) analyzed on a DB-5 and a DB-1701 fused-silica open tubular column. Temperature
program: 150°C (0.5 min hold) to 220°C at 5°C/min, then to 275°C (13 min hold) at 3°C/nrin.
8061 - 22
Revision 0
November 1990
-------
ISTARf)
METHOD 8061
PHTHALATE ESTERS BY CAPILLARY GAS CHRQMATQGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
7.1 Extraction
7.1.1 Refer to Chapter 2 for
guidance on choosing
an extraction procedure.
Reccomendations given.
7.1.2 Determine spike sample
recovery and detection limit
for each new sample matrix
and a given extraction
procedure.
7.1.3 Aqueous sample extraction
with CIS disks:
.1 Precondition disks using
solvent train.
.2 Concentrate sample
analytes on disk.
.3 Elute sample analytes
with acetonitrile.
.4 Concentrate extract:
.1 Micro-Snyder Column
Technique
.2 Nitrogen Slowdown
Technique
.1 Evaporate solvent to
desired level.
.2 Rinse tube walls
frequently and avoid
evaporating to dryness,
7.2 Solvent Exchange to Hexane
7.2.1 Evaporate extract volume to
1-2 ml. using K-D assembly.
7.2.2 Add hexane to K-D assembly
and evaporate to 1-2 ml.
7.2.3 Rinse K-D components and
adjust volume to desired
level.
1
7.3 Cleanup/Fractionation
7.3.1 Cleanup may not be necessary
for extracts w/clean sample
matrices. Fraction collection
and methods outlined for
other compd. groups of
interest.
7.3.2 Florisil Cartridge Cleanup
.1 Check each lot of Florisil
cartridge for analyte
recovery by eluting and
analyzing a composite std.
.2 Wash and adjust solvent
flow through cartridges.
.3 Place culture tubes or 5 ml.
vol. flasks for eluate
collection.
.4 Transfer appropriate extract
volume on cartridge.
.5 Elute the cartridges and
dilute to mark on flask.
Transfer eluate to glass
vials for concentration.
7.3.3 Collect 2 fractions if PCBs
and organochlorine pesticides
are known to be present.
7.4 Gas Chromatograph
| 7.4.1 Set GC operating parameters. |
7.4.2 Table 1 and Figure 1 shows
MDLs and analyte retention
times.
8061 - 23
Revision 0
November 1990
-------
METHOD 8061
m m
A
7.5 Calibration
1
7.5.1
'
See Mefhod 8000 for
calibration technique.
1
7.5.2
F
Refer to Method 8000 for
internal/external std.
procedure.
1
7.6 GC Analysis
7.6.1 Refer to Method 8000.
1
7.6.2
F
Follow Section 7.6 in
Method 8000 for instructions
on analysis sequence,
dilutions, retention time
windows, and identification
criteria.
i
7.6.3
Record injection volume and
sample peak areas.
i
7.6.4
r
Identify and quantify each
component peak using the
internal or external std.
procedure.
\
7.6.5
r
Dilute extracts which show
analyte levels outside of the
calibration range.
i
7.6.6
r
Identify compounds in the
sample by comparing
retention times in the
sample and the standard
chromatograms.
i
i
8061 - 24
Revision 0
November 1990
-------
METHOD 8080B
ORGANOCHLORINE PESTICIDES AND POLYCHLORINATED BIPHENYLS
BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8080 is used to determine the concentration of various
organochlorine pesticides and polychlorinated biphenyls (PCBs). The following
compounds can be determined by this method:
Compound Name
CAS No.4
Aldrin
a-BHC
/3-BHC
6-BHC
T-BHC (Lindane)
Chlordane (technical)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
309-00-2
319-84-6
319-85-7
319-86-8
58-89-9
12789-03-6
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33212-65-9
1031-07-8
72-20-8
7421-93-4
76-44-8
1024-57-3
72-43-5
8001-35-2
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
a Chemical Abstract Services Registry Number.
1.1 Table 1 lists the method detection limit for each compound in organic-
free reagent water. Table 2 lists the estimated quantitation limit (EQL) for
other matrices.
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2.0 SUMMARY OF METHOD
2.1 Method 8080 provides gas chromatographic conditions for the detection
of ppb concentrations of certain organochlorine pesticides and PCBs. Prior to
the use of this method, appropriate sample extraction techniques must be used.
Both neat and diluted organic liquids (Method 3580, Waste Dilution) may be
analyzed by direct injection. A 2 to 5 nl sample is injected into a gas
chromatograph (GC) using the solvent flush technique, and compounds in the GC
effluent are detected by an electron capture detector (ECD) or an electrolytic
conductivity detector (HECD).
2.2 The sensitivity of Method 8080 usually depends on the concentration
of interferences rather than on instrumental limitations. If interferences
prevent detection of the analytes, Method 8080 may also be performed on samples
that have undergone cleanup. Method 3620, Florisil Column Cleanup, by itself
or followed by Method 3660, Sulfur Cleanup, may be used to eliminate
interferences in the analysis.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Interferences by phthalate esters can pose a major problem in
pesticide determinations when using the electron capture detector. These
compounds generally appear in the chromatogram as large late-eluting peaks,
especially in the 15% and 50% fractions from the Florisil cleanup. Common
flexible plastics contain varying amounts of phthalates. These phthalates are
easily extracted or leached from such materials during laboratory operations.
Cross contamination of clean glassware routinely occurs when plastics are handled
during extraction steps, especially when solvent-wetted surfaces are handled.
Interferences from phthalates can best be minimized by avoiding contact with any
plastic materials. Exhaustive cleanup of reagents and glassware may be required
to eliminate background phthalate contamination. The contamination from
phthalate esters can be completely eliminated with a microcoulometric or
electrolytic conductivity detector.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph: Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Columns
4.1.2.1 Column 1: Supelcoport (100/120 mesh) coated with
1.5% SP-2250/1.95% SP-2401 packed in a 1.8 m x 4 mm ID glass column
or equivalent.
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4.1.2.2 Column 2: Supelcoport (100/120 mesh) coated with 3%
OV-1 in a 1.8 m x 4 mm ID glass column or equivalent.
4.1.3 Detectors: Electron capture (ECD) or electrolytic
conductivity detector (HECD).
4.2 Kuderna-Danish (K-D) apparatus:
4.2.1 Concentrator tube: 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask: 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column: Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column: Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Boiling chips: Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.4 Water bath: Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.5 Volumetric flasks, Class A: 10, 50, and 100 ml, ground-glass stopper.
4.6 Microsyringe: 10 nl.
4.7 Syringe: 5 ml.
4.8 Vials: Glass, 2, 10, and 20 ml capacity with Teflon-lined screw caps
or crimp tops.
4.9 Balances: Analytical, 0.0001 g and Top loading, 0.01 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
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5.3 Solvents
5.3.1 Hexane, C6H14 - Pesticide quality or equivalent.
5.3.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.3.3 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.3.4 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.4 Stock standard solutions:
5.4.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in isooctane
and diluting to volume in a 10 ml volumetric flask. A small volume of
toluene may be necessary to put some pesticides in solution. Larger
volumes can be used at the convenience of the analyst. When compound
purity is assayed to be 96% or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.4.2 Transfer the stock standard solutions into vials with Teflon-
lined screw caps or crimp tops. Store at 4°C and protect from light.
Stock standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.4.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.5 Calibration standards: Calibration standards at a minimum of five
concentrations for each parameter of interest are prepared through dilution of
the stock standards with isooctane. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples or should define the working range of the GC. Calibration
solutions must be replaced after six months, or sooner, if comparison with check
standards indicates a problem.
5.6 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Section 5.5.
5.6.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.6.3 Analyze each calibration standard according to Section 7.0.
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5.7 Surrogate standards: The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and organic-free reagent water blank with pesticide surrogates.
Because GC/ECD data are much more subject to interference than GC/MS, a secondary
surrogate is to be used when sample interference is apparent. Two surrogate
standards (tetrachloro-m-xylene (TCMX) and decachlorobiphenyl) are added to each
sample; however, only one need be calculated for recovery. Proceed with
corrective action when both surrogates are out of limits for a sample (Section
8.3). Method 3500 indicates the proper procedure for preparing these surrogates.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within
40 days of extraction.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510
or 3520. Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.2.2 Increase the temperature of the hot water bath to
about 90°C. Momentarily remove the Snyder column, add 50 mL of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature,
as required, to complete concentration in 5-10 min. At the proper
rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 1 mL, remove the K-D apparatus and allow it to drain and cool
for at least 10 min.
7.1.2.3 Remove the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1-2 mL of hexane. A 5 mL
syringe is recommended for this operation. Adjust the extract volume
to 10.0 mL. Stopper the concentrator tube and store refrigerated
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at 4°C, if further processing will not be performed immediately. If
the extract will be stored longer than two days, it should be
transferred to a vial with a Teflon-lined screw cap or crimp top.
Proceed with gas chromatographic analysis if further cleanup is not
required.
7.2 Gas chromatography conditions (Recommended):
7.2.1 Column 1:
Carrier gas (5% methane/95% argon) flow rate: 60 mL/min
Column temperature: 200°C isothermal
When analyzing for the low molecular weight PCBs (PCB 1221-PCB 1248),
it is advisable to set the oven temperature to 160°C.
7.2.2 Column 2:
Carrier gas (5% methane/95% argon) flow rate: 60 mL/min
Column temperature: 200°C isothermal
When analyzing for the low molecular weight PCBs (PCB 1221-PCB 1248),
it is advisable to set the oven temperature to 140°C.
7.2.3 When analyzing for most or all of the analytes in this method,
adjust the oven temperature and column gas flow so that 4,4'-DDT has a
retention time of approximately 12 min.
7.3 Calibration: Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.3.2 Because of the low concentration of pesticide standards
injected on a GC/ECD, column adsorption may be a problem when the GC has
not been used for a day. Therefore, the GC column should be primed or
deactivated by injecting a PCB or pesticide standard mixture approximately
20 times more concentrated than the mid-concentration standard. Inject
this prior to beginning initial or daily calibration.
7.4 Gas chromatographic analysis:
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /zL of internal standard to the sample prior to
injection.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
Note: A 72 hour sequence is not required with this method.
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7.4.3 Examples of GC/ECD chromatograms for various pesticides and
PCBs are shown in Figures 1 through 5.
7.4.4 Prime the column as per Section 7.3.2.
7.4.5 DDT and endrin are easily degraded in the injection port if
the injection port or front of the column is dirty. This is the result
of buildup of high boiling residue from sample injection. Check for
degradation problems by injecting a mid-concentration standard containing
only 4,4'-DDT and endrin. Look for the degradation products of 4,4'-DDT
(4,4'-DDE and 4,4'-ODD) and endrin (endrin ketone and endrin aldehyde).
If degradation of either DDT or endrin exceeds 20%, take corrective action
before proceeding with calibration, by following the GC system maintenance
outlined in of Method 8000. Calculate percent breakdown as follows:
% breakdown Total DDT degradation peak area (DDE + DDD)
for 4,4'-DDT = x 100
peak areas (DDT + DDE + DDD)
Total endrin degradation peak area
% breakdown (endrin aldehyde + endrin ketone)
for Endrin = x 100
peak areas (endrin + aldehyde + ketone)
7.4.6 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
7.4.7 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.4.8 If peak detection and identification are prevented due to
interferences, the hexane extract may need to undergo cleanup using Method
3620. The resultant extract(s) may be analyzed by GC directly or may
undergo further cleanup to remove sulfur using Method 3660.
7.5 Cleanup:
7.5.1 Proceed with Method 3620, followed by, if necessary, Method
3660, using the 10 ml hexane extracts obtained from Section 7.1.2.3.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous sections and in Method 8000.
7.6 Quantitation of Multiple Component Analytes:
7.6.1 Scope (excerpted from U.S. FDA, PAM): Residues which are
mixtures of two or more components present problems in measurement. When
they are found together, e.g., toxaphene and DDT, the problem of
quantitation becomes even more difficult. Suggestions are offered in the
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following sections for handing toxaphene, chlordane, PCB, DDT, and BHC.
7.6.2 Toxaphene: Quantitative calculation of toxaphene or strobane
is difficult, but reasonable accuracy can be obtained. To calculate
toxaphene on GC/ECD: (a) adjust the sample size so that the major
toxaphene peaks are 10-70% of full-scale deflection (FSD); (b) inject a
toxaphene standard that is estimated to be within ±10 ng of the sample;
(c) quantitate using the five major peaks or the total area of the
toxaphene pattern.
7.6.2.1 To measure total area, construct the baseline of
standard toxaphene between its extremities; and construct the
baseline under the sample, using the distances of the peak troughs
to baseline on the standard as a guide. This procedure is made
difficult by the fact that the relative heights and widths of the
peaks in the sample will probably not be identical to the standard.
7.6.2.2 A series of toxaphene residues have been calculated
using the total peak area for comparison to the standard and also
using the area of the last four peaks only, in both sample and
standard. The agreement between the results obtained by the two
methods justifies the use of the latter method for calculating
toxaphene in a sample where the early eluting portion of the
toxaphene chromatogram shows interferences from other substances
such as DDT.
7.6.3 Chlordane is a technical mixture of at least 11 major
components and 30 or more minor components. Trans- and cis-chlordane
(alpha and gamma), respectively, are the two major components of technical
chlordane. However, the exact percentage of each in the technical material
is not completely defined, and is not consistent from batch to batch.
7.6.3,1 The GC pattern of a chlordane residue may differ
considerably from that of the technical standard. Depending on the
sample substrate and its history, residues of chlordane can consist
of almost any combination of: constituents from the technical
chlordane, plant and/or animal metabolites, and products of
degradation caused by exposure to environmental factors such as
water and sunlight.
7.6.3.2 When the chlordane residue does not resemble technical
chlordane, but instead consists primarily of individual, identifiable
peaks, quantitate the peaks of alpha-chlordane, gamma-chlordane, and
heptachlor separately against the appropriate reference materials,
and report the individual residues.
7.6.3.3 When the GC pattern of the residue resembles that of
technical chlordane, the analyst may quantitate chlordane residues
by comparing the total area of the chlordane chromatogram using the
five major peaks or the total area. If the heptachlor epoxide peak
is relatively small, include it as part of the total chlordane area
for calculation of the residue. If heptachlor and/or heptachlor
epoxide are much out of proportion, calculate these separately and
subtract their areas from the total area to give a corrected
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chlordane area. (Note that octachlor epoxide, a metabolite of
chlordane, can easily be mistaken for heptachlor epoxide on a
nonpolar GC column.)
7.6.3.4 To measure the total area of the chlordane
chromatogram, proceed as in Section 7.6.2 on toxaphene. Inject an
amount of technical chlordane standard which will produce a
chromatogram in which the major peaks are approximately the same
size as those in the sample chromatograms.
7.6.4 Polychlorinated biphenyls (PCBs): Quantitation of residues
of PCB involves problems similar to those encountered in the quantitation
of toxaphene, strobane, and chlordane. In each case, the chemical is made
up of numerous compounds which generate multi-peak chromatograms. Also,
in each case, the chromatogram of the residue may not match that of the
standard.
7.6.4.1 Mixtures of PCBs of various chlorine contents were
sold for many years in the U.S. by the Monsanto Co. under the trade
name Aroclor (1200 series and 1016). Although these Aroclors are
no longer marketed, the PCBs remain in the environment and are
sometimes found as residues in foods, especially fish.
7.6.4.2 Since standards are not generally available for all
of the congeners of chlorinated biphenyl, PCB residues are
quantitated by comparison to one or more of the Aroclor materials,
depending on the chromatographic pattern of the residue. A choice
must be made as to which Aroclor or mixture of Aroclors will produce
a chromatogram most similar to that of the residue. This may also
involve a judgement about what proportion of the different Aroclors
to combine to produce the appropriate reference material.
7.6.4.3 PCB Quantitation option #1- Quantitate the PCB
residues by comparing the total area of the chlorinated biphenyl
peaks to the total area of peaks from the appropriate Aroclor(s)
reference materials. Measure the total area or height response from
the common baseline under all the peaks. Use only those peaks from
the sample that can be attributed to chlorobiphenyls. These peaks
must also be present in the chromatogram of the reference materials.
A mixture of Aroclors may be required to provide the best match of
the GC patterns of the sample and reference.
7.6.4.4 PCB Quantitation option #2- Quantitate the PCB
residues by comparing the responses of 3 to 5 major peaks in each
appropriate Aroclor standard with the peaks obtained from the
chlorinated biphenyls in the sample extract. The amount of Aroclor
is calculated using each of the major peaks, and the results of the
3 to 5 determinations are averaged. Major peaks are defined as those
peaks in the Aroclor standards that are at least 30% of the height
of the largest Aroclor peak. Later eluting Aroclor peaks are
generally the most stable in the environment.
7.6.5 Hexachlorocyclohexane (BHC, from the former name, benzene
hexachloride): Technical grade BHC is a cream-colored amorphous solid
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with a very characteristic musty odor; it consists of a mixture of six
chemically distinct isomers and one or more heptachlorocyclohexanes and
octachlorocyclohexanes. Commercial BHC preparations may show a wide
variance in the percentage of individual isomers present. Quantitate
each isomer (a, Ł, 7, and 5) separately against a standard of the
respective pure isomer.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8.2 Mandatory quality control to evaluate the GC system operation is
found in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each single-component parameter of interest at the following
concentrations in acetone: 4,4'-ODD, 10 mg/L; 4,4'-DDT, 10 mg/L;
endosulfan II, 10 mg/L; endosulfan sulfate, 10 mg/L; endrin, 10 mg/L; and
any other single-component pesticide, 2 mg/L. If this method is only to
be used to analyze for PCBs, chlordane, or toxaphene, the QC check sample
concentrate should contain the most representative multi-component
parameter at a concentration of 50 mg/L in acetone.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both Tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following are required.
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze
the sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
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8.4 GC/MS confirmation: Any compounds confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by GC/MS
as determined by the laboratory generated detection limits.
8.4.1 The GC/MS would normally require a minimum concentration of
10 ng//iL in the final extract, for each single-component compound.
8.4.2 The pesticide extract and associated blank should be analyzed
by GC/MS as per Section 7.0 of Method 8270.
8.4.3 The confirmation may be from the GC/MS analysis of the
base/neutral-acid extractables extracts (sample and blank). However, if
the compounds are not detected in the base/neutral-acid extract even though
the concentration is high enough, a GC/MS analysis of the pesticide extract
should be performed.
8.4.4 A reference standard of the compound must also be analyzed
by GC/MS. The concentration of the reference standard must be at a level
that would demonstrate the ability to confirm the pesticides/PCBs
identified by GC/ECD.
9.0 METHOD PERFORMANCE
9.1 The method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked
at six concentrations. Concentrations used in the study ranged from 0.5 to
30 M9/L for single-component pesticides and from 8.5 to 400 /xg/L for multi-
component parameters. Single operator precision, overall precision, and method
accuracy were found to be directly related to the concentration of the parameter
and essentially independent of the sample matrix. Linear equations to describe
these relationships for a flame ionization detector are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, optional cleanup techniques, and
calibration procedures used.
10.0 REFERENCES
1. U.S. EPA, "Development and Application of Test Procedures for Specific
Organic Toxic Substances in Wastewaters, Category 10: Pesticides and PCBs,"
Report for EPA Contract 68-03-2605.
2. U.S. EPA, "Interim Methods for the Sampling and Analysis of Priority
Pollutants in Sediments and Fish Tissue," Environmental Monitoring and
Support Laboratory, Cincinnati, OH 45268, October 1980.
3. Pressley, T.A., and J.E. Longbottom, "The Determination of Organohalide
Pesticides and PCBs in Industrial and Municipal Wastewater: Method 617,"
U.S. EPA/EMSL, Cincinnati, OH, EPA-600/4-84-006, 1982.
4. "Determination of Pesticides and PCB's in Industrial and Municipal
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Wastewaters, U.S. Environmental Protection Agency," Environmental Monitoring
and Support Laboratory, Cincinnati, OH 45268, EPA-600/4-82-023, June 1982.
5. Goerlitz, D.F. and L.M. Law, Bulletin for Environmental Contamination and
Toxicology, 6, 9, 1971.
6. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
7. Webb, R.G. and A.C. McCall, "Quantitative PCB Standards for Electron Capture
Gas Chromatography," Journal of Chromatographic Science, 11, 366, 1973.
8. Millar, J.D., R.E. Thomas and H.J. Schattenberg, "EPA Method Study 18,
Method 608: Organochlorine Pesticides and PCBs," U.S. EPA/EMSL, Research
Triangle Park, NC, EPA-600/4-84-061, 1984.
9. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
11. U.S. Food and Drug Administration, Pesticide Analytical Manual, Vol. 1,
June 1979.
12. Sawyer, L.D., JAOAC, 56, 1015-1023 (1973), 61 272-281 (1978), 61 282-291
(1978).
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TABLE 1.
GAS CHROMATOGRAPHY OF PESTICIDES AND PCBs"
Retention time (min)
Analyte
Aldrin
a-BHC
0-BHC
6-BHC
7-BHC (Lindane)
Chlordane (technical)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
•U.S. EPA. Method 617
Monitoring and Support
Col. 1
2.40
1.35
1.90
2.15
1.70
e
7.83
5.13
9.40
5.45
4.50
8.00
14.22
6.55
11.82
2.00
3.50
18.20
e
e
e
e
e
e
e
, e
Organochl
Col. 2
4.10
1.82
1.97
2.20
2.13
e
9.08
7.15
11.75
7.23
6.20
8.28
10.70
8.10
9.30
3.35
5.00
26.60
e
e
e
e
e
e
e
e
orine Pesticides
Laboratory, Cincinnati, Ohio
Method
Detection
limit (/ig/L)
0.004
0.003
0.006
0.009
0.004
0.014
0.011
0.004
0.012
0.002
0.014
0.004
0.066
0.006
0.023
0.003
0.083
0.176
0.24
nd
nd
nd
0.065
nd
nd
nd
and PCBs. Environmental
45268.
e = Multiple peak response.
nd = not determined.
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TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQLs) FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
Sample EQLs are highly matrix-dependent. The EQLs listed herein
are provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For
non-aqueous samples, the factor is on a wet-weight basis.
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Analyte
Aldrin
o-BHC
0-BHC
5-BHC
T-BHC
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Heptachlor
Heptachlor epoxide
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
Test
cone.
(M9/L)
2.0
2.0
2.0
2.0
2.0
50
10
2.0
10
2.0
2.0
10
10
10
2.0
2.0
50
50
50
50
50
50
50
50
Limit
for s
(M9/L)
0.42
0.48
0.64
0.72
0.46
10.0
2.8
0.55
3.6
0.76
0.49
6.1
2.7
3.7
0.40
0.41
12.7
10.0
24.4
17.9
12.2
15.9
13.8
10.4
Range
for x
(M9/L)
1.08-2.24
0.98-2.44
0.78-2.60
1.01-2.37
0.86-2.32
27.6-54.3
4.8-12.6
1.08-2.60
4.6-13.7
1.15-2.49
1.14-2.82
2.2-17.1
3.8-13.2
5.1-12.6
0.86-2.00
1.13-2.63
27.8-55.6
30.5-51.5
22.1-75.2
14.0-98.5
24.8-69.6
29.0-70.2
22.2-57.9
18.7-54.9
Range
P. PS
(%)
42-122
37-134
17-147
19-140
32-127
45-119
31-141
30-145
25-160
36-146
45-153
D-202
26-144
30-147
34-111
37-142
41-126
50-114
15-178
10-215
39-150
38-158
29-131
8-127
s = Standard deviation of four recovery measurements, in
x = Average recovery for four recovery measurements, in M9/L.
P, Ps = Percent recovery measured.
D = Detected; result must be greater than zero.
"Criteria from 40 CFR Part 136 for Method 608. These criteria are based directly
upon the method performance data in Table 4. Where necessary, the limits for
recovery have been broadened to assure applicability of the limits to
concentrations below those used to develop Table 4.
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Analyte
Aldrin
o-BHC
)3-BHC
6-BHC
7-BHC
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Heptachlor
Heptachlor epoxide
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
Accuracy, as
recovery, x'
(M9/L)
0.81C+0.04
0.84C+0.03
0.81C+0.07
0.81C+0.07
0.82C-0.05
0.82C-0.04
0.84C+0.30
0.85C+0.14
0.93C-0.13
0.90C+0.02
0.97C+0.04
0.93C+0.34
0.89C-0.37
0.89C-0.04
0.69C+0.04
0.89C+0.10
0.80C+1.74
0.81C+0.50
0.96C+0.65
0.91C+10.79
0.91C+10.79
0.91C+10.79
0.91C+10.79
0.91C+10.79
Single analyst
precision, s/
(M9/L)
0.16X-0.04
0.13x+0.04
0.22X+0.02
O.lSx+0.09
0.12x+0.06
0.13X+0.13
0.20X-0.18
O.lSx+0.06
0.17X+0.39
0.12X+0.19
O.lOx+0.07
0.41x-0.65
0.13X+0.33
0.20X+0.25
0.06X+0.13
O.lBx-0.11
0.09x+3.20
O.lSx+0.15
0.29x-0.76
0.21X-1.93
0.21X-1.93
0.21X-1.93
0.21X-1.93
0.21X-1.93
Overall
precision,
S' (M9/L)
0.20x-0.01
0.23X-0.00
0.33X-0.95
0.25X+0.03
0.22X+0.04
O.lSx+0.18
0.27X-0.14
0.28X-0.09
O.Slx-0.21
0.16X+0.16
O.lSx+0.08
0.47X-0.20
0.24X+0.35
0.24X+0.25
0.16x+0.08
0.25X-0.08
0.20x+0.22
O.lSx+0.45
0.35X-0.62
0.31X+3.50
O.Slx+3.50
O.Slx+3.50
0.31X+3.50
0.31x+3.50
x' = Expected recovery for one or more measurements of a sample containing
concentration C, in fj.g/1.
s/ = Expected single analyst standard deviation of measurements at an average
concentration of x, in /ug/L.
S' = Expected interlaboratory standard deviation of measurements at an average
concentration found of x, in
C = True value for the concentration, in M9/L-
x = Average recovery found for measurements of samples containing a concentration
of C, in
8080B - 16
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Figure 1
Gas Chromatogram of Pesticides
Column :1.5% P 3250*
1JS% SF-2401 en Suotieepen
TtmptrtTurc: XXPC
OttKter: Electron Cteturi
4 I 12
ftlTf NTION TIME (MINUTES)
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Figure 2
Gas Chromatogram of Chlordane
Column: 1.5% SP-2250*
1.MH SF 2401 en Swpcieeeoa
Ttmotrtturt 20C°C
DffUCtor: Electron C«oaurt
4 I 12
METENTION TIME (MINUTES)
16
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Figure 3
Gas Chromatogram of Toxaphene
Column: 1 J% SP-2250*
1JS% SP-2401 on Syptieooon
Ttmptrnurr 200°C
DetKtor: Iltetron Caoturi
10 14 II
RITINTION TIME (MINUTES)
22
26
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Figure 4
Gas Chromatogram of Aroclor 1254
Column: 1.5% SP 2250*
1JS% SP 2401 en Swpcieeeert
Ttmotntut: 200°C
Ocueter: Iltctron CiPTurt
I 10 U
MITINTION TIMC (MINUTIS)
It
22
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Figure 5
Gas Chromatogram of Aroclor 1260
1Jf% S*-J401 on SuMlcooo*
200°C
Itetron Caeturt
lit
10 U II
HITINTION TIMf (MINUTIS)
M
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Figure 6
J..L
Fig. e—Baseline construction for ram typical gas chromatosrapnlc peaks.
a, symmetrical separated flat baseline; b and c. overlapping flat bastllna;
d, saparaud (pan dots net rscum to baaalim bet wean peaks); a, separated
sloping baseline; f. separated (pan goes below'baseline between peaks):
g, «- andY-BHC sloping baseline; h. •-,Ł-, and 7-BHC sloping baseline;
1. chlordane flat baseline; J. bepudUor and neptaddor epoxide super-
invosed on calordane; k, cnalr-sbapad peaks, unsymmetrtcal peak; 1.
p,p'«OOT superimposed on to«sphane.
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Figure 7
Flf.7>-*Bu«llM construction for multlplt ruiduti with sundtrd
teuphtM.
far auiflpU mtdma »M>MK»-
008 Mtf «,pS Md P*'-OOT.
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Figure 8
Fit. ••—BaMllM construction (or multiple r»tldu»s: standard toxaphtn*.
Mr multiple rasiduMt tie* bran with BHC,
DOT* aatf i
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Figure 9
Fig. 0a—BaMlia* construction for multiple ratidiMi: standard chlordan*.
Flf,
for mddpto rwtduMt rtc* bnn
MdOOT.
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METHOD 8080B
ORGANOCHLORINE PESTICIDES AND POLYCHLORINATED BIPHENYLS
BY GAS CHROMATOGRAPHY
Start
7.1.1 Choose
appropriate
extraction
procedure.
7.1.2
Exchange
extraction
solvent to
hexane.
7.2 Set
chromatographic
conditions.
7.3 Refer to
Method 8000
for proper
calibration
techniques.
7.3.2 Prime or
deactivate the
CC column prior
to daily
calibration.
7.4 Perform
CC analysis .
7.4.8
Is peak
detection and
identification
prevented?
7.5.1 Cleanup
using Method
3620 and 3660
if necessary.
7.6.1 Oo
residues hav
two or more
7.6 Calculate
concentrations
components?
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METHOD 8081
ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8081 1s used to determine the concentrations of various
organochlorine pesticides, and polychlorinated biphenyls (PCBs) as Aroclors, 1n
extracts from solid and liquid matrices. A large number of compounds will give
a response in the electron capture detector (ECD) using this method; performance
data for the following compounds are provided as part of this method:
Compound Name
CAS No.1
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma- BHC (Lindane)
gamma- Chi ordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
309-00-2
319-84-6
319-85-7
319-86-8
58-89-9
57-74-9
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33212-65-9
1031-07-8
72-20-8
7421-93-4
76-44-8
1024-57-3
72-43-5
8001-35-2
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
Chemical Abstract Services Registry Number.
1.2 This capillary GC/ECD method allows the analyst the option of using
0.25-0.32 mm ID capillary columns (narrow bore) or 0.53 mm ID capillary columns
(wide bore). Performance data are provided for both options.
8081 - 1
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1.3 The use of narrow bore columns are recommended when the analyst
requires greater chromatographlc resolution and 1s analyzing a relatively clean
sample or an extract that has been prepared with one or more of the clean-up
options referenced in the method. Wider bore columns (0.53 mm) are suitable for
more complex environmental and waste matrices. The 0.53 mm ID columns can be
mounted in 1/4 inch packed column injectors.
1.4 Table 1 lists average retention times and method detection limits
(MDLs) for each compound of interest, in water and soil matrices, for the wide-
bore capillary column version of this method. Table 2 lists average retention
times and method detection limits (MDLs) for each compound of interest, in water
and soil matrices, for the narrow-bore capillary column version of this method.
The MDLs for the components of a specific sample may differ from those listed
in Tables 1 and 2 because they are dependent upon the nature of interferences
in the sample matrix. Retention time information given in Table 2 was obtained
on two wide-bore, open tubular columns connected to the injector port of a gas
chromatograph through an injection tee made of deactivated glass. Table 3 lists
the Estimated Quantitation Limits (EQLs) for other matrices.
1.5 When this method is used to analyze for any or all of the target
compounds, compound identification based on single column analysis should be
confirmed on a second column, or should be supported by at least one other
qualitative technique. This method describes analytical conditions for a second
gas chromatographic column that can be used to confirm the measurements made with
the primary column.
1.6 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph (GC) and in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.7 Extracts suitable for analysis by this method may also be analyzed
for organophosphorus pesticides (Method 8141) and triazine herbicides.
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 L for liquids,
2 g to 30 g for solids) is extracted using the appropriate sample extraction
technique specified in Methods 3510, 3520, 3540, 3541, 3550 and 3580. Liquid
samples are extracted at neutral pH with methylene chloride using either a
separatory funnel (Method 3510) or a continuous liquid-liquid extractor (Method
3520). Solid samples are extracted with hexane-acetone (1:1) or methylene
chloride-acetone (1:1) using either Soxhlet extraction (Method 3540), Automated
Soxhlet (Method 3541), or Ultrasonic Extraction (Method 3550). A variety of
cleanup steps may be applied to the extract, depending on (1) the nature of the
coextracted matrix interferences and (2) the target analytes. After cleanup,
the extract is analyzed by injecting a 1 /iL sample into a gas chromatograph with
a narrow- or wide-bore fused silica capillary column and electron capture
detector (GC/ECD).
2.2 The MDLs achievable in routine analyses of complex samples using
Method 8081 will usually be dependent on the degree of interference associated
8081 - 2 Revision 0
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with the presence of coelutlng compounds to which the ECD will respond, rather
than on the Inherent limitations in detector performance or the Irreducible
noise associated with Instrument electronics. If Interferences prevent
Identification and qualification of the analytes within quality control (QC)
limits at relevant concentrations, Method 8081 may also be performed on samples
that have undergone cleanup. Method 3630, Silica Gel Column Cleanup, by itself,
or followed by Method 3660, Sulfur Cleanup, may be used to eliminate
Interferences in the analysis. Method 3640, Gel-Permeation Cleanup, is
applicable for samples that contain high amounts of lipids, waxes and other high
molecular weight co-extractables.
3.0 INTERFERENCES
3.1 Refer to Methods 3550 (Section 3.5, in particular), 3600, and 8000.
3.2 Sources of interference in this method can be grouped into three
broad categories: contaminated solvents, reagents or sample processing hardware;
contaminated GC carrier gas, parts, column surfaces or detector surfaces; and
the presence of coelutlng compounds in the sample matrix to which the ECD will
respond. Knowledge of good laboratory practices is assumed, including steps to
be followed in routine testing and cleanup of solvents, reagents and sample
processing hardware, and Instrument maintenance. The discussion that follows
focuses on sources of interference associated with the sample matrix and compound
classes that represent common sources of interference, particularly phthalate
esters, organosulfur compounds, lipids, and waxes. Interferences coextracted
from the samples will vary considerably from waste to waste. While general
cleanup techniques are referenced or provided as part of this method, unique
samples may require additional cleanup approaches to achieve desired degrees of
discrimination and quantitation.
3.3 Interferences by phthalate esters introduced during sample preparation
can pose a major problem in pesticide determinations. These materials may be
removed prior to analysis using Gel Permeation Cleanup - pesticide option (Method
3640) or as Fraction III of the silica gel cleanup procedure (Method 3630).
Common flexible plastics contain varying amounts of phthalate esters which are
easily extracted or leached from such materials during laboratory operations.
Cross-contamination of clean glassware routinely occurs when plastics are handled
during extraction steps, especially when solvent-wetted surfaces are handled.
Interferences from phthalate esters can best be minimized by avoiding contact
with any plastic materials and checking all solvents and reagents for phthalate
contamination. Exhaustive cleanup of solvents, reagents and glassware may be
required to eliminate background phthalate ester contamination.
3.4 The presence of elemental sulfur will result in large peaks that
Interfere with the detection of later eluting organochlorine pesticides. Method
3660 1s suggested for removal of sulfur. Since the recovery of endrin aldehyde
(using the TBA procedure) 1s drastically reduced, this compound must be
determined prior to sulfur cleanup.
3.5 Waxes, lipids other high molecular weight co-extractables can be
removed by Gel-Permeation Cleanup (Method 3640).
8081 - 3 Revision 0
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3.6 Other pesticides may be interferences in this method. Table 4 lists
the names and retention times of organophosphorus pesticides which co-elute with
organochlorine pesticides on wide-bore capillary columns. Organophosphorus
pesticides are eliminated by the Gel Permeation Chromatography cleanup -
pesticide option (Method 3640).
3.7 It may be difficult to quantitate Aroclor patterns and single
component pesticides together. Pesticides can be removed by sulfuric
acid/permanganate cleanup (Method 3665) and silica fractionation (Method 3630).
Guidance on the identification of PCBs is given in Section 7.6.4.
4.0 APPARATUS AND MATERIALS
4.1 Glassware (see Methods 3510, 3520, 3540, 3541, 3550, 3630, 3640, and
3660 for specifications).
4.2 Kuderna-Danish (K-D) apparatus.
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). If extracts are stored in the concentrator tube, a ground
glass stopper is used to prevent evaporation of concentrates.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator with springs.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Springs, clips and clamps - 1/2 inch springs (Kontes K-662750
or equivalent), or any other equivalent fastener, e.g., neck standard taper
clips. Clamp (Kontes 675300 or equivalent).
4.2.5 Boiling chips - Approximately 10/40 mesh (silicon carbide or
equivalent). Prior to use, heat to 400°C for 30 minutes or Soxhlet extract
with methylene chloride.
4.3 Gas chromatograph - Analytical system complete with gas chromatograph
suitable for on-column and split-splitless injection and all required accessories
including syringes, analytical columns, gases, electron capture detector, and
recorder/integrator or data system.
4.3.1 Narrow-bore columns
4.3.1.1 Column 1 - 30 m x 0.25 or 0.32 mm internal diameter
(ID) fused silica capillary column chemically bonded with SE-54
(DB 5 or equivalent), 1 /xm film thickness.
4.3.1.2 Column 2 - 30 m x 0.25 mm ID fused silica capillary
column chemically bonded with 35 percent phenyl methylpolysiloxane
(DB 608, SPB 608, or equivalent), 25 pm coating thickness, 1 pm film
thickness.
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4.3.1.3 Narrow bore columns should be Installed in
split/splitless (Grob-type) injectors.
4.3.2 Wide-bore columns
4.3.2.1 Column 1 - 30 m x 0.53 mm ID fused silica capillary
column chemically bonded with 35 percent phenyl methylpolysiloxane
(DB 608, SPB 608, RTx-35, or equivalent), 0.5 Aim or 0.83 Mm film
thickness.
4.3.2.2 Column 2 - 30 m x 0.53 mm 10 fused silica capillary
column chemically bonded with 50 percent phenyl methylpolysiloxane
(OB 1701, or equivalent), 1.0 /xm film thickness.
4.3.2.3 Column 3 - 30 m x 0.53 mm ID fused silica capillary
column chemically bonded with SE-54 (OB 5, SPB 5, RTx, or
equivalent), 1.5 urn film thickness.
4.3.2.4 Wide-bore columns should be installed in 1/4 inch
injectors with deactivated liners designed specifically for use with
these columns.
4.4 GC injector ports can be of critical concern, especially in the
analysis of DDT and Endrin. Injectors that are contaminated, chemically active,
or too hot can cause the degradation ("breakdown") of the analytes. Endrin and
DDT breakdown to endrin aldehyde, endrin ketone, ODD, or DDE. When such
breakdown is observed, clean and deactivate the injector port, break off at least
0.5 M of the column and remount it. Check the injector temperature and lower
it to 205°C, if required. Endrin and DDT breakdown is less of a problem when
ambient on-column injectors are used.
5.0 REAGENTS
5.1 Reagent or pesticide grade chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water as defined in Chapter One.
5.3 Solvents and reagents: As appropriate for Method 3510, 3520, 3540,
3541, 3550, 3630, 3640, or 3660: n-hexane, diethyl ether, methylene chloride,
acetone, ethyl acetate, and isooctane (2,2,4-trimethylpentane). All solvents
should be pesticide quality or equivalent, and each lot of solvent should be
determined to be phthalate free.
5.4 Silica gel (optional) PR grade (100/200 mesh) - Before use, activate
at least 16 hours at 130° to 140°C. Deactivate with water (3.3 percent, by
8081 - 5 Revision 0
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weight) and equilibrate for 1 hour. Disposable silica cartridges (LC-silica or
equivalent), 1 g each, may be used in place of the deactivated silica gel.
5.5 Stock standard solutions:
5.5.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.1000 ± 0.0010 g of assayed reference material
in isooctane or hexane and diluting to volume in a 100 ml volumetric flask.
When compound purity is assayed to be 96 percent or greater, the weight
can be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.5.1.1 Beta-BHC and dieldrin are not adequately soluble in
isooctane. Acetone, or toluene should be used for the preparation
of the stock standard solutions of these compounds.
5.5.2 Transfer the stock standard solutions into bottles with
Teflon-lined screw-caps. Store at 4°C and protect from light. Stock
standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.5.3 Stock standard solutions must be replaced after six months,
or sooner if comparison with check standards indicates a problem.
5.6 Calibration standards;
5.6.1 Calibration standards, at a minimum of three concentrations
for each parameter of interest, are prepared through dilution of the stock
standards with isooctane. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations
found in real samples or should define the working range of the GC.
5.6.2 Calibration solutions must be replaced after two months, or
sooner, if comparison with check standards indicates a problem.
5.6.3 Although all single column analytes can be resolved on a new
35 percent phenyl methylpolysiloxane column, some analytes co-elute on the
other columns or on older 35 percent phenyl methylpolysiloxane columns.
Two calibration mixtures should be prepared for the single component
analytes of this method to eliminate potential resolution and quantitation
problems. Recommended low point mixtures are given in Table 9.
5.7 Internal standards (if internal standard calibration is used):
5.7.1 To use this approach, the analyst must select one or more
internal standards that are similar in analytical behavior to the compounds
of interest. The analyst must further demonstrate that the measurement
of the internal standard is not affected by method or matrix interferences.
Pentachloronitrobenzene is suggested as an internal standard.
8081 - 6 Revision 0
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5.7.2 Prepare calibration standards at a minimum of three
concentrations for each analyte of interest as described in Section 5.6.
5.7.3 To each calibration standard, add a known constant amount of
one or more internal standards.
5.7.4 Analyze each calibration standard according to Section 7.0.
5.8 Surrogate standards: The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, and the effectiveness
of the method in dealing with each sample matrix, by spiking each sample,
standard, and organic-free reagent water blank with pesticide surrogates.
Because GC/ECD analyses are more subject to interference than GC/MS analyses,
a secondary surrogate is to be used when sample interference is apparent.
Decachlorobiphenyl is the primary surrogate, and should be used whenever
possible. However, if recovery is low, or compounds interfere with
decachlorobiphenyl, then 2,4,5,6-tetrachloro-m-xylene should be evaluated for
acceptance. Proceed with corrective action when both surrogates are out of
limits for a sample (Section 8.3). Method 3500, Section 5.3.2, indicates the
proper procedure for preparing these surrogates.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Extracts must be stored under refrigeration and analyzed within 40
days of extraction.
7.0 PROCEDURE
7.1 Extraction;
7.1.1 Refer to Chapter Two for guidance in choosing the appropriate
extraction procedure. In general, water samples are extracted at a neutral
pH with methylene chloride as a solvent using a separatory funnel (Method
3510) or a continuous liquid-liquid extractor (Method 3520). Extract solid
samples with hexane-acetone (1:1) using either of the Soxhlet extraction
(Method 3540 or 3541) or ultrasonic extraction (Method 3550) procedures.
NOTE: Hexane/acetone (1:1) may be a more effective extraction solvent for
organochlorine pesticide and PCBs in some environmental and waste matrices.
The current solvent mixture recommended in Method 3550 is methylene
chloride/acetone (1:1).
7.1.2 Spiked samples are used to verify the applicability of the
chosen extraction technique to each new sample type. Each sample must be
spiked with the compounds of interest to determine the percent recovery
and the limit of detection for that sample.
7.1.2.1 Spiking of water samples should be performed by adding
appropriate amounts of pesticide and PCB compounds, dissolved in
8081 - 7 Revision 0
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methanol, to the water sample immediately prior to extraction. After
addition of the spike, mix the samples manually for 1 to 2 minutes.
Typical spiking concentrations for water samples are 1 to 10 jug/L
for samples in which pesticides and PCBs were not detected and 2 to
5 times the background concentration in those cases where pesticides
and PCBs are present (use of mixtures of Aroclors other than
1016/1260 is not recommended with this method).
7.1.2.2 Spiking of soil samples should be performed by adding
appropriate amounts of pesticide and PCB compounds, which are
dissolved in methanol, to the solid samples. The solid sample should
be wet prior to the addition of the spike (at least 20 percent
moisture) and should be mixed thoroughly with a glass rod to
homogenize the material. Allow the spike to equilibrate with the
solid for 1 hour at room temperature prior to extraction. Transfer
the entire spiked portion with the test compounds to the extraction
thimble for Soxhlet extraction (Method 3540), Automated Soxhlet
(Method 3541), or proceed with the ultrasonic extraction (Method
3550).
7.2 Cleanup/Fractionation
7.2.1 Cleanup procedures may not be necessary for a relatively
clean sample matrix, but most extracts from environmental and waste samples
will require additional preparation before analysis. The specific cleanup
procedure used will depend on the nature of the sample to be analyzed and
the data quality objectives for the measurements. General guidance for
sample extract cleanup is provided in this section.
7.2.2 If a sample is of biological origin, or contains high
molecular weight materials, the use of GPC cleanup/pesticide option (Method
3640) is recommended.
7.2.3 If only PCBs are to be measured in a sample, the sulfuric
acid/permanganate cleanup (Method 3665), followed by silica gel
fractionation (Method 3630) or Florisil cartridge cleanup (Method 3620),
is recommended.
7.2.4 If both PCBs and pesticides are to be measured in the sample,
isolation of the PCB fraction by silica gel fractionation (Method 3630)
is recommended.
7.2.5 If only pesticides are to be measured, cleanup by Method 3620
or Method 3630 is recommended.
7.2.6 Elemental sulfur, which may appear in certain sediments and
industrial wastes, interferes with the electron capture gas chromatography
of certain pesticides. Sulfur should be removed by the technique described
in Method 3660, Sulfur Cleanup.
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7.3 Gas chromatographv conditions (Recommended):
7.3.1 Narrow-bore columns:
7.3.1.1 Column 1:
Carrier gas (He) = 16 psi
Injector temperature = 225°C
Detector temperature = 300°C
Initial temeprature = 100°C, hold 2 minutes
Temperature program = 100°C to 160°C at 15°C/min, followed by;
160°C to 270°C at 5°C/nrin
Final temperature = 270°C.
7.3.1.2 Column 2:
Carrier gas (N2) = 20 psi
Injector temperature = 225°C
Detector temperature = 300°C
Initial temeprature = 160°C, hold 2 minutes
Temperature program = 160°C to 290°C at 5°C/min
Final temperature = 290°C, hold 1 minute.
7.3.1.3 Table 1 gives the retention times and MDLs that can
be achieved by this method for the organochlorine pesticides and
PCBs. Examples of the separations achieved with the SE-54 fused
silica capillary column are shown in Figures 1 through 6.
7.3.2 Wide-bore columns:
7.3.2.1 Column 1 and Column 2:
Carrier gas (He) = 5-7 mL/minute
Makeup gas (argon/methane (P-5 or P-10) or N2) = 30 mL/min
Injector temperature = 250°C
Detector temperature = 290°C
Initial temeprature = 150°C, hold 0.5 minute
Temperature program = 150°C to 270°C at 5°C/min
Final temperature = 270°C, hold 10 minutes.
7.3.2.2 Column 3:
Carrier gas (He) = 6 mL/minute
Makeup gas (argon/methane (P-5 or P-10) or N2) = 30 mL/min
Injector temperature = 205°C
Detector temperature = 290°C
Initial temeprature = 140°C, hold 2 minutes
Temperature program = 140°C to 240°C at 10°C/min,
hold 5 minutes at 240°C,
240°C to 265°C at 5°C,
Final temperature = 265°C, hold 18 minutes.
7.3.3 Additional columns - The columns listed in this section were
used to develop the method performance data; they are recommended for use
in the analysis of organochlorine pesticides and PCBs. Their specification
is not intended to prevent laboratories from using columns that are
developed after promulgation of the method. Laboratories may use other
capillary columns if they document method performance data (e.g.
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chromatographic resolution, analyte breakdown, and MDL's) equal to or
better than that provided with the method.
7.3.4 Table 2 gives the retention times and MDLs that can be achieved
by this method for the organochlorine pesticides or PCBs. Examples of the
separations achieved with the 35 percent phenyl methylpolysiloxane, 50
percent phenyl methyl polysiloxane and SE-54 fused-silica, wide-bore, open-
tubular columns are shown in Figures 1 through 6.
7.4 Calibration:
7.4.1 Refer to Method 8000 for proper calibration techniques. Use
Tables 1 and 2 for guidance on selecting the lowest point on the
calibration curve.
7.4.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4.3 Because several of the pesticides may co-elute on any single
column, two calibration mixtures are provided that minimize the problem
(Section 5.6.3). These calibration mixtures are also listed in Table 9,
along with the low point concentration of each analyte in the mixture.
The concentrations provided should be detectable on a GC/ECD suitable for
use with this method. Mixtures of Aroclors other than 1016/1260 are not
recommended for use with this method.
7.4.4 Because of the low concentration of pesticide standards
injected on a GC/ECD, column adsorption may be a problem when the GC has
not been used for a day. Therefore, the GC column should be primed or
deactivated by injecting a PCB or pesticide standard mixture approximately
20 times more concentrated than the mid-concentration standard. Inject
this standard mixture prior to beginning initial or daily calibration.
Caution: Several analytes, including Aldrin, may be observed in the injection
just following this system priming. Always run an acceptable blank
prior to running any standards or samples.
7.5 Gas chromatographic analysis;
7.5.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /xL of internal standard to the sample extract
prior to injection.
7.5.2 Follow Method 8000 for instruction on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Analysis of a mid-concentration standard after
each group of 20 samples is recommended (Section 8.3.4).
7.5.3 Examples of GC/ECD chromatograms generated by instruments
with wide- or narrow-bore columns are presented in Figures 1 through 6.
7.5.4 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
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7.5.5 If the peak response Is less than 2.5 times the baseline
noise level, the validity of the quantitative result may be questionable.
The analyst should consult with the source of the sample to determine
whether further concentration of the sample is warranted by the context
in which the result is to be used.
7.5.6 If the peak response exceeds the working range of the system,
dilute the extract and reanalyze.
7.5.7 Identification of mixtures (i.e. PCBs and toxaphene) is based
on the characteristic "fingerprint" retention time and shape of the
indicator peak(s); and quantitation is based on the area under the
characteristic peaks as compared to the area under the corresponding
calibration peak(s) of the same retention time and shape generated using
either internal or external calibration procedures (Section 7.6).
7.5.8 Identify compounds in the sample by comparing the retention
times of the peaks in the sample chromatogram with those of the peaks in
standard chromatograms. The retention time window used to make
identifications is based upon measurements of actual retention time
variations over the course of 7 to 10 consecutive injections. (Tables 5
and 6). A suggested window size can be calculated by multiplying the
standard deviation of a retention time window by three.
7.5.9 Quantitation of the compound(s) of interest is premised on:
1) a linear response of the BCD to the ranges of concentrations of the
compound(s) of interest encountered in the sample extract and the
corresponding calibration extract; and 2) a direct linear proportionality
between the magnitude of response of the ECD over the width(s) of the
retention window(s) (the area under the characteristic or "fingerprint"
peak[s]) in the sample and calibration extracts. Proper quantitation
requires the appropriate selection of a baseline from which the area under
the characteristic peak(s) can be calculated.
7.5.10 If compound identification or quantitation are precluded
due to interference (e.g., broad, rounded peaks or ill-defined baselines
are present) cleanup of the extract or replacement of the capillary column
or detector is warranted. Rerun sample on alternate instrumentation to
determine if the problem is of instrument or sample origin. Refer to
Section 7.2 for the procedures to be followed in sample cleanup.
7.6 Quantitation of Multiple Component Analvtes:
7.6.1 Scope (excerpted from U.S. FDA, RAM): Residues which are
mixtures of two or more components present problems in measurement. When
they are found together, e.g., toxaphene and DDT, the problem of
quantitation becomes even more difficult. Suggestions are offered in the
following sections for handing toxaphene, chlordane, PCB, DDT, and BHC.
7.6.2 Toxaphene: Quantitative calculation of toxaphene or strobane
is difficult, but reasonable accuracy can be obtained. To calculate
toxaphene on GC/ECD: (a) adjust the sample size so that the major
toxaphene peaks are 10-70% of full-scale deflection (FSD); (b) inject a
toxaphene standard that is estimated to be within ±10 ng of the sample;
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(c) quantitate using the five major peaks or the total area of the
toxaphene pattern.
7.6.2.1 To measure total area, construct the baseline of
standard toxaphene between Its extremities; and construct the
baseline under the sample, using the distances of the peak troughs
to baseline on the standard as a guide. This procedure Is made
difficult by the fact that the relative heights and widths of the
peaks in the sample will probably not be identical to the standard.
7.6.2.2 A series of toxaphene residues have been calculated
using the total peak area for comparison to the standard and also
using the area of the last four peaks only, in both sample and
standard. The agreement between the results obtained by the two
methods justifies the use of the latter method for calculating
toxaphene in a sample where the early eluting portion of the
toxaphene chromatogram shows interferences from other substances
such as DDT.
7.6.3 Chlordane is a technical mixture of at least 11 major
components and 30 or more minor components. Trans- and cis-chlordane
(alpha and gamma), respectively, are the two major components of technical
chlordane. However, the exact percentage of each in the technical material
is not completely defined, and is not consistent from batch to batch.
7.6.3.1 The GC pattern of a chlordane residue may differ
considerably from that of the technical standard. Depending on the
sample substrate and its history, residues of chlordane can consist
of almost any combination of: constituents from the technical
chlordane, plant and/or animal metabolites, and products of
degradation caused by exposure to environmental factors such as
water and sunlight.
7.6.3.2 When the chlordane residue does not resemble technical
chlordane, but instead consists primarily of individual, identifiable
peaks, quantitate the peaks of alpha-chlordane, gamma-chlordane, and
heptachlor separately against the appropriate reference materials,
and report the individual residues.
7.6.3.3 When the GC pattern of the residue resembles that of
technical chlordane, the analyst may quantitate chlordane residues
by comparing the total area of the chlordane chromatogram using the
five major peaks or the total area. If the heptachlor epoxide peak
is relatively small, include it as part of the total chlordane area
for calculation of the residue. If heptachlor and/or heptachlor
epoxide are much out of proportion, calculate these separately and
subtract their areas from the total area to give a corrected
chlordane area. (Note that octachlor epoxide, a metabolite of
chlordane, can easily be mistaken for heptachlor epoxide on a
nonpolar GC column.)
7.6.3.4 To measure the total area of the chlordane
chromatogram, proceed as in Section 7.6.2 on toxaphene. Inject an
amount of technical chlordane standard which will produce a
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chromatogram in which the major peaks are approximately the same
size as those in the sample chromatograms.
7.6.4 Polychlorinated biphenyls (PCBs): Quantitation of residues
of PCB involves problems similar to those encountered in the quantitation
of toxaphene, strobane, and chlordane. In each case, the chemical is made
up of numerous compounds which generate multi-peak chromatograms. Also,
in each case, the chromatogram of the residue may not match that of the
standard.
7.6.4.1 Mixtures of PCBs of various chlorine contents were
sold for many years in the U.S. by the Monsanto Co. under the trade
name Aroclor (1200 series and 1016). Although these Aroclors are
no longer marketed, the PCBs remain in the environment and are
sometimes found as residues in foods, especially fish.
7.6.4.2 Since standards are not generally available for all
of the congeners of chlorinated biphenyl, PCB residues are
quantitated by comparison to one or more of the Aroclor materials,
depending on the chromatographic pattern of the residue. A choice
must be made as to which Aroclor or mixture of Aroclors will produce
a chromatogram most similar to that of the residue. This may also
involve a judgement about what proportion of the different Aroclors
to combine to produce the appropriate reference material.
7.6.4.3 PCB Quantitation option #1- Quantitate the PCB
residues by comparing the total area of the chlorinated biphenyl
peaks to the total area of peaks from the appropriate Aroclor(s)
reference materials. Measure the total area or height response from
the common baseline under all the peaks. Use only those peaks from
the sample that can be attributed to chlorobiphenyls. These peaks
must also be present in the chromatogram of the reference materials.
A mixture of Aroclors may be required to provide the best match of
the GC patterns of the sample and reference.
7.6.4.4 PCB Quantitation option #2- Quantitate the PCB
residues by comparing the responses of 3 to 5 major peaks in each
appropriate Aroclor standard with the peaks obtained from the
chlorinated biphenyls in the sample extract. The amount of Aroclor
is calculated using each of the major peaks, and the results of the
3 to 5 determinations are averaged. Major peaks are defined as those
peaks in the Aroclor standards that are at least 30% of the height
of the largest Aroclor peak. Later eluting Aroclor peaks are
generally the most stable in the environment.
7.6.4.5 For samples where Aroclor patterns are not apparent,
but appear to contain weathered PCBs, several diagnostic peaks have
been identified in Table 10. Analysts should examine chromatographs
containing these peaks carefully, as these samples may contain PCBs.
7.6.5 Hexachlorocyclohexane (BHC, from the former name, benzene
hexachloride): Technical grade BHC is a cream-colored amorphous solid
with a very characteristic musty odor; it consists of a mixture of six
chemically distinct isomers and one or more heptachlorocyclohexanes and
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octachlorocyclohexanes. Commercial BHC preparations may show a wide
variance in the percentage of individual isomers present. Quantitate
each isomer (a, 0, 7, and 6) separately against a standard of the
respective pure isomer.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control (QC) procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If the extract cleanup was performed, follow
the QC in Method 3600 and in the specific cleanup method.
8.2 DDT and endrin are easily degraded in the injection port, if the
injection port or front of the column is contaminated with buildup of high
boiling residue from sample injection. Check for degradation problems by
injecting a mid-concentration standard containing only 4,4'-DDT and endrin.
Look for the degradation products of 4,4'-DDT (4,4'-DDE and 4,4'-ODD) and endrin
(endrin ketone and endrin aldehyde). If degradation of either DDT or endrin
exceeds 20%, take corrective action before proceeding with calibration, (refer
to Method 8000 and Section 4.4 of Method 8081). Calculate percent breakdown as
fol1ows:
% breakdown Total DDT degradation peak area (DDE + ODD)
for 4,4'-DDT = x 100
peak areas (DDT + DDE + ODD)
Total endrin degradation peak area
% breakdown (endrin aldehyde + endrin ketone)
for Endrin = x 100
peak areas (endrin + aldehyde + ketone)
8.3 Mandatory quality control to evaluate the GC system operation is
found in Method 8000. The following steps are recommended as additional method
QC.
8.3.1 The quality control (QC) reference sample concentrate
(Method 8000) should contain the organochlorine pesticides at 10 /xg/L.
If this method is to be used for analysis of PCBs, chlordane or toxaphene
only, the QC reference sample concentrate should contain the most
representative multi-component mixture at a concentration of 50 mg/L in
acetone. The frequency of the QC reference sample analysis is equivalent
to a minimum of 1 per 20 samples or 1 per batch if less than 20 samples.
If the recovery of any compound found in the QC reference sample is less
than 80 percent or greater than 120 percent of the certified value, the
laboratory performance is judged to be out of control, and the problem must
be corrected. A new set of calibration standards should be prepared and
analyzed.
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8.3.2 Calculate surrogate standard recovery on all samples, blanks,
and spikes. Determine if the recovery is within limits (limits established
by performing QC procedures outlined in Method 8000).
If recovery is not within limits, the following are required:
8.3.2.1 Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check instrument
performance.
8.3.2.2 Recalculate the data and/or reanalyze the extract if
any of the above checks reveal a problem.
8.3.2.3 Reextract and reanalyze the sample if none of the
above are a problem or flag the data as "estimated concentration."
8.3.3 The breakdown of DOT and endrin should be measured before
samples are analyzed. Injector maintenance and recalibration should be
completed if the breakdown is greater than 15% for either compound
(Section 8.2).
8.3.4 Include a mid-concentration calibration standard after each
group of 20 samples in the analysis sequence as a calibration check. The
response factors for the mid-concentration calibration should be within
30 percent of the average values for the multiconcentration calibration.
When this continuing calibration is out of this acceptance window, the
laboratory should stop analyses, clean the injector, replace the septum
and recalibrate the system.
8.3.5 Whenever quantitation is accomplished using an internal
standard, internal standards must be evaluated for acceptance. The
measured area of the internal standard must be no more than 50 percent
different from the average area calculated during calibration. When the
internal standard peak area is outside the limit, all samples that fall
outside the QC criteria must be reanalyzed.
8.4 GC/MS confirmation: Any compounds confirmed by two columns should
also be confirmed by GC/MS if the concentration is sufficient for detection by
GC/MS as determined by the laboratory generated detection limits.
8.4.1 The GC/MS would normally require a minimum concentration of
10 ng/juL in the final extract for each single-component compound.
8.4.2 The pesticide extract and associated blank should be analyzed
by GC/MS as per Section 7.0 of Method 8270.
8.4.3 The confirmation may be from the GC/MS analysis of the
base/neutral-acid extracts (sample and blank). However, if the compounds
are not detected in the base/neutral-acid extract even though the
concentrations are high enough, a GC/MS analysis of the pesticide extract
should be performed.
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8.4.4 A QC reference sample of the compound must also be analyzed
by GC/MS. The concentration of the QC reference standard must demonstrate
the ability to confirm the pesticides/Aroclors identified by GC/ECD.
8.5 Whenever silica gel cleanup is used, demonstrate that the
fractionation scheme is reproducible. Batch to batch variation in the
composition of the silica gel material may cause a change in the distribution
patterns of the organochlorine pesticides and PCBs as Aroclors. When compounds
are found in more than one fraction, add the concentrations of the various
fractions, making corrections for the final volume of the fractions. It is up
to the analyst to decide whether the cut-off point should be 5 percent or less
of the concentration in the fraction where the compound is expected to elute.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDL concentrations listed in
Tables 1 and 2 were obtained using organic-free reagent water and sandy loam
soil. Details for determining MDLs are given in Chapter One. The MDL actually
achievable in a given analysis will vary depending on detector response
characteristics, irreducible noise from instrument electronics and matrix
effects.
9.2 This method has been tested in a single laboratory by using clean
hexane and liquid and solid waste extracts that were spiked with the test
compounds at three concentrations. Single-operator precision, overall precision,
and method accuracy were found to be related to the concentration of the compound
and the type of matrix. Results of the single-laboratory method evaluation are
given in Table 4.
9.3 The accuracy and precision obtainable following this method will be
determined by the sample matrix, sample preparation technique, optional cleanup
techniques, and calibration procedures used.
10.0 REFERENCES
1. Lopez-Avila, V.; Schoen, S.; Milanes, J. "Single-Laboratory Evaluation of
Method 8080 - Organochlorine Pesticides and PCBs"; final report to the U.S.
Environmental Protection Agency on Contract 68-03-3226; Acurex Corporation,
Environmental Systems Division: Mountain View, CA, 1986. EPA-600/4-87/022.
2. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 10 - Pesticides and PCB Report for the
U.S. Environmental Protection Agency on Contract 68-03-2606.
3. Goerlitz, D.F.; Law, L.M. "Removal of Elemental Sulfur Interferences from
Sediment Extracts for Pesticide Analysis"; Bull. Environ. Contam. Toxicol.
1971, 6, 9.
4. Blumer, M. "Removal of Elemental Sulfur from Hydrocarbon Fractions"; Anal.
Chem. 1957, 29, 1039.
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5. Ahnoff, M.; Josefsson, B. "Cleanup Procedures for PCB Analysis on River
Water Extracts"; Bull. Environ. Contain. Toxicol. 1975, 13, 159.
6. Jensen, S.; Renberg, L.; Reutergardth, L. "Residue Analysis of Sediment
and Sewage Sludge for Organochlorines in the Presence of Elemental Sulfur";
Anal. Chem. 1977, 49, 316-318.
7. Wise, R.H.; Bishop, D.F.; Williams, R.T.; Austern, B.M. "Gel Permeation
Chromatography in the GC/MS Analysis of Organics in Sludges"; U.S.
Environmental Research Laboratory. Cincinnati, OH 45268.
8. Pionke, H.B.; Chesters, G.; Armstrong, D.E. "Extraction of Chlorinated
Hydrocarbon Insecticides from Soil"; Agron. J. 1968, 60, 289.
9. Burke, J.A.; Mills, P.A.; Bostwick, D.C. "Experiments with Evaporation of
Solutions of Chlorinated Pesticides"; J. Assoc. Off. Anal. Chem. 1966, 49,
999.
10. Glazer, J.A., et al. "Trace Analyses for Wastewaters"; Environ. Sci. and
Technol. 1981, 15, 1426.
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TABLE 1
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS8
USING WIDE-BORE CAPILLARY COLUMNS
Retention Time (min) MDLb Water MDLb Soil
Compound
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma-BHC (Lindane)
alpha-Chlordane
gamma-Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
Water = Organic-free
DB 608C
11.84
8.14
9.86
11.20
9.52
15.24
14.63
18.43
16.34
19.48
16.41
15.25
18.45
20.21
17.80
19.72
10.66
13.97
22.80
MR
MR
MR
MR
MR
MR
MR
MR
DB 1701C
12.50
9.46
13.58
14.39
10.84
16.48
16.20
19.56
16.76
20.10
17.32
15.96
19.72
22.36
18.06
21.18
11.56
15.03
22.34
MR
MR
MR
MR
MR
MR
MR
MR
(M9/L)
0.034
0.035
0.023
0.024
0.025
0.008
0.037
0.050
0.058
0.081
0.044
0.030
0.040
0.035
0.039
0.050
0.040
0.032
0.086
NA
0.054
NA
NA
NA
NA
NA
0.90
(M9/Kg)
2.2
1.9
3.3
1.1
2.0
1.5
4.2
2.5
3.6
NA
2.1
2.4
3.6
3.6
1.6
2.0
2.1
5.7
NA
57.0
NA
NA
NA
NA
NA
70.0
reagent water.
Soil = Sandy loam soil.
MR = Multiple peak
responses.
NA = Data not available.
U.S. EPA Method 8081. Organochlorine Pesticides and PCBs as Aroclors.
Environmental Protection Agency. Office of Research and Development,
Washington, DC 20460.
8081 - 18
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TABLE 1
(Continued)
MDL is the method detection limit. MDL was determined from the analysis of
seven replicate aliquots of each matrix processed through the entire analytical
method (extraction, silica gel cleanup, and GC/ECD analysis). MDL = t(n-l,
0.99) x SD, where t(n-l, 0.99) is the student's t value appropriate for a 99%
confidence interval and a standard deviation with n-1 degrees of freedom, and
SD is the standard deviation of the seven replicate measurements.
Temperature program: 150°C (hold 1/2 minutes) to 270°C at 5°C/min, helium head
pressure at 10 psi.
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TABLE 2
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS"
USING NARROW-BORE CAPILLARY COLUMNS
Compound
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma -BHC (Lindane)
alpha-Chlordane
gamma-Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
Retention
Col. lc
14.51
11.43
12.59
13.69
12.46
17.34
21.67
19.09
23.13
19.67
18.27
22.17
24.45
21.37
23.78
13.41
16.62
28.65
MR
MR
MR
MR
MR
MR
MR
MR
Liquid = Organic-free reagent
Time (min)
Col. ld
14.70
10.94
11.51
12.20
11.71
17.02
20.11
18.30
21.84
18.74
17.62
20.11
21.84
19.73
20.85
13.59
16.05
24.43
MR
MR
MR
MR
MR
MR
MR
MR
water.
MDLb Liquid
(M9/L)
0.034
0.035
0.023
0.024
0.025
0.037
0.050
0.058
0.081
0.044
0.030
0.040
0.035
0.039
0.050
0.040
0.032
NA
0.086
NA
0.054
NA
NA
NA
NA
0.90
Solid
(Mg/Kg)
2.2
1.9
3.3
1.1
2.0
1.5
4.2
2.5
3.6
NA
2.1
2.4
3.6
3.6
1.6
2.0
2.1
NA
5.7
NA
57.0
NA
NA
NA
NA
70.0
Solid = Sandy loam soil .
MR = Multiple
NA = Data not
peak responses.
available.
8081 - 20
Revision 0
November 1990
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TABLE 2
(Continued)
U.S. EPA Method 8081. Organochlorine Pesticides and PCBs as Aroclors.
Environmental Protection Agency. Office of Research and Development,
Washington, DC 20460.
MDL is the method detection limit. MDL was determined from the analysis of
seven replicate aliquots of each matrix processed through the entire analytical
method (extraction, cleanup, and GC/ECD analysis). MDL = t(n-l, 0.99) x SD,
where t(n-l, 0.99) is the student's t value appropriate for a 99% confidence
interval and a standard deviation with n-1 degrees of freedom, and SD is the
standard deviation of the seven replicate measurements.
30 m x 0.25 mm ID DB 608 fused silica, open-tubular column (1 pm film
thickness).
30 m x 0.25 mm ID DB 5 fused silica, open-tubular column (1 urn film thickness).
8081 - 21 Revision 0
November 1990
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TABLE 3
ESTIMATED QUANTITATION LIMITS (EQL) FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
Sample EQLs are highly matrix-dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
EQL = [Method detection limit for water (Table 1) or (Table 2) wide bore
or narrow bore options] x [Factor (Table 3)]. For nonaqueous samples,
the factor is on a wet-weight basis.
8081 - 22 Revision 0
November 1990
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TABLE 4
RETENTION TIMES OF OTHER PESTICIDES DETECTED USING METHOD 8081
Analyte DB 608 DB 1701
Trifluralin 5.16 8.58
Diallate (isomer 1) 7.15 8.05
Diallate (isomer 2) 7.42 8.58
PCNB 9.03 9.91
Dichlone 10.80 decomp.
Isodrin 13.47 13.93
Dichlorvos
Naled
Prometon
Propazlne
Atrazine
Terbuthylazine
Simazine
Dichlorofenthion
Methyl chlorphrophos
Ronnel
Captan 16.83 17.32
Chiorobenzilate 17.58 18.97
Prometryn
Ametryn
Metribuzin
Terbutryn
Chlorpyrophos
Trichlorinate
Chlorfenvinphos
Tetrachlorovinphos
Anilazine
Cynazine
Hexazinone
Captafol 22.51 23.11
Mirex 22.75 23.11
Leptophos
Coumaphos
Temperature program: 150°C (hold 1/2 minutes) to 270°C at 5°C/min, helium
head pressure at 10 psi.
8081 - 23 Revision 0
November 1990
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TABLE 5
REPRODUCIBILITY OF RETENTION TIMES OF THE ORGANOCHLORINE
PESTICIDES FOR TEN CONSECUTIVE INJECTIONS
USING THE NARROW-BORE CAPILLARY COLUMNS
Retention Time Reproducibility
Compound SD (min)a
alpha- BHC
beta-BHC
gamma -BHC
delta-BHC
Heptachlor
Aldrin
Heptachlor epoxide
alpha-Chlordane
gamma-Chlordane
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
Endosulfan II
4,4'-DDD
Endrin aldehyde
Endosulfan sulfate
4,4'-DDT
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1260
0.010
0.009
0.011
0.011
0.008
0.009
0.009
0.012
0.010
0.008
0.008
0.007
0.006
0.008
0.007
0.008
0.008
0.007
0.004-0.006"
0.042-0.104"
0.035-0.040"
SD = Standard deviation.
a Number of determinations is 10.
b
Value determined for 3 major peaks of each mixture.
8081 - 24 Revision 0
November 1990
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TABLE 6
REPRODUCIBILITY OF RETENTION TIMES OF THE ORGANOCHLORINE
PESTICIDES FOR TEN CONSECUTIVE INJECTIONS USING
THE WIDE-BORE CAPILLARY COLUMNS
Compound
Retention Time Reproducibility
SD (min)a
DB 5
DB 608
alpha-BHC
beta-BHC
gamma -BHC
delta-BHC
Heptachlor
Aldrin
Heptachlor epoxide
alpha-Chlordane
gamma-Chlordane
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
Endosulfan II
4,4'-DDD
Endrin aldehyde
Endosulfan sulfate
4,4'-DDT
4,4'-Methoxychlor
SD = Standard deviation
0.006
0.007
0.007
0.005
0.007
0.007
0.007
0.007
0.007
0.008
0.007
0.008
0.013
0.013
0.010
0.007
0.007
0.007
0.007
0.008
0.008
0.006
0.008
0.008
0.008
0.009
0.009
0.007
0.009
0.007
0.010
0.010
0.010
0.010
0.007
0.007
a Number of determinations is 9.
pairs cannot be resolved on the DB 5 wide-bore open
tubular column under the conditions listed in Section 7.3.
8081 - 25
Revision 0
November 1990
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TABLE 7
ELUTION PATTERNS AND AVERAGE RECOVERIES OF THE ORGANOCHLORINE
PESTICIDES AND AROCLOR BY METHOD 8081 WITH SILICA GEL FRACTIONATION
(LIQUID WASTE NO. 1 EXTRACT)
Compound
Average Recovery + SD (RSD)
a,b
Fraction I
hexane (80 ml)
Fraction II
hexane (50 mL)
Fraction III
methylene
chloride
(15 mL)
Total Recovery
alpha-BHC
beta-BHC
gamma -BHC
delta-BHC
Heptachlor
Aldrin
Heptachlor epoxide
alpha-Chlordane
gamma-Chlordane
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
Endosulfan II
4,4'-DDD
Endrin aldehyde
Endosulfan sulfate
4,4'-DDT
4,4'-Methoxychlor
Aroclor-1016
Aroclor-1260
57 + 2.5(4.4)
90 + 11(12)
92 ± 9.2(10)
85 + 7.2(8.5) 10 + 9.2(92)
95 ± 16(17)
33 ± 4.0(15)
88 ± 18(21)
118 + 9.8(8.3)
100 ± 18(18)
22 + 9.2(42)
90 + 3.1(3.4)
90 + 4.0(4.4)
90 ± 11(12)
89 ± 4.1(4.6)
88 + 3.8(4.3)
82 + 4.3(5.3)
65 + 3.1(4.7)
79 + 7.1(9.0)
43 + 16(37)
C
83 ± 4.0(4.8)
75 ± 4.6(6.1)
79 + 10(13)
90 + 3.1(3.4)
90 + 4.0(4.4)
90 + 11(12)
90 + 11(12)
92 + 9.2(10)
89 ± 4.1(4.6)
95 + 8.0(8.4)
88 + 3.8(4.3)
95 + 16(17)
82 + 4.3(5.3)
65 + 3.1(4.7)
79 + 7.1(9.0)
76 + 16(21)
C
83 + 4.0(4.8)
88 + 18(21)
75 + 4.6(6.1)
118 + 9.8(8.3)
100 + 18(18)
8081 - 26
Revision 0
November 1990
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TABLE 7
(continued)
The values given represent the average percent recoveries from three
replicate determination + one standard deviation. The numbers in
parentheses are the relative standard deviations.
The amounts spiked are 15,000 30,000, and 150,000 ng per 2 ml extract
per column for the organochlorine pesticides and Aroclor-1016/Aroclor-
1260, respectively.
Unable to determine recovery because of interference.
8081 - 27 Revision 0
November 1990
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TABLE 8
ELUTION PATTERNS AND AVERAGE RECOVERIES OF THE ORGANOCHLORINE
PESTICIDES AND AROCLOR BY SILICA GEL CHROMATOGRAPHY
Compound
Average Recovery + SD (RSD)
a,b
Fraction I
hexane (80 mL)
Fraction II
hexane (50 mL)
Fraction III
methylene
chloride
(15 mL)
Total Recovery
alpha-BHC
beta-BHC
gamma -BHC
delta-BHC
Heptachlor
Aldrin
Heptachlor epoxide
alpha-Chlordane
gamma-Chlordane
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
Endosulfan II
4,4'-DDD
Endrin aldehyde
Endosulfan sulfate
4,4'-DDT
4,4'-Methoxychlor
Aroclor-1016
Aroclor-1260
55 ± 6.1(11)
70 + 7.7(11)
65 ± 4.6(7.1)
71 ± 3.2(4.5) 10 ± 2.0(20)
76 ± 7.1(9.3)
36 ± 2.0(5.6)
61 ± 7.9(13)
104 + 2.5(2.4)
95 ± 7.5(7.9)
20 + 1.7(8.7)
94 + 3.0(3.2)
89 + 4.1(4.6)
92 ± 5.2(5.6)
91 ± 5.7(6.3)
88 + 5.1(5.8)
85 + 9.4(11)
87 + 6.4(7.3)
81 + 4.5(5.5)
49 + 1.2(2.4)
71 + 9.2(13)
86 ± 5.0(5.8)
99 ± 17(17)
75 + 6.0(8.0)
94 + 3.0(3.2)
89 + 4.1(4.6)
92 + 5.2(5.6)
70 + 7.7(11)
65 + 4.6(7.1)
91 ± 5.7(6.3)
81 + 4.9(6.1)
88 + 5.1(5.8)
76 + 7.1(9.3)
85 + 9.4(11)
87 + 6.4(7.3)
81 + 4.5(5.5)
85 + 3.1(3.6)
71 + 9.2(13)
86 + 5.0(5.8)
61 + 7.9(13)
99 + 17(17)
104 + 2.5(2.4)
95 + 7.5(7.9)
8081 - 28
Revision 0
November 1990
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TABLE 8
(Continued)
The values given represent the average percent recoveries from three
replicate determinations ± one standard deviation. The numbers in
parentheses are the relative standard deviations.
The amounts spiked are 3,000, 6,000, and 30,000 ng per 2 ml extract per
column for the organochlorine pesticides and Aroclor-1016/Aroclor-1260,
respectively.
8081 - 29 Revision 0
November 1990
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Table 9
Individual Standard Mixtures For Single Component Pesticides.
Individual
Standard Mix A
Low Point
Concentration
( g/L)
Individual
Standard Mix B
Low Point
Concentration
( g/L)
a-BHC 5.0
Heptachlor 5.0
7-BHC 5.0
Endosulfan I 5.0
Dieldrin 10.0
Endrin 10.0
p,p'-DDD 10.0
p,p'-DDT 10.0
Methoxychlor 50.0
Tetrachloro-m-xylene 20.0
Decachlorobiphenyl 20.0
0-BHC
6-BHC
Aldrin
Heptachlor epoxide
o-Chlordane
'Y-Chlordane
p,p'-DDE
Endosulfan sulfate
Endrin aldehyde
Endrin ketone
Endosulfan II
Tetrachloro-m-xylene
Decachlorobiphenyl
5.0
5.0
5.0
5.0
5.0
5.0
10.0
10.0
10.0
10.0
10.0
20.0
20.0
8081 - 30
Revision 0
November 1990
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TABLE 10
PEAKS DIAGNOSTIC OF PCBs OBSERVED IN 0.53 mm ID COLUMN ANALYSIS
Peak RT on RT on Pesticide
No. DB 608a DB 1701a Aroclor" Retention Window
I 4.90 4.66 1221 Before TCmX
II 7.15 6.96 1221, 1232, 1248 Before o-BHC
III 7.89 7.65 1061, 1221. 1232, 1242, Before o-BHC
IV 9.38 9.00 1016, 1232, 1242, 1248, just after o-BHC on
DB 1701;just before
7-BHC on DB 608
V 10.69 10.54 1016. 1232. 1242. 1248 o-BHC and
heptachlor on DB 1701;
just after heptachlor on
DB 608
VI 14.24 14.12 1248. 1254 T-BHC and heptachlor
epoxide on DB 1701;
heptachlor epoxide and
T-chlordane on DB 608
VII 14.81 14.77 1254 Heptachlor epoxide and
T-chlordane on DB 1701;
a- and 7-chlordane on
DB 608
VIII 16.71 16.38 1254 DDE and dieldrin on
DB 1701; dieldrin and
endrin on DB 608
IX 19.27 18.95 1254, 1260 Endosulfan II on
DB 1701; DDT on DB 608
X 21.22 21.23 1260 Endrin aldehyde and
endosulfan sulfate on
DB 1701; endosulfan
sulfate and methoxychlor
on DB 608
XI 22.89 22.46 1260 Just before endrin
ketone on DB 1701; after
endrin ketone on DB 608
5Using oven temperature program: T, = 150°C, hold 30 seconds; increase
temperature at 5°C/minutes to 275°C.
b Underlined Aroclor indicates the largest peak in the pattern.
8081 - 31 Revision 0
November 1990
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FIGURE 1.
GAS CHROMATOGRAM OF THE MIXED ORGANOCHLORINE PESTICIDE STANDARD
Start Tim : 0.00 min End Time : 33.00 nrin Lou Point : 20.00 mv High Point : 420.00 mV
Scale Factor; 0 Plot Offset: 20 mv Plot Scale: 400 mV
Response [mV]
NJ
I I I
•^f i^^r tji \^j \^t
I I I I I I I I I ill I I I I I I I I I
o cp o
I I I I I I I I I I I I I I I I I
o"
3D
fD
rT
3
,-*-
6' ->_
~*i L?l
IT
Ln
O'
=-4.68
7.99
9.93
-23.18
26.23
- -28.64
-0.95
-8.60
-30.19
Column: 30 m x 0.25 mm ID, DB 5
Temperature program: 100°C (hold 2 minutes) to J60°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 32
Revision 0
November 1990
-------
FIGURE 2.
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX A
Start Time : 0.00 min
Scale factor: 0
LI—
IT)
it"
6 -»_
D Ui
ro
End Time : 33.00 min
Plot Offset: 20 mv
Low Point : 20.00 mv
Plot Scale: 250 mv
Hign Point : 270.00 mv
Response [mV]
(_n a 01
o o o
i i i I i i i i I i i i i I i i i i I i i i i I j i
o
o
ro
u->
o
•J.3
=-7.93
-14.27
-17.08
20.22
.77
22.68
-23.73
•28.52
•s.es
-8 . 54
TCMX
-12.33
-9.86 Alpha-BHC
10.98 Gamma-BHC
13.58 Heptachlor
-17.54 Endosulfan I
—18.47 Dieldrin
•19.78 ODD
— 21.13 DDT
-19.24 Endrin
-23.08 Methoxychlor
-30.05
Column: 30 m x 0.25 mm 10, DB 5
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/m1n to 270°C; carrier He at 16 ps1.
8081 - 33
Revision 0
November 1990
-------
FIGURE 3.
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX B
St»rt ''"» : 3.30 t : 2'O.^C «v
Scale Factor: 3 'lot Ot'set: JO mv Plot Sole: 250 mV
Response [mV]
Li—
0 ->.
"3 C/l
H
l™~1 NO
3 °-
KJ
g-
_» _i hO
en a -9
--10.71 Beta-BHC
11<73 Delta-BHC
14.27
V15.24
i f. n«;
Ml
j^2"0.69
22.00
-14.84 Aldrin
-16.23 Heptachlor Epoxide
-17.08 Gamma-Chlordane
•$.93
-3.54 TCMX
-17.63 Alpha-Chlordane
— 18.31 DDE
13. ;t Endosulfan II
-20.19 Endrin • Aldehyde
-21.03 Endosulfan Sulfate
--22.68 Eftdrin Ketone
-30.04
OCB
Column: 30 m x 0.25 mm ID, DB 5
Temperature program: 100°C (hold 2 minutes) to 160°C at l5°C/m1n, then at
5°C/min to 270°C; carrier He at 16 ps1.
8081 - 34
Revision 0
November 1990
-------
FIGURE 4.
GAS CHROMATOGRAM OF THE TOXAPHENE STANDARD
Start Time : 0.00 min
Scale Factor: 0
End Tim : 33.00 min
Plot Offset: 20 mv
Low Point : 20.00 mv
Plot Scale: 60 mv
High Point : 80.00 mv
Response [rnV]
r\>
O
O O O O O
I I I I II I I I I I I I III I I I I I I I I II I I I I I I I I II I I I I I I I I II I I I I I I I I II
CD
I
24.32
Column: 30 m x 0.25 mm ID, DB 5
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 35
Revision 0
November 1990
-------
FIGURE 5.
GAS CHROMATOGRAM OF THE AROCLOR-1016 STANDARD
Start Time : 0.00 min
Sole factor: 0
End Time : 33.00 min
Plot Offset: 20 mV
Low Point : 20.00 mv
Plot Scale: 100 mV
High Point : 120.00 mV
Response [mV]
N> •*»• O) DO C
I I I I I I I I I III I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
O
O
I I I I I I I I I
Ul—
D
3
-1.81
-12.95
-1.03
Column: 30 m x 0.25 mm ID DB 5 fused silica capillary.
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 36
Revision 0
November 1990
-------
FIGURE 6.
GAS CHROMATOGRAM OF THE TECHNICAL CHLORDANE STANDARD
Start Time : 0.00 min
Scale Factor: 0
End Tim : 13.00 min
Plot Offset: 20 mv
low Point : 20.00 mV
Plot Scale: 200 mV
High Point : 220.00 mV
Response [mV]
Ol
1
10
o
1
I I
rt)
D
6' -'.J
H
3'
0)
5'
-0.97
.59
-12.92
13.60
-17.11
17.65
Column: 30 m x 0.25 mm ID DB 5 fused silica capillary.
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 37
Revision 0
November 1990
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METHOD 8081
ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
Start
711 Choose
appropriate
• x traction
technique (see
Chapter 2)
7.12 Add
specified
matrix spike
to sample.
7 2 Routine
cleanup/
fractio nation .
73 Set
chromatographic
condi tions .
7 4 Refer to
Method 8000
for proper
calibration
technique*
744 Prime or
deactivate CC
daily
cal ibra tion .
i
7 . 5 Perform
CC
analysis («ee
Method 8000)
1
Si . 5 ION.
./Any sample >. Ye*
>. f erences? /
No
/I 6 IN.
X^Do residues >. Ye»
C have >1 ) »
>. component ^r
No |
1
r .... ^
7.5.10
Additional
cleanup/
f ractionation .
(see Sect. 7.2)
7 . 6 Calculation
of toxaphene ,
chlordane ,
PCB« . DDT , and
BHC done here
8081 - 38
Revision 0
November 1990
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METHOD 8120A
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8120 is used to determine the concentration of certain
chlorinated hydrocarbons. The following compounds can be determined by this
method:
Appropriate Preparation Techniques
Compounds CAS No" 3510 3520 3540 3550 3580
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachl orobutad i ene
Hexachl orocycl ohexane
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1,2,4-Trichlorobenzene
91-58-7
95-50-1
541-73-1
106-46-7
118-74-1
87-68-3
608-73-1
77-47-4
67-72-1
120-82-1
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
a Chemical Abstract Services Registry Number.
x Greater than 70 percent recovery by this technique
ND Not determined.
1.2 Table 1 indicates compounds that may be determined by this method and
lists the method detection limit for each compound in organic-free reagent water.
Table 2 lists the estimated quantitation limit (EQL) for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8120 provides gas chromatographic conditions for the detection
of ppb concentrations of certain chlorinated hydrocarbons. Prior to use of this
method, appropriate sample extraction techniques must be used. Both neat and
diluted organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
injection. A 2 to 5 /zL aliquot of the extract is injected into a gas
chromatograph (GC) using the solvent flush technique, and compounds in the GC
effluent are detected by an electron capture detector (ECD).
8120A - 1 Revision 1
November 1990
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2.2 If interferences are encountered in the analysis, Method 8120 may also
be performed on extracts that have undergone cleanup using Method 3620.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All of these materials must be demonstrated to be free
from interferences, under the conditions of the analysis, by analyzing method
blanks. Specific selection of reagents and purification of solvents by
distillation in all glass systems may be required.
3.3 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 1.8 m x 2 mm ID glass column packed with
1% SP-1000 on Supelcoport (100/120 mesh) or equivalent.
4.1.2.2 Column 2 - 1.8 m x 2 mm ID glass column packed with
1.5% OV-1/2.4% OV-225 on Supelcoport (80/100 mesh) or equivalent.
4.1.3 Detector - Electron capture (ECD).
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation of
extracts
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
8120A - 2 Revision 1
November 1990
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4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.4 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.5 Volumetric flasks - 10, 50, and 100 ml, with ground glass stoppers.
4.6 Microsyringe - 10 /zL-
4.7 Syringe - 5 ml.
4.8 Vials - Glass, 2, 10, and 20 ml capacity with Teflon lined screw-
caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Hexane, C6H14. Pesticide quality or equivalent.
5.3.2 Acetone, CH3COCH3. Pesticide quality or equivalent.
5.3.3 Isooctane, C6H18. Pesticide quality or equivalent.
5.4 Stock standard solutions
5.4.1 Prepare stock standard solutions at a concentration of
1.00x/ig//iL by dissolving 0.0100 g of assayed reference material in
isooctane or hexane and diluting to volume in a 10 ml volumetric flask.
Larger volumes can be used at the convenience of the analyst. When
compound purity is assayed to be 96% or greater, the weight can be used
without correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
8120A - 3 Revision 1
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5.4.2 Transfer the stock standard solutions into vials with Teflon
lined screw caps or crimp tops. Store at 4°C and protect from light.
Stock standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.4.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.5 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared through dilution of the stock standards with
isooctane or hexane. One of the concentrations should be at a concentration
near, but above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples or
should define the working range of the GC. Calibration solutions must be
replaced after six months, or sooner if comparison with check standards indicates
a problem.
5.6 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Section 5.5.
5.6.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane or
hexane.
5.6.3 Analyze each calibration standard according to Section 7.0.
5.7 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and organic-free reagent water blank with one or two surrogates (e.g.
chlorinated hydrocarbons that are not expected to be in the sample) recommended
to encompass the range of the temperature program used in this method. Method
3500 details instructions on the preparation of base/neutral surrogates.
Deuterated analogs of analytes should not be used as surrogates for gas
chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Extracts must be stored under refrigeration and analyzed within 40
days of extraction.
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7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510
or 3520. Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 ml using the macro Snyder column, allow the apparatus to cool and
drain for at least 10 minutes.
7.1.2.2 Momentarily remove the Snyder column, add 50 ml of
hexane, a new boiling chip, and reattach the macro Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature,
as required, to complete concentration in 5-10 minutes. At the
proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 1 ml, remove the K-D apparatus and allow it to
drain and cool for at least 10 minutes. The extract will be handled
differently at this point, depending on whether or not cleanup is
needed. If cleanup is not required, proceed to Section 7.1.2.3.
If cleanup is needed, proceed to Section 7.1.2.4.
7.1.2.3 If cleanup of the extract is not required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 ml of hexane. A 5 ml syringe is
recommended for this operation. Adjust the extract volume to
10.0 ml. Stopper the concentrator tube and store refrigerated at
4°C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be transferred
to a vial with a Teflon lined screw cap or crimp top. Proceed with
gas chromatographic analysis.
7.1.2.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of hexane. A 5 mL syringe
is recommended for this operation. Add a clean boiling chip to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top. Place the
micro K-D apparatus on the water bath (80°C) so that the concentrator
tube is partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but the
8120A - 5 Revision 1
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chambers will not flood. When the apparent volume of liquid reaches
0.5 ml, remove the K-D apparatus and allow it to drain and cool for
at least 10 minutes.
7.1.2.5 Remove the micro Snyder column and rinse the flask
and its lower joint into the concentrator tube with 0.2 ml of hexane.
Adjust the extract volume to 2.0 ml and proceed with Method 3620.
7.2 Gas chromatographic conditions (Recommended)
7.2.1 Column 1
Carrier gas (5% methane/95% argon) flow rate = 25 mL/min
Column temperature = 65°C isothermal, unless otherwise specified (see
Table 1).
7.2.2 Column 2
Carrier gas (5% methane/95% argon) flow rate = 25 mL/min
Column temperature = 75°C isothermal, unless otherwise specified (see
Table 1).
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.3.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will validate elution patterns and the
absence of interferents from the reagents.
7.4 Gas chromatographic analysis
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /uL of internal standard to the sample prior to
injecting.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Examples of GC/ECD chromatograms for certain chlorinated
hydrocarbons are shown in Figures 1 and 2.
7.4.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.4.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Method 8000 for calculation equations.
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7.4.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using Method 3620.
7.5 Cleanup: If required, the samples may be cleaned up using the Methods
presented in Chapter 4.
7.5.1 Proceed with Method 3620 using the 2 ml hexane extracts
obtained from Section 7.1.2.5.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each parameter of interest in acetone at the following
concentrations: hexachloro-substituted hydrocarbon, 10 ng/ml't and any
other chlorinated hydrocarbon, 100 jug/mL.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both Tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures
are required.
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
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9.0 METHOD PERFORMANCE
9.1 The method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked
at six concentrations over the range 1.0 to 356 /xg/L. Single operator
precision, overall precision, and method accuracy were found to be directly
related to the concentration of the parameter and essentially independent of the
sample matrix. Linear equations to describe these relationships for a flame
ionization detector are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample preparation technique, and calibration procedures used.
10.0 REFERENCES
1. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 3 - Chlorinated Hydrocarbons, and
Category 8 - Phenols," Report for EPA Contract 68-03-2625 (in preparation).
2. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
3. "EPA Method Validation Study 22, Method 612 (Chlorinated Hydrocarbons),"
Report for EPA Contract 68-03-2625 (in preparation).
4. "Method Performance for Hexachlorocyclopentadiene by Method 612," Memorandum
from R. Slater, U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, Ohio 45268, December 7, 1983.
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
6. "Determination of Chlorinated Hydrocarbons in Industrial and Municipal
Wastewaters," Report for EPA Contract 68-03-2625 (in preparation).
8120A - 8 Revision 1
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TABLE 1.
GAS CHROMATOGRAPHY OF CHLORINATED HYDROCARBONS
Compound
Retention time (min)
Col. 1 Col. 2
ND = Not determined.
a!50°C column temperature.
b!65°C column temperature.
°100°C column temperature.
Method
Detection
limit (M9/L)
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl ohexane
Hexachl orocycl opentad i ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1,2,4-Trichlorobenzene
2.7a
6.6
4.5
5.2
5.6a
7.7
ND
4.9
15.5
3.6b
9.3
6.8
7.6
10. lb
20.0
16.5°
8.3
22.3
0.94
1.14
1.19
1.34
0.05
0.34
0.40
0.03
0.05
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TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQL) FOR VARIOUS MATRICES3
Matrix Factor"
Ground water 10
Low-concentration soil by ultrasonic extraction with GPC cleanup 670
High-concentration soil and sludges by ultrasonic extraction 10,000
Non-water miscible waste 100,000
Sample EQLs are highly matrix dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet weight basis.
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Parameter
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
1,2,4-Trichlorobenzene
Test
cone.
(M9A)
100
100
100
100
10
10
10
10
100
Limit
for s
(M9/L)
37.3
28.3
26.4
20.8
2.4
2.2
2.5
3.3
31.6
Range
for x
(M9/L)
29.5-126.9
23.5-145.1
7.2-138.6
22.7-126.9
2.6-14.8
D-12.7
D-10.4
2.4-12.3
20.2-133.7
Range
P, Ps
(%)
9-148
9-160
D-150
13-137
15-159
D-139
D-lll
8-139
5-149
s = Standard deviation of four recovery measurements, in M9/L-
x = Average recovery for four recovery measurements, in M9/L.
P,PS= Percent recovery measured.
D = Detected; result must be greater than zero.
a Criteria from 40 CFR Part 136 for Method 612. These criteria are based
directly upon the method performance data in Table 4. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 4.
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentad i enea
Hexachloroethane
1,2,4-Trichlorobenzene
Accuracy, as
recovery, x'
(M9/L)
0.75C+3.21
0.85C-0.70
0.72C+0.87
0.72C+2.80
0.87C-0.02
0.61C+0.03
0.47C
0.74C-0.02
0.76C+0.98
Single analyst
precision, s/
(M9/L)
0.28X-1.17
0.22X-2.95
0.21X-1.03
0.16X-0.48
0.14X+0.07
0.18X+0.08
0.24x
0.23X+0.07
0.23X-0.44
Overall
precision,
S' (M9A)
0.38X-1.39
0.41X-3.92
0.49X-3.98
0.35X-0.57
0.36X-0.19
0.53X-0.12
0.50x
0.36X-0.00
0.40X-1.37
x' ^Expected recovery for one or more measurements of a sample containing a
concentration of C, in p.g/1.
s/= Expected single analyst standard deviation of measurements at an average
concentration of x, in /xg/L.
S' =Expected interlaboratory standard deviation of measurements at an average
concentration found of x, in M9/L-
C =True value for the concentration, in M9/L.
x = Average recovery found for measurements of samples containing a
concentration of C, in p.g/1.
a Estimates based upon the performance in a single laboratory.
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FIGURE 1
Column: 1.5% OV-1+1.5% OV-226 on Gas Chrom Q
Ttmptriturr 76°C
Dtuctor: Eltetron Capture
»•
4 8 12 16
RETENTION TIME (MINUTES)
20
Gaschromatogram of chlorinattd hydrocarbons (low moltcular wtight compounds).
8120A - 13
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FIGURE 2
Column: 1.5% OV-1+1.5% OV-225 on GM Chrom Q
Ttmptrtturt: 160°C
Detector: Electron Capturt
4 8 12 16
RETENTION TIME (MINUTES)
Gas chromatogram of chlorinated hydrocarbons (high molecular weight compounds).
8120A - 14
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METHOD 8120A
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY
Start
71.1 Choose
appropriate
ex trac11on
procedure (see
Chapter 2|
7.1.2 Exchange
axtraction solvent
to hexane during
K-D procedures
7 2 Set gas
chr oma tography
conditions
7 3 Refer to Method
8000 for proper
calibration
techniques
Yes
7 3.2 Process a
series of standards
through cleanup
procedure: analyze
by CC
No
7 4 Perform CC
analysis (see
'Method 8000)
7.5 1 Cleanup using
Method 3620
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METHOD 8121
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the determination of certain
chlorinated hydrocarbons in water, soil/sediment and waste matrices. The
following compounds can be determined by this method:
Compound Name CAS No."
Benzal chloride 98-87-3
Benzotrichloride 98-07-7
Benzyl chloride 100-44-7
2-Chloronaphthalene 91-58-7
1,2-Dichlorobenzene 95-50-1
1,3-Dichlorobenzene 541-73-1
1,4-Dichlorobenzene 106-46-1
Hexachlorobenzene 118-74-1
Hexachlorobutadiene 87-68-3
alpha-Hexachlorocyclohexane (alpha-BHC) 319-84-6
beta-Hexachlorocyclohexane (beta-BHC) 319-85-7
gamma-Hexachlorocyclohexane (gamma-BHC) 58-89-9
delta-Hexachlorocyclohexane (delta-BHC) 319-86-8
Hexachlorocyclopentadiene 77-47-4
Hexachloroethane 67-72-1
Pentachlorobenzene 608-93-5
1,2,3,4-Tetrachlorobenzene 634-66-2
1,2,4,5-Tetrachlorobenzene 95-94-2
1,2,3,5-Tetrachlorobenzene 634-90-2
1,2,4-Trichlorobenzene 120-82-1
1,2,3-Trichlorobenzene 87-61-6
1,3,5-Trichlorobenzene 108-70-3
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists method detection limits (MDL) for each compound in an
organic-free reagent water matrix. The MDLs for the compounds of a specific
sample may differ from those listed in Table 1 because they are dependent upon
the nature of interferences in the sample matrix. Table 2 lists the estimated
quantitation limits (EQL) for other matrices.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph and in the interpretation
of gas chromatograms.
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2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample is extracted by using one of
the appropriate sample extraction techniques specified in Methods 3510, 3520,
3540, or 3550, or diluted using Method 3580. Aqueous samples are extracted at
neutral pH with methylene chloride by using either a separatory funnel (Method
3510) or a continuous liquid-liquid extractor (Method 3520). Solid samples are
extracted with hexane/acetone (1:1) by using a Soxhlet extractor (Method 3540)
or with methylene chloride/acetone (1:1) by using an ultrasonic extractor (Method
3550). After cleanup, the extract or diluted sample is analyzed by gas
chromatography with electron capture detection (GC/ECD).
2.2 When this method is used to analyze for any or all of the target
compounds, compound identification should be supported by at least one additional
qualitative technique. This method describes analytical conditions for a second
gas chromatographic column that can be used to confirm the measurements made with
the primary column. Retention time information obtained on two gas
chromatographic columns is given in Table 3. Alternatively, gas
chromatography/mass spectrometry could be used for compound confirmation if
concentration permits.
2.3 The sensitivity of Method 8121 usually depends on the level of
interferences rather than on instrumental limitations. If interferences prevent
detection of the analytes, Method 8121 may also be performed on samples that have
undergone cleanup. This method may be used in conjunction with Method 3620,
Florisil Column Cleanup, Method 3660, Sulfur Cleanup, and Method 3640, Gel
Permeation Chromatography, to aid in the elimination of interferences.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other hardware used in sample
processing may introduce artifacts which may result in elevated baselines,
causing misinterpretation of gas chromatograms. These materials must therefore
be demonstrated to be free from interferants, under the conditions of the
analysis, by analyzing method blanks. Specific selection of reagents and
purification of solvents by distillation in all-glass systems may be required.
Pesticide grade or distilled-in-glass solvents are suitable for trace analysis
without further purification. Each new batch of solvent should be checked for
possible interferants as follows: concentrate the amount of solvent equivalent
to the total volume to be used in the analysis to 1 ml. Inject 1 to 2 /xL of the
concentrate into a gas chromatograph equipped with an electron capture detector
(ECD) set at the lowest attenuation. If extraneous peaks are detected that are
greater than 10 pg on-column, the solvent must be purified either by
redistillation or by passing it through a column of highly activated alumina
(acidic or basic alumina, activated at 300°C to 400°C) or Florisil.
3.3 Interferants coextracted from the samples will vary considerably from
waste to waste. While general cleanup techniques are provided as part of this
method, specific samples may require additional cleanup steps to achieve desired
sensitivities.
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3.4 Glassware must be scrupulously clean. Clean all glassware as soon
as possible after use by rinsing with the last solvent used, followed by thorough
washing of the glassware in hot, aqueous detergent solution. Rinse with tap
water, distilled water, acetone, and finally pesticide quality hexane. Heavily
contaminated glassware may require treatment in a muffle furnace at 400°C for 2
to 4 hours. Some high boiling materials, such as PCBs, may not be eliminated
by this treatment. Volumetric glassware should not be heated in a muffle
furnace. Glassware should be sealed and stored in a clean environment
immediately after drying and cooling to prevent any accumulation of dust or other
contaminants. Store the glassware by inverting or capping with aluminum foil.
3.5 Phthalate esters, if present in a sample, will interfere only with
the BHC isomers because they elute in Fraction 2 of the Florisil procedure
described in Method 3620. The presence of phthalate esters can usually be
minimized by avoiding contact with any plastic materials.
3.6 The presence of elemental sulfur will result in large peaks, and can
often mask the region of compounds eluting after 1,2,4,5-tetrachlorobenzene
(Compound No. 18 in the gas chromatogram shown in Figure 1). The
tetrabutylammonium (TBA)-sulfite procedure (Method 3660) works well for the
removal of elemental sulfur.
3.7 Waxes and lipids can be removed by gel permeation chromatography
(Method 3640). Extracts containing high concentrations of lipids are viscous
and may even solidify at room temperature.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detector, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 30 m x 0.53 mm ID fused-silica capillary
column chemically bonded with trifluoropropyl methyl silicone (DB-
210 or equivalent).
4.1.2.2 Column 2 - 30 m x 0.53 mm ID fused-silica capillary
column chemically bonded with polyethylene glycol (DB-WAX or
equivalent).
4.1.3 Detector - electron capture detector
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation
of extracts.
8121 - 3 Revision 0
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4.3.2 Evaporation flask - 500 mi (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Glassware: See Methods 3510, 3520, 3540, 3550, 3580, 3620, 3640, and
3660 for specifications.
4.5 Boiling chips, approximately 10/40 mesh. Heat to 400°C for 30 min,
or Soxhlet-extract with methylene chloride, prior to use.
4.6 Vials - 10 ml, glass, with Teflon lined screw-caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Preservatives:
5.3.1 Sodium hydroxide, NaOH, (ACS certified), 10 N in distilled
water.
5.3.2 Sulfuric acid, H2S04, (ACS certified), mix equal volumes of
concentrated sulfuric acid and distilled water.
5.4 Solvents:
5.4.1 Acetone, CH3COCH3 - pesticide quality or equivalent.
5.4.2 Hexane, C6H14 - pesticide quality or equivalent.
5.4.3 Diethyl ether, C2H5OC2H5 - pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.4.4 Methylene chloride, CH2C12 - pesticide quality or equivalent.
5.4.5 Petroleum ether - pesticide quality or equivalent.
8121 - 4 Revision 0
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5.5 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.6 Stock standard solutions
5.6.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in hexane
and diluting to volume in a 10 ml volumetric flask. Larger volumes can
be used at the convenience of the analyst. When compound purity is assayed
to be 96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards can be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.6.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp tops. Store at 4°C and protect from
light. Stock standards should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.6.3 Stock standard solutions must be replaced after one year,
or sooner if comparison with check standards indicates a problem.
5.7 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared through dilution of the stock standards with
hexane. One of the concentrations should be at a concentration near, but above,
the method detection limit. The remaining concentrations should correspond to
the expected range of concentrations found in real samples or should define the
working range of the GC. The suggested concentrations are listed in Table 4.
Calibration solutions must be replaced after six months, or sooner if comparison
with check standards indicates a problem.
5.8 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences.
5.8.1 The suggested internal standards are: 2,5-dibromotoluene,
1,3,5-tribromobenzene, and a,a'-dibromo-m-xylene. The analyst can use any
of the three compounds provided that they are resolved from matrix
interferences.
5.8.2 Prepare an internal standard spiking solution which contains
50 mg/L of any of the compounds listed above. Addition of 10 /iL of this
solution to 1 mL of sample extract is recommended. The spiking
concentration of the internal standard should be kept constant for all
samples and calibration standards. Store the internal standard spiking
solutions at 4°C in Teflon-sealed containers. Standard solutions should
be replaced when ongoing QC (Section 8) indicates a problem.
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5.9 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, and the effectiveness
of the method in dealing with each sample matrix, by spiking each sample,
standard, and organic-free reagent water blank with the surrogate compounds.
5.9.1 Recommended surrogate compounds: a,2,6-trichlorotoluene,
1,4-dichloronaphthalene, and 2,3,4,5,6-pentachlorotoluene.
5.9.2 Prepare a surrogate standard spiking solution which contains
1 mg/L of a,2,6-trichlorotoluene and 2,3,4,5,6-pentachlorotoluene and
10 mg/L of 1,4-dichloronaphthalene. Addition of 1 ml of this solution to
1 L of a water sample or 10 g of a solid sample is equivalent to 1 /xg/L
or 100 M9/k9 of a,2,6-trichlorotoluene and 2,3,4,5,6-pentachlorotoluene
and 10 jug/L or 1000 /xg/kg of 1,4-dichloronaphthalene. The spiking
concentration of the surrogate standards may be adjusted accordingly, if
the final volume of extract is reduced below 10 ml. Store the spiking
solutions at 4°C in Teflon-sealed containers. The solutions must be
replaced after 6 months, or sooner if ongoing QC (Section 8) indicates
problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a neutral
pH with methylene chloride by using a separatory funnel (Method 3510) or
a continuous liquid-liquid extractor (Method 3520). Solid samples are
extracted with hexane/acetone (1:1 v:v) by using a Soxhlet extractor
(Method 3540) or with methylene chloride/acetone (1:1 v:v) by using an
ultrasonic extractor (Method 3550). Non-aqueous waste samples may be
diluted using Method 3580.
7.2 Solvent exchange: Prior to Florisil cleanup or gas chromatographic
analysis, the extraction solvent must be exchanged to hexane. Sample extracts
that will be subjected to gel permeation chromatography do not need solvent
exchange. The exchange is performed during the K-D procedures listed in all of
the extraction methods. The exchange is performed as follows:
7.2.1 Add one or two clean boiling chips to the flask and attach
a three ball Snyder column. Prewet the column by adding about 1.0 mL of
methylene chloride to the top of the column. Place the K-D apparatus in
a hot water bath (15-20°C above the boiling point of the solvent) so that
the concentrator tube is partially immersed in the hot water and the entire
lower rounded surface of the flask is bathed with hot vapor. Adjust the
vertical position of the apparatus and the water temperature, as required,
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to complete the concentration in 10-20 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but the
chambers will not flood. When the apparent volume of liquid reaches 1-2
ml, remove the K-D apparatus from the water bath and allow it to drain and
cool for at least 10 minutes.
7.2.2 Momentarily remove the Snyder column, add 50 ml of hexane,
a new boiling chip, and attach the macro Snyder column. Concentrate the
extract as described in Section 7.2.1, using 1 ml of hexane to prewet the
Snyder column, raising the temperature of the water bath, if necessary,
to maintain proper distillation, and completing the concentration in 10-20
minutes. When the apparent volume of liquid reaches 1-2 ml, remove the
K-D apparatus and allow it to drain and cool for at least 10 min.
7.2.3 Remove the Snyder column and rinse the flask and its lower
joint into the concentrator tube with 1 to 2 ml of hexane. A 5 mL syringe
is recommended for this operation. Adjust the extract volume to 10 mL.
Stopper the concentrator tube and store at 4"C if further processing will
be performed immediately. If the extract will be stored for two days or
longer, it should be transferred to a glass vial with a Teflon lined
screw-cap or crimp top. Proceed with the cleanup or gas chromatographic
analysis.
7.3 Cleanup/Fractionation:
7.3.1 Cleanup procedures may not be necessary for a relatively
clean matrix. If removal of interferences such as chlorinated phenols,
phthalate esters, etc., is required, proceed with the procedure outlined
in Method 3620. Collect Fraction 1 by eluting with 200 ml petroleum ether
and Fraction 2 by eluting with 200 ml of diethyl ether/petroleum ether
(1:1). Note that, under these conditions, benzal chloride and
benzotrichloride are not recovered from the Florisil column. The elution
patterns and compound recoveries are shown in Table 5.
7.3.2 Removal of waxes and lipids by gel permeation chromatography
(optional): Refer to Method 3640.
7.3.3 Elemental Sul.fur Removal (optional): refer to Method 3660,
Section 7.3.
7.4 Gas chromatographic conditions (recommended):
7.4.1 Column 1:
Carrier gas (He) =
Column temperature:
Initial temperature =
Temperature program =
Final temeprature =
Injector temperature =
Detector temperature =
10 mL/min
65°C
65°C to 175°C at 4°C/nnn
175°C, hold 20 minutes.
220°C
250°C
8121 - 7
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7.4.2 Column 2:
Carrier gas (He) = 10 mL/min
Column temperature:
Initial temperature = 60°C
Temperature program = 60°C to 170°C at 4°C/min
Final temeprature = 170°C, hold 30 minutes.
Injector temperature = 200°C
Detector temperature = 230°C
7.4.3 Tables 1 and 3 give the MDLs and the retention times for 22
chlorinated hydrocarbons. Examples of the separations achieved with the
trifluoropropyl methyl silicone and polyethylene glycol fused-silica
capillary columns are shown in Figures 1 and 2, respectively.
7.5 Calibration:
7.5.1 Refer to Method 8000 for proper calibration techniques. Use
Table 4 for guidance.
7.5.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.6 Gas chromatographic analysis:
7.6.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /iL of internal standard to the sample prior to
injection.
7.6.2 Follow Method 8000 for instructions on analysis sequence,
appropriate dilutions, daily retention time windows, and identification
criteria.
7.6.3 Record the sample volume injected and the resulting peak
areas.
7.6.4 Using either internal or external calibration procedures
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.6.5 If the response of a peak exceeds the working range of the
system, dilute the extract and reanalyze.
7.6.6 Identify compounds in the sample by comparing the retention
times of the peaks in the sample chromatogram with those of the peaks in
standard chromatograms obtained on the two columns specified in
Section 7.4. The retention time window used to make identifications should
be based upon measurements of actual retention time variations over the
course of 10 consecutive injections. Three times the standard deviation
of a retention time window can be used to calculate a suggested window
size.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
individual extraction method protocols. If extract cleanup is required, follow
the QC presented in Method 3600 and in the specific cleanup method protocols.
8.2 Mandatory quality control to evaluate the GC system operation is
found in Method 8000.
8.2.1 Analyze a quality control check standard to demonstrate that
the operation of the gas chromatograph is in control. The frequency of
the check standard analysis is equivalent to 10 percent of the samples
analyzed. If the recovery of any compound found in the check standard is
less than 80 percent of the certified value, the laboratory performance
is judged to be out of control, and the problem must be corrected. A new
set of calibration standards must be prepared and analyzed.
8.3 Calculate surrogate standard recoveries for all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If the recoveries are not within limits, the following are
required:
8.3.1.1 Check to be sure that there are no errors in
calculations, surrogate solutions, and internal standards. Also
check instrument performance.
8.3.1.2 Recalculate the data or reanalyze the extract if any
of the above checks reveals a problem.
8.3.1.3 Reextract and reanalyze the sample if none of the
above is a problem or designate the data as "estimated
concentration."
8.4 An internal standard peak area check must be performed on all samples.
The internal standard must be evaluated for acceptance by determining whether
the measured area for the internal standard deviates by more than 30 percent from
the average area for the internal standard in the calibration standards. When
the internal standard peak area is outside that limit, all samples that fall
outside the QC criteria must be analyzed.
8.5 GC/MS confirmation: Any compound confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by GC/MS
as determined by the laboratory-generated detection limits.
8.5.1 The GC/MS would normally require a minimum concentration of
in the final extract for each compound.
8.5.2 The sample extract should be analyzed by GC/MS as per
Section 7.0 of Method 8270.
8121 - 9 Revision 0
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8.5.3 A reference standard of the compound must also be analyzed
by GC/MS. The concentration of the reference standard must be at a
concentration that would demonstrate the ability to confirm the compounds
identified by GC/ECD.
8.6 Include a mid-concentration calibration standard after each group of
20 samples in the analysis sequence. The response factors for the mid-
concentration calibration must be within ±15 percent of the average values for
the multiconcentration calibration.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDLs listed in Table 1 were
obtained by using organic-free reagent water. Details on how to determine MDLs
are given in Chapter One. The MDLs actually achieved in a given analysis will
vary since they depend on instrument sensitivity and matrix effects.
9.2 This method has been tested in a single laboratory by using organic-
free reagent water, sandy loam samples and extracts which were spiked with the
test compounds at one concentration. Single-operator precision and method
accuracy were found to be related to the concentration of compound and the type
of matrix. Results of the single-laboratory method evaluation are given in
Tables 6 and 7.
9.3 The accuracy and precision obtained will be determined by the sample
matrix, sample preparation technique, optional cleanup techniques, and
calibration procedures used.
10.0 REFERENCES
1. Lopez-Avila, V., N.S. Dodhiwala, and J. Milanes, "Single Laboratory
Evaluation of Method 8120, Chlorinated Hydrocarbons", 1988, EPA Contract
Numbers 68-03-3226 and 68-03-3511.
2. Glazer, J.A., G.D. Foerst, G.D. McKee, S.A. Quave, and W.L. Budde, "Trace
Analyses for Wastewaters," Environ. Sci. and Technol. 15:1426-1431, 1981.
3. Lopez-Avila, V., Baldin, E., Benedicto, J., Milanes, J., and Beckert, W.F.,
"Application of Open-Tubular Columns to SW-846 GC Methods", EMSL-Las Vegas,
1990.
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November 1990
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Table 1.
METHOD DETECTION LIMITS FOR CHLORINATED HYDROCARBONS
ON CAPILLARY COLUMNS
MDLa
Compound name CAS no. (ng/L)
Benzal chloride 98-87-3 2-5b
Benzotrichloride 98-07-7 6.0
Benzyl chloride 100-44-7 180
2-Chloronaphthalene 91-58-7 1,300
1,2-Dichlorobenzene 95-50-1 270
1,3-Dichlorobenzene 541-73-1 250
1,4-Dichlorobenzene 106-46-1 890
Hexachlorobenzene 118-74-1 5.6
Hexachlorobutadiene 87-68-3 1.4
alpha-Hexachlorocyclohexane (alpha-BHC) 319-84-6 11
beta-Hexachlorocyclohexane (beta-BHC) 319-85-7 31
gamma-Hexachlorocyclohexane (gamma-BHC) 58-89-9 23
delta-Hexachlorocyclohexane (delta-BHC) 319-86-8 20
Hexachlorocyclopentadiene 77-47-4 240
Hexachloroethane 67-72-1 1.6
Pentachlorobenzene 608-93-5 38
1,2,3,4-Tetrachlorobenzene 634-66-2 11
1,2,4,5-Tetrachlorobenzene 95-94-2 9.5
1,2,3,5-Tetrachlorobenzene 634-90-2 8.1
1,2,4-Trichlorobenzene 120-82-1 130
1,2,3-Trichlorobenzene 87-61-6 39
1,3,5-Trichlorobenzene 108-70-3 12
MDL is the method detection limit for organic-free reagent water. MDL was
determined from the analysis of eight replicate aliquots processed through
the entire analytical method (extraction, Florisil cartridge cleanup, and
GC/ECD analysis).
MDL = t(n.1i0.99)xSD
where t(n.1099) is the student's t value appropriate for a 99 percent
confidence interval and a standard deviation with n-1 degrees of freedom,
and SD is the standard deviation of the eight replicate measurements.
Estimated from the instrument detection limit.
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Table 2.
ESTIMATED QUANTITATION LIMIT (EQL) FACTORS FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil by ultrasonic extraction 670
with GPC cleanup
High-concentration soil and sludges by ultrasonic 10,000
extraction
Waste not miscible with water 100,000
a Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
b EQL = [Method detection limit (Table 1)] x [Factor (Table 2)]. For
nonaqueous samples, the factor is on a wet-weight basis.
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Table 3
GAS CHROMATOGRAPHIC RETENTION TIMES FOR CHLORINATED HYDROCARBONS
ON CAPILLARY COLUMNS
Compound
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
Compound name
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
alpha-BHC
beta-BHC
gamma-BHC
delta-BHC
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachlorobenzene
1,2,3,4-Tetrachlorobenzene
1,2,4,5-Tetrachlorobenzene
1,2,3,5-Tetrachlorobenzene
1,2,4-Trichlorobenzene
1,2,3-Trichlorobenzene
1,3,5-Trichlorobenzene
Internal Standards
2,5-Dibromotoluene
1,3, 5-Tri bromobenzene
o,a'-Dibromo-meta-xylene
Surrogates
a,2,6-Trichlorotoluene
1,4-Dichloronaphthalene
2,3,4,5,6-Pentachlorotoluene
Retention
DB-210"
6.86
7.85
4.59
13.45
4.44
3.66
3.80
19.23
5.77
22.21
25.54
24.07
26.16
8.86
3.35
14.86
11.90
10.18
10.18
6.86
8.14
5.45
9.55
11.68
18.43
12.96
17.43
18.96
time (min)
OB-WAX"
15.91
15.44
10.37
23.75
9.58
7.73
8.49
29.16
9.98
41.62
33.84
54.30
33.79
c
8.13
23.75
21.17
17.81
17.50
13.74
16.00
10.37
18.55
22.60
35.94
22.53
26.83
27.91
GC operating conditions: 30 m x 0.53 mm ID DB-210 fused-silica capillary
column; 1 /zm film thickness; carrier gas helium at 10 mL/min; makeup gas is
nitrogen at 40 mL/min; temperature program from 65°C to 175°C (hold 20
minutes) at 4°C/min; injector temperature 220°C; detector temperature
250°C.
GC operating conditions: 30 m x 0.53 mm ID DB-WAX fused-silica capillary
column; 1 /im film thickness; carrier gas helium at 10 mL/min; makeup gas is
nitrogen at 40 mL/min; temperature program from 60°C to 170°C (hold 30
minutes) at 4°C/"nn; injector temperature 200°C; detector temperature 230°C.
Compound decomposes on-column.
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Table 4.
SUGGESTED CONCENTRATIONS FOR THE CALIBRATION SOLUTIONS8
Concentration (ng//il)
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chl oronaphthal ene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
alpha-BHC
beta-BHC
gamma- BHC
delta-BHC
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachlorobenzene
1,2,3,4-Tetrachlorobenzene
1,2,4 , 5-Tetrachl orobenzene
1,2,3 , 5-Tetrachl orobenzene
1, 2, 4-Trichl orobenzene
1 , 2 , 3-Tri chl orobenzene
1, 3, 5-Trichl orobenzene
Surrogates
a,2,6-Trichlorotoluene
1,4-Di chl oronaphthal ene
2,3,4,5,6-Pentachlorotoluene
0.1
0.1
0.1
2.0
1.0
1.0
1.0
0.01
0.01
0.1
0.1
0.1
0.1
0.01
0.01
0.01
0.1
0.1
0.1
0.1
0.1
0.1
0.02
0.2
0.02
0.2
0.2
0.2
4.0
2.0
2.0
2.0
0.02
0.02
0.2
0.2
0.2
0.2
0.02
0.02
0.02
0.2
0.2
0.2
0.2
0.2
0.2
0.05
0.5
0.05
0.5
0.5
0.5
10
5.0
5.0
5.0
0.05
0.05
0.5
0.5
0.5
0.5
0.05
0.05
0.05
0.5
0.5
0.5
0.5
0.5
0.5
0.1
1.0
0.1
0.8
0.8
0.8
16
8.0
8.0
8.0
0.08
0.08
0.8
0.8
0.8
0.8
0.08
0.08
0.08
0.8
0.8
0.8
0.8
0.8
0.8
0.15
1.5
0.15
1.0
1.0
1.0
20
10
10
10
0.1
0.1
1.0
1.0
1.0
1.0
0.1
0.1
0.1
1.0
1.0
1.0
1.0
1.0
1.0
0.2
2.0
0.2
One or more internal standards should be spiked prior to GC/ECD analysis
into all calibration solutions. The spike concentration of the internal
standards should be kept constant for all calibration solutions.
8121 - 14
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Table 5.
ELUTION PATTERNS OF CHLORINATED HYDROCARBONS
FROM THE FLORISIL COLUMN BY ELUTION WITH PETROLEUM ETHER (FRACTION 1)
AND 1:1 PETROLEUM ETHER/DIETHYL ETHER (FRACTION 2)
Recovery (percent)8
Compound
Benzal chloride"
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachlorobutadiene
alpha-BHC
beta-BHC
gamma-BHC
delta-BHC
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachlorobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2,4 , 5-Tetrachl orobenzene6
1,2,3 , 5-Tetrachl orobenzene6
1 , 2 , 4-Tri chl orobenzene
1 , 2 , 3-Tr i chl orobenzene
1, 3, 5-Tri chl orobenzene
Amount
(/xg) Fraction 1"
10
10
100
200
100
100
100
1.0
1.0
10
10
10
10
1.0
1.0
1.0
10
10
10
10
10
10
0
0
82
115
102
103
104
116
101
93
100
129
104
102
102
59
96
102
Fraction 2C
0
0
16
95
108
105
71
Values given represent average values of duplicate experiments.
1 was eluted with 200 mL petroleum ether.
Fraction 2 was eluted with 200 mL petroleum ether/diethyl ether (1:1).
This compound coelutes with 1,2,4-trichlorobenzene; separate experiments were
performed with benzal chloride to verify that this compound is not recovered
from the Florisil cleanup in either fraction.
This pair cannot be resolved on the DB-210 fused-silica capillary columns.
8121 - 15
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Table 6.
ACCURACY AND PRECISION DATA FOR METHOD 3510 AND METHOD 8121
Compounds
Benzal chloride0
Benzotrichloride
Benzyl chloride
2-Chl oronaphthal ene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachlorobutadiene
alpha-BHC
beta-BHC
gamma-BHC
delta-BHC
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachl orobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2,4 , 5-Tetrachl orobenzened
1,2,3 , 5-Tetrachl orobenzened
l,2,4-Trichlorobenzenec
1 , 2 , 3-Tri chl orobenzene
1 , 3 , 5-Tri chl orobenzene
Spike
concentration
(M9/L)
10
1.0
100
200
100
100
100
1.0
1.0
10
10
10
10
10
1.0
1.0
10
10
10
10
10
10
Average
recovery31"
(percent)
95
97
90
91
92
87
89
92
95
96
103
96
103
97
96
89
96
93
93
95
95
93
Precision
(percent RSD)
3.0
2.1
6.2
6.5
5.7
8.7
8.9
7.1
3.6
2.6
3.6
2.8
2.7
5.1
4.0
6.5
3.4
4.6
4.6
3.0
4.4
6.2
Surrogates
a,2,6-Trichlorotoluene
1,4-Dichloronaphthalene
2,3,4,5,6-Pentachlorotoluene
1.0
10
1.0
85
78
80
6.5
6.1
5.9
a The number of determinations is 5.
b Final volume of extract was 10 ml. Florisil cleanup was not performed on any
of the samples.
Cid These pairs cannot be resolved on the DB-210 fused-silica capillary column.
8121 - 16
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1990
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Table 7.
ACCURACY AND PRECISION DATA FOR METHOD 3550 AND METHOD 8121
Compounds
Spike
concentration
(ng/L)
Average
recovery
(percent)
,a,b
Precision
(percent RSD)
Benzal chloride0
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
alpha-BHC
beta-BHC
gamma-BHC
delta-BHC
Hexachlorocyclopentadi ene
Hexachloroethane
Pentachlorobenzene
1,2,3,4-Tetrachlorobenzene
1,2,4,5-Tetrachlorobenzened
1,2,3,5-Tetrachlorobenzened
1,2,4-Trichlorobenzenec
1,2,3-Trichlorobenzene
1,3,5-Trichlorobenzene
Surrogates
a,2,6-Trichlorotoluene
1,4-Dichloronaphthalene
2,3,4,5,6-Pentachlorotoluene
3,300
3,300
33,000
66,000
33,000
33,000
33,000
330
330
3,300
300
3,300
3,300
330
330
330
3,300
3,300
3,300
3,300
3,300
3,300
330
3,300
330
89
90
121
100
84
81
89
81
83
100
92
99
97
44
83
81
88
80
80
89
79
75
86
88
98
.5
.9
2.7
2.9
5.9
6.4
7.1
12.6
11.0
3.2
4.7
2.9
2.4
4.1
1
25.
4.6
3.5
2.9
4.4
4.4
2.7
4.3
5.3
2.7
4.5
11.7
a The number of determinations is 5.
b Final volume of extract was 10 ml. Florisil cleanup was not performed on any
of the samples.
Cid These pairs cannot be resolved on the DB-210 fused-silica capillary column.
8121 - 17
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3 20 2 21
10 12 11 13
19
18
14
16
10 15 20
TIME (mln)
25
30
Figure 1.
GC/ECD chromatogram of Method 8121 composite standard analyzed
on a 30 m x 0.53 mm ID DB-210 fused-silica capillary column.
GC operating conditions are given in Section 7.4. See Table
3 for compound identification.
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17
4
16
10
11
13
JL
12
0
10 15 20 25 30 35
TIME (min)
40
45
50
55
Figure 2.
GC/ECD chromatogram of Method 8121 composite standard analyzed
on a 30 m x 0.53 mm ID DB-WAX fused-silica capillary column.
GC operating conditions are given in Section 7.4. See Table
3 for compound identification.
8121 - 19
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METHOD 8121
CHLORINATED HYDROCARBONS BY GAS CHRQMATQGRAPHY.
CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose appropriate
extraction procedure
7.1.2 Add appropriate spiking
compounds to sample prior
to extraction procedure
7.2 Exchange extraction
solvent to hexane during
K-D procedures
7.2.1 Following concentration of
methylene chloride allow K-D
apparatus to drain and cool
7.2.2 Increase temperature of hot
water bath; add hexane; attach
Snyder column; place apparatus
on water bath; concentrate;
remove from water bath; cool
7.2.3 Remove column; rinse flask
and joints with hexane; adjust
extract volume
7.3 Choose approriate cleanup
technique, if necessary;
fluorosil cleanup is recommended.
Refer to Method 3620 or to
Section 7.3.2
.2.3 Will furth
processing be
performed within
two days?
7.2.3 Transfer extract to
Teflon sealed screw-cap
vials; refrigerate
7.3.4 Elemental
sulfur removal
required?
7.3.3 GPC
cleanup
required?
7.3.4 Refer to
Method 3660,
Section 7.3
7.3.3 Refer to
Method 3640
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METHOD 8121
(CONTINUED)
7.2.3 Stopper concentrator
and refrigerate
7.4.1 Set column 1 conditions
7.4.2 Set column 2 conditions
7.5.1 Refer to Method 8000 for
calibration techniques; select
lowest point on calibration curve
7.5.2 Choose and perform
internal or external calibration
(refer to Method 8000)
7.6.1 Add internal standard if
necessary
7.6.2 Establish daily retention time
windows, analysis sequence,
dilutions, and identification criteria
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METHOD 8121
(CONCLUDED)
7.6.3 Record sample volume
injected and resulting peak
sizes
7.6.4 Determine identity and
quantity of each component peak
that corresponds to compound
used for calibration
7.6.5 Dilute extract; reanalyze
7.6.6 Compare standard and
sample retention times; identify
compounds
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METHOD 8141A
ORGANOPHOSPHORUS COMPOUNDS BY GAS CHROMAT06RAPHY;
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8141 is a gas chromatographic (GC) method used to determine
the concentration of various organophosphorus compounds. The following compounds
can be determined by this method:
Compound Name
CAS No.'
Azinphos methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton, 0 and S
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion-ethyl
Parathion-methyl
Phorate
Ronnel
Sulfotep
TEPP
Stirophos (Tetrachlorovinphos)
Tokuthion (Protothiofos)
Trichloronate
86-50-0
35400-43-2
2921-88-2
56-72-4
8065-48-3
333-41-5
62-73-7
60-51-5
298-04-4
2104-64-5
13194-48-4
115-90-2
55-38-9
121-75-5
150-50-5
7786-34-7
6923-22-4
300-76-5
56-38-2
298-00-0
298-02-2
299-84-3
3689-24-5
21646-99-1
22248-79-9
34643-46-4
327-98-0
Chemical Abstract Services Registry Number.
1.2 Table 1 lists method detection limits (MDL) for each compound in a
water and a soil matrix. Table 2 lists the estimated quantitation limits (EQLs)
for other matrices.
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1.3 Analytical difficulties encountered with specific organophosphorus
compounds may include (but are not limited to) the following:
1.3.1 Tetraethyl pyrophosphate (TEPP) is an unstable diphosphate
which is readily hydrolyzed in water and is thermally labile (TEPP
decomposes at 170°C). Care must be taken to minimize loss during GC
analysis and during sample preparation. Identification of bad standard
lots is difficult since the electron impact mass spectrum of TEPP is nearly
identical to its major breakdown product, triethyl phosphate.
1.3.2 The water solubility of dichlorvos is 10 g/L at 20°C, and
recovery is poor from aqueous solution.
1.3.3 Naled is converted to dichlorvos on column by debromination.
This reaction may also occur during sample workup. The extent of
debromination will depend on the nature of the matrix being analyzed. The
analyst must consider the potential for debromination when naled is to be
determined.
1.3.4 Trichlorofon (not determined by this method) rearranges and
is dehydrochlorinated in acidic, neutral, or basic media to form dichlorvos
and hydrochloric acid. If this method is to be used for the determination
of organophosphates in the presence of trichlorofon, the analyst should
be aware of the possibility of rearrangement to dichlorvos to prevent
misidentification.
1.3.5 Demeton is a mixture of two compounds;
0,0-Diethyl 0-[2-(ethylthio)ethyl] phosphorothioate (Demeton-0) and
0,0-Diethyl S-[2-(ethylthio)ethyl]phosphorothioate(Demeton-S). Standards
for the individual isomers are no longer available through the EPA
repository, and two peaks will be observed in all mixed Demeton standards.
It is recommended that the early eluting compound (Demeton-S) be used for
quantitation.
1.3.6 Tributyl phosphorotrithioite (Merphos) is a single component
compound that is readily oxidized in the environment and during storage
to the phosphorotrithioate. The analyst may observe two peaks in the
chromatograms of merphos standards.
1.4 Recoveries for some additional organophosphorus compounds have been
determined for water. They include:
Azinphos ethyl HMPA
Carbofenthion Leptophos
Chlorfenvinphos Phosmet
Dioxathion Phosphamidion
Ethion Terbuphos
Famphur TOCP
As Method 8141 has not been fully validated for the determination of these
compounds, the analyst must demonstrate recoveries of greater than 70 percent
with precision of no more than 15 percent RSD before Method 8141 is used for
these or any additional analytes.
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1.5 When Method 8141 is used to analyze unfamiliar samples, compound
identifications should be supported by a single confirmatory analysis. Section
8.4 provides gas chromatograph/mass spectrometer (GC/MS) criteria appropriate
for the qualitative confirmation of compound identifications.
1.6 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography and in the interpretation
of chromatograms.
1.7 The use of Gel Permeation Cleanup (Method 3640) for sample cleanup
has been demonstrated to yield recoveries of less than 85 percent for many method
analytes and is therefore not recommended for use with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8141 provides gas chromatographic conditions for the detection
of ppb concentrations of organophosphorus compounds. Prior to the use of this
method, appropriate sample preparation techniques must be used. Water samples
are extracted at a neutral pH with methylene chloride as a solvent by using a
separatory funnel (Method 3510) or a continuous liquid-liquid extractor (Method
3520). Soxhlet extraction (Method 3540) or ultrasonic extraction (Method 3550)
using methylene chloride/acetone (1:1) are used for solid samples. Both neat
and diluted organic liquids (Method 3580, Waste Dilution) may be analyzed by
direct injection. Spiked samples are used to verify the applicability of the
chosen extraction technique to each new sample type. A gas chromatograph with
a flame photometric or nitrogen-phosphorus detector is used for this multiresidue
procedure.
2.2 If interferences are encountered in the analysis, Method 8141 may also
be performed on extracts that have undergone cleanup using Method 3620 or Method
3660.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 The use of Florisil cleanup materials (Method 3620) for some of the
compounds in this method has been demonstrated to yield recoveries less than
85% and is therefore not recommended for all compounds. Refer to Table 2 of
Method 3620 for recoveries of organophosphorus compounds as a function of
Florisil fractions. Use of phosphorus or halogen specific detectors, however,
often obviates the necessity for cleanup for relatively clean sample matrices.
If particular circumstances demand the use of an alternative cleanup procedure,
the analyst must determine the elution profile and demonstrate that the recovery
of each analyte is no less than 85%.
3.3 Use of a flame photometric detector in the phosphorus mode will
minimize interferences from materials that do not contain phosphorus. Elemental
sulfur, however, may interfere with the determination of certain organophosphorus
compounds by flame photometric gas chromatography. Sulfur cleanup using Method
3660 may alleviate this interference.
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3.4 A halogen specific detector (i.e. electrolytic conductivity or
microcoulometric) is very selective for the halogen containing compounds and
may be used for the determination of chlorpyrifos, ronnel, coumaphos, tokuthion,
trichloronate, dichlorvos, EPN, naled, and stirophos only.
3.5 Please note in Table 3 that a few analytes coelute on certain columns.
Therefore, select a second column for confirmation where coelution of the
analytes of interest does not occur.
3.6 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis by analyzing reagent blanks (refer to Section 8.1).
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph and all required accessories including syringes, analytical
columns, gases, detector and data system, integrator or stripchart
recorder. A data system or integrator is recommended for measuring peak
areas and/or peak heights.
4.1.2 Columns
4.1.2.1 Column 1 - 15 m x 0.53 mm wide-bore capillary column,
1.0 urn film thickness, coated with 50 percent trifluoropropyl, 50%
methyl silicone (DB-210, SP-2401, QF1, UCON, HB-280X, Triton X-100),
or equivalent.
4.1.2.2 Column 2-15mx0.53mm wide-bore capillary column,
1.5 jum film thickness, coated with 35 percent phenyl
methylpolysiloxane (DB-608, SPB-608, RTx-35), or equivalent.
4.1.2.3 Column 3 - 15 m x 0.53 mm wide-bore capillary column,
1.0 jum film thickness, coated with 5 percent phenyl, 95 percent
methyl silicone (DB-5, SE-54, SPB-5, RTx-5), or equivalent.
4.1.3 Detector - These detectors have proven effective in analysis
for all analytes listed in Table 1 and Section 1.4 and were used to develop
the accuracy and precision statements in Section 9.0.
4.1.3.1 Nitrogen Phosphorus Detector (NPD) operated in the
phosphorus specific mode is recommended.
4.1.3.2 Flame Photometric Detector (FPD) operated in the
phosphorus specific mode is recommended.
4.1.3.3 Halogen specific detectors (electrolytic conductivity
8141A - 4 Revision 1
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or microcoulometric) may be used if only halogenated or sulfur
analytes are to be determined.
4.2 Kuderna-Danish (K-D) apparatus (Kontes K-570025-0500):
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Vials - 10 ml, glass with Teflon lined screw-caps or crimp tops.
4.4 Water bath - Heated with concentric ring cover, capable of temperature
control (± 2°C). The bath should be used in a hood.
4.5 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.6 Boiling chips - Solvent extracted with methylene chloride,
approximately 10/40 mesh (silicon carbide or equivalent).
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform
to the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.4 Isooctane, C8H18 - Pesticide quality or equivalent.
5.5 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and blank with one or two surrogates (e.g. organophosphorus compounds
not expected to be present in the sample) recommended to encompass the range of
8141A - 5 Revision 1
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the temperature program used in this method. Deuterated analogs of analytes
should not be used as surrogates for gas chromatographic analysis due to
coelution problems.
5.6 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially prepared
stock standards can be used if they are verified against EPA standards. If EPA
standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable.
5.6.1 Prepare stock standard solutions by accurately weighing
0.0100 g of pure material. Dissolve the material in hexane or other
suitable solvent and dilute to known volume in a volumetric flask. If
compound purity is certified at 96% or greater, the weight can be used
without correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
the are certified by the manufacturer or by an independent source.
5.6.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at 4°C and protect from light.
Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.6.3 Stock standard solutions must be replaced after six months
or sooner if comparison with check standards indicates a problem. All
stock standards must be stored in a freezer at 4°C.
5.7 Calibration standards - A minimum of five concentrations for each
analyte of interest should be prepared through dilution of the stock standards
with hexane. One of the concentrations should be at a concentration near, but
above, the MDL. The remaining concentrations should correspond to the expected
range of concentrations found in real samples or should define the working range
of the GC. Calibration standards must be replaced after one to two months, or
sooner if comparison with check standards indicates a problem.
5.8 Internal standards should only be used on well characterized samples.
To use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.8.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Section 5.7.
5.8.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with hexane or other
suitable solvent.
5.8.3 Analyze each calibration standard according to Section 7.0.
8141A - 6 Revision 1
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Extracts are to be refrigerated at 4°C and analyzed within 40 days of
extraction.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a neutral
pH with methylene chloride, using either Method 3510 or 3520. Solid
samples are extracted using either Method 3540 or 3550 with methylene
chloride/acetone (1:1) as the extraction solvent.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
may be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The analyst must
ensure quantitative transfer of the extract concentrate. Single laboratory
data indicates that samples should not be transferred with 100 percent
hexane during sample workup as the more water soluble organophosphorus
compounds may be lost. This transfer is best accomplished with a
hexane/acetone solvent mixture. The exchange is performed as follows:
7.1.2.1 Following K-D concentration of the methylene chloride
extract to 1 mL using the macro Snyder column, allow the apparatus
to cool and drain for at least 10 minutes.
7.1.2.2 Momentarily remove the Snyder column, add 50 mL of
hexane/acetone solvent mixture, a new glass bead or boiling chip,
and attach the micro Snyder column. Concentrate the extract using
1 mL of hexane to prewet the Snyder column. Place the K-D apparatus
on the water bath so that the concentrator tube is partially immersed
in the hot water. Adjust the vertical position of the apparatus and
the water temperature, as required, to complete concentration in
5-10 minutes. At the proper rate of distillation the balls of the
column will actively chatter, but the chambers will not flood. When
the apparent volume of liquid reaches 1 mL, remove the K-D apparatus
and allow it to drain and cool for at least 10 minutes.
7.1.2.3 Remove the Snyder column and rinse the flask and its
lower joint with 1-2 mL of hexane into the concentrator tube. A 5 mL
syringe is recommended for this operation. Adjust the extract volume
to 10 mL. Stopper the concentrator tube and store refrigerated at
4°C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be transferred
to a vial with a Teflon lined screw-cap or crimp top. Proceed with
gas chromatographic analysis if further cleanup is not required.
8141A - 7 Revision 1
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7.2 Gas chromatography conditions (recommended): Three megabore capillary
columns are included for analysis of organophosphates by this method. Column 1
(DB-210 or equivalent) and Column 2 (SPB-608 or equivalent) are recommended if
a large number of organophosphorus analytes are to be determined. If the
superior resolution offered by Column 1 and Column 2 is not required, Column 3
(DB-5 or equivalent) may be used.
7.2.1 Columns 1 and 2
Carrier gas (He) flow rate = 5 mL/min
Initial temperature = 50°C, hold for 1 minute
Temperature program = 50°C to 140°C at 5°C/nrin, hold for 10 minutes,
followed by 140°C to 240°C at 10°C/min, hold
for 10 minutes (or a sufficient amount of time
for last compound to elute).
7.2.2 Column 3
Carrier gas (He) flow rate = 5 mL/min
Initial temperature = 130°C, hold for 3 minutes
Temperature program = 130°C to 180°C at 5°C/min, hold for 10 minutes,
followed by 180°C to 250°C at 2°C/min, hold
for 15 minutes (or a sufficient amount of time
for last compound to elute).
7.2.3 Retention times for all analytes on each column are presented
in Table 3. The analyst should note that several method analytes coelute
on column 3.
7.3 Calibration, refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.3.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will confirm elution patterns and the
absence of interferences from the reagents.
7.4 Gas chromatographic analysis
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 juL of internal standard to the sample prior to
injection.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.4.3 For megabore capillary columns, automatic injections of 1 /iL
are recommended. Hand injections of no more than 2 fj.1 may be used if the
analyst demonstrates quantitation precision of < 10 percent relative
8141A - 8 Revision 1
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standard deviation. The solvent flush technique may be used if the amount
of solvent is kept at a minimum.
7.4.4 Examples of chromatograms for various organophosphorus
compounds are shown in Figures 1 through 4.
7.4.5 Record the sample volume injected to the nearest 0.05 /xL and
the resulting peak sizes (in area units or peak heights).
7.4.6 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Method 8000 for calculation equations.
7.4.7 If peak detection and identification is prevented by the
presence of interferences, further cleanup is required. Before using any
cleanup procedure, the analyst must process a series of calibration
standards through the procedure to establish elution patterns and to
determine recovery of target compounds. The absence of interference from
reagents must be demonstrated by routine processing of reagent blanks
through the chosen cleanup procedure.
7.4.8 Naled has been reported to be converted to DDVP on some
columns by debromination. If this process is demonstrated on the GC system
that is used for analysis, clean the injector and break off several inches
of a megabore column or change the glass wool of a packed column prior to
analyzing samples. If subsequent injections of naled give DDVP, report
naled as DDVP, but, in this instance, both naled and DDVP may not be
reported in the same sample.
7.5 Cleanup: If required, the samples may be cleaned up using the Methods
presented in Chapter 4, Section 2.2.2.
7.5.1 Proceed with Florisil column Cleanup (Method 3620), followed
by, if necessary, Sulfur Cleanup (Method 3660), using the 10 ml hexane
extracts obtained from Section 7.1.2.3.
NOTE: The use of Gel Permeation (Method 3640) for sample cleanup has been
demonstrated to yield recoveries of less than 85 percent for many method
analytes and is therefore not recommended for use with this method.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous Sections and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
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8.2 Procedures to check the GC system operation are found in Method 8000.
8.3 GC/MS confirmation
8.3.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Follow the GC/MS
operating requirements specified in Method 8270.
8.3.2 When available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process.
8.3.3 To confirm an identification of a compound, the background
corrected mass spectrum of the compound must be obtained from the sample
extract and must be compared with a mass spectrum from a stock or
calibration standard analyzed under the same chromatographic conditions.
At least 25 ng of material should be injected into the GC/MS. The
following criteria must be met for qualitative confirmation:
The molecular ion and all other ions present above 20 percent
relative abundance in the mass spectrum of the standard must be present
in the mass spectrum of the sample with agreement to ± 10 percent. For
example, if the relative abundance of an ion is 30 percent in the mass
spectrum of the standard, the allowable limits for the relative abundances
of that ion in the mass spectrum for the sample would be 20 to 40 percent.
The retention time of the compound in the sample must be within six
seconds of the retention time for the same compound in the standard
solution.
Compounds that have very similar mass spectra can be explicitly
identified by GC/MS only on the basis of retention time data.
8.3.4 Where available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process because of the
extensive fragmentation of organophosphorus compounds during electron
impact MS processes.
8.3.5 Should the MS procedure fail to provide satisfactory results,
additional steps may be taken before reanalysis. These steps may include
the use of alternate packed or capillary GC columns or additional sample
cleanup.
9.0 METHOD PERFORMANCE
9.1 Estimated MDLs and associated chromatographic conditions for water
and clean soil (uncontaminated with synthetic organics) are listed in Table 1.
As detection limit will vary with the particular matrix to be analyzed, guidance
for estimating EQLs is given in Table 2.
9.2 Single operator accuracy and precision studies have been conducted
with spiked water and soil samples. The results of these studies are presented
in Tables 4-7.
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10.0 REFERENCES
1. Taylor, V.; Hickey, D.M.; Marsden, P.J. "Single Laboratory Validation of
EPA Method 8140"; U.S. Environmental Protection Agency. Environmental
Monitoring Systems Laboratory. Office of Research and Development, Las
Vegas, NV, 1987; EPA-600/4-87-009.
2. Pressley, T.A; Longbottom, J.E. "The Determination of Organophosphorus
Pesticides in Industrial and Municipal Wastewater: Method 614"; U.S.
Environmental Protection Agency. Environmental Monitoring and Support
Laboratory. Cincinnati, OH, 1982; EPA-600/4-82-004.
3. "Analysis of Volatile Hazardous Substances by GC/MS: Pesticide Methods
Evaluation"; Letter Reports 6, 12A, and 14 to the U.S.Environmental
Protection Agency on Contract 68-03-2697, 1982.
4. "Method 622, Organophosphorus Pesticides"; U.S. Environmental Protection
Agency. Environmental Monitoring and Support Laboratory. Cincinnati, OH
45268.
5. Chau, A.S.Y.; Afghan, B.K. Analysis of Pesticides in Water. Vol. II;
"Chlorine and Phosphorus-Containing Pesticides"; CRC: Boca Raton, FL, 1982,
pp 91-113, 238.
6. Hild, J.; Schulte, E; Thier, H.P. "Separation of Organophosphorus Pesticides
and Their Metabolites on Glass-Capillary Columns"; Chromatooraphia. 1978,
11-17.
7. Luke, M.A.; Froberg, J.E.; Doose, G.M.; Masumoto, H.T. "Improved
Multiresidue Gas Chromatographic Determination of Organophosphorus,
Organonitrogen, and Organohalogen Pesticides in Produce, Using Flame
Photometric and Electrolytic Conductivity Detectors"; jh. Assoc. Off. Anal.
Chem. 1981, 1187. 64.
8. Sherma, J.; Berzoa, M. "Analysis of Pesticide Residues in Human and
Environmental Samples"; U.S. Environmental Protection Agency. Research
Triangle Park, NC; EPA-600/8-80-038.
9. Desmarchelier, J.M.; Wustner, D.A.; Fukuto, T.R. "Mass Spectra of
Organophosphorus Esters and Their Alteration Products"; Residue Reviews.
1974, pp 63, 77.
8141A - 11 Revision 1
November 1990
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TABLE 1.
METHOD DETECTION LIMITS IN A WATER AND A SOIL
MATRIX USING A FLAME PHOTOMETRIC DETECTOR
Compound
Reagent
Water (3510)8
Soil (3540)b
(M9/Kg)
Azinphos methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton, 0, S
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Naled
Parathion-ethyl
Parathion-methyl
Phorate
Ronnel
Sulfotep
TEPPC
Tetrachl orovi nphos
Tokuthion (Protothiofos)0
Trichloronate0
0.10
0.07
0.07
0.20
0.12
0.20
0.80
0.26
0.07
0.04
0.20
0.08
0.08
0.11
0.20
0.50
0.50
0.06
0.12
0.04
0.07
0.07
0.80
0.80
0.07
0.80
5.0
3.5
5.0
10.0
6.0
10.0
40.0
13.0
3.5
2.0
10.0
4.0
5.0
5.5
10.0
25.0
25.0
3.0
6.0
2.0
3.5
3.5
40.0
40.0
5.5
40.0
Sample extracted using Method 3510, Separatory Funnel Liquid-Liquid
Extraction.
Sample extracted using Method 3540, Soxhlet Extraction.
Purity of these standards not established by the EPA Pesticides and
Industrial Chemicals Repository, RTP, NC.
8141A - 12
Revision 1
November 1990
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TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS
(EQL) FOR VARIOUS MATRICES8
Matrix Factor"
Ground water (Methods 3510 or 3520) 10
Low-concentration soil by Soxhlet and no cleanup 10
Low-concentration soil by ultrasonic extraction with GPC cleanup 6.7
High-concentration soil and sludges by ultrasonic extraction 500
Non-water miscible waste (Method 3580) 1000
c
c
1C
Sample EQLs are highly matrix dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
Multiply this factor times the soil MDL.
8141A - 13 Revision 1
November 1990
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TABLE 3.
RETENTION TIMES FOR METHOD 8141 ANALYTES
Compound
TEPP
Dichlorvos
Mevinphos
Demeton, 0 and S
Ethoprop
Naled
Phorate
Monocrotophos
Sul f otep
Dimethoate
Disulfoton
Diazinon
Merphos
Ronnel
Chlorpyrifos
Malathion
Parathion, methyl
Parathion, ethyl
Trichloronate
Tetrachlorovinphos
Tokuthion (Protothiofos)
Fensulfothion
Bolstar^ (Sulprofos)
Famphur*
EPN
Azinphos methyl
Fenthion
Coumaphos
"Method 8141 has not been fully val
Initial temperature
Initial time
Program 1 rate
Program 1 final temperature
Program 1 hold
Program 2 rate
Program 2 final temperature
Program 2 hold
Caoill
DBS
6.44
9.63
14.178
18.31
18.618
19.01
19.94
20.04
20.11
20.636
23.71
24.27
26.82
29.23
31.17
31.72
31.84
31.85
32.19
34.65
34.67
35.85
36.34
36.40
37.80
38.342
38.83
39.83
arv Column
SPB608
5.12
7.91
12.88
15.90
16.48
17.40
17.52
20.11
18.02
20.18
19.96
20.02
21.73
22.98
26.88
28.78
23.71
27.62
28.41
32.99
24.58
35.20
35.08
36.93
36.71
38.04
29.45
38.87
DB210
10.66
12.79
18.44
17.24
18.67
19.35
18.19
31.42
19.58
27.96
20.66
19.68
32.44
23.19
25.18
32.58
32.17
33.39
29.95
33.68
39.913
36.80
37.55
37.86
36.74
37.24
28.86
39.47
i dated for Famphur.
130°C
3 minutes
5°C/min
180°C
10 minutes
2°C/min
250°C
15 minutes
50°C
1 minute
5°C/min
140°C
10 minutes
10°C/min
240°C
10 minutes
50°C
1 minute
5°C/min
140°C
10 minutes
10°C/nrin
240°C
10 minutes
8141A - 14
Revision 1
November 1990
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TABLE 4.
RECOVERY OF 27 ORGANOPHOSPHATES BY SEPARATORY FUNNEL EXTRACTION
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
126
134
7
103
33
136
80
NR
48
113
82
84
NR
127
NR
NR
NR
NR
101
NR
94
67
87
96
79
NR
NR
Medium
143 + 8
141 + 8
89 + 6
90 + 6
67 + 11
121 + 9.5
79 + 11
47 + 3
92 + 7
125 + 9
90 + 6
82 + 12
48 + 10
92 + 6
79
NR
18 + 4
NR
94 + 5
46 + 4
77 + 6
97 + 5
85 + 4
55 + 72
90 + 7
45 + 3
35
High
101
101
86
96
74
82
72
101
84
97
80
96
89
86
81
55
NR
NR
86
44
73
87
83
63
80
90
94
NR = Not recovered.
8141A - 15
Revision 1
November 1990
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TABLE 5.
RECOVERY OF 27 ORGANOPHOSPHATES BY CONTINUOUS LIQUID EXTRACTION
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Dlazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Famphur
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
NR
NR
13
94
38
NR
81
NR
94
NR
39
--
90
8
105
NR
NR
NR
NR
106
NR
84
82
40
39
56
132
NR
Medium
129
126
82 + 4
79 + 1
23 + 3
128 + 37
32 + 1
10 + 8
69 + 5
104 + 18
76 + 2
63 + 15
67 + 26
32 + 2
87 + 4
80
87
30
NR
81 + 1
50 + 30
63 + 3
83 + 7
77 + 1
18 + 7
70 + 14
32 + 14
NR
High
122
128
88
89
41
118
74
102
81
119
83
--
90
86
86
79
49
1
74
87
43
74
89
85
70
83
90
21
NR = Not recovered.
8141A - 16
Revision 1
November 1990
-------
TABLE 6.
RECOVERY OF 27 ORGANOPHOSPHATES BY SOXHLET EXTRACTION
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dlmethoate
Disulfoton
EPN
Ethoprop
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
156
102
NR
93
169
87
84
NR
78
114
65
72
NR
100
62
NR
NR
NR
75
NR
75
NR
67
36
50
NR
56
Medium
110 + 6
103 + 15
66 + 17
89 + 11
64 + 6
96 + 3
39 + 21
48 + 7
78 + 6
93 + 8
70 + 7
81 + 18
43 + 7
81+8
53
71
NR
48
80 + 8
41+3
77 + 6
83 + 12
72 + 8
34 + 33
81 + 7
40 + 6
53
High
87
79
79
90
75
75
71
98
76
82
75
111
89
81
60
63
NR
NR
80
28
78
79
78
63
83
89
53
NR = Not recovered.
8141A - 17
Revision 1
November 1990
-------
TABLE 7.
RECOVERY OF 27 ORGANOPHOSPHATES BY ULTRASONIC EXTRACTION
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthlon
Trichloroate
Low
NR
NR
NR
NR
NR
NR
41
NR
30
14
19
NR
NR
55
NR
NR
NR
82
NR
63
NR
70
NR
43
NR
NR
NR
Medium
27 + 10
103 + 15
79 + 7
60
NR
90 + 14
13 + 9
67
44 + 22
86 + 38
34 + 26
37
35
67
71
NR
NR
40
74 + 13
NR
51 + 9
84 + 8
68 + 10
7
47 + 24
NR
NR
High
21
114
77
15
16
78
27
NR
69
105
35
2
84
31
155
23
NR
33
75
17
64
81
76
3
69
82
31
NR = Not recovered.
8141A - 18
Revision 1
November 1990
-------
FIGURE 1.
CHARACTERISTIC RESPONSE OF ORGANOPHOSPHATES ON DB210 WITH FPD DETECTOR
300.00
250.00
200.00
150.00
100.00
50.00
0.00
I
Q
ll
1
at g c
* 9- 1
c o
faraipinr
Tetrachlorovi
FensuM
' V... Jl
I (...) I .. | I > I | I I I | I I I | I I •) I I I ) I I I | I I I f , I I | , I ,, I <. | I . I ( !,,,...,,.,,, I I ,,,,,,.., I , I ,..!,
3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
8141A - 19
Revision 1
November 1990
-------
FIGURE 2.
CHARACTERISTIC RESPONSE OF ORGANOPHOSPHATES ON DB210 WITH NPD DETECTOR
300.00
250.00
200.00-
150.00-
100.00-
50.00-
»^ - i
0.00 -1
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
8141A - 20
Revision 1
November 1990
-------
FIGURE 3.
CHROMATOGRAM OF ORGANOPHOSPHATES ON DB210 WITH FPD DETECTOR
300.00
250.00
200.00
150.00
100.00
50.00
0.00
V
Q.
UJ
o
0.
•• i
, I , I ,,,..,, ,, , •. I,...,,,. I I. , ,11 ,, I .. I ...,..., , . I I ,, . I, , ., ,, ,, . ,,, I,..,,,,,,, ,.,,,.,
3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
8141A - 21
Revision 1
November 1990
-------
FIGURE 4.
CHROMATOGRAM OF ORGANOPHOSPHATES ON DB210 WITH NPD DETECTOR
300.00 -,
250.00-
200.00-
150.00-
100.00-
50.00 -I
DL
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
8141A - 22
Revision 1
November 1990
-------
METHOD 8141A
ORGANOPHOSPHORUS COMPOUNDS
I Start 1
1
7.1.1 Refer to
Chapter Two for
guidance on
choosing the
appropr late
ex t ract ion
pr ocedur e
7.1.2 Perform
s o 1 vent exchange
during K-D
procedures in all
extraction methods
1
7.2 Select CC
condi tions
1
7.3 Refer to Method
8000 for
ca 1 ibration
techniques
I
7.3.1 In ternal or
ex ternal
calibration may be
used
-J
-*
7.4.1 Add internal
standard to sampl e
if necessary
7.4.2 Refer to
Method 8000. Step
7.6 for
ins t ructions on
analysis sequence,
di lutions ,
retention times ,
and identification
criteria
I
7.4.3 Inject sample
1
7.4.5 Record sample
volume injected and
resulting peak
sizes
I
7.4.6 Determine
identity and
quantity of each
component peak:
refer to Method — '
8000, Step 7.8 for
calculation
equations
Yes
7.5.1 Perform
appropriate cleanup
No
7.5.2 Reanalyze by
CC
Stc
8141A - 23
Revision 1
November 1990
-------
METHOD 8150B
CHLORINATED HERBICIDES BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8150 is a gas chromatographic (GC) method for determining
certain chlorinated acid herbicides. The following compounds can be determined
by this method:
Compound Name CAS No."
2,4-D 94-75-7
2,4-DB 94-82-6
2,4,5-TP (Silvex) 93-72-1
2,4,5-T 93-76-5
Dalapon 75-99-0
Dicamba 1918-00-9
Dichlorprop 120-36-5
Dinoseb 88-85-7
MCPA 94-74-6
MCPP 93-65-2
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limit for each compound in organic-
free reagent water. Table 2 lists the estimated quantitation limit (EQL) for
other matrices.
1.3 When Method 8150 is used to analyze unfamiliar samples, compound
identifications should be supported by at least one additional qualitative
technique. This method describes analytical conditions for a second gas
chromatographic column that can be used to confirm measurements made with the
primary column. Section 8.4 provides gas chromatograph/mass spectrometer (GC/MS)
criteria appropriate for the qualitative confirmation of compound
identifications.
1.4 Only experienced analysts should be allowed to work with diazomethane
due to the potential hazards associated with its use (the compound is explosive
and carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Method 8150 provides extraction, esterification, and gas
chromatographic conditions for the analysis of chlorinated acid herbicides.
Spiked samples are used to verify the applicability of the chosen extraction
technique to each new sample type. The esters are hydrolyzed with potassium
8150B - 1 Revision 2
November 1990
-------
hydroxide, and extraneous organic material is removed by a solvent wash. After
acidification, the acids are extracted with solvent and converted to their methyl
esters using diazomethane as the derivatizing agent. After excess reagent is
removed, the esters are determined by gas chromatography employing an electron
capture detector, microcoulometric detector, or electrolytic conductivity
detector (Goerlitz and Lamar, 1967). The results are reported as the acid
equivalents.
2.2 The sensitivity of Method 8150 usually depends on the level of
interferences rather than on instrumental limitations.
3.0 INTERFERENCES
3.1 Refer to Method 8000.
3.2 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination. Phenols, including chlorophenols, may also
interfere with this procedure.
3.3 Alkaline hydrolysis and subsequent extraction of the basic solution
remove many chlorinated hydrocarbons and phthalate esters that might otherwise
interfere with the electron capture analysis.
3.4 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Therefore, glassware and
glass wool must be acid rinsed, and sodium sulfate must be acidified with
sulfuric acid prior to use to avoid this possibility.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Columns
4.1.2.1 Column la and Ib - 1.8 m x 4 mm ID glass, packed with
1.5% SP-2250/1.95% SP-2401 on Supelcoport (100/120 mesh) or
equivalent.
4.1.2.2 Column 2 - 1.8 m x 4 mm ID glass, packed with 5% 0V-
210 on Gas Chrom Q (100/120 mesh) or equivalent.
4.1.2.3 Column 3 - 1.98 m x 2 mm ID glass, packed with 0.1%
SP-1000 on 80/100 mesh Carbopack C or equivalent.
4.1.3 Detector - Electron capture (ECD).
8150B - 2 Revision 2
November 1990
-------
4.2 Erlenmeyer flasks - 250 and 500 mL Pyrex, with 24/40 ground glass
joint.
4.3 Beaker - 500 ml.
4.4 Diazomethane generator - Refer to Section 7.4 to determine which
method of diazomethane generation should be used for a particular application.
4.4.1 Diazald kit - recommended for the generation of diazomethane
using the procedure given in Section 7.4.2 (Aldrich Chemical Co., Cat. No.
210,025-2 or equivalent).
4.4.2 Assemble from two 20 x 150 mm test tubes, two Neoprene rubber
stoppers, and a source of nitrogen. Use Neoprene rubber stoppers with
holes drilled in them to accommodate glass delivery tubes. The exit tube
must be drawn to a point to bubble diazomethane through the sample extract.
The generator assembly is shown in Figure 1. The procedure for use of this
type of generator is given in Section 7.4.3.
4.5 Vials - 10 to 15 ml, amber glass, with Teflon lined screw cap or crimp
top.
4.6 Separatory funnel - 2000 mL, 125 ml, and 60 ml.
4.7 Drying column - 400 mm x 20 mm ID Pyrex chromatographic column with
Pyrex glass wool at bottom and a Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits may
be purchased. Use a small pad of Pyrex glass wool to retain the adsorbent.
Prewash the glass wool pad with 50 ml of acetone followed by 50 ml of
elution solvent prior to packing the column with adsorbent.
4.8 Kuderna-Danish (K-D) apparatus
4.8.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation of
extracts
4.8.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.8.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.8.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.8.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.9 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
8150B - 3 Revision 2
November 1990
-------
4.10 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.11 Microsyringe - 10 ^l.
4.12 Wrist shaker - Burrell Model 75 or equivalent.
4.13 Glass wool - Pyrex, acid washed.
4.14 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.15 Syringe - 5 ml.
4.16 Glass rod.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sulfuric acid solution
5.3.1 ((1:1) (v/v)) - Slowly add 50 ml H2S04 (sp. gr. 1.84) to 50 ml
of organic-free reagent water.
5.3.2 ((1:3) (v/v)) - Slowly add 25 ml H2S04 (sp. gr. 1.84) to 75 ml
of organic-free reagent water.
5.4 Hydrochloric acid ((1:9) (v/v)), HC1. Add one volume of concentrated
HC1 to 9 volumes of organic-free reagent water.
5.5 Potassium hydroxide solution (KOH) - 37% aqueous solution (w/v).
Dissolve 37 g potassium hydroxide pellets in organic-free reagent water, and
dilute to 100 ml.
5.6 Carbitol (Diethylene glycol monoethyl ether), C2H5OCH2CH2OCH2CH2OH.
Available from Aldrich Chemical Co.
5.7 Solvents
5.7.1 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.7.2 Methanol, CH3OH - Pesticide quality or equivalent.
8150B - 4 Revision 2
November 1990
-------
5.7.3 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.7.4 Hexane, C6H14 - Pesticide quality or equivalent.
5.7.5 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.8 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.9 N-Methyl-N-nitroso-p-toluenesulfonamide (Diazald), CH3C6H4S02N(CH3)NO.
Available from Aldrich Chemical Co.
5.10 Silicic acid. Chromatographic grade, nominal 100 mesh. Store at
130°C.
5.11 Stock standard solutions - Stock standard solutions can be prepared
from pure standard materials or purchased as certified solutions.
5.11.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure acids. Dissolve the acids in pesticide quality acetone
and dissolve the esters in 10% acetone/isooctane (v/v) and dilute to volume
in a 10 ml volumetric flask. Larger volumes can be used at the convenience
of the analyst. If compound purity is certified at 96% or greater, the
weight can be used without correction to calculate the concentration of
the stock standard. Commercially prepared stock standards can be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.11.2 Transfer the stock standard solutions into vials with Teflon
lined screw caps or crimp tops. Store at 4°C and protect from light. Stock
standard solutions should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards
from them.
5.11.3 Stock standard solutions must be replaced after 1 year, or
sooner if comparison with check standards indicates a problem.
5.12 Calibration standards - A minimum of five calibration standards for
each parameter of interest should be prepared through dilution of the stock
standards with diethyl ether. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples or should define the working range of the GC. Calibration
solutions must be replaced after six months, or sooner if comparison with check
standards indicates a problem.
8150B - 5 Revision 2
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5.13 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.13.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest as described in Section 5.12.
5.13.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with hexane.
5.13.3 Analyze each calibration standard according to Section 7.0.
5.14 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and organic-free reagent water blank with one or two herbicide
surrogates (e.g. herbicides that are not expected to be present in the sample).
The surrogates selected should elute over the range of the temperature program
used in this method. 2,4-Dichlorophenylacetic acid (DCAA) is recommended as a
surrogate compound. Deuterated analogs of analytes should not be used as
surrogates for gas chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within
40 days of extraction.
7.0 PROCEDURE
7.1 Preparation of waste samples
7.1.1 Extraction
7.1.1.1 Follow Method 3580 except use diethyl ether as the
dilution solvent, acidified anhydrous sodium sulfate, and acidified
glass wool.
7.1.1.2 Transfer 1.0 mL (a lesser volume or a dilution may
be required if herbicide concentrations are high) to a 250 mL ground
glass-stoppered Erlenmeyer flask. Proceed to Section 7.2.2
hydrolysis.
7.2 Preparation of soil, sediment, and other solid samples
7.2.1 Extraction
7.2.1.1 To a 500 mL, wide mouth Erlenmeyer flask add 50 g (dry
weight) of the well mixed, moist solid sample. Adjust the pH to 2
8150B - 6 Revision 2
November 1990
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with concentrated HC1 and monitor the pH for 15 minutes with
occasional stirring. If necessary, add additional HC1 until the pH
remains at 2.
7.2.1.2 Add 20 ml acetone to the flask and mix the contents
with the wrist shaker for 20 minutes. Add 80 ml diethyl ether to
the same flask and shake again for 20 minutes. Decant the extract
and measure the volume of solvent recovered.
7.2.1.3 Extract the sample twice more using 20 ml of acetone
followed by 80 ml of diethyl ether. After addition of each solvent,
the mixture should be shaken with the wrist shaker for 10 minutes
and the acetone-ether extract decanted.
7.2.1.4 After the third extraction, the volume of extract
recovered should be at least 75% of the volume of added solvent.
If this is not the case, additional extractions may be necessary.
Combine the extracts in a 2 liter separatory funnel containing 250 ml
of 5% acidified sodium sulfate. If an emulsion forms, slowly add
5 g of acidified sodium sulfate (anhydrous) until the solvent-water
mixture separates. A quantity of acidified sodium sulfate equal to
the weight of the sample may be added, if necessary.
7.2.1.5 Check the pH of the extract. If it is not at or below
pH 2, add more concentrated HC1 until stabilized at the desired pH.
Gently mix the contents of the separatory funnel for 1 minute and
allow the layers to separate. Collect the aqueous phase in a clean
beaker and the extract phase (top layer) in a 500 ml ground glass-
stoppered Erlenmeyer flask. Place the aqueous phase back into the
separatory funnel and re-extract using 25 ml of diethyl ether. Allow
the layers to separate and discard the aqueous layer. Combine the
ether extracts in the 500 ml Erlenmeyer flask.
7.2.2 Hydrolysis
7.2.2.1 Add 30 ml of organic-free reagent water, 5 ml of 37%
KOH, and one or two clean boiling chips to the flask. Place a three
ball Snyder column on the flask, evaporate the diethyl ether on a
water bath, and continue to heat for a total of 90 minutes.
7.2.2.2 Remove the flask from the water bath and allow to
cool. Transfer the water solution to a 125 mL separatory funnel and
extract the basic solutions once with 40 mL and then twice with 20 ml
of diethyl ether. Allow sufficient time for the layers to separate
and discard the ether layer each time. The phenoxy-acid herbicides
remain soluble in the aqueous phase as potassium salts.
7.2.3 Solvent cleanup
7.2.3.1 Adjust the pH to 2 by adding 5 mL cold (4°C) sulfuric
acid (1:3) to the separatory funnel. Be sure to check the pH at this
point. Extract the herbicides once with 40 mL and twice with 20 mL
of diethyl ether. Discard the aqueous phase.
8150B - 7 Revision 2
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7.2.3.2 Combine ether extracts in a 125 ml Erlenmeyer flask
containing 5-7 g of acidified anhydrous sodium sulfate. Stopper and
allow the extract to remain in contact with the acidified sodium
sulfate. If concentration and esterification are not to be performed
immediately, store the sample overnight in the refrigerator.
NOTE: The drying step is very critical to ensuring complete esterification. Any
moisture remaining in the ether will result in low herbicide recoveries.
The amount of sodium sulfate is adequate if some free flowing crystals are
visible when swirling the flask. If all the sodium sulfate solidifies in
a cake, add a few additional grams of acidified sodium sulfate and again
test by swirling. The 2 hour drying time is a minimum, however, the
extracts may be held overnight in contact with the sodium sulfate.
7.2.3.3 Transfer the ether extract, through a funnel plugged
with acid washed glass wool, into a 500 ml K-D flask equipped with
a 10 ml concentrator tube. Use a glass rod to crush caked sodium
sulfate during the transfer. Rinse the Erlenmeyer flask and column
with 20-30 ml of diethyl ether to complete the quantitative transfer.
7.2.3.4 Add one or two clean boiling chips to the flask and
attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 ml of diethyl ether to the top. Place the apparatus
on a hot water bath (60°-65°C) so that the concentrator tube is
partially immersed in the hot water and the entire lower rounded
surface of the flask is bathed in vapor. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 15-20 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus from the water bath and allow
it to drain and cool for at least 10 minutes.
7.2.3.5 Remove the Snyder column and rinse the flask and its
lower joints into the concentrator tube with 1-2 ml of diethyl ether.
A 5 ml syringe is recommended for this operation. Add a fresh
boiling chip, attach a micro Snyder column to the concentrator tube,
and prewet the column by adding 0.5 ml of ethyl ether to the top.
Place the micro K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature as
required to complete concentration in 5-10 minutes. When the
apparent volume of the liquid reaches 0.5 ml, remove the micro K-D
from the bath and allow it to drain and cool. Remove the Snyder
column and add 0.1 ml of methanol. Rinse the walls of the
concentrator tube while adjusting the extract volume to 1.0 ml with
diethyl ether. Proceed to Section 7.4 for esterification.
7.3 Preparation of aqueous samples
7.3.1 Extraction
7.3.1.1 Using a 1 liter graduated cylinder, measure 1 liter
(nominal) of sample, record the sample volume to the nearest 5 mL,
8150B - 8 Revision 2
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and transfer it to the separatory funnel. If high concentrations
are anticipated, a smaller volume may be used and then diluted with
organic-free reagent water to 1 liter. Adjust the pH to less than
2 with sulfuric acid (1:1).
7.3.1.2 Add 150 ml of diethyl ether to the sample bottle,
seal, and shake for 30 seconds to rinse the walls. Transfer the
solvent wash to the separatory funnel and extract the sample by
shaking the funnel for 2 minutes with periodic venting to release
excess pressure. Allow the organic layer to separate from the water
layer for a minimum of 10 minutes. If the emulsion interface between
layers is more than one third the size of the solvent layer, the
analyst must employ mechanical techniques to complete the phase
separation. The optimum technique depends upon the sample and may
include stirring, filtration of the emulsion through glass wool,
centrifugation, or other physical methods. Drain the aqueous phase
into a 1 liter Erlenmeyer flask. Collect the solvent extract in a
250 mL ground glass Erlenmeyer flask containing 2 ml of 37% KOH.
Approximately 80 ml of the diethyl ether will remain dissolved in
the aqueous phase.
7.3.1.3 Repeat the extraction two more times using 50 mL of
diethyl ether each time. Combine the extracts in the Erlenmeyer
flask. (Rinse the 1 liter flask with each additional aliquot of
extracting solvent.)
7.3.2 Hydrolysis
7.3.2.1 Add one or two clean boiling chips and 15 mL of
organic-free reagent water to the 250 mL flask and attach a three
ball Snyder column. Prewet the Snyder column by adding about 1 mL
of diethyl ether to the top of the column. Place the apparatus on
a hot water bath (60°-65°C) so that the bottom of the flask is bathed
with hot water vapor. Although the diethyl ether will evaporate in
about 15 minutes, continue heating for a total of 60 minutes,
beginning from the time the flask is placed in the water bath.
Remove the apparatus and let stand at room temperature for at least
10 minutes.
7.3.2.2 Transfer the solution to a 60 mL separatory funnel
using 5-10 mL of organic-free reagent water. Wash the basic solution
twice by shaking for 1 minute with 20 mL portions of diethyl ether.
Discard the organic phase. The herbicides remain in the aqueous
phase.
7.3.3 Solvent cleanup
7.3.3.1 Acidify the contents of the separatory funnel to pH 2
by adding 2 mL of cold (4°C) sulfuric acid (1:3). Test with pH
indicator paper. Add 20 mL diethyl ether and shake vigorously for
2 minutes. Drain the aqueous layer into a 250 mL Erlenmeyer flask,
and pour the organic layer into a 125 mL Erlenmeyer flask containing
about 5-7 g of acidified sodium sulfate. Repeat the extraction twice
more with 10 mL aliquots of diethyl ether, combining all solvent in
8150B - 9 Revision 2
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the 125 ml flask. Allow the extract to remain in contact with the
sodium sulfate for approximately 2 hours.
NOTE: The drying step is very critical to ensuring complete esterification. Any
moisture remaining in the ether will result in low herbicide recoveries.
The amount of sodium sulfate is adequate if some free flowing crystals are
visible when swirling the flask. If all the sodium sulfate solidifies in
a cake, add a few additional grams of acidified sodium sulfate and again
test by swirling. The 2 hour drying time is a minimum, however, the
extracts may be held overnight in contact with the sodium sulfate.
7.3.3.2 Transfer the ether extract, through a funnel plugged
with acid washed glass wool, into a 500 mL K-D flask equipped with
a 10 ml concentrator tube. Use a glass rod to crush caked sodium
sulfate during the transfer. Rinse the Erlenmeyer flask and column
with 20-30 ml of diethyl ether to complete the quantitative transfer.
7.3.3.3 Add one or two clean boiling chips to the flask and
attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 ml of diethyl ether to the top. Place the apparatus
on a hot water bath (60°-65°C) so that the concentrator tube is
partially immersed in the hot water and the entire lower rounded
surface of the flask is bathed in vapor. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 15-20 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus from the water bath and allow
it to drain and cool for at least 10 minutes.
7.3.3.4 Remove the Snyder column and rinse the flask and its
lower joints into the concentrator tube with 1-2 ml of diethyl ether.
A 5 mL syringe is recommended for this operation. Add a fresh
boiling chip, attach a micro Snyder column to the concentrator tube,
and prewet the column by adding 0.5 ml of ethyl ether to the top.
Place the micro K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature as
required to complete concentration in 5-10 minutes. When the
apparent volume of the liquid reaches 0.5 mL, remove the micro K-D
from the bath and allow it to drain and cool. Remove the Snyder
column and add 0.1 mL of methanol. Rinse the walls of the
concentrator tube while adjusting the extract volume to 1.0 mL with
diethyl ether.
7.4 Esterification
7.4.1 Two methods may be used for the generation of diazomethane:
the bubbler method (set up shown in Figure 1) and the Diazald kit method.
The bubbler method is suggested when small batches (10-15) of samples
require esterification. The bubbler method works well with samples that
have low concentrations of herbicides (e.g. aqueous samples) and is safer
to use than the Diazald kit procedure. The Diazald kit method is good for
large quantities of samples needing esterification. The Diazald kit method
8150B - 10 Revision 2
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is more effective than the bubbler method for soils or samples that may
contain high concentrations of herbicides (e.g. samples such as soils that
may result in yellow extracts following hydrolysis may be difficult to
handle by the bubbler method). The diazomethane derivatization (U.S. EPA,
1971) procedures, described below, will react efficiently with all of the
chlorinated herbicides described in this method and should be used only
by experienced analysts, due to the potential hazards associated with its
use. The following precautions should be taken:
CAUTION: Diazomethane is a carcinogen and can explode under certain
conditions.
Use a safety screen.
Use mechanical pipetting aides.
Do not heat above 90°C -- EXPLOSION may result.
Avoid grinding surfaces, ground glass joints, sleeve bearings, glass
stirrers -- EXPLOSION may result.
Store away from alkali metals -- EXPLOSION may result.
Solutions of diazomethane decompose rapidly in the presence of solid
materials such as copper powder, calcium chloride, and boiling chips.
7.4.2 Diazald kit method - Instructions for preparing diazomethane
are provided with the generator kit.
7.4.2.1 Add 2 mL of diazomethane solution and let sample stand
for 10 minutes with occasional swirling.
7.4.2.2 Rinse inside wall of ampule with several hundred pi
of diethyl ether. Allow solvent to evaporate spontaneously at room
temperature to about 2 mL.
7.4.2.3 Dissolve the residue in 5 mL of hexane. Analyze by
gas chromatography.
7.4.3 Bubbler method - Assemble the diazomethane bubbler (see
Figure 1).
7.4.3.1 Add 5 mL of diethyl ether to the first test tube.
Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5 mL of 37% KOH, and
0.1-0.2 g Diazald to the second test tube. Immediately place the
exit tube into the concentrator tube containing the sample extract.
Apply nitrogen flow (10 mL/min) to bubble diazomethane through
the extract for 10 minutes or until the yellow color of diazomethane
persists. The amount of Diazald used is sufficient for
esterification of approximately three sample extracts. An additional
0.1-0.2 g of Diazald may be added (after the initial Diazald is
consumed) to extend the generation of the diazomethane. There is
sufficient KOH present in the original solution to perform a maximum
of approximately 20 minutes of total esterification.
8150B - 11 Revision 2
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7.4.3.2 Remove the concentrator tube and seal it with a
Neoprene or Teflon stopper. Store at room temperature in a hood for
20 minutes.
7.4.3.3 Destroy any unreacted diazomethane by adding 0.1-0.2 g
silicic acid to the concentrator tube. Allow to stand until the
evolution of nitrogen gas has stopped. Adjust the sample volume to
10.0 mL with hexane. Stopper the concentrator tube and store
refrigerated if further processing will not be performed immediately.
It is recommended that the methylated extracts be analyzed
immediately to minimize the trans-esterification and other potential
reactions that may occur. Analyze by gas chromatography.
7.5 Gas chromatographic conditions (Recommended)
7.5.1 Column la
Carrier gas (5% methane/95% argon) flow rate:
Temperature program: 185°C, isothermal.
70 mL/min
7.5.2 Column Ib
Carrier gas (5% methane/95% argon) flow rate: 70 mL/min
Initial temperature: 140°C, hold for 6 minutes
Temperature program: 140°C to 200°C at 10°C/min, hold until last compound
has eluted.
7.5.3 Column 2
Carrier gas (5% methane/95% argon) flow rate:
Temperature program: 185°C, isothermal.
70 mL/min
7.5.4 Column 3
Carrier gas (ultra-high purity N2) flow rate: 25 mL/min
Initial temperature: 100°C, no hold
Temperature program: 100°C to 150°C at 10°C/min, hold until last compound
has eluted.
7.6 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.6.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.6.2 The following gas chromatographic columns are recommended for
the compounds indicated:
Analvte
Dicamba
2,4-D
2,4,5-TP
2,4,5-T
2,4-DB
Column
la,2
la,2
la,2
la,2
la
Analvte
Dalapon
MCPP
MCPA
Dichloroprop
Dinoseb
Column
3
Ib
Ib
Ib
Ib
8150B - 12
Revision 2
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7.7 Gas chromatographic analysis
7.7.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 p.1 of internal standard to the sample prior to
injection.
7.7.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration check standard after
each group of 10 samples in the analysis sequence.
7.7.3 Examples of chromatograms for various chlorophenoxy herbicides
are shown in Figures 2 through 4.
7.7.4 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
7.7.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.7.6 If calibration standards have been analyzed in the same manner
as the samples (e.g. have undergone hydrolysis and esterification), then
the calculation of concentration given in Method 8000 should be used.
However, if calibration is done using standards made from methyl ester
compounds (compounds not esterified by application of this method), then
the calculation of concentration must include a correction for the
molecular weight of the methyl ester versus the acid herbicide.
7.7.7 If peak detection and identification are prevented due to
interferences, further cleanup is required. Before using any cleanup
procedure, the analyst must process a series of standards through the
procedure to validate elution patterns and the absence of interferences
from reagents.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000.
8.2.1 Select a representative spike concentration for each compound
(acid or ester) to be measured. Using stock standards, prepare a quality
control check sample concentrate in acetone 1,000 times more concentrated
than the selected concentrations.
8150B - 13 Revision 2
November 1990
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8.2.2 Table 3 indicates Single Operator Accuracy and Precision for
this method. Compare the results obtained with the results given in
Table 3 to determine if the data quality is acceptable.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures
are required.
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4 GC/MS confirmation
8.4.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Refer to Method 8270
for the appropriate GC/MS operating conditions and analysis procedures.
8.4.2 When available, chemical ionization mass spectra may be
employed to aid the qualitative identification process.
8.4.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps may
include the use of alternate packed or capillary GC columns or additional
cleanup.
9.0 METHOD PERFORMANCE
9.1 In a single laboratory, using organic-free reagent water and effluents
from publicly owned treatment works (POTW), the average recoveries presented in
Table 3 were obtained. The standard deviations of the percent recoveries of
these measurements are also included in Table 3.
10.0 REFERENCES
1. U.S. EPA, National Pollutant Discharge Elimination System, Appendix A, Fed.
Reg., 38, No. 75, Pt. II, Method for Chlorinated Phenoxy Acid Herbicides
in Industrial Effluents, Cincinnati, Ohio, 1971.
8150B - 14 Revision 2
November 1990
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2. Goerlitz, D.G., and W.L. Lamar, "Determination of Phenoxy Acid Herbicides
in Water by Electron Capture and Microcoulometric Gas Chromatography," U.S.
Geol. Survey Water Supply Paper, 1817-C, 1967.
3. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
4. U.S. EPA, "Extraction and Cleanup Procedure for the Determination of Phenoxy
Acid Herbicides in Sediment," EPA Toxicant and Analysis Center, Bay St.
Louis, Mississippi, 1972.
5. "Pesticide Methods Evaluation," Letter Report #33 for EPA Contract No. 68-
03-2697. Available from U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, Ohio 45268.
6. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry," Analytical Chemistry, 47, 995, 1975.
7. Glaser, J.A. et.al., "Trace Analysis for Wastewaters," Environmental Science
& Technology, 15, 1426, 1981.
8. U.S. EPA, "Method 615. The Determination of Chlorinated Herbicides in
Industrial and Municipal Wastewater," Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio, 45268, June 1982.
8150B - 15 Revision 2
November 1990
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND DETECTION LIMITS
FOR CHLORINATED HERBICIDES
Compound
Retention time (min)a
Col.la Col.lb Col.2 Col.3
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQL) FOR VARIOUS MATRICES3
Method
detection
limit (M9/L)
2,4-D
2,4-DB
2,4,5-T
2,4,5-TP (Silvex)
Dalapon
Dicamba
Dichloroprop
Dinoseb
MCPA
MCPP
2.0
4.1
3.4
2.7
-
1.2
-
-
-
-
.
-
-
-
-
-
4.8
11.2
4.1
3.4
1.6
-
2.4
2.0
5.0
1.0
-
-
-
- -
1.2
0.91
0.20
0.17
5.8
0.27
0.65
0.07
249
192
Matrix
Factor15
Ground water (based on one liter sample size)
Soil/sediment and other solids
Waste samples
10
200
100,000
"Sample EQLs are highly matrix dependent. The EQLs listed herein are provided for
guidance and may not always be achievable.
bEQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-aqueous
samples, the factor is on a wet weight basis.
8150B - 16
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TABLE 3.
SINGLE OPERATOR ACCURACY AND PRECISION8
Compound
2,4-D
Dalapon
2,4-DB
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
2,4,5-T
2,4,5-TP
Sample
Type
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
Spike
(M9/L)
10.9
10.1
200
23.4
23.4
468
10.3
10.4
208
1.2
1.1
22.2
10.7
10.7
213
0.5
102
2020
2020
21400
2080
2100
20440
1.1
1.3
25.5
1.0
1.3
25.0
Mean
Recovery
(%)
75
77
65
66
96
81
93
93
77
79
86
82
97
72
100
86
81
98
73
97
94
97
95
85
83
78
88
88
72
Standard
deviation
(%)
4
4
5
8
13
9
3
3
6
7
9
6
2
3
2
4
3
4
3
2
4
3
2
6
4
5
5
4
5
"All results based upon seven replicate analyses. Esterification performed using
the bubbler method. Data obtained from reference 9.
DW = ASTM Type II
MW = Municipal water
8150B - 17
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FIGURE 1.
DIAZOMETHANE GENERATOR
glass tubing
nitrogen
rubber stopper
lube 1
tube 2
8150B - 18
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FIGURE 2.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1.5% SP-2250/1.95* SP-2401 en Suptlcoport (100/120
Ttmpcriturt: Isothermal at 18S°C
Oflttetor: Electron Capture
0 12346
RETENTION TIME (MINUTES)
8150B - 19
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FIGURE 3.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1.5% SP-2250/1.95% SP-2401 on Suptlcoport (100/120 Mttfi)
Program: 140°C for 6 Min. 10°C/Minut§ to 200°C
Octtctor: ElKtron Capturt
4 68
RETENTION TIME (MINUTES)
10
12
8150B - 20
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FIGURE 4.
GAS CHROMATOGRAM OF DALAPON, COLUMN 3
Column: 0.1% SP-1000 on 80/100 Mtsh Cartaopak C
Program: 100°C, 10°C/Min to 160°C
Ofttetor: Electron Coptura
0246
RETENTION TIME (MINUTES)
8150B - 21
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METHOD 8150B
CHLORINATED HERBICIDES
7.2.1.1
Adjust sample
pH with HC1.
Liquid
sample
7.2.1.2 Extract
sample three
times with
acetone and
diethyl ether.
7.2.1.4
Combine
extracts.
7.2.1.5 Check
pH of extract ,
adjust if
necessary,
Separate layers
7.2.1.5
Re-extract
and discard
aqueous
phase.
7.1.1.1 Follow
Method 3580 for
extraction, using
diethyl ether,
acidified anhydrous
sodium sulfate and
acidified glass
wool .
7.2.2 Proceed
with
hydrolysis.
7.1.1.2 Use
1.0 mL of
sample for
hydrolysis .
7.2.3 Proceed
with solvent
cleanup.
7.3.1 Extract
three times
with diethyl
ether.
7.3.1.3
Combine
extracts.
7.3.2 Proceed
with
hydrolysis .
7.3.3 Proceed
with solvent
cleanup.
8150B - 22
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METHOD 8150B
(Continued)
7.4.3 Assembe
diazomethane
bubbler;
generate
diazomethane
7.42 Prepare
diazomethane
according to
kit
instructions .
7.5 Set
chromatographic
condi t i ons.
7.6 Claibrate
according to
Method 8000.
7.6.2 Choose
appropriate
CC column.
7.7 Analyze
by CC (refer
to MEthod
8000) .
7.7.7 Do
interferences
prevent peak
detection?
7.7.7 Process
series of
standards
through system
cleanup.
8150B - 23
Revision 2
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METHOD 8151
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATION
DERIVATIZATION; CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8151 Is a capillary gas chromatographic (GC) method for
determining certain chlorinated acid herbicides in aqueous, soil and waste
matrices. Specifically, Method 8151 may be used to determine the following
compounds:
Compound Name CAS No."
Acifluorfen 50594-66-6
Bentazon 25057-89-0
Chloramben 133-90-4
2,4-D 94-75-7
Dalapon 75-99-0
2,4-DB 94-82-6
DCPA diacid" 2136-79-0
Dicamba 1918-00-9
3,5-Dichlorobenzoic acid 51-36-5
Dichlorprop 120-36-5
Dinoseb 88-85-7
5-Hydroxydicamba 7600-50-2
MCPA 94-74-6
MCPP 93-65-2
4-Nitrophenol 100-02-1
Pentachlorophenol 87-86-5
Picloram 1918-02-1
2,4,5-TP (Silvex) 93-72-1
2,4,5-T 93-76-5
a Chemical Abstract Services Registry Number.
b DCPA monoacid and diacid metabolites included in method scope; DCPA
diacid metabolite used for validation studies. DCPA is a dimethyl ester.
Because these compounds are produced and used in various forms (i.e., acid,
salt, ester, etc.), Method 8151 includes a hydrolysis step to convert the
herbicide to the acid form prior to analysis.
1.2 When Method 8151 is used to analyze unfamiliar samples, compound
identifications should be supported by at least one additional qualitative
technique. Section 8.4 provides gas chromatograph/mass spectrometer (GC/MS)
criteria appropriate for the qualitative confirmation of compound
identifications.
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1.3 The estimated detection limits for each of the compounds in aqueous
and soil matrices are listed in Table 1. The detection limits for a specific
waste sample may differ from those listed, depending upon the nature of the
interferences and the sample matrix.
1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.5 Only experienced analysts should be allowed to work with diazomethane
due to the potential hazards associated with its use (explosive, carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Method 8151 provides hydrolysis, extraction, derivatization and gas
chromatographic conditions for the analysis of chlorinated acid herbicides in
water, soil and waste samples.
2.1.1 Water samples are hydrolyzed in situ, extracted with diethyl
ether and then esterified with either diazomethane or pentafluorobenzyl
bromide. The derivatives are determined by gas chromatography with an
electron capture detector (GC/ECD). The results are reported as acid
equivalents.
2.1.2 Soil and waste samples are extracted, then hydrolyzed,
reextracted and esterified with either diazomethane or pentafluorobenzyl
bromide. The derivatives are determined by gas chromatography with an
electron capture detector (GC/ECD). The results are reported as acid
equivalents.
2.2 The sensitivity of Method 8151 depends on the level of interferences
in addition to instrumental limitations. Table 1 lists the GC/ECD and GC/MS
limits of detection that can be obtained in aqueous and soil matrices in the
absence of interferences. Detection limits for a typical waste sample should
be higher.
3.0 INTERFERENCES
3.1 Refer to Method 8000.
3.2 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis, by analyzing reagent blanks.
3.2.1 Glassware must be scrupulously cleaned. Clean each piece of
glassware as soon as possible after use by rinsing it with the last solvent
used in it. This should be followed by detergent washing with hot water
and rinses with tap water, then with organic-free reagent water. Glassware
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should be solvent-rinsed with acetone and pesticide-quality hexane. After
rinsing and drying, glassware should be sealed and stored in a clean
environment to prevent any accumulation of dust or other contaminants.
Store glassware inverted or capped with aluminum foil. Immediately prior
to use, glassware should be rinsed with the next solvent to be used.
3.2.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all-
glass systems may be required.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from waste to waste, depending upon the nature and diversity of
the waste being sampled.
3.4 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination by methyl ation. Phenols, including
chlorophenols, may also interfere with this procedure. The determination using
pentafluorobenzylation is more sensitive, and more prone to interferences from
the presence of organic acids or phenols than by methylation.
3.5 Alkaline hydrolysis and subsequent extraction of the basic solution
removes many chlorinated hydrocarbons and phthalate esters that might otherwise
interfere with the electron capture analysis.
3.6 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Therefore, glassware must
be acid-rinsed and then rinsed to constant pH with organic-free reagent water.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - Analytical system complete with gas
chromatograph suitable for Grob-type injection using capillary columns,
and all required accessories including detector, capillary analytical
columns, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Narrow Bore Columns
4.1.2.1.1 Primary Column 1 - 30 m x 0.25 mm, 5%
phenyl/95% methyl silicone (DB-5, J&W Scientific, or
equivalent), 0.25 urn film thickness.
4.1.2.1.2 Primary Column la (GC/MS) - 30 m x 0.32 mm,
5% phenyl/95% methyl silicone, (DB-5, J&W Scientific, or
equivalent), 1 jum film thickness.
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4.1.2.1.3 Column 2 - 30 m x 0.25 mm DB-608 (J&W
Scientific or equivalent) with a 25 ;um film thickness.
4.1.2.1.4 Confirmation Column - 30 m x 0.25 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or
equivalent), 0.25 pm film thickness.
4.1.2.2 Megabore Columns
4.1.2.2.1 Primary Column - 30 m x 0.53 mm DB-608 (J&W
Scientific or equivalent) with 0.83 /xm film thickness.
4.1.2.2.2 Confirmation Column - 30 m x 0.53 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or
equivalent), 1.0 /*m film thickness.
4.1.3 Detector - Electron Capture Detector (BCD)
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Diazomethane Generator: Refer to Section 7.5 to determine which
method of diazomethane generation should be used for a particular generation.
4.3.1 Diazald Kit - Recommended for the generation of diazomethane
(Aldrich Chemical Co., Cat No. 210,025-0, or equivalent).
4.3.2 Assemble from two 20 mm x 150 mm test tubes, two Neoprene
rubber stoppers, and a source of nitrogen. Use Neoprene rubber stoppers
with holes drilled in them to accommodate glass delivery tubes. The exit
tube must be drawn to a point to bubble diazomethane through the sample
extract. The generator assembly is shown in Figure 1. The procedure for
use of this type of generator is given in Section 7.5.1.1.
4.4 Other Glassware
4.4.1 Beaker - 400 ml, thick walled.
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4.4.2 Funnel - 75 mm diameter.
4.4.3 Separatory funnel - 500 ml, with Teflon stopcock.
4.4.4 Centrifuge bottle - 500 ml (Pyrex 1260 or equivalent).
4.4.5 Centrifuge bottle - 24/40 500 ml
4.4.6 Continuous Extractor (Hershberg-Wolfe type, Lab Glass No.
LG-6915, or equivalent)
4.4.7 Pipet - Pasteur, glass, disposable (140 mm x 5 mm ID).
4.4.8 Vials - 10 ml, glass, with Teflon lined screw-caps.
4.4.9 Volumetric flasks, Class A - 10 ml to 1000 ml.
4.5 Filter paper - 15 cm diameter (Whatman No. 1 or equivalent).
4.6 Glass Wool - Pyrex, acid washed.
4.7 Boiling chips - Solvent extracted with methylene
chloride,approximately 10/40 mesh (silicon carbide or equivalent).
4.8 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
4.9 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.10 Centrifuge.
4.11 Ultrasonic preparation - A horn-type device equipped with a titanium
tip, or a device that will give equivalent performance, shall be used.
4.11.1 Ultrasonic Disrupter - The disrupter must have a minimum
power wattage of 300 watts, with pulsing capability. A device designed
to reduce the cavitation sound is recommended. Follow the manufacturers
instructions for preparing the disrupter for extraction of samples. Use
a 3/4" horn for most samples.
4.12 Sonabox - Recommended with above disrupters for decreasing cavitation
sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.13 Filter paper - Whatman #1, or equivalent.
4.14 pH paper.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
8151 - 5 Revision 0
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such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free water, as defined in Chapter One.
5.3 Sodium hydroxide solution (0.1 N), NaOH. Dissolve 4 g NaOH in
organic-free reagent water and dilute to 1.0 L.
5.4 Potassium hydroxide solution (37% aqueous solution (w/v)), KOH.
Dissolve 37 g potassium hydroxide pellets in organic-free reagent water and
dilute to 100 ml.
5.5 Phosphate buffer pH = 2.5 (0.1 M). Dissolve 12 g sodium phosphate
(NaH2P04) in organic-free reagent water and dilute to 1.0 L. Add phosphoric
acid to adjust the pH to 2.5.
5.6 Carbitol (diethylene glycol monoethyl ether), C2H5OCH2CH2OCH2CH2OH.
5.7 N-methyl-N-nitroso-p-toluenesulfonamide (Diazald). High
purity,available from Aldrich Chemical Co. or equivalent.
5.8 Silicic acid, H2Si05. 100 mesh powder, store at 130°C.
5.9 Potassium carbonate, K2C03.
5.10 2,3,4,5,6-Pentafluorobenzyl bromide (PFBBr), C6F5CH2Br. Pesticide
quality or equivalent.
5.11 Sodium sulfate (granular, acidified, anhydrous), Na2S04. Purify by
heating at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there
is no interference from the sodium sulfate. Acidify by slurrying 100 g sodium
sulfate with enough diethyl ether to just cover the solid; then add 0.1 mL of
concentrated sulfuric acid and mix thoroughly. Remove the ether under vacuum.
Mix 1 g of the resulting solid with 5 ml of organic-free reagent water and
measure the pH of the mixture. It must be below a pH of 4. Store the remaining
solid at 130°C.
5.12 Solvents
5.12.1 Methylene chloride, CH2C12. Pesticide quality or equivalent.
5.12.2 Acetone, CH3COCH3. Pesticide quality or equivalent.
5.12.3 Methanol, CH3OH. Pesticide quality or equivalent.
5.12.4 Toluene, C6H5CH3. Pesticide quality or equivalent.
5.12.5 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
8151 - 6 Revision 0
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test strips. After cleanup, 20 mL of ethyl alcohol preservative must be
added to each liter of ether.
5.12.6 Isooctane, (CH3)3CH2CH(CH3)2. Pesticide quality or equivalent.
5.12.7 Hexane, C6H14. Pesticide quality or equivalent.
5.13 Stock standard solutions (1000 mg/L) - Can be prepared from pure
standard materials or can be purchased as certified solutions. Commercially
prepared stock standards can be used if they are verified against EPA standards.
If EPA standards are not available for verification, then standards certified
by the manufacturer and verified against a standard made from pure material is
acceptable.
5.13.1 Prepare stock standard solutions by accurately weighing
about 0.010 g of pure acid. Dissolve the material in pesticide quality
acetone and dilute to volume in a 10 ml volumetric flask. Stocks prepared
from pure methyl esters are dissolved in 10% acetone/isooctane (v/v).
Larger volumes may be used at the convenience of the analyst. If compound
purity is certified at 96% or greater, the weight may be used without
correction to calculate the concentration of the stock standard.
5.13.2 Transfer the stock standard solutions to vials with Teflon
lined screw-caps. Store at 4°C, protected from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially immediately prior to preparing calibration
standards from them.
5.13.3 Stock standard solutions of the derivatized acids must be
replaced after 1 year, or sooner, if comparison with check standards
indicates a problem. Stock standard solutions of the free acids degrade
more quickly and should be replaced after two months, or sooner if
comparison with check standards indicates a problem.
5.14 Internal Standard Spiking Solution (if internal standard calibration
is used) - To use this approach, the analyst must select one or more internal
standards that are similar in analytical behavior to the compounds of interest.
The analyst must further demonstrate that the measurement of the internal
standard is not affected by method or matrix interferences. The compound 4,4'-
dibromooctafluorobiphenyl (DBOB) has been shown to be an effective internal
standard, but other compounds, such as 1,4-dichlorobenzene, may be used.
5.14.1 Prepare an internal standard spiking solution by accurately
weighing approximately 0.0025 g of pure DBOB. Dissolve the DBOB in acetone
and dilute to volume in a 10 ml volumetric flask. Transfer the internal
standard spiking solution to a vial with a Teflon lined screw-cap, and
store at room temperature. Addition of 10 nl of the internal standard
spiking solution to 10 ml of sample extract results in a final internal
standard concentration of 0.25 M9/L. The solution should be replaced if
there is a change in internal standard response greater than 20 percent
of the original response recorded.
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5.15 Calibration standards - Calibration standards, at a minimum of five
concentrations for each parameter of interest, should be prepared through
dilution of the stock standards with diethyl ether. One of the concentrations
should be at a concentration near, but above, the method detection limit. The
remaining concentrations should correspond to the expected range of
concentrations found in real samples or should define the working range of the
GC. Calibration solutions must be replaced after six months, or sooner if
comparison with check standards indicates a problem.
5.15.1 Derivatize each calibration standard prepared from free
acids in a 10 ml K-D concentrator tube, according to the procedures
beginning at Section 7.5. If the calibration standards are prepared from
salts or other esters, begin with the hydrolysis step 7.2.1.6, using a 250
ml Erlenmeyer flask.
5.15.2 Add a known constant amount of one or more internal standards
to each derivatized calibration standard, and dilute to volume with the
solvent indicated in the derivative option used.
5.16 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and determinative step, and the
effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two herbicide surrogates (e.g.,
herbicides that are not expected to be present in the sample) recommended to
encompass the range of the temperature program used in this method. Deuterated
analogs of analytes should not be used as surrogates in gas chromatographic
analysis due to coelution problems. The surrogate standard recommended for use
is 2,4-Dichlorophenylacetic acid (DCAA).
5.16.1 Prepare a surrogate standard spiking solution by accurately
weighing approximately 0.001 g of pure DCAA. Dissolve the DCAA in acetone,
and dilute to volume in a 10 ml volumetric flask. Transfer the surrogate
standard spiking solution to a vial with a Teflon lined screw-cap, and
store at room temperature. Addition of 50 /iL of the surrogate standard
spiking solution to 1 L of sample, prior to extraction, results in a final
concentration in the extract of 0.5 mg/L.
5.17 pH Adjustment Solutions
5.17.1 Sodium hydroxide, NaOH, 6 N.
5.17.2 Sulfuric acid, H2S04, 12 N.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Extracts must be stored under refrigeration (4°C).
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7.0 PROCEDURE
7.1 Preparation of High Concentration Waste Samples
7.1.1 Extraction
7.1.1.1 Follow Method 3580, Waste Dilution, with the following
exceptions:
o use diethyl ether as the dilution solvent,
o use acidified anhydrous sulfate, and acidified glass wool,
o spike the sample with surrogate compound(s) according to
Section 5.16.1.
7.1.1.2 Transfer 1.0 ml (a smaller volume or a dilution may
be required if herbicide concentrations are large) to a 250 ml ground
glass Erlenmeyer flask. Proceed to Section 7.2.1.7 (hydrolysis).
7.2 Preparation of Soil, Sediment, and Other Solid Samples
7.2.1 Extraction
7.2.1.1 To a 400 mL, thick-wall beaker add 30 g (dry weight)
of the well-mixed solid sample. Acidify solids in each beaker with
85 mL of 0.1 M phosphate buffer (pH =2.5) and thoroughly mix the
contents with a glass stirring rod. Spike the sample with surrogate
compound(s) according to Section 5.16.1.
7.2.1.2 The ultrasonic extraction of solids must be optimized
for each type of sample. In order for the ultrasonic extractor to
efficiently extract solid samples, the sample must be free flowing
when the solvent is added. Acidified anhydrous sodium sulfate should
be added to clay type soils, or any other solid that is not a free
flowing sandy texture, until a free flowing mixture is obtained.
7.2.1.3 Add 100 ml of methylene chloride to the beaker.
Perform ultrasonic extraction for 3 minutes, with output control
knob set at 10 (full power) and with mode switch on Pulse (pulsing
energy rather than continuous energy) and percent-duty cycle knob
set at 50% (energy on 50% of time and off 50% of time). Allow the
solids to settle. Transfer the organic layer into a 500 ml
centrifuge bottle.
7.2.1.4 Ultrasonically extract the sample twice more using
100 mL of methylene chloride and the same ultrasonic condition.
7.2.1.5 Combine the three organic extracts from the sample
in the centrifuge bottle and centrifuge 10 minutes to settle the
fine particles. Filter the combined extract through filter paper
(Whatman #1, or equivalent) into 500 mL 24/40 Erlenmeyer flask.
7.2.1.6 Add boiling chips and attach the macro Snyder column.
Evaporate the methylene chloride on the water bath to a volume of
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approximately 25 ml. Remove the flasks from the water bath and allow
them to cool.
7.2.1.7 Add 5 mL of 37% aqueous potassium hydroxide, 30 mL
of water and 40 ml of methanol to the extract. Add additional
boiling chips to the flask. Reflux the mixture on a water bath at
60-65°C for 2 hours. Remove the flasks from the water bath and cool
to room temperature.
7.2.1.8 Transfer the hydrolyzed aqueous solution to a 500 mL
separatory funnel and extract the solution three times with 100 ml
portions of methylene chloride. Discard the methylene chloride
phase. At this point the basic solution contains the herbicide
salts.
7.2.1.9 Adjust the pH of the solution to <2 with cold (4°C)
sulfuric acid (1+3) and extract three times with 100 ml portions of
methylene chloride. Combine the extracts and pour them through a
pre-rinsed drying column containing 7 to 10 cm of acidified anhydrous
sodium sulfate. Collect the dried extracts in a 500 mL K-D flask
fitted with a 10 mL concentrator tube. Proceed to section 7.4 for
extract concentration.
7.3 Preparation of Aqueous Samples
7.3.1 Separatory Funnel
7.3.1.1 Using a graduated cylinder, measure out a 1 liter of
sample and transfer it into a 2 L separatory funnel. Spike the
sample with surrogate compound(s) according to Section 5.16.1.
7.3.1.2 Add 250 g of NaCl to the sample, seal, and shake to
dissolve the salt.
7.3.1.3 Add 17 mL of 6 N NaOH to the sample, seal, and shake.
Check the pH of the sample with pH paper; if the sample does not have
a pH greater than or equal to 12, adjust the pH by adding more 6 N
NaOH. Let the sample sit at room temperature for 1 hour, shaking
the separatory funnel and contents periodically.
7.3.1.4 Add 60 mL of methylene chloride to the sample bottle
to rinse the bottle. Transfer the methylene chloride to the
separatory funnel and extract the sample by vigorously shaking the
funnel for 2 minutes, with periodic venting to release excess
pressure. Allow the organic layer to separate from the water phase
for a minimum of 10 minutes. If the emulsion interface between the
layers is more than one-third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete the phase
separation. The optimum technique depends upon the sample, but may
include stirring, filtration through glass wool, centrifugation, or
other physical methods. Discard the methylene chloride phase.
7.3.1.5 Add a second 60 mL volume of methylene chloride to
the sample bottle and repeat the extraction procedure a second time,
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discarding the methylene chloride layer. Perform a third extraction
in the same manner.
7.3.1.6 Add 17 ml of cold (4°C) 12 N sulfuric acid to the
sample, seal, and shake to mix. Check the pH of the sample with pH
paper: if the sample does not have a pH less than or equal to 2,
adjust the pH by adding more acid.
7.3.1.7 Add 120 ml diethyl ether to the sample, seal, and
extract the sample by vigorously shaking the funnel for 2 min with
periodic venting to release excess pressure. Allow the organic
layer to separate from the water phase for a minimum of 10 min. If
the emulsion interface between layers is more than one third the
volume of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. The optimum techniques
to complete the phase separation depends upon the sample, but may
include stirring, filtration through glass wool, centrifugation, or
other physical methods. Remove the aqueous phase to a 2 L Erlenmeyer
flask and collect the ether phase in a 500 ml Erlenmeyer flask
containing approximately 10 g of acidified anhydrous sodium sulfate.
Periodically, vigorously shake the extract and drying agent.
7.3.1.8 Return the aqueous phase to the separatory funnel,
add 60 ml of diethyl ether to the sample, and repeat the extraction
procedure a second time, combining the extracts in the 500 ml
Erlenmeyer flask. Perform a third extraction with 60 ml diethyl
ether in the same manner. Allow the extract to remain in contact
with the sodium sulfate for approximately 2 hours.
Note: The drying step is very critical to ensuring complete esterification.
Any moisture remaining in the ether will result in low herbicide
recoveries. The amount of sodium sulfate is adequate if some free flowing
crystals are visible when swirling the flask. If all of the sodium sulfate
solidifies in a cake, add a few additional grams of acidified sodium
sulfate and again test by swirling. The 2 hour drying time is a minimum,
however, the extracts may be held in contact with the sodium sulfate
overnight.
7.3.1.9 Pour the dried extract through a funnel plugged with
acid washed glass wool, and collect the extract in the K-D
concentrator. Use a glass rod to crush any caked sodium sulfate
during the transfer. Rinse the round bottom flask and funnel with
20 to 30 ml of diethyl ether to complete the quantitative transfer.
Proceed to section 7.4 for extract concentration.
7.4 Extract Concentration
7.4.1 Add one or two clean boiling chips to the flask and attach
a three ball Snyder column. Prewet the Snyder column by adding about 1
ml of diethyl ether to the top of the column. Place the K-D apparatus on
a hot water bath (15-20°C above the boiling point of the solvent) so that
the concentrator tube is partially immersed in the hot water and the entire
lower rounded surface of the flask is bathed with hot vapor. Adjust the
vertical position of the apparatus and the water temperature, as required,
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to complete the concentration in 10-20 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but the
chambers will not flood. When the apparent volume of liquid reaches 1 ml,
remove the K-D apparatus from the water bath and allow it to drain and cool
for at least 10 minutes.
7.4.2 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 ml of diethyl ether. The
extract may be further concentrated by using either the micro Snyder column
technique (Section 7.4.3) or nitrogen blowdown technique (Section 7.4.4).
7.4.3 Micro Snyder Column Technique
7.4.3.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 ml of diethyl ether to the top of the
column. Place the K-D apparatus in a hot water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature,
as required, to complete the concentration in 5-10 minutes. At the
proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes. Remove
the Snyder column and rinse the flask and its lower joints with about
0.2 ml of diethyl ether and add to the concentrator tube. Proceed
to Section 7.4.5.
7.4.4 Nitrogen Blowdown Technique
7.4.4.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the required
level using a gentle stream of clean, dry nitrogen (filtered through
a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon trap and the sample.
7.4.4.2 The internal wall of the tube must be rinsed down
several times with diethyl ether during the operation. During
evaporation, the solvent level in the tube must be positioned to
prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become
dry. Proceed to Section 7.4.5.
7.4.5 Dilute the extract with 1 ml of isooctane and 0.5 ml of
methanol. Dilute to a final volume of 4 ml with diethyl ether. The sample
is now ready for methylation with diazomethane. If PFB derivation is being
performed, dilute to 4 mL with acetone.
7.5 Esterification - For diazomethane derivatization proceed with Section
7.5.1. For PFB derivatization proceed with Section 7.5.2.
8151 - 12 Revision 0
November 1990
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7.5.1 Diazomethane Derivatization - Two methods may be used for
the generation of diazomethane: the bubbler method (see Figure 1), Section
7.5.1.1, and the Diazald kit method, Section 7.5.1.2.
CAUTION: Diazomethane is a carcinogen and can explode under certain conditions.
The bubbler method is suggested when small batches of samples (10-15)
require esterification. The bubbler method works well with samples that
have low concentrations of herbicides (e.g., aqueous samples) and is safer
to use than the Diazald kit procedure. The Diazald kit method is good for
large quantities of samples needing esterification. The Diazald kit method
is more effective than the bubbler method for soils or samples that may
contain high concentrations of herbicides (e.g., samples such as soils that
may result in yellow extracts following hydrolysis may be difficult to
handle by the bubbler method). The diazomethane derivatization (U.S.EPA,
1971) procedures, described below, will react efficiently with all of the
chlorinated herbicides described in this method and should be used only
by experienced analysts, due to the potential hazards associated with its
use. The following precautions should be taken:
o Use a safety screen.
o Use mechanical pipetting aides.
o Do not heat above 90°C - EXPLOSION may result.
o Avoid grinding surfaces, ground-glass joints, sleeve bearings,
and glass stirrers - EXPLOSION may result.
o Store away from alkali metals - EXPLOSION may result.
o Solutions of diazomethane decompose rapidly in the presence of
solid materials such as copper powder, calcium chloride, and
boiling chips.
7.5.1.1 Bubbler method - Assemble the diazomethane bubbler
(see Figure 1).
7.5.1.1.1 Add 5 mL of diethyl ether to the first test
tube. Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5 mL of
37% KOH, and 0.1-0.2 g of Diazald to the second test tube.
Immediately place the exit tube into the concentrator tube
containing the sample extract. Apply nitrogen flow (10 mL/min)
to bubble diazomethane through the extract for 10 minutes or
until the yellow color of diazomethane persists. The amount
of Diazald used is sufficient for esterification of
approximately three sample extracts. An additional 0.1-0.2
g of Diazald may be added (after the initial Diazald is
consumed) to extend the generation of the diazomethane. There
is sufficient KOH present in the original solution to perform
a maximum of approximately 20 minutes of total esterification.
7.5.1.1.2 Remove the concentrator tube and seal it with
a Neoprene or Teflon stopper. Store at room temperature in
a hood for 20 minutes.
7.5.1.1.3 Destroy any unreacted diazomethane by adding
0.1-0.2 g of silicic acid to the concentrator tube. Allow to
stand until the evolution of nitrogen gas has stopped. Adjust
8151 - 13 Revision 0
November 1990
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the sample volume to 10.0 ml with hexane. Stopper the
concentrator tube or transfer 1 ml of sample to a GC vial, and
store refrigerated if further processing will not be performed
immediately. It is recommended that the methylated extracts
be analyzed immediately to minimize the trans-esterification
and other potential reactions that may occur. Analyze by gas
chromatography.
7.5.1.1.4 Extracts should be stored at 4°C away from
light. Preservation study results indicate that most analytes
are stable for 28 days; however, it is recommended that the
methylated extracts be analyzed immediately to minimize the
trans-esterification and other potential reactions that may
occur. Analyze by gas chromatography.
7.5.1.2 Diazald kit method - Instructions for preparing
diazomethane are provided with the generator kit.
7.5.1.2.1 Add 2 ml of diazomethane solution and let
the sample stand for 10 minutes with occasional swirling.
The yellow color of diazomethane should be evident and should
persist for this period.
7.5.1.2.2 Rinse the inside wall of the ampule with 700
/iL of di ethyl ether. Reduce the sample volume to
approximately 2 ml to remove excess diazomethane by allowing
the solvent to evaporate spontaneously at room temperature.
Alternatively, 10 mg of silicic acid can be added to destroy
the excess diazomethane.
7.5.1.2.3 Dilute the sample to 10.0 ml with hexane.
Analyze by gas chromatography.
7.5.2 PFB Method
7.5.2.1 Add 30 /*L of 10% K2C03 and 200 /iL of 3% PFBBr in
acetone. Close the tube with a glass stopper and mix on a vortex
mixer. Heat the tube at 60°C for 3 hours.
7.5.2.2 Evaporate the solution to 0.5 ml with a gentle stream
of nitrogen. Add 2 ml of hexane and repeat evaporation just to
dryness at ambient temperature.
7.5.2.3 Redissolve the residue in 2 ml of toluene:hexane
(1:6) for column cleanup.
7.5.2.4 Top the silica column with 0.5 cm of anhydrous sodium
sulfate. Prewet the column with 5 ml hexane and let the solvent
drain to the top of the adsorbent. Quantitatively transfer the
reaction residue to the column with several rinsings of the
toluene:hexane solution (total 2-3 ml).
8151 - 14 Revision 0
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7.5.2.5 Elute the column with sufficient toluene:hexane to
collect 8 ml of eluent. Discard this fraction which contains excess
reagent.
7.5.2.6 Elute the column with toluenerhexane (1:9) to collect
8 mL of eluent containing PFB derivatives in a 10 ml volumetric
flask. Dilute to 10 ml with hexane. Analyze by 6C/ECD.
7.6 Gas chromatographic conditions (recommended):
7.6.1 Narrow Bore
7.6.1.1 Primary Column 1:
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /ul_, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.1.2 Primary Column la:
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /nL. splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.1.3 Column 2:
Temperature program: 60°C to 300PC, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /itL, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.1.4 Confirmation Column:
Temperature program: 60°C to 300°C, at 4°C/irin
Helium carrier flow: 30 cm/sec
Injection volume: 2 pi, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.2 Megabore
7.6.2.1 Primary Column:
Temperature program: 0.5 minute at 150°C, 150°C to 270°C at 5°C/min
Helium carrier flow: 7 mL/min
Injection volume: 1 ^L
7.6.2.2 Confirmatory Column:
Temperature program: 0.5 minute at 150°C, 150°C to 270°C at 5°C/min
Helium carrier flow: 7 mL/min
Injection volume: 1 pi
8151 - 15 Revision 0
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7.7 Calibration
7.7.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
Use Table 1 for guidance on selecting the lowest point on the calibration
curve.
7.8 Gas chromatographic analysis
7.8.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 nl of internal standard to the sample prior to
injection.
7.8.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.8.3 An example of a chromatogram for a methylated chlorophenoxy
herbicide is shown in Figure 2. Tables 2 and 3 present retention times
for the target analytes after esterification, using the diazomethane
derivatization procedure and the PFB derivatization procedure,
respectively.
7.8.4 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
7.8.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.8.6 If calibration standards have been analyzed in the same manner
as the samples (e.g. have undergone hydrolysis and esterification), then
the calculation of concentration given in Method 8000 should be used.
However, if calibration is performed using standards made from methyl ester
compounds (compounds not esterified by application of this method), then
the calculation of concentration must include a correction for the
molecular weight of the methyl ester versus the acid herbicide.
7.8.7 If peak detection and identification are prevented due to
interferences, further cleanup is required. Before using any cleanup
procedure, the analyst must process a series of standards through the
procedure to validate elution patterns and the absence of interferences
from reagents.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8151 - 16 Revision 0
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8.2 Procedures to check the GC system operation are found in Method 8000.
8.2.1 Select a representative spike concentration for each compound
(acid or ester) to be measured. Using stock standards, prepare a quality
control check sample concentrate, in acetone, that is 1000 times more
concentrated than the selected concentrations. Use this quality control
check sample concentrate to prepare quality control check samples.
8.2.2 Tables 4 and 5 present bias and precision data for water and
clay matrices, using the diazomethane derivatization procedure. Table 6
presents relative recovery data generated using the PFB derivatization
procedure and water samples. Compare the results obtained with the results
given in these Tables to determine if the data quality is acceptable.
8.3 Calculate surrogate standard recovery on all standards, samples,
blanks, and spikes. Determine if the recovery is within limits (limits
established by performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures
are required:
8.3.1.1 Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check instrument
performance.
8.3.1.2 Recalculate the data and/or reanalyze the extract if
any of the above checks reveal a problem.
8.3.1.3 Reextract and reanalyze the sample if none of the above
are a problem or flag the data as "estimated concentration."
8.4 GC/MS confirmation
8.4.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Refer to Method 8270
for the appropriate GC/MS operating conditions and analysis procedures.
8.4.2 When available, chemical ionization mass spectra may be
employed to aid the qualitative identification process.
8.4.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps
may include the use of alternate packed or capillary GC columns or
additional cleanup.
9.0 METHOD PERFORMANCE
9.1 In single laboratory studies using organic-free reagent water and
clay/still bottom samples, the mean recoveries presented in Tables 4 and 5 were
obtained for diazomethane derivatization. The standard deviations of the percent
recoveries of these measurements are also in Tables 4 and 5.
8151 - 17 Revision 0
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9.2 Table 6 presents relative recoveries of the target analytes obtained
using the PFB derivatization procedure with spiked water samples.
10.0 REFERENCES
1. Fed. Reg. 1971, 38, No. 75, Pt. II.
2. Goerlitz, D. G.; Lamar, W.L., "Determination of Phenoxy Acid Herbicides in
Water by Electron Capture and Microcoulometric Gas Chromatography,". U.S.
Geol. Survey Water Supply Paper 1967, 1817-C.
3. Burke, J. A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects, J. Assoc. Off Anal. Chem. 1965, 48, 1037.
4. "Extraction and Cleanup Procedures for the Determination of Phenoxy Acid
Herbicides in Sediment"; U.S. Environmental Protection Agency. EPA Toxicant
and Analysis Center: Bay St. Louis, MS, 1972.
5. Shore, F.L.; Amick, E.N.; Pan, S. T. "Single Laboratory Validation of EPA
Method 8151 for the Analysis of Chlorinated Herbicides in Hazardous Waste";
U.S. Environmental Protection Agency. Environmental Monitoring Systems
Laboratory. Office of Research and Development, Las Vegas, NV, 1985;
EPA-60014-85-060.
6. Method 515.1, "Determination of Chlorinated Acids in Water by Gas
Chromatography with an Electron Capture Detector", Revision 4.0, USEPA,
Office of Research and Development, Environmental Monitoring Systems
Laboratory, Cincinnati, Ohio.
7. Method 1618, "Organo-halide and Organo-phosphorus Pesticides and Phenoxy-
acid Herbicides by Wide Bore Capillary Column Gas Chromatography with
Selective Detectors", Revision A, July 1989, USEPA, Office of Water
Regulations and Standards, Washington, DC.
8151 - 18 Revision 0
November 1990
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Figure 1
DIAZOMETHANE GENERATOR
nitrogen
rubber stopper
gloss tubing
tube 1
tube 2
8151 - 19
Revision 0
November 1990
-------
TABLE 1
ESTIMATED METHOD DETECTION LIMITS FOR METHOD 8151,
DIAZOMETHANE DERIVATIZATION
Analyte
Aqueous Samples
GC/ECD
Estimated
Detection
Limit8
(Mg/L)
Soil
GC/ECD
Estimated
Detection
Limitb
(WJ/Kg)
Samples
GC/MS
Estimated
Identification
Limit0
(ng)
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacid6
Dicamba
3,5-Dichlorobenzoic acid
Dichlorprop
Dinoseb
5-Hydroxydicamba
MCPP
MCPA
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-T
2,4,5-TP
0.096
0.2
0.093
0.2
1.3
0.8
0.02
0.081
0.061
0.26
0.19
0.04
0.09d
0.056d
0.13
0.076
0.14
0.08
0.075
4.0
0.11
0.12
0.38
66
43
0.34
0.16
0.28
1.7
1.25
0.5
0.65
0.43
0.3
0.44
1.3
4.5
a EDL = estimated detection limit; defined as either the MDL (40 CFR Part 136,
Appendix B, Revision 1.11 ), or a concentration of analyte in a sample
yielding a peak in the final extract with signal-to-noise ratio of
approximately 5, whichever value is higher.
b Detection limits determined from standard solutions corrected back to 50 g
samples, extracted and concentrated to 10 ml, with 5 /xL injected.
Chromatography using narrow bore capillary column, 0.25 urn film,
5% phenyl/95% methyl silicone.
c The minimum amount of analyte to give a Finnigan INCOS FIT value of 800 as
the methyl derivative vs. the spectrum obtained from 50 ng of the respective
free acid herbicide.
40 CFR Part 136, Appendix B (49 FR 43234).
capillary column.
Chromatography using megabore
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 20
Revision 0
November 1990
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TABLE 2
RETENTION TIMES (MINUTES) OF METHYL DERIVATIVES OF CHLORINATED HERBICIDES
Analyte
Dalapon
3,5-Dichlorobenzoic
4-Nitrophenol
DCAA (surrogate)
Dicamba
Dichlorprop
2,4-D
DBOB (internal std.)
Pentachlorophenol
Chloramben
2,4,5-TP
5-Hydroxydicamba
2,4,5-T
2,4-DB
Dinoseb
Bentazon
Picloram
DCPA diacidc
Acifluorfen
MCPP
MCPA
Narrow
Primary8
Column
3.4
acid 18.6
18.6
22.0
22.1
25.0
25.5
27.5
28.3
29.7
29.7
30.0
30.5
32.2
32.4
33.3
34.4
35.8
41.5
Bore Columns
Confirmation3
Col umn
4.7
17.7
20.5
14.9
22.6
25.6
27.0
27.6
27.0
32.8
29.5
30.7
30.9
32.2
34.1
34.6
37.5
37.8
42.8
Megabore
Primary"
Col umn
4.39
5.15
5.85
6.97
7.92
8.74
4.24
4.74
Columns
Confirmation"
Col umn
4.39
5.46
6.05
7.37
8.20
9.02
4.55
4.94
8151 - 21
Revision 0
November 1990
-------
TABLE 2 (continued)
Primary Column: 5% phenyl/95% methyl silicone
Confirmation Column: 14% cyanopropyl phenyl silicone
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /iL, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
Primary Column: DB-608
Confirmatory Column: 14% cyanopropyl phenyl silicone
Temperature program: 0.5 minute at 150°C,
150°C to 270°C, at 5°C/nrin
Helium carrier flow: 7 mL/min
Injection volume: 1 nl
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 22 Revision 0
November 1990
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TABLE 3
RETENTION TIMES (MINUTES) OF PFB DERIVATIVES OF CHLORINATED HERBICIDES
Gas chromatoqraphic column
Herbicide Thin-film DB-5aSP-2250"Thick-film DB-5C
Dalapon
MCPP
Dicamba
MCPA
Dichlorprop
2,4-D
Silvex
2,4,5-T
Dinoseb
2,4-DB
10.41
18.22
18.73
18.88
19.10
19.84
21.00
22.03
22.11
23.85
12.94
22.30
23.57
23.95
24.10
26.33
27.90
31.45
28.93
35.61
13.54
22.98
23.94
24.18
24.70
26.20
29.02
31.36
31.57
35.97
DB-5 capillary column, 0.25 jum film thickness, 0.25 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 17 minutes.
SP-2550 capillary column, 0.25 pm film thickness, 0.25 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 10 minutes.
DB-5 capillary column, 1.0 urn film thickness, 0.32 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 10 minutes.
8151 - 23 Revision 0
November 1990
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TABLE 4
ACCURACY AND PRECISION FOR METHOD 8151
DIAZOMETHANE DERIVATIZATION, ORGANIC-FREE REAGENT WATER MATRIX
Spike
Concentration
Analyte (M9/L)
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacidb
Dicamba
3,5-Dichlorobenzoic Acid
Dichlorprop
Dinoseb
5-Hydroxydicamba
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-TP
2,4,5-T
0.2
1
0.4
1
10
4
0.2
0.4
0.6
2
0.4
0.2
1
0.04
0.6
0.4
0.2
Mean8 Standard
Percent Deviation of
Recovery Percent Recovery
121
120
111
131
100
87
74
135
102
107
42
103
131
130
91
117
134
15.7
16.8
14.4
27.5
20.0
13.1
9.7
32.4
16.3
20.3
14.3
16.5
23.6
31.2
15.5
16.4
30.8
Mean percent recovery calculated from 7-8 determinations of spiked
organic-free reagent water.
DCPA monoacid and diacid metabolites included in method scope; DCPA
diacid metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 24
Revision 0
November 1990
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TABLE 5
ACCURACY AND PRECISION FOR METHOD 8151
DIAZOMETHANE DERIVATIZATION, CLAY MATRIX
Analyte
Mean8
Percent Recovery
Linear"
Concentration
Range
(ng/g)
Percent
Relative0
Standard Deviation
(n-20)
Dicamba
MCPP
MCPA
Dichlorprop
2,4-D
2,4,5-TP
2,4,5-T
2,4-DB
Dinoseb
95.7
98.3
96.9
97.3
84.3
94.5
83.1
90.7
93.7
0.52
620
620
1.5
1.2
0.42
0.42
4.0
0.82
- 104
- 61,800
- 61,200
- 3,000
- 2,440
- 828
- 828
- 8,060
- 1,620
7.5
3.4
5.3
5.0
5.3
5.7
7.3
7.6
8.7
Mean percent recovery calculated from 10 determinations of spiked clay
and clay/still bottom samples over the linear concentration range.
Linear concentration range was determined on standard solutions and
corrected to 50 g solid samples.
Percent relative standard deviation was calculated on standard solutions,
10 samples high in the linear concentration range, and 10 samples low in
the range.
8151 - 25
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November 1990
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TABLE 6
RELATIVE RECOVERIES OF PFB DERIVATIVES OF HERBICIDES8
Std.concn,
Analyte
MCPP
Dicamba
MCPA
Dichlorprop
2,4-D
Si 1 vex
2,4,5-T
2,4-DB
Mean
Mg/mL
5
3
10
6
9
10
12
20
.1
.9
.1
.0
.8
.4
.8
.1
1
95.6
91.4
89.6
88.4
55.6
95.3
78.6
99.8
86.8
2
88.8
99.2
79.7
80.3
90.3
85.8
65.6
96.3
85.7
Relative recoveries, %
3
97.1
100
87.0
89.5
100
91.5
69.2
100
91.8
4
100
92.7
100
100
65.9
100
100
88.4
93.4
5
95.5
84.0
89.5
85.2
58.3
91.3
81.6
97.1
85.3
6
97.2
93.0
84.9
87.9
61.6
95.0
90.1
92.4
89.0
7
98.1
91.1
92.3
84.5
60.8
91.1
84.3
91.6
87.1
8
98.2
90.1
98.6
90.5
67.6
96.0
98.5
91.6
91.4
Mean
96.3
92.7
90.2
88.3
70.0
93.3
83.5
95.0
Percent recovery determinations made using spiked water samples.
8151 - 26
Revision 0
November 1990
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METHOD 8151
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATIQN
PERIVATIZATION; CAPILLARY COLUMN TECHNIQUE
7.1.1 1 Follow
method 3580
waste dilution
unrig the given
exceptions
Soil
7.2 1.1 Heigh
sample and add
to beaker ; add
buffer and
spike ; mix wel 1
7.3.1.1 Measure
1L of sample
and transfer to
a 2L sap.
funnel.
7.3.1.2 Add
2SOg NaCI to
•ample and
shake to
dissolve
7.1.1.2
Transfer 1 0 ml
to flask and
proceed to step
7.2.1.7
7.2.1.2
Opti-
mize ultrasonic
solid OK trac-
tion for each
ma t r i K .
,
7.2.1
HeCl to
3 Add
sample
and extract 3
mm ; let settle
and decant MeCl
72.1
4 & 5
Ul trasonically
extract
2 more
7.2.1.5
sample
times
Combine
organic
ex tracts ,
centrifuge, and
filter ex t rac t .
7 2.1.7 Add
KOH. water, and
for 2 hours and
allow to cool
7 2
1 6
Concentrate
MeCl to
25 ml with
Snyder column.
8151 - 27
7.3.1.3 Add 6N
NaOH to sample
and shake. Add
until pH>12.
Let stand 1 hr.
7.3.1.4 Add
MeCl and ex-
tract by sha-
king for 2 min.
Discard MeCl.
No
7 3.1.5
Repeat
ex tract ion
twice more .
Discard MeCl .
7.3 1.6 Add
12N sulfuric
acid and
shake. Add
until pH<2.
Yes
Employ mechanical
techniques to
complete phase
separation (e.g.
stirring, filteration
through glass wool,
centrifugalion, or
other physical meth-
ods). Discard MeCl.
Revision 0
November 1990
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METHOD 8151
(continued)
7 3.1
7 Add
die thy 1 ether
to sampl e and
ex t rac
L Save
both phases .
/
/
I
X
/Does \
/ difficult X. Ye
emulsion j—
\ form? /
\
\
X.
No
7.3.1.8
/
/
/
Re turn
aqueous phase to
separator/
ex tracts ,
extract to
funnel and
and allow
remain in
contact with sodium
sulfate for 2 hours.
73.1.9 Pour
extract through
proseed to Step
7 .1
1.
7.2.18
Trans-
f er to sep fun-
nel and
extract
3x with MeCl
Discard MeCl .
Employ mechanical
techniques to com-
plete phase separ-
s ation (e.g. sti r r ing ,
> filtration through
glass wool, centfi-
f uga tion. or other
physical methods )
Save both phases.
•
7.2.1.9 Adjust to
pH<2 »ith cold
sul fur ic acid ,
extract
3x tilth
MeCl, dry MeCl on
sodium sulfate
column, transfer to
K-D apparatus for
concentration .
1
I
741 Place K-D
apparatus in
concentrate ,
and cool .
7.4.2
74.4
Complete
concentration vith
micro Snyder column
or nitrogen bloM
dot
*n .
•
7 45 Dilute
extract
• ith 1
ml isooctane
and 0
S ml
methanol
8151 - 28
Revision 0
November 1990
-------
METHOD 8151
(continued)
7 4 5 Diluta
axtract to 4
ml »ith
ma than* diathy1
ether
7 5 1 1 1 Add 5ml to
1st test tub* Add 1
ml diethyl •that. 1ml
carbitol. 1 5 ml of
37% KOH. and 0 1-0 2
9 of Diaxald to the
2nd tub* Bubble -ith
nitrogan for 10 mm
or yal1ow persists.
ml diaiomathana
7 5 1 2 2 Rins
ampule with
ether and evaporat
to 2 ml to remove
diazomethane
Alterna 1iv»ly,
silicic acid may b
added
7 5 2 6 Discard 1st
elution vith enough
toluene hexane (1:9)
to codlect 8nl more
aluent Transfer to a
10ml volumatric flask
and dilute to the
mark with Havana
7 S 1.1.3 Add silicic
to concentrator tuba
and let stand until
nitrogen evolution
has stopped Adjust
sample volume to 10
ml with hexana Stop-
per Immediate analy-
8151 - 29
Revision 0
November 1990
-------
METHOD 8151
(continued)
7.7 Internal or
• M terna.1
calibration may
be used. (See
Method 8000)
7.8.1 Add lOul
internal
standard to the
sample prior to
infection
78.2 See Method 8000
for analysis se-
quence, appropriate
dilution*, establish-
ing daily retenion
time windoMS, and
identification
criteria. Check stds
every 10 samples
7 8,4 Record
volume
injected and
the resulting
peak sizes
7.8.S Determine
the identity
and quantify
component
peaks.
Calculate the
correction for
molecular wt.
of methyl ester
vs herbicide
7 86 Calculate
concentration
using procedure
in Method 8000
7.8.7 Perform
further
cleanup if
necessary.
Stop
8151 - 30
Revision
November
0
1990
-------
METHOD 8240B
VOLATILE ORGANICS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS):
PACKED COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8240 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Analyte
CAS No.
Appropriate Technique
Direct
Purge-and-Trap Injection
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Ally! alcohol
Allyl chloride
Benzene
Benzyl chloride
Bromoacetone
Bromochloromethane (I.S.)
Bromodi chl oromethane
4-Bromofluorobenzene (surr.)
Bromoform
Bromomethane
2-Butanone
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chlorobenzene
Chlorobenzene-ds (I.S.)
Chl orodi bromomethane
Chloroethane
2-Chloroethanol
bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Chloroprene
3-Chloropropionitrile
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Di bromomethane
67-64-1
75-05-8
107-02-8
107-13-1
107-18-6
107-05-1
71-43-2
100-44-7
598-31-2
74-97-5
75-27-4
460-00-4
75-25-2
74-83-9
78-93-3
75-15-0
56-23-5
75-87-6
108-90-7
3114-55-4
124-48-1
75-00-3
107-07-3
505-60-2
110-75-8
67-66-3
74-87-3
126-99-8
542-76-7
96-12-8
106-93-4
74-95-3
PP
PP
PP
PP
PP
a
a
PP
PP
a
a
a
a
a
PP
PP
a
PP
a
a
a
a
PP
PP
a
a
a
a
ND
PP
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
pc
a
a
a
8240B - 1
Revision 2
November 1990
-------
Appropriate Technique
Analyte
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4(surr.)
1,1-Dichloroethene
trans- 1,2-Di chl oroethene
1 , 2-Di chl oropropane
l,3-Dichloro-2-propanol
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
l,2:3,4-Diepoxybutane
1,4-Difluorobenzene (I.S.)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl benzene
Ethyl ene oxide
Ethyl methacryl ate
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Malononitrile
Methacrylonitrile
Methyl ene chloride
Methyl iodide
Methyl methacryl ate
4-Methyl -2-pentanone
Pentachloroethane
2-Picoline
Propargyl alcohol
b-Propiolactone
Propionitrile
n-Propylamine
Pyridine
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
Toluene-dB (surr.)
1 , 1 , 1-Tri chl oroethane
1 , 1 , 2-Tri chl oroethane
Tri chl oroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
CAS No.b
764-41-0
75-71-8
75-34-3
107-06-2
107-06-2
75-35-4
156-60-5
78-87-5
96-23-1
10061-01-5
10061-02-6
1464-53-5
540-36-3
123-91-1
106-89-8
64-17-5
100-41-4
75-21-8
97-63-2
591-78-6
78-97-7
74-88-4
78-83-1
109-77-3
126-98-7
75-09-2
74-88-4
80-62-6
108-10-1
76-01-7
109-06-8
107-19-7
57-57-8
107-12-0
107-10-8
110-86-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
2037-26-5
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
Purge-and-Trap
PP
a
a
a
a
a
a
a
PP
a
a
a
a
PP
i
i
a
PP
a
PP
ND
a
PP
PP
PP
a
a
a
PP
i
PP
PP
PP
PP
a
i
a
a
a
a
a
a
a
a
a
a
a
Direct
Injection
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
a
a
a
a
a
a
a
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
8240B - 2
Revision 2
November 1990
-------
Appropriate Technique
Direct
Analyte CAS No." Purge-and-Trap Injection
Vinyl acetate
Vinyl chloride
Xylene (Total)
108-05-4
75-01-4
1330-20-7
a
a
a
a
a
a
a Adequate response by this technique.
b Chemical Abstract Services Registry Number.
pp Poor purging efficiency resulting in high EQLs.
i Inappropriate technique for this analyte.
pc Poor chromatographic behavior.
1.2 Method 8240 can be used to quantitate most volatile organic compounds
that have boiling points below 200°C and that are insoluble or slightly soluble
in water. Volatile water-soluble compounds can be included in this analytical
technique. However, for the more soluble compounds, quantitation limits are
approximately ten times higher because of poor purging efficiency. The method
is also limited to compounds that elute as sharp peaks from a GC column packed
with graphitized carbon lightly coated with a carbowax. Such compounds include
low molecular weight halogenated hydrocarbons, aromatics, ketones, nitriles,
acetates, acrylates, ethers, and sulfides. See Table 1 for a list of compounds,
retention times, and their characteristic ions that have been evaluated on a
purge-and-trap GC/MS system.
1.3 The estimated quantitation limit (EQL) of Method 8240 for an
individual compound is approximately 5 jig/Kg (wet weight) for soil/sediment
samples, 0.5 mg/Kg (wet weight) for wastes, and 5 /xg/L for ground water (see
Table 2). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 Method 8240 is based upon a purge-and-trap, gas chromatographic/mass
spectrometric (GC/MS) procedure. This method is restricted to use by, or under
the supervision of, analysts experienced in the use of purge-and-trap systems
and gas chromatograph/mass spectrometers, and skilled in the interpretation of
mass spectra and their use as a quantitative tool.
1.5 To increase purging efficiencies of acrylonitrile and acrolein, refer
to Methods 5030 and 8030 for proper purge-and-trap conditions.
2.0 SUMMARY OF METHOD
2.1 The volatile compounds are introduced into the gas chromatograph by
the purge-and-trap method or by direct injection (in limited applications).
The components are separated via the gas chromatograph and detected using a mass
spectrometer, which is used to provide both qualitative and quantitative
information. The chromatographic conditions, as well as typical mass
8240B - 3 Revision 2
November 1990
-------
spectrometer operating parameters, are given.
2.2 If the above sample introduction techniques are not applicable, a
portion of the sample is dispersed in methanol to dissolve the volatile organic
constituents. A portion of the methanolic solution is combined with organic-
free reagent water in a specially designed purging chamber. It is then analyzed
by purge-and-trap GC/MS following the normal water method.
2.3 The purge-and-trap process - An inert gas is bubbled through the
solution at ambient temperature, and the volatile components are efficiently
transferred from the aqueous phase to the vapor phase. The vapor is swept
through a sorbent column where the volatile components are trapped. After
purging is completed, the sorbent column is heated and backflushed with inert
gas to desorb the components onto a gas chromatographic column. The gas
chromatographic column is heated to elute the components, which are detected
with a mass spectrometer.
3.0 INTERFERENCES
3.1 Interferences purged or coextracted from the samples will vary
considerably from source to source, depending upon the particular sample or
extract being tested. The analytical system, however, should be checked to
ensure freedom from interferences, under the analysis conditions, by analyzing
method blanks.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal into
the sample during shipment and storage. A trip blank, prepared from organic-
free reagent water and carried through the sampling and handling protocol, can
serve as a check on such contamination.
3.3 Cross contamination can occur whenever high-concentration and low-
concentration samples are analyzed sequentially. Whenever an unusually
concentrated sample is analyzed, it should be followed by the analysis of
organic-free reagent water to check for cross contamination. The purge-and-trap
system may require extensive bake-out and cleaning after a high-concentration
sample.
3.4 The laboratory where volatile analysis is performed should be
completely free of solvents.
3.5 Impurities in the purge gas and from organic compounds out-gassing
from the plumbing ahead of the trap account for the majority of contamination
problems. The analytical system must be demonstrated to be free from
contamination under the conditions of the analysis by running calibration and
reagent blanks. The use of non-TFE plastic coating, non-TFE thread sealants,
or flow controllers with rubber components in the purging device should be
avoided.
8240B - 4 Revision 2
November 1990
-------
4.0 APPARATUS AND MATERIALS
4.1 Microsyringes - 10 /A, 25 p.L, 100 jitL, 250 /iL, 500 juL, and 1,000 ni.
These syringes should be equipped with a 20 gauge (0.006 In. ID) needle having
a length sufficient to extend from the sample Inlet to within 1 cm of the glass
frit In the purging device. The needle length will depend upon the dimensions
of the purging device employed.
4.2 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.3 Syringe - 5 ml, gas-tight with shutoff valve.
4.4 Balances - Analytical, 0.0001 g, and top-loading, 0.1 g.
4.5 Glass scintillation vials - 20 ml, with screw caps and Teflon liners
or glass culture tubes with a screw cap and Teflon liner.
4.6 Volumetric flasks, Class A - 10 ml and 100 ml, with ground-glass
stoppers.
4.7 Vials - 2 ml, for GC autosampler.
4.8 Spatula - Stainless steel.
4.9 Disposable pipets - Pasteur.
4.10 Heater or heated oil bath - Should be capable of maintaining the
purging chamber to within 1°C over the temperature range of ambient to 100°C.
4.11 Purge-and-trap device - The purge-and-trap device consists of three
separate pieces of equipment: the sample purger, the trap, and the desorber.
Several complete devices are commercially available.
4.11.1 The recommended purging chamber is designed to accept 5 ml
samples with a water column at least 3 cm deep. The gaseous headspace
between the water column and the trap must have a total volume of less than
15 ml. The purge gas must pass through the water column as finely divided
bubbles with a diameter of.less than 3 mm at the origin. The purge gas
must be introduced no more than 5 mm from the base of the water column.
The sample purger, illustrated in Figure 1, meets these design criteria.
Alternate sample purge devices may be utilized, provided equivalent
performance is demonstrated.
4.11.2 The trap must be at least 25 cm long and have an inside
diameter of at least 0.105 in. Starting from the inlet, the trap must
contain the following amounts of adsorbents: 1/3 of 2,6-diphenylene oxide
polymer, 1/3 of silica gel, and 1/3 of coconut charcoal. It is recommended
that 1.0 cm of methyl silicone coated packing be inserted at the inlet to
extend the life of the trap (see Figure 2). If it is not necessary to
analyze for dichlorodifluoromethane or other fluorocarbons of similar
volatility, the charcoal can be eliminated and the polymer increased to
fill 2/3 of the trap. If only compounds boiling above 35°C are to be
8240B - 5 Revision 2
November 1990
-------
analyzed, both the silica gel and charcoal can be eliminated and the
polymer increased to fill the entire trap. Before initial use, the trap
should be conditioned overnight at 180°C by backflushing with an inert gas
flow of at least 20 mL/min. Vent the trap effluent to the room, not to
the analytical column. Prior to daily use, the trap should be conditioned
for 10 minutes at 180°C with backflushing. The trap may be vented to the
analytical column during daily conditioning. However, the column must be
run through the temperature program prior to analysis of samples.
*
4.11.3 The desorber should be capable of rapidly heating the trap
to 180°C for desorption. The polymer section of the trap should not be
heated higher than 180°C, and the remaining sections should not exceed
220°C during bake out mode. The desorber design illustrated in Figure 2
meets these criteria.
4.11.4 The purge-and-trap device may be assembled as a separate
unit or may be coupled to a gas chromatograph, as shown in Figures 3 and 4.
4.11.5 Trap Packing Materials
4.11.5.1 2,6-Diphenylene oxide polymer - 60/80 mesh,
chromatographic grade (Tenax GC or equivalent).
4.11.5.2 Methyl silicone packing - OV-1 (3%) on Chromosorb-W,
60/80 mesh or equivalent.
4.11.5.3 Silica gel - 35/60 mesh, Davison, grade 15 or
equivalent.
4.11.5.4 Coconut charcoal - Prepare from Barnebey Cheney,
CA-580-26, lot #M-2649, by crushing through 26 mesh screen (or
equivalent).
4.12 Gas chromatograph/mass spectrometer system
4.12.1 Gas chromatograph - An analytical system complete with a
temperature programmable gas chromatograph and all required accessories
including syringes, analytical columns, and gases.
4.12.2 Column - 6 ft x 0.1 in. ID glass, packed with 1% SP-1000
on Carbopack-B (60/80 mesh) or equivalent.
4.12.3 Mass spectrometer - Capable of scanning from 35-260 amu
every 3 seconds or less, using 70 volts (nominal) electron energy in the
electron impact mode and producing a mass spectrum that meets all the
criteria in Table 3 when 50 ng of 4-bromofluorobenzene (BFB) are injected
through the gas chromatograph inlet.
4.12.4 GC/MS interface - Any GC-to-MS interface that gives
acceptable calibration points at 50 ng or less per injection for each of
the analytes and achieves all acceptable performance criteria (see Table 3)
may be used. GC-to-MS interfaces constructed entirely of glass or of glass-
lined materials are recommended. Glass can be deactivated by silanizing
with dichlorodimethylsilane.
8240B - 6 Revision 2
November 1990
-------
4.12.5 Data system - A computer system that allows the continuous
acquisition and storage on machine readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows searching any GC/MS data file for ions of a specified mass and
plotting such ion abundances versus time or scan number. This type of plot
is defined as an Extracted Ion Current Profile (EICP). Software must also
be available that allows integrating the abundances in any EICP between
specified time or scan number limits. The most recent version of the
EPA/NIST Mass Spectral Library should also be available.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock solutions - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standard solutions
in methanol, using assayed liquids or gases, as appropriate.
5.3.1 Place about 9.8 ml of methanol in a 10 mL tared ground-glass-
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all alcohol wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
5.3.2 Add the assayed reference material, as described below.
5.3.2.1 Liquids - Using a 100 /iL syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.3.2.2 Gases - To prepare standards for any compounds that
boil below 30°C (e.g. bromomethane, chloroethane, chloromethane, or
vinyl chloride), fill a 5 mL valved gas-tight syringe with the
reference standard to the 5.0 mL mark. Lower the needle to 5 mm
above the methanol meniscus. Slowly introduce the reference standard
above the surface of the liquid. The heavy gas will rapidly dissolve
in the methanol. Standards may also be prepared by using a lecture
bottle equipped with a Hamilton Lecture Bottle Septum (#86600).
Attach Teflon tubing to the side-arm relief valve and direct a gentle
stream of gas into the methanol meniscus.
5.3.3 Reweigh, dilute to vofume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
8240B - 7 Revision 2
November 1990
-------
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.3.4 Transfer the stock standard solution into a Teflon sealed
screw cap bottle. Store, with minimal headspace, at -10°C to -20°C and
protect from light.
5.3.5 Prepare fresh standards every two months for gases. Reactive
compounds such as 2-chloroethylvinyl ether and styrene may need to be
prepared more frequently. All other standards must be replaced after six
months. Both gas and liquid standards must be monitored closely by
comparison to the initial calibration curve and by comparison to QC check
standards. It may be necessary to replace the standards more frequently
if either check exceeds a 25% difference.
5.4 Secondary dilution standards - Using stock standard solutions, prepare
in methanol, secondary dilution standards containing the compounds of interest,
either singly or mixed together. Secondary dilution standards must be stored
with minimal headspace and should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards from
them.
5.5 Surrogate standards - The surrogates recommended are toluene-da,
4-bromofluorobenzene, and l,2-dichloroethane-d4. Other compounds may be used
as surrogates, depending upon the analysis requirements. A stock surrogate
solution in methanol should be prepared as described in Section 5.3, and a
surrogate standard spiking solution should be prepared from the stock at a
concentration of 250 /ug/10 ml in methanol. Each sample undergoing GC/MS
analysis must be spiked with 10 /iL of the surrogate spiking solution prior to
analysis.
5.6 Internal standards - The recommended internal standards are
bromochloromethane, 1,4-difluorobenzene, and chlorobenzene-ds. Other compounds
may be used as internal standards as long as they have retention times similar
to the compounds being detected by GC/MS. Prepare internal standard stock and
secondary dilution standards in methanol using the procedures described in
Sections 5.3 and 5.4. It is recommended that the secondary dilution standard
should be prepared at a concentration of 25 mg/L of each internal standard
compound. Addition of 10 p.1 of this standard to 5.0 ml of sample or calibration
standard would be the equivalent of 50 M9/L-
5.7 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng/jiL of BFB in methanol should be prepared.
5.8 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared from the secondary dilution of stock standards
(see Sections 5.3 and 5.4). Prepare these solutions in organic-free reagent
water. One of the concentrations should be at a concentration near, but above,
the method detection limit. The remaining concentrations should correspond to
the expected range of concentrations found in real samples but should not exceed
8240B - 8 Revision 2
November 1990
-------
the working range of the GC/MS system. Each standard should contain each analyte
for detection by this method (e.g. some or all of the target analytes may be
included). Calibration standards must be prepared daily.
5.9 Matrix spiking standards - Matrix spiking standards should be prepared
from volatile organic compounds which will be representative of the compounds
being investigated. The suggested compounds are 1,1-dichloroethene,
trichloroethene, chlorobenzene, toluene, and benzene. The standard should be
prepared in methanol, with each compound present at a concentration of
250 jug/10.0 ml.
5.10 Great care must be taken to maintain the integrity of all standard
solutions. It is recommended that all standards in methanol be stored at -10°C
to -20°C in screw cap amber bottles with Teflon liners.
5.11 Methanol, CH3OH. Pesticide quality or equivalent. Store apart from
other solvents.
5.12 Reagent Tetraglyme - Reagent tetraglyme is defined as tetraglyme in
which interference is not observed at the method detection limit of compounds
of interest.
5.12.1 Tetraglyme (tetraethylene glycol dimethyl ether, Aldrich
#17, 240-5 or equivalent), C8H1805. Purify by treatment at reduced pressure
in a rotary evaporator. The tetraglyme should have a peroxide content of
less than 5 ppm as indicated by EM Quant Test Strips (available from
Scientific Products Co., Catalog No. P1126-8 or equivalent).
CAUTION; Glycol ethers are suspected carcinogens. All solvent handling should
be done in a hood while using proper protective equipment to minimize
exposure to liquid and vapor.
Peroxides may be removed by passing the tetraglyme through a column
of activated alumina. The tetraglyme is placed in a round bottom flask
equipped with a standard taper joint, and the flask is affixed to a rotary
evaporator. The flask is immersed in a water bath at 90-100°C and a vacuum
is maintained at < 10 mm Hg for at least two hours using a two stage
mechanical pump. The vacuum system is equipped with an all glass trap,
which is maintained in a dry ice/methanol bath. Cool the tetraglyme to
ambient temperature and add 0.1 mg/mL of 2,6-di-tert-butyl-4-methyl-phenol
to prevent peroxide formation. Store the tetraglyme in a tightly sealed
screw cap bottle in an area that is not contaminated by solvent vapors.
5.12.2 In order to demonstrate that all interfering volatiles have
been removed from the tetraglyme, an organic-free reagent water/tetraglyme
blank must be analyzed.
5.13 Polyethylene glycol, H(OCH2CH2)nOH. Free of interferences at the
detection limit of the analytes.
8240B - 9 Revision 2
November 1990
-------
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the Introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Direct Injection - In very limited applications (e.g. aqueous process
wastes), direct Injection of the sample Into the GC/MS system with a 10 ML
syringe may be appropriate. One such application is for verification of the
alcohol content of an aqueous sample prior to determining if the sample is
ignitable (Methods 1010 or 1020). In this case, it is suggested that direct
injection be used. The detection limit is very high (approximately 10,000 M9/L);
therefore, 1t is only permitted when concentrations in excess of 10,000 ng/L are
expected or for water soluble compounds that do not purge. The system must be
calibrated by direct injection (bypassing the purge-and-trap device).
7.2 Initial calibration for purge-and-trap procedure
7.2.1 Recommended GC/MS operating conditions
Electron energy: 70 volts (nominal).
Mass range: 35-260 amu.
Scan time: To give 5 scans/peak, but not to exceed 7
sec/scan.
Initial column temperature: 45°C.
Initial column holding time: 3 minutes.
Column temperature program: 8°C/minute.
Final column temperature: 220°C.
Final column holding time: 15 minutes.
Injector temperature: 200-225°C.
Source temperature: According to manufacturer's specifications.
Transfer line temperature: 250-300°C.
Carrier gas: Hydrogen at 50 cm/sec or helium at 30 cm/sec.
7.2.2 Each GC/MS system must be hardware tuned to meet the criteria
in Table 3 for a 50 ng injection or purging of 4-bromofluorobenzene (2 pi
injection of the BFB standard). Analyses must not begin until these
criteria are met.
7.2.3 Assemble a purge-and-trap device that meets the specification
in Section 4.11. Condition the trap overnight at 180°C in the purge mode
with an inert gas flow of at least 20 mL/min. Prior to use, condition the
trap daily for 10 min while backflushing at 180°C with the column at 220°C.
7.2.4 Connect the purge-and-trap device to a gas chromatograph.
7.2.5 Prepare the final solutions containing the required
concentrations of calibration standards, including surrogate standards,
directly in the purging device (use freshly prepared stock solutions when
preparing the calibration standards for the initial calibration.) Add
5.0 mL of organic-free reagent water to the purging device. The organic-
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free reagent water is added to the purging device using a 5 ml glass
syringe fitted with a 15 cm, 20 gauge needle. The needle Is Inserted
through the sample Inlet shown In Figure 1. The Internal diameter of the
14 gauge needle that forms the sample Inlet will permit Insertion of the
20 gauge needle. Next, using a 10 nl or 25 ML mlcrosyrlnge equipped with
a long needle (Section 4.1), take a volume of the secondary dilution
solution containing appropriate concentrations of the calibration standards
(Section 5.6). Add the aliquot of calibration solution directly to the
organic-free reagent water in the purging device by inserting the needle
through the sample inlet. When discharging the contents of the
microsyringe, be sure that the end of the syringe needle is well beneath
the surface of the organic-free reagent water. Similarly, add 10 /uL of
the internal standard solution (Section 5.4). Close the 2 way syringe
valve at the sample inlet.
7.2.6 Carry out the purge-and-trap analysis procedure as described
in Section 7.4.1.
7.2.7 Tabulate the area response of the characteristic ions (see
Table 1) against concentration for each compound and each internal
standard. Calculate response factors (RF) for each compound relative to
one of the internal standards. The internal standard selected for the
calculation of the RF for a compound should be the internal standard that
has a retention time closest to the compound being measured (Section
7.5.2). The RF is calculated as follows:
RF = (AXC1S)/(A1SCX)
where:
AX = Area of the characteristic ion for the compound being measured.
A,s = Area of the characteristic ion for the specific internal standard.
C,s = Concentration of the specific internal standard.
Cx = Concentration of the compound being measured.
7.2.8 The average RF must be calculated for each compound. A system
performance check should be made before this calibration curve is used.
Five compounds (the System Performance Check Compounds, or SPCCs) are
checked for a minimum average response factor. These compounds are
chloromethane, 1,1-dichloroethane, bromoform, 1,1,2,2-tetrachloroethane,
and chlorobenzene. The minimum acceptable average RF for these compounds
should be 0.300 (0.250 for bromoform). These compounds typically have RFs
of 0.4-0.6 and are used to check compound instability and to check for
degradation caused by contaminated lines or active sites in the system.
Examples of these occurrences are:
7.2.8.1 Chloromethane - This compound is the most likely
compound to be lost if the purge flow is too fast.
7.2.8.2 Bromoform - This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in the transfer lines may adversely
affect response. Response of the quantitation ion (m/z 173) is
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directly affected by the tuning of BFB at ions m/z 174/176.
Increasing the m/z 174/176 ratio may improve bromoform response.
7.2.8.3 Tetrachloroethane and 1,1-dichloroethane - These
compounds are degraded by contaminated transfer lines in purge-and-
trap systems and/or active sites in trapping materials.
7.2.9 Using the RFs from the initial calibration, calculate the
percent relative standard deviation (%RSD) for Calibration Check Compounds
(CCCs).
SD
%RSD x 100
where:
RSD = relative standard deviation.
x = mean of 5 initial RFs for a compound.
SD = standard deviation of average RFs for a compound.
SD
N (x, -
i=l N - 1
The %RSD for each individual CCC should be less than 30 percent.
This criterion must be met in order for the individual calibration to be
valid. The CCCs are:
1,1-Dichloroethene,
Chloroform,
1,2-Dichloropropane,
Toluene,
Ethyl benzene, and
Vinyl chloride.
7.3 Daily GC/MS calibration
7.3.1 Prior to the analysis of samples, inject or purge 50 ng of
the 4-bromofluorobenzene standard. The resultant mass spectra for the
BFB must meet all of the criteria given in Table 3 before sample analysis
begins. These criteria must be demonstrated each 12 hour shift.
7.3.2 The initial calibration curve (Section 7.2) for each compound
of interest must be checked and verified once every 12 hours of analysis
time. This is accomplished by analyzing a calibration standard that is
at a concentration near the midpoint concentration for the working range
of the GC/MS by checking the SPCC (Section 7.3.3) and CCC (Section 7.3.4).
7.3.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made each 12 hours. If the SPCC criteria are
met, a comparison of response factors is made for all compounds. This is
the same check that is applied during the initial calibration. If the
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minimum response factors are not met, the system must be evaluated, and
corrective action must be taken before sample analysis begins. The minimum
response factor for volatile SPCCs is 0.300 (0.250 for Bromoform). Some
possible problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
and active sites in the column or chromatographic system.
7.3.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Section 7.2.9 are used to check
the validity of the initial calibration. Calculate the percent difference
using:
RF, - RF
% Difference = —= - x 100
RF,
where:
RF, = average response factor from initial calibration.
RFC = response factor from current verification check standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. If the percent difference
for each CCC is less than 25%, the initial calibration is assumed to be
valid. If the criterion is not met (> 25% difference), for any one CCC,
corrective action MUST be taken. Problems similar to those listed under
SPCCs could affect this criterion. If no source of the problem can be
determined after corrective action has been taken, a new five point
calibration MUST be generated. This criterion MUST be met before
quantitative sample analysis begins.
7.3.5 The internal standard responses and retention times in the
check calibration standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last check calibration (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (- 50% to + 100%) from the last daily
calibration standard check, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate. When
corrections are made, reanalysis of samples analyzed while the system was
malfunctioning are necessary.
7.4 GC/MS analysis
7.4.1 Water samples
7.4.1.1 Screening of the sample prior to purge-and-trap
analysis will provide guidance on whether sample dilution is
necessary and will prevent contamination of the purge-and-trap
system. Two screening techniques that can be used are: the
headspace sampler (Method 3810) using a gas chromatograph (GC)
equipped with a photo ionization detector (PID) in series with an
electrolytic conductivity detector (HECD); and extraction of the
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sample with hexadecane and analysis of the extract on a GC with a
FID and/or an ECD (Method 3820).
7.4.1.2 All samples and standard solutions must be allowed
to warm to ambient temperature before analysis.
7.4.1.3 Set up the GC/MS system as outlined in Section 7.2.1.
7.4.1.4 BFB tuning criteria and daily GC/MS calibration
criteria must be met (Section 7.3) before analyzing samples.
7.4.1.5 Adjust the purge gas (helium) flow rate to 25-
40 mL/min on the purge-and-trap device. Optimize the flow rate to
provide the best response for chloromethane and bromoform, if these
compounds are analytes. Excessive flow rate reduces chloromethane
response, whereas insufficient flow reduces bromoform response (see
Section 7.2.8).
7.4.1.6 Remove the plunger from a 5 ml syringe and attach a
closed syringe valve. Open the sample or standard bottle, which
has been allowed to come to ambient temperature, and carefully pour
the sample into the syringe barrel to just short of overflowing.
Replace the syringe plunger and compress the sample. Open the
syringe valve and vent any residual air while adjusting the sample
volume to 5.0 ml. This process of taking an aliquot destroys the
validity of the liquid sample for future analysis; therefore, if
there is only one VOA vial, the analyst should fill a second syringe
at this time to protect against possible loss of sample integrity.
This second sample is maintained only until such time when the
analyst has determined that the first sample has been analyzed
properly. Filling one 20 ml syringe would allow the use of only one
syringe. If a second analysis is needed from a syringe, it must be
analyzed within 24 hours. Care must be taken to prevent air from
leaking into the syringe.
7.4.1.7 The following procedure is appropriate for diluting
purgeable samples. All steps must be performed without delays until
the diluted sample is in a gas tight syringe.
7.4.1.7.1 Dilutions may be made in volumetric flasks
(10 to 100 ml). Select the volumetric flask that will allow
for the necessary dilution. Intermediate dilutions may be
necessary for extremely large dilutions.
7.4.1.7.2 Calculate the approximate volume of organic-
free reagent water to be added to the volumetric flask selected
and add slightly less than this quantity of organic-free
reagent water to the flask.
7.4.1.7.3 Inject the proper aliquot of samples from
the syringe prepared in Section 7.4.1.6 into the flask.
Aliquots of less than 1 ml are not recommended. Dilute the
sample to the mark with organic-free reagent water. Cap the
flask, invert, and shake three times. Repeat above procedure
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for additional dilutions.
7.4.1.7.4 Fill a 5 ml syringe with the diluted sample
as in Section 7.4.1.6.
7.4.1.8 Add 10.0 /iL of surrogate spiking solution (Section
5.3) and 10 /iL of internal standard spiking solution (Section 5.4)
through the valve bore of the syringe; then close the valve. The
surrogate and internal standards may be mixed and added as a single
spiking solution. The addition of 10 juL of the surrogate spiking
solution to 5 ml of sample is equivalent to a concentration of
50 /itg/L of each surrogate standard.
7.4.1.9 Attach the syringe-syringe valve assembly to the
syringe valve on the purging device. Open the syringe valves and
inject the sample into the purging chamber.
7.4.1.10 Close both valves and purge the sample for 11.0 ±
0.1 minutes at ambient temperature.
7.4.1.11 At the conclusion of the purge time, attach the
trap to the chromatograph, adjust the device to the desorb mode,
and begin the gas chromatographic temperature program and GC/MS data
acquisition. Concurrently, introduce the trapped materials to the
gas chromatographic column by rapidly heating the trap to 180°C while
backflushing the trap with inert gas between 20 and 60 mL/min for
4 minutes. If this rapid heating requirement cannot be met, the gas
chromatographic column must be used as a secondary trap by cooling
it to 30°C (or subambient, if problems persist) instead of the
recommended initial program temperature of 45°C.
7.4.1.12 While the trap is being desorbed into the gas
chromatograph, empty the purging chamber. Wash the chamber with a
minimum of two 5 ml flushes of organic-free reagent water (or
methanol followed by organic-free reagent water) to avoid carryover
of pollutant compounds into subsequent analyses.
7.4.1.13 After desorbing the sample for 4 minutes,
recondition the trap by returning the purge-and-trap device to the
purge mode. Wait 15 seconds; then close the syringe valve on the
purging device to begin gas flow through the trap. The trap
temperature should be maintained at 180°C. Trap temperatures up to
220°C may be employed; however, the higher temperature will shorten
the useful life of the trap. After approximately 7 minutes, turn
off the trap heater and open the syringe valve to stop the gas flow
through the trap. When cool, the trap is ready for the next sample.
7.4.1.14 If the initial analysis of a sample or a dilution
of the sample has a concentration of analytes that exceeds the
initial calibration range, the sample must be reanalyzed at a higher
dilution. Secondary ion quantitation is allowed only when there are
sample interferences with the primary ion. When a sample is analyzed
that has saturated ions from a compound, this analysis must be
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followed by a blank organic-free reagent water analysis. If the
blank analysis is not free of interferences, the system must be
decontaminated. Sample analysis may not resume until a blank can
be analyzed that is free of interferences.
7.4.1.15 For matrix spike analysis, add 10 /nL of the matrix
spike solution (Section 5.7) to the 5 ml of sample to be purged.
Disregarding any dilutions, this is equivalent to a concentration
of 50 M9/L of each matrix spike standard.
7.4.1.16 All dilutions should keep the response of the major
constituents (previously saturated peaks) in the upper half of the
linear range of the curve. Proceed to Sections 7.5.1 and 7.5.2 for
qualitative and quantitative analysis.
7.4.2 Water miscible liquids
7.4.2.1 Water miscible liquids are analyzed as water samples
after first diluting them at least 50 fold with organic-free reagent
water.
7.4.2.2 Initial and serial dilutions can be prepared by
pipetting 2 ml of the sample to a 100 ml volumetric flask and
diluting to volume with organic-free reagent water. Transfer
immediately to a 5 ml gas tight syringe.
7.4.2.3 Alternatively, prepare dilutions directly in a 5 ml
syringe filled with organic-free reagent water by adding at least
20 MU but not more than 100 /iL of liquid sample. The sample is
ready for addition of internal and surrogate standards.
7.4.3 Sediment/soil and waste samples - It is highly recommended
that all samples of this type be screened prior to the purge-and-trap
GC/MS analysis. The headspace method (Method 3810) or the hexadecane
extraction and screening method (Method 3820) may be used for this purpose.
These samples may contain percent quantities of purgeable organics that
will contaminate the purge-and-trap system, and require extensive cleanup
and instrument downtime. Use the screening data to determine whether to
use the low-concentration method (0.005-1 mg/Kg) or the high-concentration
method (> 1 mg/Kg).
7.4.3.1 Low-concentration method - This is designed for
samples containing individual purgeable compounds of < 1 mg/Kg. It
is limited to sediment/soil samples and waste that is of a similar
consistency (granular and porous). The low-concentration method is
based on purging a heated sediment/soil sample mixed with organic-
free reagent water containing the surrogate and internal standards.
Analyze all reagent blanks and standards under the same conditions
as the samples. See Figure 5 for an illustration of a low soils
impinger.
7.4.3.1.1 Use a 5 g sample if the expected concentration
is < 0.1 mg/Kg or a 1 g sample for expected concentrations
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between 0.1 and 1 mg/Kg.
7.4.3.1.2 The GC/MS system should be set up as in
Sections 7.4.1.2-7.4.1.4. This should be done prior to the
preparation of the sample to avoid loss of volatiles from
standards and samples. A heated purge calibration curve must
be prepared and used for the quantitation of all samples
analyzed with the low-concentration method. Follow the initial
and daily calibration instructions, except for the addition
of a 40°C purge temperature.
7.4.3.1.3 Remove the plunger from a 5 ml Luerlock type
syringe equipped with a syringe valve and fill until
overflowing with water. Replace the plunger and compress the
water to vent trapped air. Adjust the volume to 5.0 ml. Add
10 /uL each of surrogate spiking solution (Section 5.3) and
internal standard solution (Section 5.4) to the syringe through
the valve. (Surrogate spiking solution and internal standard
solution may be mixed together.) The addition of 10 /uL of the
surrogate spiking solution to 5 g of sediment/soil is
equivalent to 50 M9/K9 of each surrogate standard.
7.4.3.1.4 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not discard
any supernatant liquids. Mix the contents of the sample
container with a narrow metal spatula. Weigh the amount
determined in Section 7.4.3.1.1 into a tared purge device.
Note and record the actual weight to the nearest 0.1 g.
7.4.3.1.5 Determine the percent dry weight of the
soil/sediment sample. This includes waste samples that are
amenable to percent dry weight determination. Other wastes
should be reported on a wet-weight basis.
7.5.3.1.5.1 Immediately after weighing the sample
for extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by
drying overnight at 105°C. Allow to cool in a desiccator
before re-weighing. Concentrations of individual
analytes are reported relative to the dry weight of
sample.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.
% dry weight = q of dry sample x 100
g of sample
7.4.3.1.6 Add the spiked water to the purge device,
which contains the weighed amount of sample, and connect the
device to the purge-and-trap system.
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NOTE; Prior to the attachment of the purge device, the procedures in Sections
7.4.3.1.4 and.7.4.3.1.6 must be performed rapidly and without interruption
to avoid loss of volatile organics. These steps must be performed in a
laboratory free of solvent fumes.
7.4.3.1.7 Heat the sample to 40°C ± 1°C and purge the
sample for 11.0 ± 0.1 minute.
7.4.3.1.8 Proceed with the analysis as outlined in
Sections 7.4.1.11-7.4.1.16. Use 5 mL of the same organic-free
reagent water as in the reagent blank. If saturated peaks
occurred or would occur if a 1 g sample were analyzed, the
high-concentration method must be followed.
7.4.3.1.9 For low-concentration sediment/soils add
10 /iL of the matrix spike solution (Section 5.7) to the 5 ml
of organic-free reagent water (Section 7.4.3.1.3). The
concentration for a 5 g sample would be equivalent to 50 M9/Kg
of each matrix spike standard.
7.4.3.2 High-concentration method - The method is based on
extracting the sediment/soil with methanol. A waste sample is either
extracted or diluted, depending on its solubility in methanol.
Wastes (i.e. petroleum and coke wastes) that are insoluble in
methanol are diluted with reagent tetraglyme or possibly polyethylene
glycol (PEG). An aliquot of the extract is added to organic-free
reagent water containing internal standards. This is purged at
ambient temperature. All samples with an expected concentration of
> 1.0 mg/Kg should be analyzed by this method.
7.4.3.2.1 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not discard
any supernatant liquids. Mix the contents of the sample
container with a narrow metal spatula. For sediment/soil and
solid wastes that are insoluble in methanol, weigh 4 g (wet
weight) of sample into a tared 20 ml vial. Use a top loading
balance. Note and record the actual weight to 0.1 gram and
determine the percent dry weight of the sample using the
procedure in Section 7.4.3.1.5. For waste that is soluble in
methanol, tetraglyme, or PEG, weigh 1 g (wet weight) into a
tared scintillation vial or culture tube or a 10 ml volumetric
flask. (If a vial or tube is used, it must be calibrated
prior to use. Pipet 10.0 ml of solvent into the vial and mark
the bottom of the meniscus. Discard this solvent.)
7.4.3.2.2 Quickly add 9.0 ml of appropriate solvent;
then add 1.0 ml of the surrogate spiking solution to the vial.
Cap and shake for 2 minutes.
NOTE; Sections 7.4.3.2.1 and 7.4.3.2.2 must be performed rapidly and without
interruption to avoid loss of volatile organics. These steps must be
performed in a laboratory free from solvent fumes.
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7.4.3.2.3 Pi pet approximately 1 mi of the extract to
a GC vial for storage, using a disposable pipet. The remainder
may be disposed of. Transfer approximately 1 ml of appropriate
solvent to a separate GC vial for use as the method blank for
each set of samples. These extracts may be stored at 4°C in
the dark, prior to analysis. The addition of a 100 /iL aliquot
of each of these extracts in Section 7.4.3.2.6 will give a
concentration equivalent to 6,200 M9/Kg of each surrogate
standard.
7.4.3.2.4 The GC/MS system should be set up as in
Sections 7.4.1.2-7.4.1.4. This should be done prior to the
addition of the solvent extract to organic-free reagent water.
7.4.3.2.5 Table 4 can be used to determine the volume
of solvent extract to add to the 5 ml of organic-free reagent
water for analysis. If a screening procedure was followed
(Method 3810 or 3820), use the estimated concentration to
determine the appropriate volume. Otherwise, estimate the
concentration range of the sample from the low-concentration
analysis to determine the appropriate volume. If the sample
was submitted as a high-concentration sample, start with 100
p.1. All dilutions must keep the response of the major
constituents (previously saturated peaks) in the upper half
of the linear range of the curve.
7.4.3.2.6 Remove the plunger from a 5.0 ml Luerlock
type syringe equipped with a syringe valve and fill until
overflowing with water. Replace the plunger and compress the
water to vent trapped air. Adjust the volume to 4.9 ml. Pull
the plunger back to 5.0 ml to allow volume for the addition
of the sample extract and of standards. Add 10 pi of internal
standard solution. Also add the volume of solvent extract
determined in Section 7.4.3.2.5 and a volume of extraction or
dissolution solvent to total 100 /LiL (excluding methanol in
standards).
7.4.3.2.7 Attach the syringe-syringe valve assembly to
the syringe valve on the purging device. Open the syringe
valve and inject the organic-free reagent water/methanol sample
into the purging chamber.
7.4.3.2.8 Proceed with the analysis as outlined in
Section 7.4.1.11-7.4.1.16. Analyze all reagent blanks on the
same instrument as that use for the samples. The standards
and blanks should also contain 100 n\. of solvent to simulate
the sample conditions.
7.4.3.2.9 For a matrix spike in the high-concentration
sediment/soil samples, add 8.0 mL of methanol, 1.0 ml of
surrogate spike solution (Section 5.3), and 1.0 ml of matrix
spike solution (Section 5.7) as in Section 7.4.3.2.2. This
results in a 6,200 M9/Kg concentration of each matrix spike
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standard when added to a 4 g sample. Add a 100 /iL aliquot of
this extract to 5 ml of organic-free reagent water for purging
(as per Section 7.4.3.2.6).
7.5 Data interpretation
7.5.1 Qualitative analysis
7.5.1.1 The qualitative identification of compounds determined
by this method is based on retention time, and on comparison of the
sample mass spectrum, after background correction, with
characteristic ions in a reference mass spectrum. The reference mass
spectrum must be generated by the laboratory using the conditions
of this method. The characteristic ions from the reference mass
spectrum are defined to be the three ions of greatest relative
intensity, or any ions over 30% relative intensity if less than three
such ions occur in the reference spectrum. Compounds should be
identified as present when the criteria below are met.
7.5.1.1.1 The intensities of the characteristic ions of a
compound maximize in the same scan or within one scan of each
other. Selection of a peak by a data system target compound
search routine where the search is based on the presence of
a target chromatographic peak containing ions specific for the
target compound at a compound-specific retention time will be
accepted as meeting this criterion.
7.5.1.1.2 The RRT of the sample component is within ± 0.06
RRT units of the RRT of the standard component.
7.5.1.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions
in the reference spectrum. (Example: For an ion with an
abundance of 50% in the reference spectrum, the corresponding
abundance in a sample spectrum can range between 20% and 80%.)
7.5.1.1.4 Structural isomers that produce very similar mass
spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
7.5.1.1.5'Identification is hampered when sample components
are not resolved chromatographically and produce mass spectra
containing ions contributed by more than one analyte. When
gas chromatographic peaks obviously represent more than one
sample component (i.e., a broadened peak with shoulder(s) or
a valley between two or more maxima), appropriate selection
of analyte spectra and background spectra is important.
Examination of extracted ion current profiles of appropriate
ions can aid in the selection of spectra, and in qualitative
8240B - 20 Revision 2
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identification of compounds. When analytes coelute (i.e., only
one chromatographic peak is apparent), the identification
criteria can be met, but each analyte spectrum will contain
extraneous ions contributed by the coeluting compound.
7.5.1.2 For samples containing components not associated with
the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the type of analyses
being conducted. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within + 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from
the sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison
of sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
7.5.2 Quantitative analysis
7.5.2.1 When a compound has been identified, the quantitation
of that compound will be based on the integrated abundance from the
EICP of the primary characteristic ion. Quantitation will take place
using the internal standard technique. The internal standard used
shall be the one nearest the retention time of that of a given
analyte (e.g. see Table 5).
7.5.2.2 Calculate the concentration of each identified analyte
in the sample as follows:
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Water
(AJdJ
concentration (M9/L) =
(Als)(RF)(V0)
where:
A,, = Area of characteristic ion for compound being measured.
Is = Amount of internal standard injected (ng).
Als= Area of characteristic ion for the internal standard.
RF = Response factor for compound being measured (Section 7.3.3).
V0 = Volume of water purged (ml), taking into consideration any
dilutions made.
Sediment/Soil Sludge (on a dry-weight basis) and Waste (normally on
a wet-weight basis)
(AJ(Is)(Vt)
concentration (Mg/Kg) =
(AJ(RF)(V,)(H.)(D)
where:
A,,, Is, Als, RF, = Same as for water.
V, = Volume of total extract (pi) (use 10,000 /xL or a factor of this
when dilutions are made).
V, = Volume of extract added (/nL) for purging.
Ws = Weight of sample extracted or purged (g).
D = % dry weight of sample/100, or 1 for a wet-weight basis.
7.6.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
given above should be used with the following modifications: The
areas A,, and Als should be from the total ion chromatograms, and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Required instrument QC is found in the following sections:
8.2.1 The GC/MS system must be tuned to meet the BFB specifications
in Section 7.2.2.
8.2.2 There must be an initial calibration of the GC/MS system as
8240B - 22 Revision 2
November 1990
-------
specified in Section 7.2.
8.2.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.3.3 and the CCC criteria in Section 7.3.4, each 12 hours.
8.3 To establish the ability to generate acceptable accuracy and precision,
the analyst must perform the following operations.
8.3.1 A quality control (QC) reference sample concentrate is
required containing each analyte at a concentration of 10 mg/L in methanol.
The QC reference sample concentrate may be prepared from pure standard
materials or purchased as certified solutions. If prepared by the
laboratory, the QC reference sample concentrate must be made using stock
standards prepared independently from those used for calibration.
8.3.2 Prepare a QC reference sample to contain 20 M9/L of each
analyte by adding 200 /LtL of QC reference sample concentrate to 100 ml of
organic-free reagent water.
8.3.3 Four 5 ml aliquots of the well mixed QC reference sample are
analyzed according to the method beginning in Section 7.4.1.
8.3.4 Calculate the average recovery (x) in p.g/1, and the standard
deviation of the recovery (s) in M9/U for each analyte using the four
results.
8.3.5 For each analyte compare s and x with the corresponding
acceptance criteria_for precision and accuracy, respectively, found in
Table 6. If s and x for all analytes meet the acceptance criteria, the
system performance is acceptable and analysis of actual samples can_begin.
If any individual s exceeds the precision limit or any individual x falls
outside the range for accuracy, then the system performance is unacceptable
for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial probability
that one or more will fail at least one of the acceptance criteria when
all analytes of a given method are determined.
8.3.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Section
8.3.6.1 or 8.3.6.2.
8.3.6.1 Locate and correct the source of the problem and
repeat the test for all analytes beginning with Section 8.3.2.
8.3.6.2 Beginning with Section 8.3.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Section
8.3.2.
8240B - 23 Revision 2
November 1990
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8.4 For aqueous and soil matrices, laboratory established surrogate
control limits should be compared with the control limits listed in Table 8.
The limits given in Table 8 are multilaboratory performance based limits for
soil and aqueous samples, and therefore, the single laboratory limits must fall
within those given in Table 8 for these matrices.
8.4.1 If recovery is not within limits, the following procedures
are required.
8.4.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.4.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.4.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.4.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4.2 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL concentrations listed in
Table 1 were obtained using organic-free reagent water. Similar results were
achieved using representative wastewaters. The MDL actually achieved in a given
analysis will vary depending on instrument sensitivity and matrix effects.
9.2 This method was tested by 15 laboratories using organic-free reagent
water, drinking water, surface water, and industrial wastewaters spiked at six
concentrations over the range 5-600 M9/L- Single operator precision, overall
precision, and method accuracy were found to be directly related to the
concentration of the analyte and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 7.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 624,"
October 26, 1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
8240B - 24 Revision 2
November 1990
-------
3. Bellar, T.A., and J.J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
739-744, 1974.
4. Bellar, T.A., and J.J. Lichtenberg, "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds," in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
5. Budde, W.L. and J.W. Eichelberger, "Performance Tests for the Evaluation
of Computerized Gas Chromatography/Mass Spectrometry Equipment and
Laboratories," EPA-600/4-79-020, U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268,
April 1980.
6. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems," Analytical Chemistry, 47, 995-1000, 1975.
7. "Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
8. "Interlaboratory Method Study for EPA Method 624-Purgeables," Final Report
for EPA Contract 68-03-3102.
9. "Method Performance Data for Method 624," Memorandum from R. Slater and T.
Pressley, U.S. Environmental Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio 45268, January 17, 1984.
10. Gebhart, J.E.; Lucas, S.V.; Naber, S.J.; Berry, A.M.; Danison, T.H.;
Burkholder, H.M. "Validation of SW-846 Methods 8010, 8015, and 8020"; U.S.
Environmental Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, Old 45268, July 1987, Contract No. 68-03-1760.
11. Lucas, S.V.; Kornfeld, R.A. "GC-MS Suitability Testing of RCRA Appendix
VIII and Michigan List Analytes "; U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
February 20, 1987, Contract No. 68-03-3224.
8240B - 25 Revision 2
November 1990
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TABLE 1.
RETENTION TIMES AND CHARACTERISTIC IONS FOR VOLATILE COMPOUNDS
Compound
Retention
Time (minutes)
Primary Ion Secondary Ion(s)
Ethyl ene oxide
Chloromethane
Di chl orodi f 1 uoromethane
Bromomethane
Vinyl chloride
Acetonitrile
Chloroethane
Methyl iodide
Methyl ene chloride
Carbon disulfide
Tri chl orofl uoromethane
Propionitrile
Ally! chloride
1,1-Dichloroethene
Bromochloromethane (I.S.)
Allyl alcohol
trans- 1,2-Di chl oroethene
1,2-Dichloroethane
Propargyl alcohol
Chloroform
1,2-Di chl oroethane-d4(surr)
2-Butanone
Methacrylonitrile
Dibromomethane
2-Chloroethanol
b-Propiolactone
Epichlorohydrin
1,1,1-Trichloroethane
Carbon tetrachloride
1,4-Dioxane
Isobutyl alcohol
Bromodichloromethane
Chloroprene
l,2:3,4-Diepoxybutane
1,2-Dichloropropane
Chloral hydrate (b)
cis-l,3-Dichloropropene
Bromoacetone
Tri chl oroethene
Benzene
trans - 1 , 3 -Di chl oropropene
1 , 1 , 2-Trichl oroethane
3-Chloropropionitrile
1,2-Di bromoethane
Pyridine
1.30
2.30
2.47
3.10
3.80
3.97
4.60
5.37
6.40
7.47
8.30
8.53
8.83
9.00
9.30
9.77
10.00
10.10
10.77
11.40
12.10
12.20
12.37
12.53
12.93
13.00
13.10
13.40
13.70
13.70
13.80
14.30
14.77
14.87
15.70
15.77
15.90
16.33
16.50
17.00
17.20
17.20
17.37
18.40
18.57
44
50
85
94
62
41
64
142
84
76
101
54
76
96
128
57
96
62
55
83
65
72
41
93
49
42
57
97
117
88
43
83
53
55
63
82
75
136
130
78
75
97
54
107
79
44, 43, 42
52, 49
85, 87, 101, 103
96, 79
64, 61
41, 40, 39
66, 49
142, 127, 141
49, 51, 86
76, 78, 44
103, 66
54, 52, 55, 40
76, 41, 39, 78
61, 98
49, 130, 51
57, 58, 39
61, 98
64, 98
55, 39, 38, 53
85, 47
102
43, 72
41, 67, 39, 52, 66
93, 174, 95, 172, 176
49, 44, 43, 51, 80
42, 43, 44
57, 49, 62, 51
99, 117
119, 121
88, 58, 43, 57
43, 41, 42, 74
85, 129
53, 88, 90, 51
55, 57, 56
62, 41
44, 84, 86, 111
77, 39
43, 136, 138, 93, 95
95, 97, 132
52, 71
77, 39
83, 85, 99
54, 49, 89, 91
107, 109, 93, 188
79, 52, 51, 50
8240B - 26
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Compound
Retention
Time (minutes)
Primary Ion Secondary Ion(s)
2-Chloroethyl vinyl ether
2-Hydroxypropi oni tri 1 e
1,4-Difluorobenzene (I.S.)
Malononitrile
Methyl methacrylate
Bromoform
1,1,1 , 2-Tetrachl oroethane
l,3-Dichloro-2-propanol
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
1,2, 3 -Tri chl oropropane
l,4-Dichloro-2-butene
n-Propylamine
2-Picoline
Toluene
Ethyl methacrylate
Chlorobenzene
Pentachl oroethane8
Ethyl benzene
1 , 2-Di bromo-3-chl oropropane
4-Bromofluorobenzene (surr.)
Benzyl chloride
Styrene
bis-(2-Chloroethyl) sulfide(b)
Acetone
Acrolein
Acrylonitrile
Chlorobenzene-d5 (I.S.)
Chl orodi bromomethane
1,1-Dichloroethane
Ethanol
O llnir-i^n^ir-i
fc-nexanone
lodomethane
4-Methyl -2-pentanone
Toluene-d8 (surr.)
Vinyl acetate
Xylene (Total)
18.60
18.97
19.60
19.60
19.77
19.80
20.33
21.83
22.10
22.20
22.20
22.73
23.00
23.20
23.50
23.53
24.60
24.83
26.40
27.23
28.30
29.50
30.83
33.53
--
--
--
--
--
--
--
--
--
--
--
--
~ ••
63
44
114
66
69
173
131
79
83
164
75
75
59
93
92
69
112
167
106
157
95
91
104
109
43
56
53
117
129
63
31
43
142
43
98
43
106
65,106
44,43,42,53
63,88
66,39,65,38
69,41,100,39
171,175,252
131,133,117,119,95
79,43,81,49
85,131,133
129,131,166
75,77,110,112,97
75,53,77,124,89
59,41,39
93,66,92,78
91,65
69,41,99,86,114
114,77
167,130,132,165,169
91
157,75,155,77
174,176
91,126,65,128
104,103,78,51,77
111, 158, 160
58
55,58
52,51
82,119
208,206
65,83
45,27,46
58,57, 100
127,141
58,57,100
70,100
86
91
a The base peak at m/e 117 was not used due to an interference at that mass
with a nearly coeluting internal standard, chlorobenzene-d5.
b Response factor judged to be too low (less than 0.02) for practical use.
8240B - 27
Revision 2
November 1990
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQL) FOR VOLATILE ORGANICS"
Estimated
Quantitation
Limits"
Ground water
Volatiles ng/l
Acetone
Acetonitrile
Allyl chloride
Benzene
Benzyl chloride
Bromodichloromethane
Bromoform
Bromomethane
2-Butanone
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chi orodi bromomethane
Chloroethane
2-Chloroethyl vinyl ether
Chloroform
Chloromethane
Chloroprene
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Oi bromomethane
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1 Dichloroethene
trans- 1, 2-Di chl oroethene
1 , 2 -Di chl oropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
Ethyl benzene
Ethyl methacrylate
2-Hexanone
Isobutyl alcohol
Methacrylonitrile
Methyl ene chloride
Methyl iodide
Methyl methacrylate
4-Methyl -2-pentanone
Pentachloroethane
100
100
5
5
100
5
5
10
100
100
5
5
5
10
10
5
10
5
100
5
5
100
5
5
5
5
5
5
5
5
5
5
50
100
100
5
5
5
50
10
Low Soil/Sediment
M9/K9
100
100
5
5
100
5
5
10
100
100
5
5
5
10
10
5
10
5
100
5
5
100
5
5
5
5
5
5
5
5
5
5
50
100
100
5
5
50
50
10
8240B - 28
Revision 2
November 1990
-------
TABLE 2.
(Continued)
Estimated
Quantltatlon
Limits"
Ground water Low Soil/Sediment
Volatlles M9/L M9/Kg
Propionitrile
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,1,1 -Tr1 chl oroethane
1 , 1 ,2-Tr1chloroethane
Trl chl oroethene
1,2,3-Trichloropropane
Vinyl acetate
Vinyl chloride
Xylene (Total)
100
5
5
5
5
5
5
5
5
5
50
10
5
100
5
5
5
5
5
5
5
5
5
50
10
5
a Sample EQLs are highly matrix dependent. The EQLs listed herein are provided
for guidance and may not always be achievable. See the following Information
for further guidance on matrix dependent EQLs.
b EQLs listed for soil/sediment are based on wet weight. Normally data Is
reported on a dry weight basis; therefore, EQLs will be higher, based on the
percent dry weight of each sample.
Other Matrices Factor0
Water miscible liquid waste 50
High-concentration soil and sludge 125
Non-water miscible waste 500
°EQL = [EQL for low soil sediment (Table 2)] X [Factor]. For non-aqueous
samples, the factor is on a wet weight basis.
8240B - 29 Revision 2
November 1990
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TABLE 3.
BFB KEY ION ABUNDANCE CRITERIA
Mass Ion Abundance Criteria
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95% but less than 101% of mass 174
177 5 to 9% of mass 176
TABLE 4.
QUANTITY OF METHANOL EXTRACT REQUIRED FOR ANALYSIS
OF HIGH-CONCENTRATION SOILS/SEDIMENTS
Approximate Volume of
Concentration Range Methanol Extract8
500- 10,000 M9/Kg 100 ML
1,000- 20,000 M9/Kg 50 juL
5,000-100,000 M9/Kg 10 pi
25,000-500,000 M9/Kg 100 /jL of 1/50 dilution"
Calculate appropriate dilution factor for concentrations exceeding this
table.
a The volume of methanol added to 5 mL of water being purged should be kept
constant. Therefore, add to the 5 mL syringe whatever volume of methanol
is necessary to maintain a volume of 100 /iL added to the syringe.
b Dilute and aliquot of the methanol extract and then take 100 /xL for
analysis.
8240B - 30 Revision 2
November 1990
-------
TABLE 5.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES ASSIGNED
FOR QUANTITATION
Bromochloromethane
Acetone
Acrolein
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chioromethane
Di chlorodi f1uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
trans-1,2-Di chloroethene
lodomethane
Methylene chloride
Tri chlorof1uoromethane
Vinyl chloride
1.4-Di f1uorobenzene
Benzene
Bromodi chloromethane
Bromoform
2-Butanone
Carbon tetrachloride
Chlorodi bromomethane
2-Chloroethyl vinyl ether
Dibromomethane
1,4-Dichloro-2-butene
1,2-Di chloropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1,1,1-Tri chloroethane
1,1,2-Trichloroethane
Trichloroethene
Vinyl acetate
Chlorobenzene-ds
Bromofluorobenzene (surrogate)
Chlorobenzene
Ethyl benzene
Ethyl methacrylate
2-Hexanone
4-Methyl-2-pentanone
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
Toluene-d8 (surrogate)
1,2,3-Trichloropropane
Xylene
8240B - 31
Revision 2
November 1990
-------
TABLE 6.
CALIBRATION AND QC ACCEPTANCE CRITERIA8
Parameter
Benzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
2-Chloroethylvinyl ether
Chloroform
Chl oromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichl oroethane
1,1-Dichloroethene
trans- 1,2-Dichl oroethene
1,2-Dichloropropane
cis-1, 3-Di chl oropropene
trans-l,3-Dichloropropene
Ethyl benzene
Methyl ene chloride
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,1,1-Trichloroethane
1 , 1 ,2-Tri chl oroethane
Tri chl oroethene
Tri chl orof 1 uoromethane
Vinyl chloride
Range
for Q
(M9/L)
12.8-27.2
13.1-26.9
14.2-25.8
2.8-37.2
14.6-25.4
13.2-26.8
D-44.8
13.5-26.5
D-40.8
13.5-26.5
12.6-27.4
14.6-25.4
12.6-27.4
14.5-25.5
13.6-26.4
10.1-29.9
13.9-26.1
6.8-33.2
4.8-35.2
10.0-30.0
11.8-28.2
12.1-27.9
12.1-27.9
14.7-25.3
14.9-25.1
15.0-25.0
14.2-25.8
13.3-26.7
9.6-30.4
0.8-39.2
Limit
for s
6.9
6.4
5.4
17.9
5.2
6.3
25.9
6.1
19.8
6.1
7.1
5.5
7.1
5.1
6.0
9.1
5.7
13.8
15.8
10.4
7.5
7.4
7.4
5.0
4.8
4.6
5.5
6.6
10.0
20.0
Range
for x
(M9/L)
15.2-26.0
10.1-28.0
11.4-31.1
D-41.2
17.2-23.5
16.4-27.4
D-50.4
13.7-24.2
D-45.9
13.8-26.6
11.8-34.7
17.0-28.8
11.8-34.7
14.2-28.4
14.3-27.4
3.7-42.3
13.6-28.4
3.8-36.2
1.0-39.0
7.6-32.4
17.4-26.7
D-41.0
13.5-27.2
17.0-26.6
16.6-26.7
13.7-30.1
14.3-27.1
18.5-27.6
8.9-31.5
D-43.5
Range
P,PS
37-151
35-155
45-169
D-242
70-140
37-160
D-305
51-138
D-273
53-149
18-190
59-156
18-190
59-155
49-155
D-234
54-156
D-210
D-227
17-183
37-162
D-221
46-157
64-148
47-150
52-162
52-150
71-157
17-181
D-251
Q = Concentration measured in QC check sample, in M9/L.
s = Standard deviation of four recovery measurements, in M9/L-
x = Average recovery for four recovery measurements, in M9/L.
p, ps = Percent recovery measured.
D = Detected; result must be greater than zero.
a Criteria from 40 CFR Part 136 for Method 624 and were calculated assuming a
QC check sample concentration of 20 M9/L. These criteria are based directly
upon the method performance data in Table 7. Where necessary, the limits for
recovery have been broadened to assure applicability of the limits to
concentrations below those used to develop Table 7.
8240B - 32
Revision 2
November 1990
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Accuracy, as
recovery, x'
(M9/L)
Single analyst
precision, s/
(Mg/L)
Overall
precision,
S' (M9/L)
Benzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethyl vinyl ether8
Chloroform
Chl oromethane
Di bromochl oromethane
1 , 2-Di chl orobenzene"
1,3-Dichlorobenzene
l,4-Dichlorobenzeneb
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 , 2 , -Di chl oroethene
1 , 2-Di chl oropropane8
cis-l,3-Dichloropropene8
trans -1,3-Di chl oropropene8
Ethyl benzene
Methyl ene chloride
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1 , 1 , 1-Trichloroethane
1 , 1 , 2-Tri chl oroethane
Tri chl oroethene
Tr i chl orof 1 uoromethane
Vinyl chloride
0.93C+2.00
1.03C-1.58
1.18C-2.35
l.OOC
1.10C-1.68
0.98C+2.28
1.18C+0.81
l.OOC
0.93C+0.33
1.03C-1.81
1.01C-0.03
0.94C+4.47
1.06C+1.68
0.94C+4.47
1.05C+0.36
1.02C+0.45
1.12C+0.61
- 1.05C+0.03
l.OOC
l.OOC
l.OOC
0.98C+2.48
0.87C+1.88
0.93C+1.76
1.06C+0.60
0.98C+2.03
1.06C+0.73
0.95C+1.71
1.04C+2.27
0.99C+0.39
l.OOC
0.26X-1.74
0.15x+0.59
0.12X+0.34
0.43x
0.12X+0.25
0.16X-0.09
0.14X+2.78
0.62x
0.16X+0.22
0.37X+2.14
0.17X-0.18
0.22X-1.45
0.14X-0.48
0.22X-1.45
0.13X-0.05
0.17X-0.32
0.17X+1.06
0.14X+0.09
0.33x
0.38x
0.25x
0.14X+1.00
0.15X+1.07
0.16X+0.69
0.13X-0.18
0.15X-0.71
0.12X-0.15
0.14X+0.02
0.13X+0.36
0.33X-1.48
0.48x
0.25X-1.33
0.20X+1.13
O.Ux+1.38
0.58x
O.llx+0.37
0.26x-1.92
0.29X+1.75
0.84x
O.lSx+0.16
0.58X+0.43
0.17X+0.49
0.30X-1.20
O.lSx-0.82
0.30X-1.20
O.lSx+0.47
0.21X-0.38
0.43X-0.22
0.19X+0.17
0.45x
0.52x
0.34x
0.26X-1.72
0.32X+4.00
0.20X+0.41
0.16X-0.45
0.22X-1.71
0.21X-0.39
0.18X+0.00
0.12X+0.59
0.34X-0.39
0.65x
x' = Expected recovery for one or more measurements of a sample containing a
concentration of C, in M9/L.
sr' = Expected single analyst standard deviation of measurements at an average
concentration of x, in ng/L.
S1 = Expected interlaboratory_ standard deviation of measurements at an average
concentration found of x, in /ng/L.
C = True value for the concentration, in /ig/L.
x = Average recovery found for measurements of samples containing a
concentration of C, in M9/L-
a Estimates based upon the performance in a single laboratory.
b Due to chromatographic resolution problems, performance statements for these
isomers are based upon the sums of their concentrations.
8240B - 33
Revision 2
November 1990
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High
Surrogate Compound Water Soil/Sediment
4-Bromofluorobenzene 86-115 74-121
l,2-Dichloroethane-d4 76-114 70-121
Toluene-d8 88-110 81-117
8240B - 34 Revision 2
November 1990
-------
FIGURE 1.
PURGING CHAMBER
OVt W M 04
t7 CM » OMAC STWNOC NCIOU
_ 00
/^ STAMUU STHk
8240B - 35
Revision 2
November 1990
-------
FIGURE 2.
TRAP PACKINGS AND CONSTRUCTION TO INCLUDE
DESORB CAPABILITY FOR METHOD 8240
CONSTRUCTION OCTA«.
TMFMLffT
8240B - 36
Revision 2
November 1990
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE FOR METHOD 8240
CAJWCftOAS
FlOW COMTMQL
uouo
- OOUJMNOVfN
jinrP
COUJMM
AMAimCAL COLUMN
OPTIONAL+*OffT COLUMN
SCLfCTWN VALVf
TftAPMlfT
13X
PUMOINQ
ocvcc
NOTI.
AU UNO •CTWflN TWA*
AMO QC SHOULD M MCATO
8240B - 37
Revision 2
November 1990
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE FOR METHOD 8240
CAJWfUQAt
ANALYTICAL COLUMN
OPTIONAL*
SfLfCnON VALVt
NOT1;
ALL UNfS ITOMEN
ANOOCSMOUL0M
TOVO
8240B - 38
Revision 2
November 1990
-------
FIGURE 5.
LOW SOILS IMPINGER
PURGE INLET FITTING
SAMPLE OUTLET HTTING
3 • 6mm o o GLASS TUBING
40ml VIAL
SEPTUM
CAP
8240B - 39
Revision 2
November 1990
-------
METHOD 8240B
GAS CHROMATOGRAPHY/MASS SPECTROMETRY FOR VOLATILE ORGANICS
Purge-and-trap
7.1
Select
procedure for
introducing
sample into
CC/MS.
7.2.1 Set
CC/MS
operating
conditions
7.2.4 Connect
purge-and-trap
device to CC.
7.2.6 Perform
purge-and-trap
analysis.
72.8
Calculate RFs
for 5 SPCCs.
7.3 Perform
daily
calibration
using SPCCs
and CCCs.
7
8240B - 40
Revision 2
November 1990
-------
METHOD 8240
(continued)
Soil/sediment
and waste
7421
Dilute sample
at least 50
fold with
water
7 4.1 1
Screen sample
us ing Method
3810 or 3820
7417
Per form
seconda ry
dilutions
7 4.1.8 Add
internal standard
and surrogate
spiking solutions.
7 4.1.10
Perform
purge-and- trap
procedure
7.4.3.1.3 Add
interna1 standard
and surrogate
spiking solutions
7 4.3.1.5
Determine
percent dry
weight of
sample.
7.4.3.1.7
Perf o rm
purge-and-trap
procedure.
sample us ing
Method 3810
or 3820
7 4.3 I*
concentration
mg/Kg?
9ize based on
es tima ted
concentra tion
7 4 1 11
Attach trap
to CC and
per fo rm
analysis
7432 Choose
solvent for
ex t raction or
dilution Weigh
sample
7511 Identify
analytes by
comparing the
sample retention
time and samp 1e
mass spect ra
7.4.32 2 Add
sol vent.
shake
752.2 Calculate
the concentration
of each identified
ana 1y te
7 4 3 2.7
Perform
purge-and - trap
pr ocedure.
7.5 2 4
Report all
results
Stop
8240B - 41
Revision 2
November 1990
-------
METHOD 8250A
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS):
PACKED COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8250 is used to determine the concentration of semivolatile
organic compounds in extracts prepared from all types of solid waste matrices,
soils, and ground water. Direct injection of a sample may be used in limited
applications. The following compounds can be determined by this method:
Compounds
CAS No"
Appropriate Preparation Techniques
3510 3520 3540 3550 3580
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
Aldrin
4-Aminobiphenyl
Aniline
Anthracene
Aroclor - 1016
Aroclor - 1221
Aroclor - 1232
Aroclor - 1242
Aroclor - 1248
Aroclor - 1254
Aroclor - 1260
Benzidine
Benzoic acid
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(g,h,i)perylene
Benzo(a)pyrene
Benzyl alcohol
a-BHC
0-BHC
6-BHC
T-BHC (Lindane)
Bi s (2-chl oroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
83-32-9
208-96-8
98-86-2
309-00-2
92-67-1
62-53-3
120-12-7
12674-11-2
11104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
92-87-5
65-85-0
56-55-3
205-99-2
207-08-9
191-24-2
50-32-8
100-51-6
319-84-6
319-85-7
319-86-8
58-89-9
111-91-1
111-44-4
108-60-1
117-81-7
101-55-3
85-68-7
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
X
X
X
X
X
X
X
X
CP
ND
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8250A - 1
Revision 1
November 1990
-------
Appropriate Preparation Techniques
Compounds
Chlordane
4-Chloroaniline
1-Chloronaphthalene
2-Chl oronaphthal ene
4-Chl oro-3-methyl phenol
2-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Chrysene-d12 (I.S.)
4,4'-DDD
4,4'-DDT
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dieldrin
Diethyl phthalate
Dimethyl ami noazobenzene
7,12-Dimethylbenz(a)-
anthracene
a,a-Dimethylphenethylamine
2,4-Dimethylphenol
Dimethyl phthalate
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di phenyl ami ne
1 , 2-Di phenyl hydrazi ne
Di-n-octyl phthalate
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
Fluoranthene
Fluorene
CAS Noa
57-74-9
106-47-8
90-13-1
91-58-7
59-50-7
95-57-8
7005-72-3
218-01-9
72-54-8
50-29-3
224-42-0
53-70-3
132-64-9
84-74-2
95-50-1
541-73-1
106-46-7
3855-82-1
91-94-1
120-83-2
87-65-0
60-57-1
84-66-2
60-11-7
57-97-6
122-09-8
105-67-9
131-11-3
534-52-1
51-28-5
121-14-2
606-20-2
122-39-4
122-66-7
117-84-0
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
62-50-0
206-44-0
86-73-7
3510
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP(45)
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
3520
X
ND
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
3540
X
ND
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
X
X
3550
X
ND
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
3580
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8250A - 2
Revision 1
November 1990
-------
AooroDriate Preparation Techniaues
Compounds
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Methoxychlor
3-Methyl chol anthrene
Methyl methanesulfonate
2-Methyl naphthalene
2-Methyl phenol
4-Methyl phenol
Naphthalene
Naphthalene-de (I.S.)
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
Nitrobenzene-d5 (surr.)
2-Nitrophenol
4-Nitrophenol
N-Nitrosodi butyl ami ne
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi -n-propylamine
N-Nitrosopiperidine
Pentachl orobenzene
Pentachl oroni trobenzene
Pentachl orophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenol
Phenol -d6 (surr.)
2-Picoline
Pronamide
Pyrene
Terphenyl-d14(surr.)
1,2,4, 5-Tetrachl orobenzene
2,3,4,6-Tetrachlorophenol
CAS No"
321-60-8
367-12-4
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
193-39-5
78-59-1
72-43-5
56-49-5
66-27-3
91-57-6
95-48-7
106-44-5
91-20-3
1146-65-2
134-32-7
91-59-8
88-74-4
99-09-2
100-01-6
98-95-3
4165-60-0
88-75-5
100-02-7
924-16-3
62-75-9
86-30-6
621-64-7
100-75-4
608-93-5
82-68-8
87-86-5
198-55-0
62-44-2
85-01-8
108-95-2
13127-88-3
109-06-8
23950-58-5
129-00-0
u95-94-3
58-90-2
3510
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
OS(44)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
DC(28)
DC(28)
ND
X
X
X
X
X
3520
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
X
X
ND
ND
X
X
X
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
X
ND
ND
3540
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
ND
ND
ND
3550
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
X
X
ND
ND
X
X
X
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
X
ND
ND
3580
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
8250A - 3
Revision 1
November 1990
-------
Appropriate Preparation Techniques
Compounds CAS Noa 3510 3520 3540 3550 3580
Toxaphene
2,4,6-Tribromophenol (surr.)
1,2,4-Trichlorobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
8001-35-2
118-79-6
120-82-1
95-95-4
88-06-2
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
a Chemical Abstract Service Registry Number.
CP = Nonreproducible chromatographic performance.
DC = Unfavorable distribution coefficient (number in parenthesis is percent
recovery).
ND = Not determined.
OS = Oxidation during storage (number in parenthesis is percent stability).
X = Greater than 70 percent recovery by this technique.
Percent Stability = Average Recovery (Day 7) x 100/Average Recovery (Day 0).
1.2 Method 8250 can be used to quantitate most neutral, acidic, and basic
organic compounds that are soluble in methylene chloride and capable of being
eluted without derivatization as sharp peaks from a gas chromatographic packed
column. Such compounds include polynuclear aromatic hydrocarbons, chlorinated
hydrocarbons and pesticides, phthalate esters, organophosphate esters,
nitrosamines, halpethers, aldehydes, ethers, ketpnes, anilines, pyridines,
quinolines, aromatic nitro compounds, and phenols, including nitrophenols. See
Table 1 for a list of compounds and their characteristic ions that have been
evaluated on the specified GC/MS system.
1.3 The following compounds may require special treatment when being
determined by this method. Benzidine can be subject to oxidative losses during
solvent concentration. Also, chromatography is poor. Under the alkaline
conditions of the extraction step, a-BHC, T-BHC, endosulfan I and II, and endrin
are subject to decomposition. Neutral extraction should be performed if these
compounds are expected and are not being determined by Method 8080.
Hexachlorocyclopentadiene is subject to thermal decomposition in the inlet of
the gas chromatograph, chemical reaction in acetone solution, and photochemical
decomposition. N-nitrosodimethylamine is difficult to separate from the solvent
under the chromatographic conditions described. N-nitrosodiphenylamine
decomposes in the gas chromatographic inlet and cannot be separated from
diphenylamine. Pentachlorophenol,2,4-dinitrophenol, 4-nitrophenol, 4,6-dinitro-
2-methylphenol, 4-chloro-3-methylphenol, benzoic acid, 2-nitroaniline, 3-
nitroaniline, 4-chloroaniline, and benzyl alcohol are subject to erratic
chromatographic behavior, especially if the GC system is contaminated with high
boiling material.
8250A - 4 Revision 1
November 1990
-------
1.4 The estimated quantitation limit (EQL) of Method 8250 for determining
an individual compound is approximately 1 mg/Kg (wet weight) for soil/sediment
samples, 1-200 mg/Kg for wastes (dependent on matrix and method of preparation),
and 10 /zg/L for ground water samples (see Table 2). EQLs will be
proportionately higher for sample extracts that require dilution to avoid
saturation of the detector.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate
the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract.
3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, and spikes must be evaluated
for interferences. Determine if the source of interference is in the preparation
and/or cleanup of the samples and take corrective action to eliminate the
problem.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system
4.1.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless injection
and all required accessories, including syringes, analytical columns, and
gases.
4.1.2 Columns
4.1.2.1 For base/neutral compound detection - 2 m x 2 mm ID
stainless or glass, packed with 3% SP-2250-DB on 100/120 mesh
Supelcoport or equivalent.
4.1.2.2 For acid compound detection - 2 m x 2 mm ID glass,
packed with 1% SP-1240-DA on 100/120 mesh Supelcoport or equivalent.
8250A - 5 Revision 1
November 1990
-------
4.1.3 Mass spectrometer - Capable of scanning from 35 to 500 amu
every 1 second or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable
of producing a mass spectrum for decafluorotriphenylphosphine (DFTPP)
which meets all of the criteria in Table 3 when 1 /iL of the GC/MS tuning
standard is injected through the GC (50 ng of DFTPP).
4.1.4 GC/MS interface - Any GC-to-MS interface that gives acceptable
calibration points at 50 ng per injection for each compound of interest
and achieves acceptable tuning performance criteria may be used. GC-to-
MS interfaces constructed entirely of glass or glass-lined materials are
recommended. Glass may be deactivated by silanizing with
dichlorodimethylsi 1ane.
4.1.5 Data system - A computer system must be interfaced to the mass
spectrometer. The system must allow the continuous acquisition and storage
on machine-readable media of all mass spectra obtained throughout the
duration of the chromatographic program. The computer must have software
that can search any GC/MS data file for ions of a specific mass and that
can plot such ion abundances versus time or scan number. This type of plot
is defined as an Extracted Ion Current Profile (EICP). Software must also
be available that allows integrating the abundances in any EICP between
specified time or scan-number limits. The most recent version of the
EPA/NIH Mass Spectral Library should also be available.
4.2 Syringe - 10 /iL.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock standard solutions (1.00 M9/ML) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 10 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96% or greater, the weight
may be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
concentration if they are certified by the manufacturer or by an
independent source.
8250A - 6 Revision 1
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5.3.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at 4°C and protect from light.
Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.3.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10,
chrysene-d12, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Section 7.3.2 are met. Dissolve 200 mg of
each compound with a small volume of carbon disulfide. Transfer to a 50 mL
volumetric flask and dilute to volume with methylene chloride so that the final
solvent is approximately 20% carbon disulfide. Most of the compounds are also
soluble in small volumes of methanol, acetone, or toluene, except for perylene-
d12. The resulting solution will contain each standard at a concentration of
4,000 ng//iL. Each 1 ml sample extract undergoing analysis should be spiked with
10 ML of the internal standard solution, resulting in a concentration of 40
ng//nL of each internal standard. Store at 4°C or less when not being used.
5.5 GC/MS tuning standard - A methylene chloride solution containing
50 ng//iL of decafluorotriphenylphosphine (DFTPP) should be prepared. The
standard should also contain 50 ng/jiL each of 4,4'-DDT, pentachlorophenol, and
benzidine to verify injection port inertness and GC column performance. Store
at 4°C or less when not being used.
5.6 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared. One of the calibration standards should be
at a concentration near, but above, the method detection limit; the others should
correspond to the range of concentrations found in real samples but should not
exceed the working range of the GC/MS system. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Table 1 may be included). Each 1 ml aliquot of calibration standard should
be spiked with 10 /xL of the internal standard solution prior to analysis. All
standards should be stored at -10°C to -20°C and should be freshly prepared once
a year, or sooner if check standards indicate a problem. The daily calibration
standard should be prepared weekly and stored at 4°C.
5.7 Surrogate standards - The recommended surrogate standards are
phenol-d6, 2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-d5, 2-
fluorobiphenyl, and p-terphenyl-d14. See Method 3500 for the instructions on
preparing the surrogate standards. Determine what concentration should be in
the blank extracts after all extraction, cleanup, and concentration steps.
Inject this concentration into the GC/MS to determine recovery of surrogate
standards in all blanks, spikes, and sample extracts. Take into account all
dilutions of sample extracts.
5.8 Matrix spike standards - See Method 3500 for instructions on preparing
the matrix spike standard. Determine what concentration should be in the blank
extracts after all extraction, cleanup, and concentration steps. Inject this
8250A - 7 Revision 1
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concentration into the GC/MS to determine recovery of standards in all matrix
spikes. Take into account all dilutions of sample extracts.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample preparation - Samples must be prepared by one of the following
methods prior to GC/MS analysis.
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3550
Waste 3540, 3550, 3580
7.1.1 Direct injection - In very limited applications direct
injection of the sample into the GC/MS system with a 10 juL syringe may
be appropriate. The detection limit is very high (approximately
10,OOOM9/L); therefore, it is only permitted where concentrations in
excess of 10,000 ng/l are expected. The system must be calibrated by
direct injection.
7.2 Extract cleanup - Extracts may be cleaned up by any of the following
methods prior to GC/MS analysis.
Compounds Methods
Phenols 3630, 3640, 8040a
Phthalate esters 3610, 3620, 3640
Nitrosamines 3610, 3620, 3640
Organochlorine pesticides & PCBs 3620, 3640, 3660
Nitroaromatics and cyclic ketones 3620, 3640
Polynuclear aromatic hydrocarbons 3611, 3630, 3640
Haloethers 3620, 3640
Chlorinated hydrocarbons 3620, 3640
Organophosphorus pesticides 3620
Petroleum waste 3611, 3650
All basic, neutral, and acidic
Priority Pollutants 3640
aMethod 8040 includes a derivatization technique followed by GC/ECD analysis,
if interferences are encountered on GC/FID.
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7.3 Recommended GC/MS operating conditions
Electron energy: 70 volts (nominal)
Mass range: 35-500 amu
Scan time: 1 sec/scan
Injector temperature: 250-300°C
Transfer line temperature: 250-300°C
Source temperature: According to manufacturer's specifications
Injector: Grob-type, splitless
Sample volume: 1-2 juL
Carrier gas: Helium at 30 mL/min
Conditions for base/neutral analysis (3% SP-2250-DB)
Initial column temperature and hold time: 50°C for 4 minutes
Column temperature program: 50-300°C at 8°C/min
Final column temperature hold: 300°C for 20 minutes
Conditions for acid analysis (1% SP-1240-DA)
Initial column temperature and hold time: 70°C for 2 minutes
Column temperature program: 70-200°C at 8°C/min
Final column temperature hold: 200°C for 20 minutes
7.4 Initial calibration
7.4.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 3 for a 50 ng injection of DFTPP. Analyses should not begin until
all these criteria are met. Background subtraction should be
straightforward and designed only to eliminate column bleed or instrument
background ions. The GC/MS tuning standard should also be used to assess
GC column performance and injection port inertness. Degradation of DDT
to DDE and DDD should not exceed 20%. Benzidine and pentachlorophenol
should be present at their normal responses, and no peak tailing should
be visible. If degradation is excessive and/or poor chromatography is
noted, the injection port may require cleaning.
7.4.2 The internal standards selected in Section 5.1 should permit
most of the components of interest in a chromatogram to have retention
times of 0.80-1.20 relative to one of the internal standards. Use the
base peak ion from the specific internal standard as the primary ion for
quantitation (see Table 1). If interferences are noted, use the next most
intense ion as the quantitation ion (i.e. for l,4-dichlorobenzene-d4 use
m/z 152 for quantitation).
7.4.3 Analyze 1 /uL of each calibration standard (containing internal
standards) and tabulate the area of the primary characteristic ion against
concentration for each compound (as indicated in Table 1). Calculate
response factors (RFs) for each compound as follows:
RF = (AXC|S)/(A1SCX)
8250A - 9 Revision 1
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where:
Ax = Area of the characteristic ion for the compound being measured.
Als = Area of the characteristic ion for the specific internal
standard.
Cx = Concentration of the compound being measured (ng//ul_).
Cls = Concentration of the specific internal standard (ng//il_).
7.4.4 The average RF should be calculated for each compound. The
percent relative standard deviation (%RSD = 100[SD/RF]) should also be
calculated for each compound. The %RSD should be less than 30% for each
compound. However, the %RSD for each individual Calibration Check Compound
(CCC) (see Table 4) must be less than 30%. The relative retention times
of each compound in each calibration run should agree within 0.06 relative
retention time units. Late-eluting compounds usually have much better
agreement.
7.4.5 A system performance check must be performed to ensure that
minimum average response factors are met before the calibration curve is
used. For semivolatiles, the System Performance Check Compounds (SPCCs)
are: N-nitroso-di-n-propylamine; hexachlorocyclopentadiene; 2,4-
dinitrophenol; and 4-nitrophenol. The minimum acceptable average RF for
these is 0.050. These SPCCs typically have very low RFs (0.1-0.2) and
tend to decrease in response as the chromatographic system begins to
deteriorate or the standard material begins to deteriorate. They are
usually the first to show poor performance. Therefore, they must meet
the minimum requirement when the system is calibrated.
7.5 Daily GC/MS calibration
7.5.1 Prior to analysis of samples, the GC/MS tuning standard must
be analyzed. A 50 ng injection of DFTPP must result in a mass spectrum
for DFTPP which meets the criteria given in Table 3. These criteria must
be demonstrated during each 12 hour shift.
7.5.2 A calibration standard(s) at mid-concentration containing all
semivolatile analytes, including all required surrogates, must be performed
every 12 hours during analysis. Compare the response factor data from the
standards every 12 hours with the average response factor from the initial
calibration for a specific instrument as per SPCC (Section 7.4.3) and CCC
(Section 7.4.4) criteria.
7.5.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made during every 12 hour shift. If the SPCC
criteria are met, a comparison of response factors is made for all
compounds. This is the same check that is applied during the initial
calibration. If the minimum response factors are not met, the system must
be evaluated, and corrective action must be taken before sample analysis
begins. The minimum RF for semivolatile SPCCs is 0.050. Some possible
problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
and active sites in the column or chromatographic system. This check must
be met before analysis begins.
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7.5.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Table 4 are used to check the
validity of the initial calibration. Calculate the percent difference
using:
RF, - RFC
% Difference = x 100
where:
RF,
RF, = Average response factor from initial calibration.
RFC = Response factor from current verification check standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. If the percent difference
for each CCC is less than 30%, the initial calibration is assumed to be
valid. If the criterion is not met (> 30% difference) for any one CCC,
corrective action must be taken. Problems similar to those listed under
SPCCs could affect these criteria. If no source of the problem can be
determined after corrective action has been taken, a new five-point
calibration must be generated. This criterion must be met before sample
analysis begins.
7.5.5 The internal standard responses and retention times in the
calibration check standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last check calibration (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration standard check, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate.
7.6 GC/MS analysis
7.6.1 It is highly recommended that the extract be screened on a
GC/FID or GC/PID using the same type of column. This will minimize
contamination of the GC/MS system from unexpectedly high concentrations
of organic compounds.
7.6.2 Spike the 1 ml extract obtained from sample preparation with
10 pi of the internal standard solution just prior to analysis.
7.6.3 Analyze the 1 ml extract by GC/MS using the appropriate column
(as specified in Section 4.1.2). The recommended GC/MS operating
conditions to be used are specified in Section 7.3.
7.6.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the GC/MS system, extract dilution must take
place. Additional internal standard must be added to the diluted extract
to maintain the required 40 ng//iL of each internal standard in the
extracted volume. The diluted extract must be reanalyzed.
8250A - 11 Revision 1
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7.6.5 Perform all qualitative and quantitative measurements as
described in Section 7.7. Store the extracts at 4°C, protected from
light in screw-cap vials equipped with unpierced Teflon lined septa.
7.7 Data interpretation
7.7.1 Qualitative analysis
7.7.1.1 The qualitative identification of compounds determined
by this method is based on retention time, and on comparison of the
sample mass spectrum, after background correction, with
characteristic ions in a reference mass spectrum. The reference mass
spectrum must be generated by the laboratory using the conditions
of this method. The characteristic ions from the reference mass
spectrum are defined to be the three ions of greatest relative
intensity, or any ions over 30% relative intensity if less than three
such ions occur in the reference spectrum. Compounds should be
identified as present when the criteria below are met.
7.7.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.7.1.1.2 The RRT of the sample component is within ±
0.06 RRT units of the RRT of the standard component.
7.7.1.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions
in the reference spectrum. (Example: For an ion with an
abundance of 50% in the reference spectrum, the corresponding
abundance in a sample spectrum can range between 20% and 80%.)
7.7.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
7.7.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background spectra
is important. Examination of extracted ion current profiles
of appropriate ions can aid in the selection of spectra, and
in qualitative identification of compounds. When analytes
8250A - 12 Revision 1
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coelute (I.e., only one chromatographic peak Is apparent), the
identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting
compound.
7.7.1.2 For samples containing components not associated with
the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search routines
should not use normalization routines that would misrepresent the
library or unknown spectra when compared to each other. For example,
the RCRA permit or waste delisting requirements may require the
reporting of nontarget analytes. Only after visual comparison of
sample spectra with the nearest library searches will the mass
spectral interpretation specialist assign a tentative identification.
Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference spectrum
(ions > 10% of the most abundant ion) should be present in the sample
spectrum.
(2) The relative intensities of the major ions should agree within
+ 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should be
present in sample the spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination
or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the sample
spectrum because of background contamination or coeluting peaks.
Data system library reduction programs can sometimes create these
discrepancies.
7.7.2 Quantitative analysis
7.7.2.1 When a compound has been identified, the quantitation
of that compound will be based on the integrated abundance from the
EICP of the primary characteristic ion. Quantitation will take place
using the internal standard technique. The internal standard used
shall be the one nearest the retention time of that of a given
analyte (e.g. see Table 5).
7.7.2.2 Calculate the concentration of each identified analyte
in the sample as follows:
8250A - 13 Revision 1
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Water
(AJ(Is)(Vt)
concentration (M9/L) =
(Als)(RF)(V0}(V,)
where:
Ax = Area of characteristic ion for compound being measured.
Is = Amount of internal standard injected (ng).
Vt = Volume of total extract, taking into account dilutions (i.e.
a l-to-10 dilution of a 1 ml extract will mean Vt = 10,000 /zL.
If half the base/neutral extract and half the acid extract are
combined, V, = 2,000).
A,s= Area of characteristic ion for the internal standard.
RF = Response factor for compound being measured (Section 7.3.3).
V0 = Volume of water extracted (ml).
V, = Volume of extract injected
Sediment/Soil Sludge (on a dry-weight basis) and Waste (normally on
a wet-weight basis
(Ax)(IJ(Vt)
concentration (jug/Kg)
(A|S)(RF)(V,)(WS)(D)
where:
A,,, Is, Vt, Als, RF, V, = Same as for water.
Ws = Weight of sample extracted or diluted in grams.
D = % dry weight of sample/100, or 1 for a wet-weight basis.
7.7.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulas
given above should be used with the following modifications: The
areas Ax and Als should be from the total ion chromatograms and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
7.7.2.4 Quantitation of multicomponent compounds (e.g.
Aroclors) is beyond the scope of Method 8250. Normally, quantitation
is performed using a GC/ECD by Method 8080.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Required instrument QC is found in the following section:
8250A - 14 Revision 1
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8.2.1 The GC/MS system must be tuned to meet the DFTPP specifications
in Sections 7.4.1 and 7.4.1.
8.2.2 There must be an initial calibration of the GC/MS system as
specified in Section 7.4.
8.2.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.5.3 and the CCC criteria in Section 7.5.4, each 12 hours.
8.3 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.3.1 A quality (QC) reference sample concentrate is required
containing each analyte at a concentration of 100 nq/ml in acetone. The
QC reference sample concentrate may be prepared from pure standard
materials or purchased as certified solutions. If prepared by the
laboratory, the QC reference sample concentrate must be made using stock
standards prepared independently from those used for calibration.
8.3.2 Using a pipet, prepare QC reference samples at a concentration
of 100 /xg/L by adding 1.00 ml of QC reference sample concentrate to each
of four 1 L aliquots of water.
8.3.3 Analyze the well-mixed QC reference samples according to the
method beginning in Section 7.1 with extraction of the samples.
8.3.4 Calculate the average recovery (x) in p.g/1, and the standard
deviation of the recovery(s) in M9/U for each analyte of interest using
the four results.
8.3.5 For each analyte compare s and x with the corresponding
acceptance criteria _for precision and accuracy, respectively, found in
Table 6. If s and x for all analytes of interest meet the acceptance
criteria, the system performance is acceptable and analysis of actual
samples can begin. If any individual s exceeds the precision limit or
any individual x falls outside the range for accuracy, then the system
performance is unacceptable for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial probability
that one or more will fail at least one of the acceptance criteria when
all analytes of a given method are analyzed.
8.3.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Section
8.3.6.1 or 8.3.6.2.
8.3.6.1 Locate and correct the source of the problem and
repeat the test for all analytes of interest beginning with Section
8.3.2.
8.3.6.2 Beginning with Section 8.3.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
8250A - 15 Revision 1
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If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Section
8.3.2.
8.4 For aqueous and soil matrices, laboratory established surrogate
control limits should be compared with the control limits listed in Table 8.
The limits given in Table 8 are multilaboratory performance based limits for
soil and aqueous samples, and therefore, the single laboratory limits must fall
within those given in Table 8 for these matrices.
8.4.1 If recovery is not within limits, the following procedures
are required.
8.4.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.4.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.4.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.4.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4.2 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
9.0 METHOD PERFORMANCE
9.1 Method 8250 was tested by 15 laboratories using organic-free reagent
water, drinking water, surface water, and industrial wastewaters spiked at six
concentrations over the range 5-1,300 /ig/L. Single operator accuracy and
precision, and method accuracy were found to be directly related to the
concentration of the analyte and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 7.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 625," October 26,
1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems," Analytical Chemistry, 47, 995-1000, 1975.
8250A - 16 Revision 1
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"Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
"Inter!aboratory Method Study for EPA Method 625-Base/Neutrals, Acids, and
Pesticides," Final Report for EPA Contract 68-03-3102 (in preparation).
Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
8250A - 17 Revision 1
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LIMITS, AND
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Retention
Compound Time (min)
Acenaphthene 17.8
Acenaphthene-d10 (I.S.)
Acenaphthylene 17.4
Acetophenone
Aldrin 24.0
4-Aminobiphenyl
Aniline
Anthracene 22.8
Aroclor-1016b 18-30
Aroclor-1221b 15-30
Aroclor-1232b 15-32
Aroclor-1242b 15-32
Aroclor-1248b 12-34
Aroclor-1254b 22-34
Aroclor-1260b 23-32
Benzidine" 28.8
Benzoic acid
Benzo(a)anthracene 31.5
Benzo(b)fluoranthene 34.9
Benzo(k)fluoranthene 34.9
Benzo(g,h,i)perylene 45.1
Benzo(a)pyrene 36.4
Benzyl alcohol
«-BHCa 21.1
6-BHC 23.4
*-BHC 23.7
T-BHC (Lindane)8 22.4
Bis(2-chloroethoxy)methane 12.2
Bis(2-chloroethyl) ether 8.4
Bis(2-chloroisopropyl) ether 9.3
Bis(2-ethylhexyl) phthalate 30.6
4-Bromophenyl phenyl ether 21.2
Butyl benzyl phthalate 29.9
Chlordane" 19-30
4-Chloroaniline
1-Chloronaphthalene
2-Chloronaphthalene 15.9
4-Chloro-3-methylphenol 13.2
2-Chlorophenol 5.9
4-Chlorophenyl phenyl ether 19.5
Chrysene 31.5
Chrysene-d12 (I.S.)
4,4'-DDD 28.6
4,4'-DDT 29.3
Method
detection
limit (/ag/L)
1.9
--
3.5
--
1.9
--
--
1.9
--
30
--
--
--
36
--
44
--
7.8
4.8
2.5
4.1
2.5
--
--
4.2
3.1
--
5.3
5.7
5.7
2.5
1.9
2.5
--
--
--
1.9
3.0
3.3
4.2
2.5
--
2.8
4.7
Primary
Ion
154
164
152
105
66
169
93
178
222
190
190
222
292
292
360
184
122
228
252
252
276
252
108
183
181
183
183
93
93
45
149
248
149
373
127
162
162
107
128
204
228
240
235
235
Secondary
Ion(s)
153, 152
162, 160
151, 153
77, 51
263, 220
168, 170
66, 65
176, 179
260, 292
224, 260
224, 260
256, 292
362, 326
362, 326
362, 394
92, 185
105, 77
229, 226
253, 125
253, 125
138, 277
253, 125
79, 77
181, 109
183, 109
181, 109
181, 109
95, 123
63, 95
77, 121
167, 279
250, 141
91, 206
375, 377
129
127, 164
127, 164
144, 142
64, 130
206, 141
226, 229
120, 236
237, 165
237, 165
8250A - 18
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TABLE 1.
(Continued)
Retention
Compound Time (min)
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
--
43.2
--
24.7
8.4
7.4
7.8
Method
detection
Primary
limit (jug/L) Ion
--
2.5
--
2.5
1.9
1.9
4.4
l,4-Dichlorobenzene-d4 (I.S.) --
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dieldrin
Diethyl phthalate
p-Dimethyl aminoazobenzene
32.2
9.8
--
27.2
20.1
--
16.5
2.7
--
2.5
1.9
--
7,12-0imethylbenz(a)anthracene --
a-, a -Dimethyl phenethyl ami ne
2, 4-Dimethyl phenol
Dimethyl phthalate
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
1 , 2-Di phenyl hydrazi ne
Di-n-octyl phthalate
Endosulfan I"
Endosulfan IIa
Endosulfan sulfate
Endrin8
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi enea
Hexachl oroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Methoxychlor
--
9.4
18.3
16.2
15.9
19.8
18.7
--
--
32.5
26.4
28.6
29.8
27.9
--
--
--
26.5
19.5
--
--
23.4
25.6
21.0
11.4
13.9
8.4
42.7
11.9
--
--
2.7
1.6
24
42
5.7
1.9
--
--
2.5
--
--
5.6
--
--
--
--
2.2
1.9
--
--
1.9
2.2
1.9
0.9
--
1.6
3.7
2.2
--
279
278
168
149
146
146
146
152
252
162
162
79
149
120
256
58
122
163
198
184
165
165
169
77
149
195
337
272
263
67
317
79
202
166
172
112
100
353
284
225
237
117
276
82
227
Secondary
Ion(s)
280,
139,
139
150,
148,
148,
148,
150,
254,
164,
164,
263,
177,
225,
241,
91,
107,
194,
51,
63,
63,
63,
168,
105,
167,
339,
339,
387,
82,
345,
67,
109,
101,
165,
171
64
272,
355,
142,
223,
235,
201,
138,
95,
228
111
279
104
111
111
111
115
126
98
98
279
150
77
257
42
121
164
105
154
89
89
167
182
43
341
341
422
81
250
319
97
203
167
274
351
249
227
272
199
227
138
8250A - 19
Revision 1
November 1990
-------
TABLE 1.
(Continued)
Retention
Compound Time (min)
3-Methylcholanthrene
Methyl methanesulfonate
2-Methyl naphthalene
2-Methyl phenol
4-Methyl phenol
Naphthalene
Naphthalene-dB (I.S.)
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
Nitrobenzene-d5 (surr.)
2-Nitrophenol
4-Nitrophenol
N-Nitroso-di-n-butylamine
N-Nitrosodimethylaminea
N-Nitrosodiphenylaminea
N-Nitroso-di-N-propylamine
N-Nitrosopiperidine
Pentachlorobenzene
Pentachloronitrobenzene
Pentachlorophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenol
Phenol -d6 (surr.)
2-Picoline
Pronamide
Pyrene
Terphenyl-d14 (surr.)
1,2,4,5-Tetrachlorobenzene
2,3,4,6-Tetrachlorophenol
--
--
--
--
--
12.1
--
--
--
--
--
--
11.1
--
6.5
20.3
--
--
20.5
--
--
--
--
17.5
--
--
22.8
--
8.0
--
--
--
27.3
--
--
--
Method
detection
Primary
limit (/ig/L) Ion
--
--
--
--
--
1.6
--
--
--
--
--
--
1.9
--
3.6
2.4
--
--
1.9
--
--
--
--
3.6
--
--
5.4
--
1.5
--
--
--
1.9
--
--
--
Toxaphene" 25-34
2,4,6-Tribromophenol (surr,
1,2,4-Trichlorobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
. ) --
11.6
--
11.8
--
1.9
--
2.7
268
80
142
108
108
128
136
143
143
65
138
138
77
82
139
139
84
42
169
70
42
250
295
266
264
108
178
188
94
99
93
173
202
244
216
232
159
330
180
196
196
Secondary
Ion(s)
253,
79,
141
107,
107,
129,
68
115,
115,
92,
108,
108,
123,
128,
109,
109,
57,
74,
168,
130,
114,
252,
237,
264,
260,
109,
179,
94,
65,
42,
66,
175,
200,
122,
214,
230,
231,
332,
182,
198,
198,
267
65
79
79
127
116
116
138
92
92
65
54
65
65
41
44
167
42
55
248
142
268
265
179
176
80
66
71
92
145
203
212
218
131
233
141
145
200
200
aSee Section 1.3
"These compounds are mixtures of various isomers.
8250A - 20
Revision 1
November 1990
-------
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix Factor"
Ground water 10
Low-concentration soil by ultrasonic extraction with GPC cleanup 670
High-concentration soil and sludges by ultrasonic extraction 10,000
Non-water miscible waste 100,000
a Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
b EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
8250A - 21 Revision 1
November 1990
-------
TABLE 3.
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA8
Mass Ion Abundance Criteria
51 30-60% of mass 198
68 < 2% of mass 69
70 < 2% of mass 69
127 40-60% of mass 198
197 < 1% of mass 198
198 Base peak, 100% relative abundance
199 5-9% of mass 198
275 10-30% of mass 198
365 > 1% of mass 198
441 Present but less than mass 443
442 > 40% of mass 198
443 17-23% of mass 442
aSee Reference 4.
8250A - 22 Revision 1
November 1990
-------
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction Acid Fraction
Acenaphthene 4-Chloro-3-methylphenol
1,4-Dichlorobenzene 2,4-Dichlorophenol
Hexachlorobutadiene 2-Nitrophenol
N-Nitroso-di-n-phenylamine Phenol
Di-n-octyl phthalate Pentachlorophenol
Benzo(a)pyrene 2,4,6-Trichlorophenol
8250A - 23 Revision 1
November 1990
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
Phenanthrene-d
10
Chrysene-d12
Perylene-d12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl ether
Di-n-butyl phthalate
4,6-Dinitro-2-methylphenol
Diphenylamine
1,2-Di phenylhydrazi ne
Fluoranthene
Hexachlorobenzene
N-Nitrosodiphenylamine
Pentachlorophenol
Pentachloroni trobenzene
Phenacetln
Phenanthrene
Pronamide
Benzidine
Benzo(a)anthracene
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethylami noazobenzene
Pyrene
Terphenyl-d14 (surr.)
Benzo(b)fluor-
anthene
Benzo(k)fluor-
anthene
Benzo(g,h,i)-
perylene
Benzo(a)pyrene
Dibenz(a,j)acridine
Dibenz(a,h)-
anthracene
7,12-Dimethylbenz-
(a)anthracene
Di-n-octyl phthalate
Indeno(l,2,3-cd)-
pyrene
3-Methylchol-
anthrene
(surr.) = surrogate
8250A - 24
Revision 1
November 1990
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
(Continued)
l,4-Dichlorobenzene-D4
Naphthalene-d8
Acenaphthene-d
10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether 4-Chloroaniline
Acetophenone Acenaphthene
Benzoic acid Acenaphthylene
Bis(2-chloroethoxy)methanel-Chloronaphthalene
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Di chlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propylamine
Phenol
Phenol-d6 (surr.)
2-Picoline
4-Chloro-3-methylphenol
2,4-Dichlorophenol
2,6-Dichlorophenol
o,a-Dimethyl-
phenethylamine
2,4-Dimethylphenol
Hexachlorobutadiene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Chloronaphthalene
4-Chlorophenyl
phenyl ether
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Fluorene
2-Fluorobiphenyl
(surr.)
Hexachlorocyclo-
pentadiene
2-Nitrophenol
N-Nitroso-di-n-butylamine 1-Naphthylamine
N-Nitrosopiperi dine 2-Naphthylamine
1,2,4-Trichlorobenzene 2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
4-Nitrophenol
Pentachlorobenzene
1,2,4,5-Tetra-
chlorobenzene
(surr.) = surrogate
8250A - 25
Revision 1
November 1990
-------
TABLE 6.
QC ACCEPTANCE CRITERIA3
Test
cone.
Compound (M9/L)
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benzo(a)anthracene
Benzo(b)fl uoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Butyl benzyl phthalate
6-BHC
i-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Bis(Z-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
.100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
27.6
40.2
39.0
32.0
27.6
38.8
32.3
39.0
58.9
23.4
31.5
21.6
55.0
34.5
46.3
41.1
23.0
13.0
33.4
48.3
31.0
32.0
61.6
70.0
16.7
30.9
41.7
32.1
71.4
30.7
26.5
23.2
21.8
29.6
31.4
16.7
32.5
32.8
20.7
37.2
54.7
24.9
26.3
24.5
Range
for x
(M9/L)
60.1-132.3
53.5-126.0
7.2-152.2
43.4-118.0
41.8-133.0
42.0-140.4
25.2-145.7
31.7-148.0
D-195.0
D-139.9
41.5-130.6
D-100.0
42.9-126.0
49.2-164.7
62.8-138.6
28.9-136.8
64.9-114.4
64.5-113.5
38.4-144.7
44.1-139.9
D-134.5
19.2-119.7
D-170.6
D-199.7
8.4-111.0
48.6-112.0
16.7-153.9
37.3-105.7
8.2-212.5
44.3-119.3
D-100.0
D-100.0
47.5-126.9
68.1-136.7
18.6-131.8
D-103.5
D-188.8
42.9-121.3
71.6-108.4
D-172.2
70.9-109.4
7.8-141.5
37.8-102.2
55.2-100.0
Range
P» Ps
(%)
47-145
33-145
D-166
27.133
33-143
24-159
11-162
17-163
D-219
D-152
24-149
D-110
12-158
33-184
36-166
8-158
53-127
60-118
25-158
17-168
D-145
4-136
D-203
D-227
1-118
32-129
D-172
20-124
D-262
29-136
D-114
D-112
39-139
50-158
4-146
D-107
D-209
26-137
59-121
D-192
26.155
D-152
24-116
40-113
8250A - 26
Revision 1
November 1990
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
(Continued)
Compound
Test
cone.
(M9/L)
Limit
for s
(M9/L)
Range
for x
(M9/L)
Range
P. Ps
(%)
Indeno(l,2,3-cd)pyrene 100 44.6
Isophorone 100 63.3
Naphthalene 100 30.1
Nitrobenzene 100 39.3
N-Nitroso-di-n-propylamine 100 55.4
PCB-1260 100 54.2
Phenanthrene 100 20.6
Pyrene 100 25.2
1,2,4-Trichlorobenzene 100 28.1
4-Chloro-3-methylphenol 100 37.2
2-Chlorophenol 100 28.7
2,4-Chlorophenol 100 26.4
2,4-Dimethylphenol 100 26.1
2,4-Dinitrophenol 100 49.8
2-Methyl-4,6-dinitrophenol 100 93.2
2-Nitrophenol 100 35.2
4-Nitrophenol 100 47.2
Pentachlorophenol 100 48.9
Phenol 100 22.6
2,4,6-Trichlorophenol 100 31.7
D-150.9
46.6-180.2
35.6-119.6
54.3-157.6
13.6-197.9
19.3-121.0
65.2-108.7
69.6-100.0
57.3-129.2
40.8-127.9
36.2-120.4
52.5-121.7
41.8-109.0
D-172.9
53.0-100.0
45.0-166.7
13.0-106.5
38.1-151.8
16.6-100.0
52.4-129.2
D-171
21-196
21-133
35-180
D-230
D-164
54-120
52-115
44-142
22-147
23-134
39-135
32-119
D-191
D-181
29-182
D-132
14-176
5-112
37-144
s =
X =
P» Ps
D =
Standard deviation of four recovery measurements, in M9/L.
Average recovery for four recovery measurements, in M9/L.
Percent recovery measured.
Detected; result must be greater than zero.
Criteria from 40 CFR Part 136 for Method 625. These criteria are based
directly on the method performance data in Table 7. Where necessary, the
limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 7.
8250A - 27
Revision 1
November 1990
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Accuracy, as
recovery, x'
Single analyst Overall
precision, s/ precision,
(M9/L) S' (M9/L)
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benzo(a)anthracene
Chloroethane
Benzo(b)f 1 uoranthene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Butyl benzyl phthalate
B-BHC
«-BHC
Bis(2-chloroethyl) ether
Bi s (2-chl oroethoxy)methane
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo(a,h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Hexachl oroethane
0.96C+0.19
0.89C+0.74
0.78C+1.66
0.80C+0.68
0.88C-0.60
0.99C-1.53
0.93C-1.80
0.87C-1.56
0.90C-0.13
0.98C-0.86
0.66C-1.68
0.87C-0.94
0.29C-1.09
0.86C-1.54
1.12C-5.04
1.03C-2.31
0.84C-1.18
0.91C-1.34
0.89C+0.01
0.91C+0.53
0.93C-1.00
0.56C-0.40
0.70C-0.54
0.79C-3.28
0.88C+4.72
0.59C+0.71
0.80C+0.28
0.86C-0.70
0.73C-1.47
1.23C-12.65
0.82C-0.16
0.43C+1.00
0.20C+1.03
0.92C-4.81
1.06C-3.60
0.76C-0.79
0.39C+0.41
0.76C-3.86
0.81C+1.10
0.90C-0.00
0.87C-2.97
0.92C-1.87
0.74C+0.66
0.71C-1.01
0.73C-0.83
0.15X-0.12
0.24X-1.06
0.27X-1.28
0.21X-0.32
0.15X+0.93
0.14X-0.13
0.22X+0.43
0.19X+1.03
0.22X+0.48
0.29x+2.40
O.lSx+0.94
0.20X-0.58
0.34X+0.86
0.35X-0.99
0.16X+1.34
0.24X+0.28
0.26X+0.73
O.lSx+0.66
0.07X+0.52
0.20X-0.94
0.28X+0.13
0.29X-0.32
0.26X-1.17
0.42X+0.19
0.30X+8.51
0.13x+1.16
0.20X+0.47
0.25X+0.68
0.24X+0.23
0.28X+7.33
0.20X-0.16
0.28X+1.44
0.54X+0.19
0.12X+1.06
0.14X+1.26
0.21X+1.19
0.12X+2.47
0.18x+3.91
0.22X-0.73
0.12X+0.26
0.24X-0.56
0.33X-0.46
O.lSx-0.10
0.19X+0.92
0.17x+0.67
0.21X-0.67
0.26X-0.54
0.43X+1.13
0.27X-0.64
0.26X-0.21
0.17X-0.28
0.29X+0.96
0.35X+0.40
0.32X+1.35
0.51X-0.44
0.53X+0.92
0.30X+1.94
0.93X-0.17
0.35X+0.10
0.26X-I-2.01
0.25X+1.04
0.36X+0.67
0.16X+0.66
0.13X+0.34
0.30X-0.46
0.33X-0.09
0.66X-0.96
0.39X-1.04
0.65X-0.58
0.59X+0.25
0.39X+0.60
0.24X+0.39
0.41X+0.11
0.29X+0.36
0.47X+3.45
0.26X-0.07
0.52X+0.22
l.OSx-0.92
0.21X+1.50
0.19X+0.35
0.37X+1.19
0.63X-1.03
0.73X-0.62
0.28X-0.60
0.13X+0.61
0.50X-0.23
0.28X+0.64
0.43X-0.52
0.26X+0.49
0.17X+0.80
8250A - 28
Revision 1
November 1990
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
(Continued)
Parameter
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitroso-di-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
4-Chl oro-3-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2, 4-Dimethyl phenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Accuracy, as
recovery, x'
(M9/L)
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.22
0.81C-10.86
0.87C+0.06
0.84C-0.16
0.94C-0.79
0.84C+0.35
0.78C+0.29
0.87C-0.13
0.71C+4.41
0.81C-18.04
1.04C-28.04
0.07C-1.15
0.61C-1.22
0.93C+1.99
0.43C+1.26
0.91C-0.18
Single analyst
precision, s/
(M9/L)
0.29X+1.46
0.27X+0.77
0.21X-0.41
0.19X+0.92
0.27X+0.68
0.35X+3.61
0.12X+0.57
0.16x+0.06
O.lBx+0.85
0.23X+0.75
O.lSx+1.46
O.lSx+1.25
0.16X+1.21
0.38X+2.36
O.lOx+42.29
O.lSx+1.94
0.38X+2.57
0.24X+3.03
0.26X+0.73
0.16X+2.22
Overall
precision,
S' (M9/L)
O.BOx-0.44
0.33X+0.26
0.30X-0.68
0.27X+0.21
0.44X+0.47
0.43X+1.82
O.lSx+0.25
O.lBx+0.31
0.21X+0.39
0.29X+1.31
0.28X+0.97
0.21x+1.28
0.22X+1.31
0.42X+26.29
0.26X+23.10
0.27X+2.60
0.44X+3.24
O.SOx+4.33
0.35X+0.58
0.22X+1.81
X' =
s,'-
S' =
Expected recovery for one or more measurements of a sample containing a
concentration of C, in M9/L.
Expected single analyst standard deviation of measurements at an average
concentration of x, in M9/L.
Expected interlaboratory_standard deviation of measurements at an average
concentration found of x, in M9/L.
True value for the concentration, in jug/l.
Average recovery found for measurements of samples containing a
concentration of C, in
8250A - 29
Revision 1
November 1990
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TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/Medium Low/Medium
Surrogate Compound Water Soil/Sediment
Nitrobenzene-d5 35-114 23-120
2-Fluorobiphenyl 43-116 30-115
p-Terphenyl -d14 33-141 18-137
Phenol-d6 10-94 24-113
2-Fluorophenol 21-100 25-121
2,4,6-Tribromophenol 10-123 19-122
8250A - 30 Revision 1
November 1990
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METHOD 8250A
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY fGC/HSl:
PACKED COLUMN TECHNIQUE
7.1 Prepare
sample using
Method 3540
or 3550.
7 . 1 Prepare
sample using
Method 3510
or 3520.
7.1 Prepare
sample using
Method 3540,
3550, or
3580.
7 . 2 Cleanup
ex tract.
7.3
Recommended
CC/MS
operating
condi tions.
7.4 Initial
calibration.
7.5 Daily
calibration-
Tune CC/MS with
TFTPP and check
SPCC & CCC.
8250A - 31
Revision 1
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METHOD 8250A
continued
7 6 1 Scrt«n
••tract in CC/FID
or CC/PID to
eliminat* too-high
concentration*
7 7.1 Identify
compound* by
comparing sample
retention time and
•ample mass spectra
to standards
8250A - 32
Revision 1
November 1990
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METHOD 8260A
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY fGC/MSl:
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8260 Is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Analyte
CAS No.b
Appropriate Technique
Direct
Purge-and-Trap Injection
Benzene
Bromobenzene
Bromochl oromethane
Bromodichloromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chi oromethane
2-Chlorotoluene
4-Chlorotoluene
Di bromochl oromethane
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Dibromomethane
1 , 2-Dichl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1 -Di chloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans- 1 , 2-Di chl oroethene
1,2-Dichloropropane
1,3-Dichloropropane
71-43-2
108-86-1
74-97-5
75-27-4
75-25-2
74-83-9
104-51-8
135-98-8
98-06-6
56-23-5
108-90-7
75-00-3
67-66-3
74-87-3
95-49-8
106-43-4
124-48-1
96-12-8
106-93-4
74-95-3
95-50-1
541-73-1
106-46-7
75-71-8
75-34-3
107-06-2
75-35-4
156-59-2
156-60-5
78-87-5
142-28-9
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
PP
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
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a
a
a
a
a
a
8260A - 1
Revision 1
November 1990
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Appropriate Technique
Direct
Analyte CAS No.b Purge-and-Trap Injection
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachl orobutadi ene
Isopropyl benzene
p-Isopropyl toluene
Methylene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1, 2 -Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tr i chl orof 1 uoromethane
1,2,3-Trichloropropane
1,2, 4-Trimethyl benzene
1, 3, 5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
a Adequate response by thi
594-20-7
563-58-6
100-41-4
87-68-3
98-82-8
99-87-6
75-09-2
91-20-3
103-65-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
87-61-6
120-82-1
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
95-63-6
108-67-8
75-01-4
95-47-6
108-38-3
106-42-3
s technique.
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
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a
a
a
a
a
a
a
a
a
a
a
a
b Chemical Abstract Services Registry Number.
pp Poor purging efficiency
i Inappropriate technique
resulting in high EQLs.
for this analyte.
pc Poor chromatographic behavior.
1.2 Method 8260 can be used to quantitate most volatile organic compounds
that have boiling points below 200°C and that are insoluble or slightly soluble
in water. Volatile water-soluble compounds can be included in this analytical
technique. However, for the more soluble compounds, quantitation limits are
approximately ten times higher because of poor purging efficiency. Such
compounds include low-molecular-weight halogenated hydrocarbons, aromatics,
ketones, nitriles, acetates, acrylates, ethers, and sulfides. See Tables 1 and
2 for lists of analytes and retention times that have been evaluated on a purge-
and-trap GC/MS system. Also, the method detection limits for 25 ml sample
volumes are presented.
1.3 The estimated quantitation limit (EQL) of Method 8260 for an
8260A - 2 Revision 1
November 1990
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individual compound is approximately 5 /ig/Kg (wet weight) for soil/sediment
samples, 0.5 mg/Kg (wet weight) for wastes, and 5 /ig/L for ground water (see
Table 3). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 Method 8260 is based upon a purge-and-trap, gas chromatographic/mass
spectrometric (GC/MS) procedure. This method is restricted to use by, or under
the supervision of, analysts experienced in the use of purge-and-trap systems
and gas chromatograph/mass spectrometers, and skilled in the interpretation of
mass spectra and their use as a quantitative tool.
2.0 SUMMARY OF METHOD
2.1 The volatile compounds are introduced into the gas chromatograph by
the purge-and-trap method or by direct injection (in limited applications).
Purged sample components are trapped in a tube containing suitable sorbent
materials. When purging is complete, the sorbent tube is heated and backflushed
with helium to desorb trapped sample components. The analytes are desorbed
directly to a large bore capillary or cryofocussed on a capillary precolumn
before being flash evaporated to a narrow bore capillary for analysis. The
column is temperature programmed to separate the analytes which are then detected
with a mass spectrometer (MS) interfaced to the gas chromatograph. Wide bore
capillary columns require a jet separator, whereas narrow bore capillary columns
can be directly interfaced to the ion source.
2.2 If the above sample introduction techniques are not applicable, a
portion of the sample is dispersed in solvent to dissolve the volatile organic
constituents. A portion of the solution is combined with organic-free reagent
water in the purge chamber. It is then analyzed by purge-and-trap GC/MS
following the normal water method.
2.3 Qualitative identifications are confirmed by analyzing standards
under the same conditions used for samples and comparing resultant mass spectra
and GC retention times. Each identified component is quantitated by relating
the MS response for an appropriate selected ion produced by that compound to the
MS response for another ion produced by an internal standard.
3.0 INTERFERENCES
3.1 Major contaminant sources are volatile materials in the laboratory
and impurities in the inert purging gas and in the sorbent trap. The use of
non-polytetrafluoroethylene (PTFE) thread sealants, plastic tubing, or flow
controllers with rubber components should be avoided since such materials out-
gas organic compounds which will be concentrated in the trap during the purge
operation. Analyses of calibration and reagent blanks provide information about
the presence of contaminants. When potential interfering peaks are noted in
blanks, the analyst should change the purge gas source and regenerate the
molecular sieve purge gas filter (Figure 1). Subtracting blank values from
sample results is not permitted. If reporting values not corrected for blanks
result in what the laboratory feels is a false positive for a sample, this should
be fully explained in text accompanying the uncorrected data.
8260A - 3 Revision 1
November 1990
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3.2 Interfering contamination may occur when a sample containing low
concentrations of volatile organic compounds is analyzed immediately after a
sample containing high concentrations of volatile organic compounds. The
preventive technique is rinsing of the purging apparatus and sample syringes
with two portions of organic-free reagent water between samples. After analysis
of a sample containing high concentrations of volatile organic compounds, one
or more calibration blanks should be analyzed to check for cross contamination.
For samples containing large amounts of water soluble materials, suspended
solids, high boiling compounds or high concentrations of compounds being
determined, it may be necessary to wash the purging device with a soap solution,
rinse it with organic-free reagent water, and then dry the purging device in an
oven at 105°C. In extreme situations, the whole purge and trap device may
require dismantling and cleaning. Screening of the samples prior to purge and
trap GC/MS analysis is highly recommended to prevent contamination of the system.
This is especially true for soil and waste samples. Screening may be
accomplished with an automated headspace technique or by Method 3820 (Hexadecane
Extraction and Screening of Purgeable Organics).
3.3 Special precautions must be taken to analyze for methylene chloride.
The analytical and sample storage area should be isolated from all atmospheric
sources of methylene chloride. Otherwise random background levels will result.
Since methylene chloride will permeate through PTFE tubing, all gas
chromatography carrier gas lines and purge gas plumbing should be constructed
from stainless steel or copper tubing. Laboratory clothing worn by the analyst
should be clean since clothing previously exposed to methylene chloride fumes
during liquid/liquid extraction procedures can contribute to sample
contamination.
3.4 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal into
the sample during shipment and storage. A trip blank prepared from organic-free
reagent water and carried through the sampling and handling protocol can serve
as a check on such contamination.
4.0 APPARATUS AND MATERIALS
4.1 Purge-and-trap device - The purge-and-trap device consists of three
separate pieces of equipment: the sample purger, the trap, and the desorber.
Several complete devices are commercially available.
4.1.1 The recommended purging chamber is designed to accept 5 mL
(and 25 mL if the lowest detection limit is required) samples with a water
column at least 3 cm deep. The gaseous headspace between the water column
and the trap must have a total volume of less than 15 mL. The purge gas
must pass through the water column as finely divided bubbles with a
diameter of less than 3 mm at the origin. The purge gas must be introduced
no more than 5 mm from the base of the water column. The sample purger,
illustrated in Figure 1, meets these design criteria. Alternate sample
purge devices (i.e. needle spargers), may be utilized, provided equivalent
performance is demonstrated.
4.1.2 The trap must be at least 25 cm long and have an inside
diameter of at least 0.105 in. Starting from the inlet, the trap must
8260A - 4 Revision 1
November 1990
-------
contain the following amounts of adsorbents: 1/3 of 2,6-diphenylene oxide
polymer, 1/3 of silica gel, and 1/3 of coconut charcoal. It is recommended
that 1.0 cm of methyl silicone-coated packing be inserted at the inlet to
extend the life of the trap (see Figure 2). If it is not necessary to
analyze for dichlorodifluoromethane or other fluorocarbons of similar
volatility, the charcoal can be eliminated and the polymer increased to
fill 2/3 of the trap. If only compounds boiling above 35°C are to be
analyzed, both the silica gel and charcoal can be eliminated and the
polymer increased to fill the entire trap. Before initial use, the trap
should be conditioned overnight at 180°C by backflushing with an inert gas
flow of at least 20 mL/min. Vent the trap effluent to the room, not to
the analytical column. Prior to daily use, the trap should be conditioned
for 10 minutes at 180°C with backflushing. The trap may be vented to the
analytical column during daily conditioning; however, the column must be
run through the temperature program prior to analysis of samples. Traps
normally last 2-3 months when used daily. Some signs of a deteriorating
trap are: uncharacteristic recoveries of surrogates, especially toluene-d8;
a loss of the response of the internal standards during a 12 hour shift;
and/or a rise in the baseline in the early portion of the scan.
4.1.3 The desorber should be capable of rapidly heating the trap
to 180°C for desorption. The trap bake-out temperature should not exceed
220°C. The desorber design illustrated in Figure 2 meets these criteria.
4.1.4 The purge-and-trap device may be assembled as a separate unit
or may be coupled to a gas chromatograph, as shown in Figures 3 and 4.
4.1.5 Trap Packing Materials
4.1.5.1 2,6-Diphenylene oxide polymer - 60/80 mesh,
chromatographic grade (Tenax GC or equivalent).
4.1.5.2 Methyl silicone packing - OV-1 (3%) on Chromosorb-W,
60/80 mesh or equivalent.
4.1.5.3 Silica gel - 35/60 mesh, Davison, grade 15 or
equivalent.
4.1.5.4 Coconut charcoal - Prepare from Barnebey Cheney, CA-
580-26 lot #M-2649 by crushing through a 26 mesh screen (or
equivalent).
4.2 Heater or heated oil bath - Should be capable of maintaining the
purging chamber to within 1°C over the temperature range of ambient to 100°C.
4.3 Gas chromatography/mass spectrometer/data system
4.3.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless injection
and all required accessories, including syringes, analytical columns, and
gases. The GC should be equipped with variable constant differential flow
controllers so that the column flow rate will remain constant throughout
desorption and temperature program operation. For some column
configuration, the column oven must be cooled to < 30°C, therefore, a
8260A - 5 Revision 1
November 1990
-------
subambient oven controller may be required. The capillary column should
be directly coupled to the source.
4.3.1.1 Capillary precolumn interface when using cryogenic
cooling - This device interfaces the purge and trap concentrator to
the capillary gas chromatograph. The interface condenses the
desorbed.sample components and focuses them into a narrow band on
an uncoated fused silica capillary precolumn. When the interface
is flash heated, the sample is transferred to the analytical
capillary column.
4.3.1.1.1 During the cryofocussing step, the temperature
of the fused silica in the interface is maintained at -150°C
under a stream of liquid nitrogen. After the desorption
period, the interface must be capable of rapid heating to 250°C
in 15 seconds or less to complete the transfer of analytes.
4.3.2 Gas chromatographic columns
4.3.2.1 Column 1 - 60 m x 0.75 mm ID capillary column coated
with VOCOL (Supelco), 1.5 urn film thickness, or equivalent.
4.3.2.2 Column 2 - 30 m x 0.53 mm ID capillary column coated
with DB-624 (J&W Scientific) or VOCOL (Supelco), 3 /im film
thickness, or equivalent.
4.3.2.3 Column 3 - 30 m x 0.32 mm ID capillary column coated
with DB-5 (J&W Scientific) or SE-54 (Supelco), 1 /xm film thickness,
or equivalent.
4.3.3 Mass spectrometer - Capable of scanning from 35 to 300 amu
every 2 sec or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable of
producing a mass spectrum for Bromofluorobenzene (BFB) which meets all of
the criteria in Table 4 when 50 ng of the GC/MS tuning standard (BFB) is
injected through the GC. To ensure sufficient precision of mass spectral
data, the desirable MS scan rate allows acquisition of at least five
spectra while a sample component elutes from the GC.
4.3.4 GC/MS interface - The GC is interfaced to the MS with an all
glass enrichment device and an all glass transfer line, but any enrichment
device or transfer line can be used if the performance specifications
described in Section 8.2 can be achieved. Any GC-to-MS interface that gives
acceptable calibration points at 50 ng or less per injection for each of
the analytes and achieves all acceptable performance criteria (see Table
4) may be used. GC-to-MS interfaces constructed entirely of glass or of
glass-lined materials are recommended. Glass can be deactivated by
silanizing with dichlorodimethylsilane. This interface is only needed for
the wide bore columns (> 0.53 mm ID).
4.3.5 Data system - A computer system that allows the continuous
acquisition and storage on machine-readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
8260A - 6 Revision 1
November 1990
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interfaced to the mass spectrometer. The computer must have software that
allows searching any 6C/MS data file for ions of a specified mass and
plotting such ion abundances versus time or scan number. This type of plot
is defined as an Extracted Ion Current Profile (EICP). Software must also
be available that allows integrating the abundances in any EICP between
specified time or scan-number limits. The most recent version of the
EPA/NIST Mass Spectral Library should also be available.
4.5 Microsyringes - 10, 25, 100, 250, 500, and 1,000 nl.
4.6 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.7 Syringes - 5, 10, or 25 ml, gas-tight with shutoff valve.
4.8 Balance - Analytical, 0.0001 g, and top-loading, 0.1 g.
4.9 Glass scintillation vials - 20 ml, with Teflon lined screw-caps or
glass culture tubes with Teflon lined screw-caps.
4.10 Vials - 2 ml, for GC autosampler.
4.11 Disposable pipets - Pasteur.
4.12 Volumetric flasks, Class A - 10 ml and 100 ml, with ground-glass
stoppers.
4.13 Spatula - Stainless steel.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform
to the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to
be free of analytes. Store apart from other solvents.
5.4 Reagent Tetraglyme - Reagent tetraglyme is defined as tetraglyme in
which interference is not observed at the method detection limit of compounds
of interest.
CAUTION; Glycol ethers are suspected carcinogens. All solvent handling should
be done in a hood while using proper protective equipment to minimize
exposure to liquid and vapor.
8260A - 7 Revision 1
November 1990
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5.4.1 Tetraglyme (tetraethylene glycol dimethyl ether, Aldrich #17,
240-5 or equivalent), C8H1805 - Purify by treatment at reduced pressure in
a rotary evaporator. The tetraglyme should have a peroxide content of less
than 5 ppm as indicated by EM Quant Test Strips (available from Scientific
Products Co., Catalog No. P1126-8 or equivalent).
5.4.1.1 Peroxides may be removed by passing the tetraglyme
through a column of activated alumina. The tetraglyme is placed in
a round bottom flask equipped with a standard taper joint, and the
flask is affixed to a rotary evaporator. The flask is immersed in
a water bath at 90-100°C and a vacuum is maintained at < 10 mm Hg for
at least two hours using a two-stage mechanical pump. The vacuum
system is equipped with an all-glass trap, which is maintained in
a dry ice/methanol bath. Cool the tetraglyme to ambient temperature
and add 100 mg/L of 2,6-di-tert-butyl-4-methyl-phenol to prevent
peroxide formation. Store the tetraglyme in a tightly sealed screw-
cap bottle in an area that is not contaminated by solvent vapors.
5.4.2 In order to demonstrate that all interfering volatiles have
been removed from the tetraglyme, an organic-free reagent water/tetraglyme
blank must be analyzed.
5.5 Polyethylene glycol, H(OCH2CH2)nOH - Free of interferences at the
detection limit of the target analytes.
5.6 Hydrochloric acid (1:1 v/v), HC1 - Carefully add a measured volume
of concentrated HC1 to an equal volume of organic-free reagent water.
5.7 Stock solutions - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standard solutions
in methanol, using assayed liquids or gases, as appropriate.
5.7.1 Place about 9.8 mL of methanol in a 10 ml tared ground-glass-
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all alcohol-wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
5.7.2 Add the assayed reference material, as described below.
5.7.2.1 Liquids - Using a 100 nl syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.7.2.2 Gases - To prepare standards for any compounds that
boil below 30°C (e.g. bromomethane, chloroethane, chloromethane, or
vinyl chloride), fill a 5 ml valved gas-tight syringe with the
reference standard to the 5.0 ml mark. Lower the needle to 5 mm
above the methanol meniscus. Slowly introduce the reference standard
above the surface of the liquid. The heavy gas will rapidly dissolve
in the methanol. Standards may also be prepared by using a lecture
bottle equipped with a Hamilton Lecture Bottle Septum (#86600).
Attach Teflon tubing to the side arm relief valve and direct a gentle
8260A - 8 Revision 1
November 1990
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stream of gas Into the methanol meniscus.
5.7.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.7.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.7.5 Prepare fresh standards for gases every two months or sooner
if comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months, or
sooner if comparison with check standards indicates a problem. Both gas
and liquid standards must be monitored closely by comparison to the initial
calibration curve and by comparison to QC check standards. It may be
necessary to replace the standards more frequently if either check exceeds
a 25% difference.
5.8 Secondary dilution standards - Using stock standard solutions, prepare
in methanol, secondary dilution standards containing the compounds of interest,
either singly or mixed together. Secondary dilution standards must be stored
with minimal headspace and should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards from
them. Store in a vial with no headspace for one week only.
5.9 Surrogate standards - The surrogates recommended are toluene-de,
4-bromofluorobenzene, and dibromofluoromethane. Other compounds may be used as
surrogates, depending upon the analysis requirements. A stock surrogate solution
in methanol should be prepared as described in Section 5.7, and a surrogate
standard spiking solution should be prepared from the stock at a concentration
of 50-250 M9/10 ml in methanol. Each sample undergoing GC/MS analysis must be
spiked with 10 /iL of the surrogate spiking solution prior to analysis.
5.10 Internal standards - The recommended internal standards are
chlorobenzene-d5, 1,4-difluorobenzene, l,4-dichlorobenzene-d4, and
pentafluorobenzene. Other compounds may be used as internal standards as long
as they have retention times similar to the compounds being detected by GC/MS.
Prepare internal standard stock and secondary dilution standards in methanol
using the procedures described in Sections 5.7 and 5.8. It is recommended that
the secondary dilution standard should be prepared at a concentration of 25 mg/L
of each internal standard compound. Addition of 10 /xL of this standard to
5.0 ml of sample or calibration standard would be the equivalent of 50 M9/L.
5.11 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng/jLtL of BFB in methanol should be prepared.
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5.12 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared from the secondary dilution of stock standards
(see Sections 5.7 and 5.8). Prepare these solutions in organic-free reagent
water. One of the concentrations should be at a concentration near, but above,
the method detection limit. The remaining concentrations should correspond to
the expected range of concentrations found in real samples but should not exceed
the working range of the GC/MS system. Each standard should contain each analyte
for detection by this* method (e.g. some or all of the compounds listed in Table
1 may be included). Calibration standards must be prepared daily.
5.13 Matrix spiking standards - Matrix spiking standards should be
prepared from volatile organic compounds which will be representative of the
compounds being investigated. At a minimum, the matrix spike should include
1,1-dichloroethene, trichloroethene, chlorobenzene, toluene, and benzene. It
is desirable to perform a matrix spike using compounds found in samples. Some
permits may require spiking specific compounds of interest, especially if they
are polar and would not be represented by the above listed compounds. The
standard should be prepared in methanol, with each compound present at a
concentration of 250 /ig/10.0 mL.
5.14 Great care must be taken to maintain the integrity of all standard
solutions. It is recommended all standards in methanol be stored at -10°C to
-20°C in amber bottles with Teflon lined screw-caps.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Direct injection - In very limited applications (e.g. aqueous process
wastes) direct injection of the sample into the GC/MS system with a 10 ;xL
syringe may be appropriate. One such application is for verification of the
alcohol content of an aqueous sample prior to determining if the sample 1s
ignitable (Methods 1010 or 1020). In this case, it is suggested that direct
injection be used. The detection limit is very high (approximately
10,000 Mg/L). Therefore, it is only permitted when concentrations in excess
of 10,000 jig/L are expected, or for water-soluble compounds that do not purge.
The system must be calibrated by direct injection using the same solvent (e.g.
water) for standards as the sample matrix (bypassing the purge-and-trap device).
7.2 Chromatographic conditions
7.2.1 General:
Injector temperature: 200-225°C
Transfer line temperature: 250-300°C
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7.2.2 Column 1 (A sample chromatogram is presented in Figure 5)
Carrier gas (He) flow rate: 15 mL/min
10°C, hold for 5 minutes
6°C/min to 160°C
160°C, hold until all expected compounds have
eluted.
Initial temperature:
Temperature program:
Final temperature:
7.2.3 Column 2, Cryogenic cooling (A sample chromatogram is
presented in Figure 6)
Carrier gas (He) flow rate: 15 mL/min
10°C, hold for 5 minutes
6°C/min to 160°C
160°C, hold until all expected compounds have
eluted.
Initial temperature:
Temperature program:
Final temperature:
7.2.4 Column 2, Non-cryogenic cooling (A sample chromatogram is
presented in Figure 7)
Carrier gas flow rate: It is recommended that carrier gas flow and split
and make-up gases be set using performance of
standards as guidance. Set the carrier gas head
pressure to » 10 psi and the split to » 30 mL/min.
Optimize the make-up gas flow for the separator
(approximately 30 mL/min) by injecting BFB, and
determining the optimum response when varying the
make-up gas. This will require several injections
of BFB. Next, make several injections of the
volatile working standard with all analytes of
interest. Adjust the carrier and split to provide
optimum chromatography and response. This is an
especially critical adjustment for the volatile
gas analytes. The head pressure should optimize
between 8-12 psi and the split between 20-60
mL/min. The use of the splitter is important to
minimize the effect of water on analyte response,
to allow the use of a larger volume of helium
during trap desorption, and to slow column flow.
Initial temperature:
Temperature program:
Final temperature:
45°C, hold for 2 minutes
8°C/min to 200°C
200°C, hold for 6 minutes,
A trap preheated to 150°C prior to trap desorption is required to provide
adequate chromatography of the gas analytes.
7.2.5 Column 3 (A sample chromatogram is presented in Figure 8)
Carrier gas (He) flow rate: 4 mL/min
10°C, hold for 5 minutes
6°C/min to 70°C, then 15°C/min to 145°C
145°C, hold until all expected compounds have
eluted.
Initial temperature:
Temperature program:
Final temperature:
7.3 Initial calibration for purge-and-trap procedure
7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
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in Table 4 for a 50 ng injection or purging of 4-bromofluorobenzene (2 /xL
injection of the BFB standard). Analyses must not begin until these
criteria are met.
7.3.2 Assemble a purge-and-trap device that meets the specification
in Section 4.1. Condition the trap overnight at 180°C in the purge mode
with an inert g*s flow of at least 20 mL/min. Prior to use, condition the
trap daily for 10 minutes while backflushing at 180°C with the column at
220°C.
7.3,3 Connect the purge-and-trap device to a gas chromatograph.
7.3.4 A set of at least five calibration standards containing the
method analytes is needed. One calibration standard should contain each
analyte at a concentration approaching but greater than the method
detection limit (Table 1) for that compound; the other calibration
standards should contain analytes at concentrations that define the range
of the method. The purging efficiency for 5 ml of water is greater than
for 25 mL. Therefore, develop the standard curve with whichever volume
of sample that will be analyzed. To prepare a calibration standard, add
an appropriate volume of a secondary dilution standard solution to an
aliquot of organic-free reagent water in a volumetric flask. Use a
microsyringe and rapidly inject the alcoholic standard into the expanded
area of the filled volumetric flask. Remove the needle as quickly as
possible after injection. Mix by inverting the flask three times only.
Discard the contents contained in the neck of the flask. Aqueous standards
are not stable and should be prepared daily. Transfer 5.0 ml (or 25 ml
if lower detection limits are required) of each standard to a gas tight
syringe along with 10 /iL of internal standard. Then transfer the contents
to a purging device.
7.3.5 Carry out the purge-and-trap analysis procedure as described
in Section 7.5.1.
7.3.6 Tabulate the area response of the characteristic ions (see
Table 5) against concentration for each compound and each internal
standard. Calculate response factors (RF) for each compound relative to
one of the internal standards. The internal standard selected for the
calculation of the RF for a compound should be the internal standard that
has a retention time closest to the compound being measured (Section
7.6.2). The RF is calculated as follows:
RF = (AXC,S)/(A,SCX)
where:
A, = Area of the characteristic ion for the compound being
measured.
Als = Area of the characteristic ion for the specific internal
standard.
Cls = Concentration of the specific internal standard.
Cx = Concentration of the compound being measured.
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7.3.7 The average RF must be calculated and recorded for each
compound. A system performance check should be made before this
calibration curve is used. Five compounds (the System Performance Check
Compounds, or SPCCs) are checked for a minimum average response factor.
These compounds are chloromethane; 1,1-dichloroethane; bromoform; 1,1,2,2-
tetrachloroethane; and chlorobenzene. The minimum acceptable average RF
for these compounds should be 0.300 (0.250 for bromoform). These compounds
typically have RFs of 0.4-0.6 and are used to check compound instability
and to check for degradation caused by contaminated lines or active sites
in the system. Examples of these occurrences are:
7.3.7.1 Chloromethane - This compound is the most likely
compound to be lost if the purge flow is too fast.
7.3.7.2 Bromoform - This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in the transfer lines may adversely
affect response. Response of the quantitation ion (m/z 173) is
directly affected by the tuning of BFB at ions m/z 174/176.
Increasing the m/z 174/176 ratio relative to m/z 95 may improve
bromoform response.
7.3.7.3 Tetrachloroethane and 1,1-dichloroethane - These
compounds are degraded by contaminated transfer lines in purge-and-
trap systems and/or active sites in trapping materials.
7.3.8 Using the RFs from the initial calibration, calculate the
percent relative standard deviation (%RSD) for Calibration Check Compounds
(CCCs). Record the %RSDs for all compounds. The percent RSD is calculated
as follows:
SD
%RSD = —^x 100
x
where:
RSD = Relative standard deviation.
x = Mean of 5 initial RFs for a compound.
SD = Standard deviation of average RFs for a compound.
N (x, - x)5
SD
i=l N - 1
The %RSD for each individual CCC must be less than 30 percent. This
criterion must be met for the individual calibration to be valid. The
CCCs are:
1,1-Dichloroethene,
Chloroform,
1,2-Dichloropropane,
Toluene,
Ethyl benzene, and
Vinyl chloride.
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7.4 Daily GC/MS calibration
7.4.1 Prior to the analysis of samples, inject or purge 50 ng of
the 4-bromofluorobenzene standard. The resultant mass spectra for the
BFB must meet all of the criteria given in Table 4 before sample analysis
begins. These criteria must be demonstrated each 12-hour shift.
7.4.2 The initial calibration curve (Section 7.3) for each compound
of interest must be checked and verified once every 12 hours of analysis
time. This is accomplished by analyzing a calibration standard that is
at a concentration near the midpoint concentration for the working range
of the GC/MS by checking the SPCC (Section 7.4.3) and CCC (Section 7.4.4).
7.4.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made each 12 hours. If the SPCC criteria are
met, a comparison of response factors is made for all compounds. This is
the same check that is applied during the initial calibration. If the
minimum response factors are not met, the system must be evaluated, and
corrective action must be taken before sample analysis begins. The minimum
response factor for volatile SPCCs is 0.300 (0.250 for Bromoform). Some
possible problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
and active sites in the column or chromatographic system.
7.4.4 Calibration Check Compounds (CCCs) - After the system
performance check is met, CCCs listed in Section 7.3.8 are used to check
the validity of the initial calibration. Calculate the percent difference
using the following equation:
RF, - RF
c
% Difference = - x 100
where:
RF,
RF, = Average response factor from initial calibration.
RFC = Response factor from current verification check standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. If the percent difference
for each CCC is less than 25%, the initial calibration is assumed to be
valid. If the criterion is not met (> 25% difference), for any one CCC,
corrective action must be taken. Problems similar to those listed under
SPCCs could affect this criterion. If no source of the problem can be
determined after corrective action has been taken, a new five-point
calibration must be generated. This criterion must be met before
quantitative sample analysis begins. If the CCCs are not required analytes
by the permit, then all required analytes must meet the 25% difference
criterion.
7.4.5 The internal standard responses and retention times in the
check calibration standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last check calibration (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
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must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration standard check, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate. When
corrections are made, reanalysis of samples analyzed while the system was
malfunctioning are necessary.
7.5 GC/MS analysis
7.5.1 Water samples
7.5.1.1 Screening of the sample prior to purge-and-trap
analysis will provide guidance on whether sample dilution is
necessary and will prevent contamination of the purge-and-trap
system. Two screening techniques that can be used are the headspace
sampler (Method 3810) using a gas chromatograph (GC) equipped with
a photo ionization detector (PID) in series with an electrolytic
conductivity detector (HECD), and extraction of the sample with
hexadecane and analysis of the extract on a GC with a FID and/or an
ECD (Method 3820).
7.5.1.2 All samples and standard solutions must be allowed
to warm to ambient temperature before analysis.
7.5.1.3 Set up the GC/MS system as outlined in Sections 4.3
and 7.2.
7.5.1.4 BFB tuning criteria and daily GC/MS calibration
criteria must be met (Section 7.4) before analyzing samples.
7.5.1.5 Adjust the purge gas (helium) flow rate to 25-
40 mL/min on the purge-and-trap device. Optimize the flow rate to
provide the best response for chloromethane and bromoform, if these
compounds are analytes. Excessive flow rate reduces chloromethane
response, whereas insufficient flow reduces bromoform response (see
Section 7.3.7).
7.5.1.6 Remove the plunger from a 5 ml syringe and attach a
closed syringe valve. If lower detection limits are required, use
a 25 ml syringe. Open the sample or standard bottle, which has been
allowed to come to ambient temperature, and carefully pour the sample
into the syringe barrel to just short of overflowing. Replace the
syringe plunger and compress the sample. Open the syringe valve and
vent any residual air while adjusting the sample volume to 5.0 ml.
This process of taking an aliquot destroys the validity of the liquid
sample for future analysis; therefore, if there is only one VOA vial,
the analyst should fill a second syringe at this time to protect
against possible loss of sample integrity. This second sample is
maintained only until such time when the analyst has determined that
the first sample has been analyzed properly. Filling one 20 ml
syringe would allow the use of only one syringe. If a second
analysis is needed from a syringe, it must be analyzed within 24
hours. Care must be taken to prevent air from leaking into the
syringe.
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7.5.1.7 The following procedure is appropriate for diluting
purgeable samples. All steps must be performed without delays until
the diluted sample is in a gas-tight syringe.
7.5.1.7.1 Dilutions may be made in volumetric flasks
(10 to 100 ml). Select the volumetric flask that will allow
foe the necessary dilution. Intermediate dilutions may be
necessary for extremely large dilutions.
7.5.1.7.2 Calculate the approximate volume of organic-
free reagent water to be added to the volumetric flask selected
and add slightly less than this quantity of organic-free
reagent water to the flask.
7.5.1.7.3 Inject the proper aliquot of sample from the
syringe prepared in Section 7.5.1.6 into the flask. Aliquots
of less than 1 ml are not recommended. Dilute the sample to
the mark with organic-free reagent water. Cap the flask,
invert, and shake three times. Repeat above procedure for
additional dilutions.
7.5.1.7.4 Fill a 5 ml syringe with the diluted sample
as in Section 7.5.1.6.
7.5.1.8 Compositing samples prior to GC/MS analysis
7.5.1.8.1 Add 5 ml or equal larger amounts of each
sample (up to 5 samples are allowed) to a 25 ml glass syringe.
Special precautions must be made to maintain zero headspace
in the syringe.
7.5.1.8.2 The samples must be cooled at 4°C during this
step to minimize volatilization losses.
7.5.1.8.3 Mix well and draw out a 5 ml aliquot for
analysis.
7.5.1.8.4 Follow sample introduction, purging, and
desorption steps described in the method.
7.5.1.8.5 If less than five samples are used for
compositing, a proportionately smaller syringe may be used
unless a 25 ml sample is to be purged.
7.5.1.9 Add 10.0 nl of surrogate spiking solution
(Section 5.9) and 10 p,L of internal standard spiking solution
(Section 5.10) through the valve bore of the syringe; then close
the valve. The surrogate and internal standards may be mixed and
added as a single spiking solution. The addition of 10 /uL of the
surrogate spiking solution to 5 ml of sample is equivalent to a
concentration of 50 /ig/L of each surrogate standard.
7.5.1.10 Attach the syringe-syringe valve assembly to the
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syringe valve on the purging device. Open the syringe valves and
inject the sample into the purging chamber.
7.5.1.11 Close both valves and purge the sample for 11.0 ±
0.1 minutes at ambient temperature. Be sure the trap is cooler than
25°C.
«
7.5.1.12 Sample desorption - The mode of sample desorption
is determined by the type of capillary column employed for the
analysis. When using a wide bore capillary column, follow the
desorption conditions of Section 7.5.1.13. The conditions for using
narrow bore columns are described in Section 7.5.1.14.
7.5.1.13 Sample desorption for wide bore capillary column.
Under most conditions, this type of column must be interfaced to
the MS through an all glass jet separator.
7.5.1.13.1 After the 11 minute purge, attach the trap
to the chromatograph, adjust the purge and trap system to the
desorb mode (Figure 4) and initiate the temperature program
sequence of the gas chromatograph and start data acquisition.
Introduce the trapped materials to the GC column by rapidly
heating the trap to 180°C while backflushing the trap with an
inert gas at 15 mL/min for 4 minutes. If the non-cryogenic
cooling technique is followed, the trap must be preheated to
150°C just prior to trap desorption at 180°C. While the purged
analytes are being introduced into the gas chromatograph, empty
the purging device using the sample syringe and wash the
chamber with two 5 ml or 25 ml portions of organic-free reagent
water depending on the size of the purge device. After the
purging device has been emptied, leave the syringe valve open
to allow the purge gas to vent through the sample introduction
needle.
7.5.1.13.2 Hold the column temperature at 10°C for
5 minutes, then program at 6°C/min to 160°C and hold until all
analytes elute.
7.5.1.13.3 After desorbing the sample for 4 minutes,
condition the trap by returning the purge-and-trap system to
the purge mode. Wait 15 seconds, then close the syringe valve
on the purging device to begin gas flow through the trap.
Maintain the trap temperature at 180°C. After approximately
7 minutes, turn off the trap heater and open the syringe valve
to stop the gas flow through the trap. When the trap is cool,
the next sample can be analyzed.
7.5.1.14 Sample desorption for narrow bore capillary column.
Under normal operating conditions, most narrow bore capillary columns
can be interfaced directly to the MS without a jet separator.
7.5.1.14.1 After the 11 minute purge, attach the trap
to the cryogenically cooled interface at -150°C and adjust
the purge-and-trap system to the desorb mode (Figure 4).
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Introduce the trapped materials to the interface by rapidly
heating the trap to 180°C while backflushing the trap with an
inert gas at 4 mL/min for 5 minutes. While the extracted
sample is being introduced into the interface, empty the
purging device using the sample syringe and rinse the chamber
with two 5 ml or 25 ml portions of organic-free reagent water
depending on the size of the purging device. After the purging
device has been emptied, leave the syringe valve open to allow
the purge gas to vent through the sample introduction needle.
After desorbing for 5 minutes, flash heat the interface to
250°C and quickly introduce the sample on the chromatographic
column. Start the temperature program sequence, and initiate
data acquisition.
7.5.1.14.2 Hold the column temperature at 10°C for
5 minutes, then program at 6°C/min to 70°C and then at 15°C/min
to 145°C. After desorbing the sample for 5 minutes,
recondition the trap by returning the purge-and-trap system
to the purge mode. Wait 15 seconds, then close the syringe
valve on the purging device to begin gas flow through the trap.
Maintain the trap temperature at 180°C. After approximately
15 minutes, turn off the trap heater and open the syringe valve
to stop the gas flow through the trap. When the trap is cool,
the next sample can be analyzed.
7.5.1.15 If the initial analysis of sample or a dilution of
the sample has a concentration of analytes that exceeds the initial
calibration range, the sample must be reanalyzed at a higher
dilution. Secondary ion quantitation is allowed only when there are
sample interferences with the primary ion. When a sample is analyzed
that has saturated ions from a compound, this analysis must be
followed by a blank organic-free reagent water analysis. If the
blank analysis is not free of interferences, the system must be
decontaminated. Sample analysis may not resume until the blank
analysis is demonstrated to be free of interferences.
7.5.1.16 For matrix spike analysis, add 10 /xL of the matrix
spike solution (Section 5.13) to the 5 ml of sample to be purged.
Disregarding any dilutions, this is equivalent to a concentration
of 50 ng/l of each matrix spike standard.
7.5.1.17 All dilutions should keep the response of the major
constituents (previously saturated peaks) in the upper half of the
linear range of the curve. Proceed to Sections 7.6.1 and 7.6.2 for
qualitative and quantitative analysis.
7.5.2 Water-miscible liquids
7.5.2.1 Water-miscible liquids are analyzed as water samples
after first diluting them at least 50 fold with organic-free reagent
water.
7.5.2.2 Initial and serial dilutions can be prepared by
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pipetting 2 ml of the sample to a 100 ml volumetric flask and
diluting to volume with organic-free reagent water. Transfer
immediately to a 5 ml gas-tight syringe.
7.5.2.3 Alternatively, prepare dilutions directly in a 5 ml
syringe filled with organic-free reagent water by adding at least
20 ML, but not more than 100 /iL of liquid sample. The sample is
ready for addition of internal and surrogate standards.
7.5.3 Sediment/soil and waste samples - It is highly recommended
that all samples of this type be screened prior to the purge-and-trap
GC/MS analysis. The headspace method (Method 3810) or the hexadecane
extraction and screening method (Method 3820) may used for this purpose.
These samples may contain percent quantities of purgeable organics that
will contaminate the purge-and-trap system, and require extensive cleanup
and instrument downtime. Use the screening data to determine whether to
use the low-concentration method (0.005-1 mg/Kg) or the high-concentration
method (> 1 mg/Kg).
7.5.3.1 Low-concentration method - This is designed for
samples containing individual purgeable compounds of < 1 mg/Kg. It
is limited to sediment/soil samples and waste that is of a similar
consistency (granular and porous). The low-concentration method is
based on purging a heated sediment/soil sample mixed with organic-
free reagent water containing the surrogate and internal standards..
Analyze all blanks and standards under the same conditions as the
samples. See Figure 9 for an illustration of a low soils impinger.
7.5.3.1.1 Use a 5 g sample if the expected concentration
is < 0.1 mg/Kg or a 1 g sample for expected concentrations
between 0.1 and 1 mg/Kg.
7.5.3.1.2 The GC/MS system should be set up as in
Sections 7.5.1.3-7.5.1.4. This should be done prior to the
preparation of the sample to avoid loss of volatile* from
standards and samples. A heated purge calibration curve must
be prepared and used for the quantitation of all samples
analyzed with the low-concentration method. Follow the initial
and daily calibration instructions, except for the addition
of a 40°C purge temperature.
7.5.3.1.3 Remove the plunger from a 5 mL Luerlock type
syringe equipped with a syringe valve and fill until
overflowing with water. Replace the plunger and compress the
water to vent trapped air. Adjust the volume to 5.0 mL. Add
10 /iL each of surrogate spiking solution (Section 5.9) and
internal standard solution (Section 5.10) to the syringe
through the valve (surrogate spiking solution and Internal
standard solution may be mixed together). The addition of
10 juL of the surrogate spiking solution to 5 g of
sediment/soil is equivalent to 50 M9/Kg of each surrogate
standard.
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7.5.3.1.4 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not discard
any supernatant liquids. Mix the contents of the sample
container with a narrow metal spatula. Weigh the amount
determined in Section 7.5.3.1.1 into a tared purge device.
Note and record the actual weight to the nearest 0.1 g.
7.5.3.1.5 Determine the percent dry weight of the
soil/sediment sample. This includes waste samples that are
amenable to percent dry weight determination. Other wastes
should be reported on a wet-weight basis.
7.5.3.1.5.1 Immediately after weighing the sample
for extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by
drying overnight at 105°C. Allow to cool in a desiccator
before re-weighing. Concentrations of individual
analytes are reported relative to the dry weight of
sample.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.
% dry weight = q of dry sample x 100
g of sample
7.5.3.1.6 Add the spiked organic-free reagent water to
the purging device, which contains the weighed amount of
sample, and connect the device to the purge-and-trap system.
NOTE: Prior to the attachment of the purge device, the procedures in Sections
7.5.3.1.4 and 7.5.3.1.6 must be performed rapidly and without interruption
to avoid loss of volatile organics. These steps must be performed in a
laboratory free of solvent fumes.
7.5.3.1.7 Heat the sample to 40°C ± 1°C and purge the
sample for 11.0 ± 0.1 minutes. Be sure the trap is cooler
than 25°C.
7.5.3.1.8 Proceed with the analysis as outlined in
Sections 7.5.1.12-7.5.1.17. Use 5 ml of the same organic-free
reagent water as in the blank. If saturated peaks occurred
or would occur if a 1 g sample were analyzed, the high-
concentration method must be followed.
7.5.3.1.9 For low-concentration sediment/soils, add
10 /iL of the matrix spike solution (Section 5.7) to the 5 ml
of organic-free reagent water (Section 7.5.3.1.3). The
concentration for a 5 g sample would be equivalent to 50 M9/Kg
of each matrix spike standard.
7.5.3.2 High-concentration method - The method is based on
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extracting the sediment/soil with methanol. A waste sample Is either
extracted or diluted, depending on its solubility in methanol. Wastes
(i.e. petroleum and coke wastes) that are insoluble in methanol are
diluted with tetraglyme or possibly polyethylene glycol (PEG). An
aliquot of the extract is added to organic-free reagent water
containing surrogate and internal standards. This is purged at
ambient temperature. All samples with an expected concentration of
> 1.0 mg/Kg should be analyzed by this method.
7.5.3.2.1 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not discard
any supernatant liquids. Mix the contents of the sample
container with a narrow metal spatula. For sediment/soil and
solid wastes that are insoluble in methanol weigh 4 g (wet
weight) of sample into a tared 20 ml vial. Use a top-loading
balance. Note and record the actual weight to 0.1 gram and
determine the percent dry weight of the sample using the
procedure in Section 7.5.3.1.5. For waste that is soluble in
methanol, tetraglyme, or PEG, weigh 1 g (wet weight) into a
tared scintillation vial or culture tube or a 10 ml volumetric
flask. (If a vial or tube is used, it must be calibrated
prior to use. Pipet 10.0 ml of solvent into the vial and mark
the bottom of the meniscus. Discard this solvent.)
7.5.3.2.2 Quickly add 9.0 ml of appropriate solvent;
then add 1.0 ml of the surrogate spiking solution to the vial.
Cap and shake for 2 minutes.
NOTE: Sections 7.5.3.2.1 and 7.5.3.2.2 must be performed rapidly and without
interruption to avoid loss of volatile organics. These steps must be
performed in a laboratory free from solvent fumes.
7.5.3.2.3 Pipet approximately 1 ml of the extract to
a GC vial for storage, using a disposable pipet. The remainder
may be disposed. Transfer approximately 1 ml of appropriate
solvent to a separate GC vial for use as the method blank for
each set of samples. These extracts may be stored at 4°C in
the dark, prior to analysis. The addition of a 100 pi aliquot
of each of these extracts in Section 7.5.3.2.6 will give a
concentration equivalent to 6,200 jug/Kg of each surrogate
standard.
7.5.3.2.4 The GC/MS system should be set up as in
Sections 7.5.1.3-7.5.1.4. This should be done prior to the
addition of the solvent extract to organic-free reagent water.
7.5.3.2.5 The information in Table 10 can be used to
determine the volume of solvent extract to add to the 5 ml of
organic-free reagent water for analysis. If a screening
procedure was followed (Method 3810 or 3820), use the estimated
concentration to determine the appropriate volume. Otherwise,
estimate the concentration range of the sample from the low-
concentration analysis to determine the appropriate volume.
8260A - 21 Revision 1
November 1990
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If the sample was submitted as a high-concentration sample,
start with 100 ML. All dilutions must keep the response of
the major constituents (previously saturated peaks) in the
upper half of the linear range of the curve.
7.5.3.2.6 Remove the plunger from a 5.0 ml Luerlock
type syringe equipped with a syringe valve and fill until
overflowing with water. Replace the plunger and compress the
water to vent trapped air. Adjust the volume to 4.9 ml. Pull
the plunger back to 5.0 ml to allow volume for the addition
of the sample extract and of standards. Add 10 /uL of internal
standard solution. Also add the volume of solvent extract
determined in Section 7.5.3.2.5 and a volume of extraction or
dissolution solvent to total 100 juL (excluding solvent in
standards).
7.5.3.2.7 Attach the syringe-syringe valve assembly to
the syringe valve on the purging device. Open the syringe
valve and inject the water/solvent sample into the purging
chamber.
7.5.3.2.8 Proceed with the analysis as outlined in
Sections 7.5.1.12-7.5.1.17. Analyze all blanks on the same
instrument as that used for the samples. The standards and
blanks should also contain 100 /*L of the dilution solvent to.
simulate the sample conditions.
7.5.3.2.9 For a matrix spike in the high-concentration
sediment/soil samples, add 8.0 ml of methanol, 1.0 ml of
surrogate spike solution (Section 5.9), and 1.0 ml of matrix
spike solution (Section 5.13) as in Section 7.5.3.2.2. This
results in a 5,200 jug/Kg concentration of each matrix spike
standard when added to a 4 g sample. Add a 100 pi aliquot of
this extract to 5 ml of organic-free reagent water for purging
(as per Section 7.5.3.2.6).
7.6 Data interpretation
7.6.1 Qualitative analysis
7.6.1.1 The qualitative identification of compounds determined
by this method is based on retention time, and on comparison of the
sample mass spectrum, after background correction, with
characteristic ions in a reference mass spectrum. The reference mass
spectrum must be generated by the laboratory using the conditions
of thi-s method. The characteristic ions from the reference mass
spectrum are defined to be the three ions of greatest relative
intensity, or any ions over 30% relative intensity if less than three
such ions occur in the reference spectrum. Compounds should be
identified as present when the criteria below are met.
7.6.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
8260A - 22 Revision 1
November 1990
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each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographlc peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.6.1.1.2 The RRT of the sample component is within ±
0.06 RRT units of the RRT of the standard component.
7.6.1.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions
in the reference spectrum. (Example: For an ion with an
abundance of 50% in the reference spectrum, the corresponding
abundance in a sample spectrum can range between 20% and 80%.)
7.6.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
7.6.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce.
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background spectra
is important. Examination of extracted ion current profiles
of appropriate ions can aid in the selection of spectra, and
in qualitative identification of compounds. When analytes
coelute (i.e., only one chromatographic peak is apparent), the
identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting
compound.
7.6.1.2 For samples containing components not associated with
the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the type of analyses
being conducted. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
8260A - 23 Revision 1
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be present In the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from
the sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison
of sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
7.6.2 Quantitative analysis
7.6.2.1 When a compound has been identified, the quantitation
of that compound will be based on the integrated abundance from the
EICP of the primary characteristic ion. Quantitation will take place
using the internal standard technique. The internal standard used
shall be the one nearest the retention time of that of a given
analyte (e.g. see Table 6).
7.6.2.2 Calculate the concentration of each identified analyte
in the sample as follows:
Water
(AJd.)
concentration (jug/L) =
(Als)(RF)(V0)
where:
A,, = Area of characteristic ion for compound being measured.
Is = Amount of internal standard injected (ng).
Als= Area of characteristic ion for the internal standard.
RF = Response factor for compound being measured (Section 7.3.3).
V0 = Volume of water purged (ml), taking into consideration any
dilutions made.
Sediment/Soil Sludge (on a dry-weight basis) and Waste (normally on
a wet-weight basis)
(Ax)(Is)(Vt)
concentration (M9/Kg) =
(AJ(RF)(Y,)(H.)(D)
8260A - 24 Revision 1
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where:
A,,, I8, Als, RF, = Same as for water.
Vt = Volume of total extract (/iL) (use 10,000 /iL or a factor of this
when dilutions are made).
V, = Volume of extract added (/iL) for purging.
W8 = Weight of sample extracted or purged (g).
D = % dry weight of sample/100, or 1 for a wet-weight basis.
7.6.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
given above should be used with the following modifications: The
areas A,, and A,s should be from the total ion chromatograms, and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Required instrument QC is found in the following sections:
8.2.1 The 6C/MS system must be tuned to meet the BFB specifications
in Section 7.3.1.
,8.2.2 There must be an initial calibration of the GC/MS system as
specified in Section 7.3.
8.2.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.4.3 and the CCC criteria in Section 7.4.4, each 12 hours.
8.3 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.3.1 A quality control (QC) reference sample concentrate is
required containing each analyte at a concentration of 10 mg/L in methanol.
The QC reference sample concentrate may be prepared from pure standard
materials or purchased as certified solutions. If prepared by the
laboratory, the QC reference sample concentrate must be made using stock
standards prepared independently from those used for calibration.
8.3.2 Prepare a QC reference sample to contain 20 /ig/L of each
analyte by adding 200 /iL of QC reference sample concentrate to 100 mL of
organic-free reagent water.
8.3.3 Four 5 mL aliquots of the well mixed QC reference sample are
analyzed according to the method beginning in Section 7.5.1.
8260A - 25 Revision 1
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8.3.4 Calculate the average recovery (x) in M9/U and the standard
deviation of the recovery (s) in M9/U for each analyte using the four
results.
8.3.5 Tables 7 and 8 provide single laboratory recovery and
precision data obtained for the method analytes from water. Similar
results from dosed water should be expected by any experienced laboratory.
Compare s and x (Section 8.3.4) for each analyte to the single laboratory
recovery and precision data. Results are comparable if the calculated
standard deviation of the recovery does not exceed 2.6 times the single
laboratory RSD or 20%, whichever is greater, and the mean recovery lies
within the interval x ± 3S or x ± 30%, whichever is greater.
NOTE: The large number of analytes in Tables 7 and 8 present a substantial
probability that one or more will fail at least one of the acceptance
criteria when all analytes of a given method are determined.
8.3.6 When one or more of the analytes tested are not comparable
to the data in Table 7 or 8, the analyst must proceed according to Section
8.3.6.1 or 8.3.6.2.
8.3.6.1 Locate and correct the source of the problem and
repeat the test for all analytes beginning with Section 8.3.2.
8.3.6.2 Beginning with Section 8.3.2, repeat the test only
for those analytes that are not comparable. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Section
8.3.2.
8.4 For aqueous and soil matrices, laboratory established surrogate
control limits should be compared with the control limits listed in Table 9.
8.4.1 If recovery is not within limits, the following procedures
are required.
8.4.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.4.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.4.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.4.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4.2 At a minimum, each laboratory should update surrogate recovery
8260A - 26 Revision 1
November 1990
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limits on a matrix-by-matrix basis, annually.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL actually achieved in a given
analysis will vary depending on instrument sensitivity and matrix effects.
9.2 This method has been tested in a single laboratory using spiked
water. Using a wide-bore capillary column, water was spiked at concentrations
between 0.5 and 10 M9/L. Single laboratory accuracy and precision data are
presented for the method analytes in Table 7. Calculated MDLs are presented in
Table 1.
9.3 The method was tested using water spiked at 0.1 to 0.5 M9/L and
analyzed on a cryofocussed narrow-bore column. The accuracy and precision data
for these compounds are presented in Table 8. MDL values were also calculated
from these data and are presented in Table 2.
10.0 REFERENCES
1. Methods for the Determination of Organic Compounds in Finished Drinking
Water and Raw Source Water Method 524.2; U.S. Environmental Protection
Agency. Office of Research Development. Environmental Monitoring and Support
Laboratory: Cincinnati, OH 1986.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Bellar, T.A.; Lichtenberg, J.J. jh Amer. Water Works Assoc. 1974, 66(12).
739-744.
4. Bellar, T.A.; Lichtenberg, J.J. "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds"; in Van Hall, Ed.; Measurement of Organic Pollutants in Water
and Wastewater. ASTM STP 686, pp 108-129, 1979.
5. Budde, W.L.; Eichelberger, J.W. "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass Spectrometry Equipment and
Laboratories"; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory. Cincinnati, OH 45268, April 1980; EPA-
600/4-79-020.
6. Eichelberger, J.W.; Harris, L.E.; Budde, W.L. "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems"; Analytical Chemistry 1975, 47, 995-1000.
7. Olynyk, P.; Budde, W.L.; Eichelberger, J.W. "Method Detection Limit for
Methods 624 and 625"; Unpublished report, October 1980.
8260A - 27 Revision 1
November 1990
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8. Non Cryogenic Temperatures Program and Chromatogram, Private Communications;
Myron Stephenson and Frank Allen, EPA Region IV Laboratory, Athens, GA.
8260A - 28 Revision 1
November 1990
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TABLE 1.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON WIDE BORE CAPILLARY COLUMNS
ANALYTE
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene chloride
trans- 1 , 2-Dichl oroethene
1,1-Dichl oroethane
2,2-Dichloropropane
cis-1, 2-Dichl oroethene
Chloroform
Bromochl oromethane
1,1,1 -Tri chl oroethane
Carbon tetrachloride
1,1-Dichloropropene
Benzene
1,2-Dichloroethane
Tri chl oroethene
1,2-Dichloropropane
Bromodi chl oromethane
Dibromomethane
trans- 1,3-Di chl oropropene
Toluene
cis- 1,3-Di chl oropropene
1,1,2-Tri chl oroethane
Tetrachl oroethene
1,3-Dichloropropane
Di bromochl oromethane
1,2-Dibromoethane
1-Chlorohexane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
o-Xylene
Styrene
Bromoform
Isopropyl benzene
1,1,2, 2-Tetrachl oroethane
RETENTION TIME
(minutes)
Column la
1.55
1.63
1.71
2.01
2.09
2.27
2.89
3.60
3.98
4.85
6.01
6.19
6.40
6.74
7.27
7.61
7.68
8.23
8.40
9.59
10.09
10.59
10.65
--
12.43
--
13.41
13.74
14.04
14.39
14.73
15.46
15.76
15.94
15.99
16.12
16.17
17.11
17.31
17.93
18.06
18.72
Column 2b
0.70
0.73
0.79
0.96
1.02
1.19
1.57
2.06
2.36
2.93
3.80
3.90
4.80
4.38
4.84
5.26
5.29
5.67
5.83
7.27
7.66
8.49
7.93
--
10.00
--
11.05
11.15
11.31
11.85
11.83
13.29
13.01
13.33
13.39
13.69
13.68
14.52
14.60
14.88
15.46
16.35
Column 2'
3.13
3.40
3.93
4.80
--
6.20
7.83
9.27
9.90
10.80
11.87
11.93
12.60
12.37
12.83
13.17
13.10
13.50
13.63
14.80
15.20
15.80
15.43
16.70
17.40
17.90
18.30
18.60
18.70
19.20
19.40
--
20.67
20.87
21.00
21.30
21.37
22.27
22.40
22.77
23.30
24.07
MDLd
(WJ/L)
c
0.10
0.13
0.17
0.11
0.10
0.08
0.12
0.03
0.06
0.04
0.35
0.12
0.03
0.04
0.08
0.21
0.10
0.04
0.06
0.19
0.04
0.08
0.24
--
0.11
--
0.10
0.14
0.04
0.05
0.06
0.05
0.04
0.05
0.06
0.13
0.05
0.11
0.04
0.12
0.15
0.04
8260A - 29
Revision 1
November 1990
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TABLE 1.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
Column la Column 2b
Column 2'
MDL"
Bromobenzene 18.95 15.86 24.00
1,2,3-Trichloropropane 19.02 16.23 24.13
n-Propylbenzene 19.06 16.41 24.33
2-Chlorotoluene 19.34 16.42 24.53
1,3,5-Trimethylbenzene 19.47 16.90 24.83
4-Chlorotoluene 19.50 16.72 24.77
tert-Butylbenzene 20.28 17.57 26.60
1,2,4-Trimethylbenzene 20.34 17.70 31.50
sec-Butyl benzene 20.79 18.09 26.13
p-Isopropyltoluene 21.20 18.52 26.50
1,3-Dichlorobenzene 21.22 18.14 26.37
1,4-Dichlorobenzene 21.55 18.39 26.60
n-Butylbenzene 22.22 19.49 27.32
1,2-Dichlorobenzene 22.52 19.17 27.43
l,2-Dibromo-3-chloropropane 24.53 21.08
1,2,4-Trichlorobenzene 26.55 23.08 31.50
Hexachlorobutadiene 26.99 23.68 32.07
Naphthalene 27.17 23.52 32.20
1,2,3-Trichlorobenzene 27.78 24.18 32.97
INTERNAL STANDARDS/SURROGATES
4-Bromofluorobenzene 18.63 15.71 23.63
0.03
0.32
0.04
0.04
0.05
0.06
0.14
0.13
0.13
0.12
0.12
0.03
0.11
.03
.26
0.04
0.11
0.04
0.03
0.
0.
Column 1 - 60 meter x 0.75 mm ID VOCOL capillary. Hold at 10°C for 5 minutes,
then program to 160°C at 6°/min.
Column 2-30 meter x 0.53 mm ID DB-624 wide-bore capillary using cryogenic
oven. Hold at 10°C for 5 minutes, then program to 160°C at 6°/min.
Column 2' - 30 meter x 0.53 mm ID DB-624 wide-bore capillary, cooling GC oven
to ambient temperatures. Hold at 10°C for 6 minutes, program to 70°C at
107min> program to 120°C at 5°/nnn, then program to 180°C at 8°/nrin.
MDL based on a 25 ml sample volume.
8260A - 30
Revision 1
November 1990
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TABLE 2.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON NARROW BORE CAPILLARY COLUMNS
ANALYTE
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl chloride
Bromomethane
Chloroethane
Tr i chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene chloride
trans- 1,2-Di chl oroethene
1,1-Dichloroethane
cis- 1,2-Di chl oroethene
2,2-Dichloropropane
Chloroform
Bromochl oromethane
1,1,1 -Tri chl oroethane
1,2-Dichloroethane
1 , 1 -Di chl oropropene
Carbon tetrachloride
Benzene
1 , 2-Di chl oropropane
Tri chl oroethene
Dibromomethane
Bromodi chl oromethane
Toluene
1,1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Tetrachl oroethene
1,2-Dibromoethane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
Bromoform
o-Xylene
Styrene
1,1,2, 2-Tetrachl oroethane
1, 2, 3-Tri chl oropropane
I sopropyl benzene
RETENTION TIME
(minutes)
Column 3a
0.88
0.97
1.04
1.29
1.45
1.77
2.33
2.66
3.54
4.03
5.07
5.31
5.55
5.63
6.76
7.00
7.16
7.41
7.41
8.94
9.02
9.09
9.34
11.51
11.99
12.48
12.80
13.20
13.60
14.33
14.73
14.73
15.30
15.30
15.70
15.78
15.78
15.78
16.26
16.42
MDLb
(M9/L)
0.11
0.05
0.04
0.06
0.02
0.07
0.05
0.09
0.03
0.03
0.06
0.08
0.04
0.09
0.04
0.02
0.12
0.02
0.03
0.02
0.02
0.01
0.03
0.08
0.08
0.08
0.07
0.05
0.10
0.03
0.07
0.03
0.06
0.03
0.20
0.06
0.27
0.20
0.09
0.10
8260A - 31
Revision 1
November 1990
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TABLE 2.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
Column 3a
MDLb
(M9/L)
Bromobenzene
2-Chlorotoluene
n-Propyl benzene
4-Chlorotoluene
1,3, 5-Trimethyl benzene
tert-Butyl benzene
1, 2, 4-Trimethyl benzene
sec-Butyl benzene
1,3-Oichlorobenzene
p-Isopropyl toluene
1,4-Dichlorobenzene
1 , 2-Di chl orobenzene
n-Butyl benzene
1 , 2-Di bromo-3-chl oropropane
1,2, 4-Tri chl orobenzene
Naphthalene
Hexachlorobutadiene
1, 2, 3-Tri chl orobenzene
16.42
16.74
16.82
16.82
16.99
17.31
17.31
17.47
17.47
17:63
17.63
17.79
17.95
18.03
18.84
19.07
19.24
19.24
0.11
0.08
0.10
0.06
0.06
0.33
0.09
0.12
0.05
0.26
0.04
0.05
0.10
0.50
0.20
0.10
0.10
0.14
a Column 3-30 meter x 0.32 mm ID DB-5 capillary with 1 urn film thickness.
b MDL based on a 25 ml sample volume.
8260A - 32
Revision 1
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TABLE 3.
ESTIMATED QUANTITATION LIMITS FOR VOLATILE ANALYTES8
Estimated
Quantitation
Limits
Ground water Low Soil/Sediment"
M9/L Aig/Kg
Volume of water purged
All analytes in Table 1
5 ml
5
25 mL
1
5
Estimated Quantitation Limit (EQL) - The lowest concentration that can be
reliably achieved within specified limits of precision and accuracy during
routine laboratory operating conditions. The EQL is generally 5 to 10 times
the MDL. However, it may be nominally chosen within these guidelines to
simplify data reporting. For many analytes the EQL analyte concentration.
is selected for the lowest non-zero standard in the calibration curve.
Sample EQLs are highly matrix-dependent. The EQLs listed herein are
provided for guidance and may not always be achievable. See the following
information for further guidance on matrix-dependent EQLs.
EQLs listed for soil/sediment are based on wet weight. Normally data is
reported on a dry weight basis; therefore, EQLs will be higher, based on
the percent dry weight in each sample.
Other Matrices Factor0
Water miscible liquid waste 50
High-concentration soil and sludge 125
Non-water miscible waste 500
°EQL = [EQL for low soil sediment (Table 3)] X [Factor]. For non-aqueous
samples, the factor is on a wet-weight basis.
8260A - 33 Revision 1
November 1990
-------
TABLE 4.
BFB MASS - INTENSITY SPECIFICATIONS (4-BROMOFLUOROBENZENE)
Mass Intensity Required (relative abundance)
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95% but less than 101% of mass 174
177 5 to 9% of mass 176
8260A - 34 Revision 1
November 1990
-------
TABLE 5.
CHARACTERISTIC MASSES (M/Z) FOR PURGEABLE ORGANIC COMPOUNDS
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
Benzene
Bromobenzene
Bromochloromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Di chlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans- 1, 2-Di chl oroethene
1,2-Dichloropropane
1,3-Di chl oropropane
2 , 2-Di chl oropropane
1,1-Dichloropropene
Ethyl benzene
Hexachl orobutadi ene
Isopropyl benzene
p-Isopropyl toluene
Methylene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
78
156
128
83
173
94
91
105
119
117
112
64
83
50
91
91
75
129
107
93
146
146
146
85
63
62
96
96
96
63
76
77
75
91
225
105
119
84
128
91
104
131
77,
49,
85,
175,
96
92,
134
91,
119
77,
66
85
52
126
126
155,
127
109,
95,
111,
111,
111,
87
65,
98
61,
61,
61,
112
78
97
110,
106
223,
120
134,
86,
-
120
78
133,
158
130
127
254
134
134
114
157
188
174
148
148
148
83
63
98
98
77
227
91
49
119
8260A - 35
Revision 1
November 1990
-------
TABLE 5.
(Continued)
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
1 , 1 , 1 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1 , 2 ,3-Tri chl orobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1 , 2 , 4-Trimethyl benzene
1,3, 5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
83
166
92
180
180
97
83
95
101
75
105
105
62
106
106
106
131,
168,
91
182,
182,
99,
97,
130,
103
77
120
120
64
91
91
91
85
129
145
145
61
85
132
INTERNAL STANDARDS/SURROGATES
4-Bromof1uorobenzene
Di bromofl uoromethane
Toluene-d8
Pentaf1uorobenzene
1,4-Di f1uorobenzene
Chlorobenzene-d5
l,4-Dichlorobenzene-d4
95
113
98
168
114
117
152
174, 176
8260A - 36
Revision 1
November 1990
-------
TABLE 6.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
Pentafluorobenzene ,
Acetone
Acrolein
Acrylonitrile
Bromochloromethane
Bromomethane
2-Butanone
Carbon disulfide
Chloroethane
Chloroform
Chioromethane
D1chlorodi fluoromethane
1,1-01chloroethane
1,1-Dichloroethene
cis-l,2-D1chloroethene
trans-1, 2-Di chloroethene
2,2-Di chloropropane
lodomethane
Methylene chloride
1,1,1-Tri chloroethane
Tri chlorof1uoromethane
Vinyl acetate
Vinyl chloride
Chlorobenzene-d;
Bromoform
Chlorodi bromomethane
Chlorobenzene
1,3-Dichloropropane
Ethyl benzene
2-Hexanone
Styrene
1,1,1,2-Tetrachloroethane
Tetrachloroethene
Xylene
1.4-Difluorobenzene
Benzene
Bromodichloromethane
Bromofluorobenzene (surrogate)
Carbon tetrachloride
2-Chloroethyl vinyl ether
1,2-Dibromoethane
Dibromomethane
1,2-Dichloroethane
1,2-Dichloroethane-d4 (surrogate)
1,2-Dichloropropane
1,1-Dichloropropene
cis-1,3-Dichloropropene
trans-1,3-Di chloropropene
4-Methyl-2-pentanone
Toluene
Toluene-dB (surrogate)
1,1,2-Trichloroethane
Trichloroethene
1,4-Dichlorobenzene-d^
Bromobenzene
n-Butylbenzene
sec-Butyl benzene
tert-Butylbenzene
2-Chlorotoluene
4-Chlorotoluene
1,2-Di bromo-3-chloropropane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Di chlorobenzene
Hexachlorobutadiene
Isopropyl benzene
p-Isopropyltoluene
Naphthalene
n-Propylbenzene
1,1,2,2-Tetrachloroethane
1,2,3-Trichlorobenzene
1,2,4-Tri chlorobenzene
1,2,3-Tri chloropropane
1,2,4-Trimethylbenzene
1,3,5-Trimethylbenzene
8260A - 37
Revision 1
November 1990
-------
TABLE 7.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR VOLATILE
ORGANIC COMPOUNDS IN WATER DETERMINED WITH A WIDE
BORE CAPILLARY COLUMN
Analyte
Benzene
Bromobenzene
Bromochl oromethane
Bromodichloromethane
Bromoform
Bromomethane
n- Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chi oromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-Chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1, 2-DI chlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1 -Di chlorobenzene
1, 2-Di chlorobenzene
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans- 1, 2-Di chl oroethene
1,2-Dichl oropropane
1,3-Di chl oropropane
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachl orobutadi ene
I sopropyl benzene
p- I sopropyl toluene
Methyl ene chloride
Naphthalene
n-Propyl benzene
Cone. Number
Range, of Recovery,8
/ig/L Samples %
0.1
0.1
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.1
0.5
0.5
0.5
0.1
0.1
0.5
0.1
0.5
0.5
0.1
0.5
0.2
0.5
0.5
0.1
0.1
0.5
0.1
0.1
0.1
0.5
0.5
0.1
0.5
0.5
0.1
0.1
0.1
0.1
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 20
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
-100
- 10
31
30
24
30
18
18
18
16
18
24
31
24
24
23
31
31
24
31
24
24
31
24
31
18
24
31
34
18
30
30
31
12
18
31
18
16
23
30
31
31
97
100
90
95
101
95
100
100
102
84
98
89
90
93
90
99
83
92
102
100
93
99
103
90
96
95
94
101
93
97
96
86
98
99
100
101
99
95
104
100
Standard Percent
Deviation tel. Std.
of Recovery" Dev.
6.5
5.5
5.7
5.7
6.4
7.8
7.6
7.6
7.4
7.4
5.8
8.0
5.5
8.3
5.6
8.2
16.6
6.5
4.0
5.6
5.8
6.8
6.6
6.9
5.1
5.1
6.3
6.7
5.2
5.9
5.7
14.6
8.7
8.4
6.8
7.7
6.7
5.0
8.6
5.8
5.7
5.5
6.4
6.1
6.3
8.2
7.6
7.6
7.3
8.8
5.9
9.0
6.1
8.9
6.2
8.3
19.9
7.0
3.9
5.6
6.2
6.9
6.4
7.7
5.3
5.4
6.7
6.7
5.6
6.1
6.0
16.9
8.9
8.6
6.8
7.6
6.7
5.3
8.2
5.8
8260A - 38
Revision 1
November 1990
-------
TABLE 7.
(Continued)
Analyte
Cone. Number
Range, of
M9/L Samples
Recovery,8
%
Standard
Deviation
of Recovery"
Percent
Rel. Std.
Dev.
Styrene
1,1,1, 2 -Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1 , 2 , 3 -Tr i chl orobenzene
1,2,4-Trichlorobenzene
1,1,1 -Tri chl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1, 2, 4-Trimethyl benzene
1 , 3 , 5-Tri methyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
0.1
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.5
-100
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 31
- 10
- 10
39
24
30
24
18
18
18
18
18
24
24
16
18
23
18
18
31
18
102
90
91
89
102
109
108
98
104
90
89
108
99
92
98
103
97
104
7.3
6.1
5.7
6.0
8.1
9.4
9.0
7.9
7.6
6.5
7.2
15.6
8.0
6.8
6.5
7.4
6.3
8.0
7.2
6.8
6.3
6.8
8.0
8.6
8.3
8.1
7.3
7.3
8.1
14.4
8.1
7.4
6.7
7.2
6.5
7.7
a Recoveries were calculated using internal standard method. Internal standard
was fluorobenzene.
b Standard deviation was calculated by pooling data form three concentrations.
8260A - 39
Revision 1
November 1990
-------
TABLE 8.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR
VOLATILE ORGANIC COMPOUNDS IN WATER DETERMINED
WITH A NARROW BORE CAPILLARY COLUMN
Analyte
Benzene
Bromobenzene
Bromochloromethane
Bromodichloromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chloromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Di bromomethane
1, 2-Di Chlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans- 1, 2-Di chl oroethene
1,2-Dichloropropane
1,3-Dichloropropane
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
I sopropyl benzene
p-Isopropyl toluene
Methyl ene chloride
Naphthalene
n-Propyl benzene
Cone.
M9/L
0.1
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.1
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
Number
of
Samples
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Recovery,"
%
99
97
97
100
101
99
94
110
110
108
91
100
105
101
99
96
92
99
97
93
97
101
106
99
98
100
95
100
98
96
99
99
102
99
100
102
113
97
98
99
Standard
Deviation
of Recovery
6.2
7.4
5.8
4.6
5.4
7.1
6.0
7.1
2.5
6.8
5.8
5.8
3.2
4.7
4.6
7.0
10.0
5.6
5.6
5.6
3.5
6.0
6.5
8.8
6.2
6.3
9.0
3.7
7.2
6.0
5.8
4.9
7.4
5.2
6.7
6.4
13.0
13.0
7.2
6.6
Percent
Rel. Std.
Dev.
6.3
7.6
6.0
4.6
5.3
7.2
6.4
6.5
2.3
6.3
6.4
5.8
3.0
4.7
4.6
7.3
10.9
5.7
5.8
6.0
3.6
5.9
6.1
8.9
6.3
6.3
9.5
3.7
7.3
6.3
5.9
4.9
7.3
5.3
6.7
6.3
11.5
13.4
7.3
6.7
8260A - 40
Revision 1
November 1990
-------
TABLE 8.
(Continued)
Analyte
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1,1,2-Trichl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1,2, 4-Trimethyl benzene
1,3, 5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
Cone.
M9/L
0.5
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.5
0.5
0.5
0.1
0.5
0.5
0.5
Number
of
Samples
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Recovery,8
%
96
100
100
96
100
102
91
100
102
104
97
96
96
101
104
106
106
97
Standard
Deviation
of Recovery
19.0
4.7
12.0
5.0
5.9
8.9
16.0
4.0
4.9
2.0
4.6
6.5
6.5
4.2
0.2
7.5
4.6
6.1
Percent
Rel. Std.
Dev.
19.8
4.7
12.0
5.2
5.9
8.7
17.6
4.0
4.8
1.9
4.7
6.8
6.8
4.2
0.2
7.1
4.3
6.3
Recoveries were calculated using internal standard method. Internal standard
was fluorobenzene.
8260A - 41
Revision 1
November 1990
-------
TABLE 9.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High
Surrogate Compound Water Soil/Sediment
4-Bromofluorobenzenea 86-115 74-121
Dibromofluoromethane3 86-118 80-120
Toluene-d8a 88-110 81-117
Single laboratory data for guidance only.
TABLE 10.
QUANTITY OF EXTRACT REQUIRED FOR ANALYSIS OF
HIGH-CONCENTRATION SAMPLES
Approximate Volume of
Concentration Range Extract8
500 - 10,000 Aig/Kg 100 nl
1,000 - 20,000 jug/Kg 50 nl
5,000 - 100,000 jug/Kg 10 nl
25,000 - 500,000 ^g/Kg 100 ML of 1/50 dilution"
Calculate appropriate dilution factor for concentrations exceeding this table.
a The volume of solvent added to 5 ml of water being purged should be kept
constant. Therefore, add to the 5 mL syringe whatever volume of solvent
is necessary to maintain a volume of 100 /iL added to the syringe.
b Dilute an aliquot of the solvent extract and then take 100 pi for
analysis.
8260A - 42 Revision 1
November 1990
-------
FIGURE 1.
PURGING DEVICE
EXIT 1M IN. O.O
EXIT 1M IN OJ>.
» U MM 0.0.
INLET IM IN. 0.0.
10 MM GLASS FNT
MEDIUM PONosny
SAMPLE INLET
*WAV SYNNQC VALVt
17 CM » OAUQC STWNQE NEEDLE
6 MM 0.0 RUeSEN SEPTUM
INLET 1M IN. 0.0.
me IN. 0.0.
/^STAINLESS STEEL
IK
MOLECULAR SIEVE
PUHOE GAS HLTER
n.OWOONTIVX
8260A - 43
Revision 1
November 1990
-------
FIGURE 2.
TRAP PACKING AND CONSTRUCTION TO INCLUDE DESORB CAPABILITY
PACKING DETAIL
Z^- 5 MM QLA38 WOOL
CONSTRUCTION OTTAJL
FTTTMQNVr
AND I
77 CM SIUCA GCL
19 CM TENAX QC
•- 1 CM 3% OV-1
9MMOLAMWOOL
u FT rn/FOOT
ncssTANceww
MWAPPCDSOUO
THEMMCOUPLB
CONTROLLER
SCN80M
ELECTMOMC
TIMPBMTUMi
OOMTMOLANO
a 106 M. LO
0.1B M OA
ST,
8260A - 44
Revision 1
November 1990
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE
CARWERGAS
F10W CONTROL
PRESSURE
REGULATOR
SELECTION VALVE
PURGE QAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
UOWO INJECTION PORTS
COLUMN OVEN
.A/IT--,
JUUV
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
TRAP INLET
PURGING
DEVICE
NOTE:
ALL LINES BETWEEN TRAP
AND OC SHOULD BE HEATED
TOWC.
8260A - 45
Revision 1
November 1990
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE
CARRCRQAB
FLO* CONTROL
PRESSURE
REGULATOR
LIQUID INJECTION PORTS
r- COLUMN OVEN
CONFIRMATORY COLUMN
JUUvP
TO DETECTOR
ANALYTICAL COLUMN
PURGE GAS
FLOWCONT
13X MOLECULAR
SIEVE FILTER
OPTIONAL 4PORT COLUMN
SELECTION VALVE
TRAP INLET
TRAP
200-C
NOTE
ALL LINES BETWEEN TRAP
AND OC SHOULD BE HEATED
TO «TC.
8260A - 46
Revision 1
November 1990
-------
FIGURE 5.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
o_
T
M
• » »1
3N3;N3aO«01M3 *. -C'3
3N3TOMlMdt»N
3N3ZN380«OTH3IM1 -» '<
S 5
o r
n
c o
UJ (k
^ u
2 2
3NVH13UOM01H3 I aOWOUf
t 'T 'T
3NOHl30U01H3ia -t '
3QXW01H3 3N31AM13W
-I 'T
•\ANXA
J
n
(VI
•
-------
FIGURE 6.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
a
c
®
M
3N3ZKG8CM01H3XU1 -Ł**'<
3N3ZN3gOUGlH3UU -«*J'
«
<«
3N3ZN380H01H3XO -3 I
-«••
3N3WA1S * JN1-UX -O
»
- -2
II i*
2s J?
?- ' u tf
OS
^ «• y
I •
i
3N3H130M01H3IU1
3N3HJ.30M01H3IQ -«':
30IMOTM3 1ANIA
i
!
8260A - 48
Revision 1
November 1990
-------
00
ro
o»
vo
Z 3D
O*
10 <
to
o
F.1C
: 40"SCW2987 »843
CALI: 40».'S042987 13
RIC DATA: 40U5CW2987 »843 SCkllS 125 TO 900
04/23/87 9:28:00
SAMPLE: 40UOASTD04
COHDS.: F4000.40-160X8.12,F4,38MLPURGE,TEt6ILGEL.G6624,SWEEP35,18FSI
RANGE: G 1.1200 LABEL:.H 8, 4.0 GUAM: A 0, 1.0 J 0 BASE: U 20, 3
CM
^
u
in
(N
^
U
264132.
260
"I
3*8
5:06
6:40
"1
500
8:20
16:08
11:43
I
SCO
13:20
CO
r>
30
oc=
-n 30
m
o —i
i— •
o
30
CT>
SCHH
15:08 TIME
-------
I. I.I
U
US MM. HI
00
ro
en
o
on
O
$
M
M riMMOWXIM (Mt I1»)
cr>
CO
o
'00
X
-H
70
•• « t /
ULi
B
U
V
JL-
SCMf
IIP!
-------
FIGURE 9.
LOU SOILS IMPINGER
PURGE INLET FITTING
SAMPLE OUTLET FITTING
• 6r*m 00 GLASS TUBING
SEPTUM
CAP
40ml VIAL
u
8260A - 51
Revision 1
November 1990
-------
METHOD 8260A
GAS CHROMATOGRAPHY/MASS SPECTROMETRY FOR VOLATILE ORGANICS
CAPILLARY COLUMN TECHNIQUE
Start
Direct
7 1
Select
procedure for
introducing
sample into
CC/MS
and - trap
7.2 Set CC/MS
. operating
condi tions .
7.3.2 Tune
CC/MS system
xith BFB
73.2/733
Assemble
purge-and-trap
device. Connect
device to CC.
7.3.4 Prepare
calibration
standards.
7.3.6 Perform
purge-and-trap
analysis.
7.37
Calculate RFs
for 5 SPCCs.
738
Calculate
XRSD of RF
for CCCs.
7.4 Perform
daily
calibration.
8260A - 52
Revision 1
November 1990
-------
METHOD 8260A
(Continued)
High concentration
soil/sediment
7
wa ter and wate
miacible liquids
75313 Prepare
aqueous solution of
sur roga te and
internal standard*
1
751/752 Screen
sample using Method
3810 or 3820
(Dilute
«a tar-miscible
liquids at l«*lt SO
fold )
7.5.1.9 Add
internal standard
and surrogate
•piking solutions.
7.5.1.10/7.5 1.11
Perform
purge-and-trap
procedure.
7 5 3 2 2 Add
solvent, internal
standard and
surrogate spiking
solutions. Shake.
7532 Store
portion of extract
for re*analysis
Prepare method
blank.
7 5 1 12 Desorb trap
onto column Analyze
sample
chroma tographically
7611 Identify
analytes by
comparing the
sample and standard
mass spectra
7622 Calculate
the concentration
of each identified
analyte
8260A - 53
Revision 1
November 1990
-------
METHOD 8270B
SEMIVOLATILE ORGANIC COMPOUNDS BY
GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS1; CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8270 is used to determine the concentration of semivolatile
organic compounds in extracts prepared from all types of solid waste matrices,
soils, and ground water. Direct injection of a sample may be used in limited
applications. The following compounds can be determined by this method:
Compounds
CAS No8
Appropriate Preparation Techniques
3510 3520 3540 3550 3580
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
2-Acetyl ami nof 1 uorene
l-Acetyl-2-thiourea
Aldrin
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
3-Amino-9-ethylcarbazole
Anilazine
Aniline
o-Anisidine
Anthracene
Aramite
Aroclor - 1016
Aroclor - 1221
Aroclor - 1232
Aroclor - 1242
Aroclor - 1248
Aroclor - 1254
Aroclor - 1260
Azinphos-methyl
Barban
Benzidine
Benzoic acid
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo (g , h , i ) peryl ene
Benzo(a)pyrene
p-Benzoquinone
Benzyl alcohol
83-32-9
208-96-8
98-86-2
53-96-3
591-08-2
309-00-2
117-79-3
60-09-3
92-67-1
132-32-1
101-05-3
62-53-3
90-04-0
120-12-7
140-57-8
12674-11-2
11104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
86-50-0
101-27-9
92-87-5
65-85-0
56-55-3
205-99-2
207-08-9
191-24-2
50-32-8
106-51-4
100-51-6
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
HS(43)
X
X
X
X
X
X
X
HS(62)
LR
CP
X
X
X
X
X
X
OE
X
X
X
X
ND
ND
ND
X
ND
ND
ND
X
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
ND
X
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
ND
X
X
X
X
X
ND
ND
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
ND
X
X
X
X
X
X
LR
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
CP
X
X
X
X
X
X
X
X
8270B - 1
Revision 2
November 1990
-------
Appropriate Preparation Techniques
Compounds CAS Noa
o-BHC
0-BHC
-------
ADorooriate Preoaration Techniaues
Compounds
Dibenzofuran
Dibenzo(a,e)pyrene
1 , 2-Di bromo-3-chl oropropane
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Dieldrin
Di ethyl phthalate
Di ethyl stilbestrol
Di ethyl sulfate
Dihydrosaffrole
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl ami noazobenzene
7,12-Dimethylbenz(a)-
anthracene
3, 3* -Dimethyl benzi dine
a , a-Dimethyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
Dioxathion
Diphenylamine
5,5-Diphenylhydantoin
1 , 2 -Di phenyl hydrazi ne
Di-n-octyl phthalate
Disulfoton
Endosulfan I
Endosulfan II
Endosulfan sulfate
CAS Noa
132-64-9
192-65-4
96-12-8
84-74-2
117-80-6
95-50-1
541-73-1
106-46-7
91-94-1
120-83-2
87-65-0
62-73-7
141-66-2
60-57-1
84-66-2
56-53-1
64-67-5
56312-13-1
60-51-5
119-90-4
60-11-7
57-97-6
119-93-7
122-09-8
105-67-9
131-11-3
528-29-0
99-65-0
100-25-4
534-52-1
51-28-5
121-14-2
606-20-2
39300-45-3
88-85-7
78-34-2
122-39-4
57-41-0
122-66-7
117-84-0
298-04-4
959-98-8
33213-65-9
1031-07-8
3510
X
ND
X
X
OE
X
X
X
X
X
X
X
X
X
X
X
AW,OS(67)
LR
ND
HE,HS(31)
X
X
CP(45)
X
ND
X
X
X
X
HE(14)
X
X
X
X
CP,HS(28)
X
ND
X
X
X
X
X
X
X
X
3520
X
ND
X
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
X
3540
ND
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
X
3550
X
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
X
3580
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
X
LR
X
CP
X
X
X
X
X
X
X
X
X
X
X
CP
X
ND
X
X
X
X
X
X
X
X
8270B - 3
Revision 2
November 1990
-------
ADDrooriate Preoaration Techniaues
Compounds
Endrin
Endrin aldehyde
Endrin ketone
EPN
Ethion
Ethyl carbamate
Ethyl methanesulfonate
Ethyl parathion
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl orophene
Hexachl oropropene
Hexamethyl phosphoramide
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methylenebis(2-chloranil
4,4'-Methylenebis
(N,N-dimethylaniline)
Methyl methanesulfonate
2-Methyl naphthal ene
2-Methyl-5-nitroaniline
Methyl parathion
2-Methyl phenol
3-Methyl phenol
CAS No"
72-20-8
7421-93-4
53494-70-5
2104-64-5
563-12-2
51-79-6
62-50-0
56-38-2
52-85-7
115-90-2
55-38-9
33245-39-5
206-44-0
86-73-7
321-60-8
367-12-4
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
70-30-4
1888-71-7
680-31-9
123-31-9
193-39-5
465-73-6
78-59-1
120-58-1
143-50-0
21609-90-5
121-75-5
108-31-6
72-33-3
91-80-5
72-43-5
56-49-5
ine) 101-14-4
101-61-1
66-27-3
91-57-6
99-55-8
298-00-0
95-48-7
108-39-4
3510
X
X
X
X
X
DC(28)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
AW,CP(62)
X
X
ND
X
X
X
DC(46)
X
X
HS(5)
HE
X
X
X
X
OE,OS(0)
X
X
X
X
X
X
X
3520
X
X
X
ND
ND
ND
ND
X
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
3540
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
3550
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
3580
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
X
X
ND
X
X
X
8270B - 4
Revision 2
November 1990
-------
ADorooriate Preoaration Techniaues
Compounds
4-Methyl phenol
2-Methylpyridine
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
Naphthalene
Naphthalene -de (I.S.)
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
Nitrobenzene-d5 (surr.)
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
Nitroquinol ine-1-oxide
N-Ni trosodi butyl ami ne
N-Ni trosodi ethyl ami ne
N-Ni trosodimethyl ami ne
N-Ni trosomethyl ethyl ami ne
N-Ni trosodi phenyl ami ne
N-Ni trosodi -n-propyl ami ne
N-Nitrosomorphol ine
N-Nitrosopiperidine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachlorobenzene
Pentachloronitrobenzene
Pentachlorophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenobarbital
Phenol
CAS No"
106-44-5
109-06-8
7786-34-7
315-18-4
2385-85-5
6923-22-4
300-76-5
91-20-3
130-15-4
134-32-7
91-59-8
54-11-5
602-87-9
88-74-4
99-09-2
100-01-6
99-59-2
98-95-3
92-93-3
1836-75-5
88-75-5
100-02-7
99-55-8
56-57-5
924-16-3
55-18-5
62-75-9
10595-95-6
86-30-6
621-64-7
59-89-2
100-75-4
930-55-2
152-16-9
101-80-4
56-38-2
608-93-5
82-68-8
87-86-5
62-44-2
85-01-8
50-06-6
108-95-2
3510
X
X
X
HE,HS(68)
X
HE
X
X
X
X
OS(44)
X
DE(67)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
LR
X
X
X
X
X
X
X
X
X
X
DC(28)
3520
ND
X
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
ND
X
3540
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
ND
X
3550
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
ND
X
3580
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
X
X
8270B - 5
Revision 2
November 1990
-------
Appropriate Preparation Techniques
Compounds
Phenol -d6 (surr.)
1 , 4-Phenyl enedi ami ne
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
Propylthiouracil
Pyrene
Pyridine
Resorcinol
Safrole
Strychnine
Sul fall ate
Terbufos
Terphenyl-d14(surr.)
1,2,4 , 5-Tetrachl orobenzene
2,3,4,6-Tetrachlorophenol
Tetrachlorvinphos
Tetraethyl dithiopyrophosphate
Tetraethyl pyrophosphate
Thionazine
Thiophenol (Benzenethiol)
Toluene diisocyanate
o-Toluidine
Toxaphene
2,4,6-Tribromophenol (surr.)
1 , 2 , 4-Tri chl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Trifluralin
2,4,5-Trimethylaniline
Trimethyl phosphate
1, 3, 5-Tri nitrobenzene
Tris(2,3-dibromopropyl) phosphate
Tri-p-tolyl phosphate
0,0,0-Triethyl phosphorothioate
CAS Noa
106-50-3
298-02-2
2310-17-0
732-11-6
13171-21-6
85-44-9
109-06-8
120-62-7
23950-58-5
51-52-5
129-00-0
110-86-1
108-46-3
94-59-7
60-41-3
95-06-7
13071-79-9
95-94-3
58-90-2
961-11-5
3689-24-5
107-49-3
297-97-2
108-98-5
584-84-9
95-53-4
8001-35-2
120-82-1
95-95-4
88-06-2
1582-09-8
137-17-7
512-56-1
99-35-4
126-72-7
78-32-0
126-68-1
3510
DC(28)
X
X
HS(65)
HS(15)
HE(63)
CP,HE(1)
ND
X
X
LR
X
ND
DC,OE(10)
X
AW,OS(55)
X
X
X
X
X
X
X
X
X
X
HE(6)
X
X
X
X
X
X
X
X
HE(60)
X
X
X
X
3520
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
3540
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
ND
X
ND
ND
ND
ND
ND
ND
ND
3550
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
3580
X
X
X
X
X
X
CP
ND
X
X
LR
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
Chemical Abstract Service Registry Number.
8270B - 6
Revision 2
November 1990
-------
AW = Adsorption to walls of glassware during extraction and storage.
CP = Nonreproducible chromatographic performance.
DC = Unfavorable distribution coefficient (number in parenthesis is percent
recovery).
HE = Hydrolysis during extraction accelerated by acidic or basic conditions
(number in parenthesis is percent recovery).
HS = Hydrolysis "during storage (number in parenthesis is percent stability).
LR = Low response.
NO = Not determined.
OE = Oxidation during extraction accelerated by basic conditions (number in
parenthesis is percent recovery).
OS = Oxidation during storage (number in parenthesis is percent stability).
X = Greater than 70 percent recovery by this technique.
Percent Stability = Average Recovery (Day 7) x 100/Average Recovery (Day 0).
1.2 Method 8270 can be used to quantitate most neutral, acidic, and basic
organic compounds that are soluble in methylene chloride and capable of being
eluted without derivatization as sharp peaks from a gas chromatographic fused-
silica capillary column coated with a slightly polar silicone. Such compounds
include polynuclear aromatic hydrocarbons, chlorinated hydrocarbons and
pesticides, phthalate esters, organophosphate esters, nitrosamines, halpethers,
aldehydes, ethers, ketones, anilines, pyridines, quinolines, aromatic nitro
compounds, and phenols, including nitrophenols. See Table 1 for a list of
compounds and their characteristic ions that have been evaluated on the specified
GC/MS system.
1.3 The following compounds may require special treatment when being
determined by this method. Benzidine can be subject to oxidative losses during
solvent concentration. Also, chromatography is poor. Under the alkaline
conditions of the extraction step, a-BHC, 7-BHC, endosulfan I and II, and endrin
are subject to decomposition. Neutral extraction should be performed if these
compounds are expected. Hexachlorocyclopentadiene is subject to thermal
decomposition in the inlet of the gas chromatograph, chemical reaction in acetone
solution, and photochemical decomposition. N-nitrosodimethylamine is difficult
to separate from the solvent under the chromatographic conditions described.
N-nitrosodiphenylamine decomposes in the gas chromatographic inlet and cannot
be separated from diphenylamine. Pentachlorophenol, 2,4-dinitrophenol,
4-nitrophenol,4,6-dinitro-2-methylphenol,4-chloro-3-methylphenol, benzoicacid,
2-nitroaniline, 3-nitroaniline, 4-chloroaniline, and benzyl alcohol are subject
to erratic chromatographic behavior, especially if the GC system is contaminated
with high boiling material.
1.4 The estimated quantitation limit (EQL) of Method 8270 for determining
an individual compound is approximately 1 mg/Kg (wet weight) for soil/sediment
samples, 1-200 mg/Kg for wastes (dependent on matrix and method of preparation),
and 10 jug/L for ground water samples (see Table 2). EQLs will be
proportionately higher for sample extracts that require dilution to avoid
saturation of the detector.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
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skilled in the interpretation of mass spectra. Each analyst must demonstrate
the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract.
3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, and spikes must be evaluated
for interferences. Determine if the source of interference is in the preparation
and/or cleanup of the samples and take corrective action to eliminate the
problem.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system
4.1.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for spl itless injection
and all required accessories, including syringes, analytical columns, and
gases. The capillary column should be directly coupled to the source.
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID) 1 urn film
thickness silicone-coated fused-silica capillary column (J&W Scientific
DB-5 or equivalent).
4.1.3 Mass spectrometer - Capable of scanning from 35 to 500 amu
every 1 sec or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable
of producing a mass spectrum for decafluorotriphenylphosphine (DFTPP)
which meets all of the criteria in Table 3 when 1 pi of the GC/MS tuning
standard is injected through the GC (50 ng of DFTPP).
4.1.4 GC/MS interface - Any GC-to-MS interface that gives acceptable
calibration points at 50 ng per injection for each compound of interest
and achieves acceptable tuning performance criteria may be used.
4.1.5 Data system - A computer system must be interfaced to the
mass spectrometer. The system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
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the duration of the chromatographic program. The computer must have
software that can search any GC/MS data file for ions of a specific mass
and that can plot such ion abundances versus time or scan number. This
type of plot is defined as an Extracted Ion Current Profile (EICP).
Software must also be available that allows integrating the abundances in
any EICP between specified time or scan-number limits. The most recent
version of the EPA/NIST Mass Spectral Library should also be available.
4.2 Syringe - 10 /nL.
4.3 Volumetric flasks, Class A - 10 ml to 1000 ml.
4.4 Balance - Analytical, 0.0001 g.
4.5 Bottles - glass with Teflon-lined screw caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock standard solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 10 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96% or greater, the weight
may be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.3.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps. Store at 4°C and protect from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.3.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-ds, acenaphthene-d10, phenanthrene-d10,
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chrysene-d12, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Section 7.3.2 are met. Dissolve 0.200 g
of each compound with a small volume of carbon disulfide. Transfer to a 50 mL
volumetric flask and dilute to volume with methylene chloride so that the final
solvent is approximately 20% carbon disulfide. Most of the compounds are also
soluble in small volumes of methanol, acetone, or toluene, except for
perylene-d12. The resulting solution will contain each standard at a
concentration of 4,000 ng/jitL. Each 1 ml sample extract undergoing analysis
should be spiked with 10 /*L of the internal standard solution, resulting in a
concentration of 40 ng//iL of each internal standard. Store at 4°C or less when
not being used.
5.5 GC/MS tuning standard - A methylene chloride solution containing
50 ng//iL of decafluorotriphenylphosphine (DFTPP) should be prepared. The
standard should also contain 50 ng//uL each of 4,4'-DDT, pentachlorophenol, and
benzidine to verify injection port inertness and GC column performance. Store
at 4°C or less when not being used.
5.6 Calibration standards - A minimum of five calibration standards should
be prepared. One of the calibration standards should be at a concentration near,
but above, the method detection limit; the others should correspond to the range
of concentrations found in real samples but should not exceed the working range
of the GC/MS system. Each standard should contain each analyte for detection
by this method (e.g. some or all of the compounds listed in Table 1 may be
included). Each 1 ml aliquot of calibration standard should be spiked with
10 /iL of the internal standard solution prior to analysis. All standards should
be stored at -10°C to -20°C and should be freshly prepared once a year, or sooner
if check standards indicate a problem. The daily calibration standard should
be prepared weekly and stored at 4°C.
5.7 Surrogate standards - The recommended surrogate standards are
phenol-d6, 2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-d5,
2-fluorobiphenyl, and p-terphenyl-d14. See Method 3500 for the instructions on
preparing the surrogate standards. Determine what concentration should be in
the blank extracts after all extraction, cleanup, and concentration steps.
Inject this concentration into the GC/MS to determine recovery of surrogate
standards in all blanks, spikes, and sample extracts. Take into account all
dilutions of sample extracts.
5.8 Matrix spike standards - See Method 3500 for instructions on preparing
the matrix spike standard. Determine what concentration should be in the blank
extracts after all extraction, cleanup, and concentration steps. Inject this
concentration into the GC/MS to determine recovery of surrogate standards in all
matrix spikes. Take into account all dilutions of sample extracts.
5.9 Acetone, hexane, methylene chloride, isooctane, carbon disulfide,
toluene, and other appropriate solvents - Pesticide quality or equivalent
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
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7.0 PROCEDURE
7.1 Sample preparation - Samples must be prepared by one of the following
methods prior to GC/MS analysis.
Matrix
Water
Soil/sediment
Waste
Methods
3510, 3520
3540, 3550
3540, 3550, 3580
7.1.1 Direct injection - In very limited applications direct
injection of the sample into the GC/MS system with a 10 p,l syringe may
be appropriate. The detection limit is very high (approximately
10,000 Mg/L); therefore, it is only permitted where concentrations in
excess of 10,000 ng/l are expected. The system must be calibrated by
direct injection.
7.2 Extract cleanup - Extracts may be cleaned up by any of the following
methods prior to GC/MS analysis.
Compounds
Phenols
Phthalate esters
Nitrosamines
Organochlorine pesticides & PCBs
Nitroaromatics and cyclic ketones
Polynuclear aromatic hydrocarbons
Haloethers
Chlorinated hydrocarbons
Organophosphorus pesticides
Petroleum waste
All priority pollutant base,
neutral, and acids
Methods
3630, 3640, 8040a
3610, 3620, 3640
3610, 3620, 3640
3620, 3660
3620, 3640
3611, 3630, 3640
3620, 3640
3620, 3640
3620
3611, 3650
3640
Method 8040 includes a derivatization technique followed by GC/ECD analysis,
if interferences are encountered on GC/FID.
7.3 Initial calibration - The recommended GC/MS operating conditions:
Mass range:
Scan time:
Initial temperature:
Temperature program:
Final temperature:
Injector temperature:
Transfer line temperature:
Source temperature:
Injector:
Sample volume:
Carrier gas:
has
35-500 amu
1 sec/scan
40°C, hold for 4 minutes
40-270°C at 10°C/min
270°C, hold until benzo[g,h,i]perylene
eluted
250-300°C
250-300°C
According to manufacturer's specifications
Grob-type, splitless
1-2 juL
Hydrogen at 50 cm/sec or helium at 30 cm/sec
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7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 3 for a 50 ng injection of DFTPP. Analyses should not begin until
all these criteria are met. Background subtraction should be
straightforward and designed only to eliminate column bleed or instrument
background ions. The GC/MS tuning standard should also be used to assess
GC column performance and injection port inertness. Degradation of DDT
to DDE and ODD should not exceed 20%. Benzidine and pentachlorophenol
should be present at their normal responses, and no peak tailing should
be visible. If degradation is excessive and/or poor chromatography is
noted, the injection port may require cleaning. It may also be necessary
to break off the first 6-12 in. of the capillary column.
7.3.2 The internal standards selected in Section 5.1 should permit
most of the components of interest in a chromatogram to have retention
times of 0.80-1.20 relative to one of the internal standards. Use the base
peak ion from the specific internal standard as the primary ion for
quantitation (see Table 1). If interferences are noted, use the next most
intense ion as the quantitation ion (i.e. for l,4-dichlorobenzene-d4 use
m/z 152 for quantitation).
7.3.3 Analyze 1 ML of each calibration standard (containing
internal standards) and tabulate the area of the primary characteristic
ion against concentration for each compound (as indicated in Table 1).
Figure 1 shows a chromatogram of a calibration standard containing
base/neutral and acid analytes. Calculate response factors (RFs) for each
compound as follows:
RF = (AxCis)/(AisCJ
where:
Ax = Area of the characteristic ion for the compound being measured.
Ais = Area of the characteristic ion for the specific internal standard.
Cis = Concentration of the specific internal standard (ng//iL).
Cx = Concentration of the compound being measured
7.3.4 The average RF should be calculated for each compound. The
percent relative standard deviation (%RSD = 100[SD/RF]) should also be
calculated for each compound. The %RSD should be less than 30% for each
compound. However, the %RSD for each individual Calibration Check Compound
(CCC) (see Table 4) must be less than 30%. The relative retention times
of each compound in each calibration run should agree within 0.06 relative
retention time units. Late-eluting compounds usually have much better
agreement.
7.3.5 A system performance check must be performed to ensure that
minimum average RFs are met before the calibration curve is used. For
semivolatiles, the System Performance Check Compounds (SPCCs) are: N-
nitroso-di-n-propylamine; hexachlorocyclopentadiene; 2,4-dinitro-phenol ;
and 4-nitrophenol . The minimum acceptable average RF for these compounds
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SPCCs is 0.050. These SPCCs typically have very low RFs (0.1-0.2) and tend
to decrease in response as the chromatographic system begins to deteriorate
or the standard material begins to deteriorate. They are usually the first
to show poor performance. Therefore, they must meet the minimum
requirement when the system is calibrated.
7.4 Daily GC/MS calibration
7.4.1 Prior to analysis of samples, the GC/MS tuning standard must
be analyzed. A 50 ng injection of DFTPP must result in a mass spectrum
for DFTPP which meets the criteria given in Table 3. These criteria must
be demonstrated during each 12 hour shift.
7.4.2 A calibration standard(s) at mid-concentration, containing
each compound of interest, including all required surrogates, must be
performed every 12 hours during analysis. Compare the response factor
data from the standards every 12 hours with the average response factor
from the initial calibration for a specific instrument as per the SPCC
(Section 7.4.3) and CCC (Section 7.4.4) criteria.
7.4.3 System Performance Check Compounds (SPCCs): A system
performance check must be made during every 12 hour shift. If the SPCC
criteria are met, a comparison of response factors is made for all
compounds. This is the same check that is applied during the initial
calibration. If the minimum response factors are not met, the system must
be evaluated, and corrective action must be taken before sample analysis
begins. The minimum RF for semivolatile SPCCs is 0.050. Some possible
problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
and active sites in the column or chromatographic system. This check must
be met before analysis begins.
7.4.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Table 4 are used to check the
validity of the initial calibration. Calculate the percent difference
using:
RF, - RFC
% Difference = —— x 100
RF,
where:
RF, = Average response factor from initial calibration.
RFC = Response factor from current verification check standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. If the percent difference
for each CCC is less than 30%, the initial calibration is assumed to be
valid. If the criterion is not met (> 30% difference) for any one CCC,
corrective action must be taken. Problems similar to those listed under
SPCCs could affect this criterion. If no source of the problem can be
determined after corrective action has been taken, a new five-point
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calibration must be generated. This criterion must be met before sample
analysis begins.
7.4.5 The internal standard responses and retention times in the
calibration check standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last check calibration (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration standard check, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate.
7.5 GC/MS analysis
7.5.1 It is highly recommended that the extract be screened on a
GC/FID or GC/PID using the same type of capillary column. This will
minimize contamination of the GC/MS system from unexpectedly high
concentrations of organic compounds.
7.5.2 Spike the 1 ml extract obtained from sample preparation with
10 pi of the internal standard solution just prior to analysis.
7.5.3 Analyze the 1 ml extract by GC/MS using a 30 m x 0.25 mm (or
0.32 mm) silicone-coated fused-silica capillary column. The volume to be
injected should ideally contain 100 ng of base/neutral and 200 ng of acid
surrogates (for a 1 /xL injection). The recommended GC/MS operating
conditions to be used are specified in Section 7.3.
7.5.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the GC/MS system, extract dilution must take
place. Additional internal standard must be added to the diluted extract
to maintain the required 40 ng/^L of each internal standard in the
extracted volume. The diluted extract must be reanalyzed.
7.5.5 Perform all qualitative and quantitative measurements as
described in Section 7.6. Store the extracts at 4°C, protected from
light in screw-cap vials equipped with unpierced Teflon lined septa.
7.6 Data interpretation
7.6.1 Qualitative analysis
7.6.1.1 The qualitative identification of compounds determined
by this method is based on retention time, and on comparison of the
sample mass spectrum, after background correction, with
characteristic ions in a reference mass spectrum. The reference mass
spectrum must be generated by the laboratory using the conditions
of this method. The characteristic ions from the reference mass
spectrum are defined to be the three ions of greatest relative
intensity, or any ions over 30% relative intensity if less than three
such ions occur in the reference spectrum. Compounds should be
identified as present when the criteria below are met.
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7.6.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.6.1.1.2 The RRT of the sample component is within ±
0.06 RRT units of the RRT of the standard component.
7.6.1.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions
in the reference spectrum. (Example: For an ion with an
abundance of 50% in the reference spectrum, the corresponding
abundance in a sample spectrum can range between 20% and 80%.)
7.6.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
7.6.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background spectra
is important. Examination of extracted ion current profiles
of appropriate ions can aid in the selection of spectra, and
in qualitative identification of compounds. When analytes
coelute (i.e., only one chromatographic peak is apparent), the
identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting
compound.
7.6.1.2 For samples containing components not associated with
the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search routines
should not use normalization routines that would misrepresent the
library or unknown spectra when compared to each other. For example,
the RCRA permit or waste deli sting requirements may require the
reporting of nontarget analytes. Only after visual comparison of
sample spectra with the nearest library searches will the mass
spectral interpretation specialist assign a tentative identification.
Guidelines for making tentative identification are:
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(1) Relative intensities of major ions in the reference spectrum
(ions > 10% of the most abundant ion) should be present in the sample
spectrum.
(2) The relative intensities of the major ions should agree within
± 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should be
present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination
or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the sample
spectrum because of background contamination or coeluting peaks.
Data system library reduction programs can sometimes create these
discrepancies.
7.6.2 Quantitative analysis
7.6.2.1 When a compound has been identified, the quantitation
of that compound will be based on the integrated abundance from the
EICP of the primary characteristic ion. Quantitation will take place
using the internal standard technique. The internal standard used
shall be the one nearest the retention time of that of a given
analyte (e.g. see Table 5).
7.6.2.2 Calculate the concentration of each identified analyte
in the sample as follows:
Water
(AJ(I.)(Vt)
concentration (/ig/L)
(AJ(RF)(V0)(V,)
where:
Ax = Area of characteristic ion for compound being measured.
Is = Amount of internal standard injected (ng).
V, = Volume of total extract, taking into account dilutions (i.e.
a l-to-10 dilution of a 1 ml extract will mean V, = 10,000 pi.
If half the base/neutral extract and half the acid extract are
combined, Vt = 2,000 juL).
A|S = Area of characteristic ion for the internal standard.
RF = Response factor for compound being measured (Section 7.3.3).
V0 = Volume of water extracted (ml).
V, = Volume of extract injected (ML).
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Sediment/Soil Sludge (on a dry-weight basis) and Waste (normally on
a wet-weight basis
(Ax)(Is)(Vt)
concentration (jig/Kg) =
(A,S)(RF)(V,)(WS)(D)
where:
A,, Is, Vt, A,.., RF, V, = Same as for water.
Ws = Weight of sample extracted or diluted in grams.
D = % dry weight of sample/100, or 1 for a wet-weight basis.
7.6.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulas
given above should be used with the following modifications: The
areas A,, and A. should be from the total ion chromatograms and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
7.6.2.4 Quantitation of multicomponent compounds (e.g. Aroclors)
is beyond the scope of Method 8270. Normally, quantitation is
performed using a GC/ECD by Method 8080.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Required instrument QC is found in the following sections:
8.2.1 The GC/MS system must be tuned to meet the DFTPP specifications
in Sections 7.3.1 and 7.4.1.
8.2.2 There must be an initial calibration of the GC/MS system as
specified in Section 7.3.
8.2.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.4.3 and the CCC criteria in Section 7.4.4, each 12 hours.
8.3 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.3.1 A quality control (QC) reference sample concentrate is required
containing each analyte at a concentration of 100 mg/L in methanol. The
QC reference sample concentrate may be prepared from pure standard
materials or purchased as certified solutions. If prepared by the
laboratory, the QC reference sample concentrate must be made using stock
standards prepared independently from those used for calibration.
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8.3.2 Using a pipet, prepare QC reference samples at a concentration
of 100 /-ig/L by adding 1.00 ml of QC reference sample concentrate to each
of four 1 L aliquots of organic-free reagent water.
8.3.3 Analyze the well-mixed QC reference samples according to the
method beginning in Section 7.1 with extraction of the samples.
8.3.4 Calculate the average recovery (x) in M9/U and the standard
deviation of the recovery (s) in ng/l, for each analyte of interest using
the four results.
8.3.5 For each analyte, compare s and x with the corresponding
acceptance criteria_for precision and accuracy, respectively, found in
Table 6. If s and x for all analytes meet the acceptance criteria, the
system performance is acceptable and analysis of actual samples can_begin.
If any individual s exceeds the precision limit or any individual x falls
outside the range for accuracy, then the system performance is unacceptable
for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial probability
that one or more will fail at least one of the acceptance criteria when
all analytes of a given method are determined.
8.3.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Section
8.3.6.1 or 8.3.6.2.
8.3.6.1 Locate and correct the source of the problem and repeat
the test for all analytes beginning with Section 8.3.2.
8.3.6.2 Beginning with Section 8.3.2, repeat the test only for
those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Section
8.3.2.
8.4 For aqueous and soil matrices, laboratory established surrogate
control limits should be compared with the control limits listed in Table 8.
The limits given in Table 8 are multilaboratory performance based limits for
soil and aqueous samples, and therefore, the single laboratory limits must fall
within those given in Table 8 for these matrices.
8.4.1 If recovery is not within limits, the following procedures
are required.
8.4.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.4.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
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8.4.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.4.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4.2 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
9.0 METHOD PERFORMANCE
9.1 Method 8250 (the packed column version of Method 8270) was tested by
15 laboratories using Organic-free reagent water, drinking water, surface water,
and industrial wastewaters spiked at six concentrations over the range 5-
1,300 /xg/L. Single operator accuracy and precision, and method accuracy were
found to be directly related to the concentration of the analyte and essentially
independent of the sample matrix. Linear equations to describe these
relationships are presented in Table 7. Method performance data for Method 8270
is being developed.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 625," October 26,
1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems," Analytical Chemistry, 47, 995-1000, 1975.
4. "Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
5. "Interlaboratory Method Study for EPA Method 625-Base/Neutrals, Acids, and
Pesticides," Final Report for EPA Contract 68-03-3102 (in preparation).
6. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
7. Lucas, S.V.; Kornfeld, R.A. "GC-MS Suitability Testing of RCRA Appendix
VIII and Michigan List Analytes "; U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
February 20, 1987, Contract No. 68-03-3224.
8. Engel, T.M.; Kornfeld, R.A.; Warner, J.S.; Andrews, K.D. "Screening of
Semi volatile Organic Compounds for Extractability and. Aqueous Stability by
SW-846, Method 3510"; U.S. Environmental Protection Agency, Environmental
8270B - 19 Revision 2
November 1990
-------
Monitoring and Support Laboratory, Cincinnati, OH 45268, June 5, 1987,
Contract 68-03-3224.
8270B - 20 Revision 2
November 1990
-------
TABLE 1.
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Compound
Retention Primary Secondary
Time (min.) Ion Ion(s)
2-Picoline
Aniline
Phenol
Bis(2-chloroethyl) ether
2-Chlorophenol
1,3-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S.)
1,4-Dichlorobenzene
Benzyl alcohol
1,2-Dichlorobenzene
N-Nitrosomethylethyl ami ne
Bis(2-chloroisopropyl) ether
Ethyl carbamate
Thiophenol (Benzenethiol)
Methyl methanesulfonate
N-Nitrosodi-n-propylamine
Hexachloroethane
Maleic anhydride
Nitrobenzene
Isophorone*
N-Nitrosodiethylamine
2-Nitrophenol
2,4-Dimethylphenol
p-Benzoquinone
Bis(2-chloroethoxy)methane
Benzole acid
2,4-Dichlorophenol
Trimethyl phosphate
Ethyl methanesulfonate
1,2,4-Tri chlorobenzene
Naphthalene-d8 (I.S.)
Naphthalene
Hexachlorobutadi ene
Tetraethyl pyrophosphate
Diethyl sulfate
4-Chloro-3-methylphenol
2-Methyl naphtha!ene
2-Methylphenol
Hexachloropropene
Hexachlorocyclopentadiene
N-Nitrosopyrrolidine
Acetophenone
4-Methylphenol
2,4,6-Trichlorophenol
o-Toluidine
3-Methylphenol
2-Chloronaphthalene
,75a
,68
,77
,82
,97
,27
,35
,40
,78
6.85
6.97
22
27
42
48
55
65
65
87
8.53
8.70
8.75
9.
9.
9.
9.
9.
9.
9.
.03
13
,23
.38
.48
,53
.62
9.67
9.75
9.82
10.43
11.07
11.37
11.68
11.87
12.40
12.45
12.60
12.65
12.67
12.82
12.85
12.87
12.93
13.30
8270B - 21
93 66,92
93 66,65
94 65,66
93 63,95
128 64,130
146 148,111
152 150,115
146 148,111
108 79,77
146 148,111
88 42,88,43,56
45 77,121
62 62,44,45,74
110 110,66,109,84
80 80,79,65,95
70 42,101,130
117 201,199
54 54,98,53,44
77 123,65
82 95,138
102 102,42,57,44,56
139 109,65
122 107,121
108 54,108,82,80
93 95,123
122 105,77
162 164,98
110 110,79,95,109,140
79 79,109,97,45,65
180 182,145
136 68
128 129,127
225 223,227
99 99,155,127,81,109
139 139,45,59,99,111,125
107 144,142
142 141
107 107,108,77,79,90
213 213,211,215,117,106,141
237 235,272
100 100,41,42,68,69
105 71,105,51,120
107 107,108,77,79,90
196 198,200
106 106,107,77,51,79
107 107,108,77,79,90
162 127,164
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary
Ion
Secondary
Ion(s)
N-Nitrosopiperidine
1,4-Phenylenediamine
1-Chloronaphthalene
2-Nitroaniline
5-Chloro-2-methylaniline
Dimethyl phthalate
Acenaphthylene
2,6-Dinitrotoluene
Phthalic anhydride
o-Anisidine
3-Nitroaniline
Acenaphthene-d10 (I.S.)
Acenaphthene
2,4-Dinitrophenol
2,6-Dinitrophenol
4-Chloroaniline
Isosafrole
Dibenzofuran
2,4-Diaminotoluene
2,4-Dinitrotoluene
4-Nitrophenol
2-Naphthylamine
1,4-Naphthoquinone
p-Cresidine
Dichlorovos
Diethyl phthalate
Fluorene
2,4,5-Trimethylaniline
N-Nitrosodibutylamine
4-Chlorophenyl phenyl ether
Hydroquinone
4,6-Dinitro-2-methylphenol
Resorcinol
N-Nitrosodiphenylamine
Safrole
Hexamethyl phosphoramide
3-(Chloromethyl)pyridine hydrochloride
Diphenylamine
1,2,4,5-Tetrachlorobenzene
1-Naphthylamine
l-Acetyl-2-thiourea
4-Bromophenyl phenyl ether
Toluene diisocyanate
2,4,5-Trichlorophenol
Hexachlorobenzene
Nicotine
Pentachlorophenol
13.55 114 42,114,55,56,41
13.62 108 108,80,53,54,52
13.65a 162 127,164
13.75 65 92,138
14.28 106 106,141,140,77,89
14.48 163 194,164
14.57 152 151,153
14.62 165 63,89
14.62 104 104,76,50,148
15.00 108 80,108,123,52
15.02 138 108,92
15.05 164 162,160
15.13 154 153,152
15.35 184 63,154
15.47 162 162,164,126,98,63
15.50 127 127,129,65,92
15.60 162 162,131,104,77,51
15.63 168 139
15.78 121 121,122,94,77,104
15.80 165 63,89
15.80 139 109,65
16.00* 143 115,116
16.23 158 158,104,102,76,50,130
16.45 122 122,94,137,77,93
16.48 109 109,185,79,145
16.70 149 177,150
16.70 166 165,167
16.70 120 120,135,134,91,77
16.73 84 84,57,41,116,158
16.78 204 206,141
16.93 110 110,81,53,55
17.05 198 51,105
17.13 110 110,81,82,53,69
17.17 169 168,167
17.23 162 162,162,104,77,103,135
17.33 135 135,44,179,92,42
17.50 92 92,127,129,65,39
17.54a 169 168,167
17.97 216 216,214,179,108,143,218
18.20 143 143,115,89,63
18.22 118 43,118,42,76
18.27 248 250,141
18.42 174 174,145,173,146,132,91
18.47 196 196,198,97,132,99
18.65 284 142,249
18.70 84 84,133,161,162
19.25 266 264,268
8270B - 22
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
5-Nitro-o-toluidine
Thionazine
4-Nitroaniline
Phenanthrene-d10(i.s.)
Phenanthrene
Anthracene
1,4-Dinitrobenzene
Mevinphos
Naled
1,3-Dinitrobenzene
Diallate (cis or trans)
1,2-Dinitrobenzene
Diallate (trans or cis)
Pentachlorobenzene
5-Nitro-o-anisidine
Pentachloron i trobenzene
4-Nitroquinoline-1-oxide
Di-n-butyl phthalate
2,3,4,6-Tetrachlorophenol
Dihydrosaffrole
Demeton-o
Fluoranthene
1,3,5-Trinitrobenzene
Dicrotophos
Benzidine
Trifluralin
Bromoxynil
Pyrene
Monocrotophos
Phorate
Sulfall ate
Demeton-s
Phenacetin
Dimethoate
Phenobarbital
Carbofuran
Octamethyl pyrophosphoramide
4-Aminobiphenyl
Dioxathion
Terbufos
o,o-Dimethylphenylamine
Pronamide
Aminoazobenzene
Dichlone
Dinoseb
Disulfoton
19.27 152 77,152,79,106,94
19.35 107 96,107,97,143,79,68
19.37 138 138,65,108,92,80,39
19.55 188 94,80
19.62 178 179,176
19.77 178 176,179
19.83 168 168,75,50,76,92,122
19.90 127 127,192,109,67,164
20.03 109 109,145,147,301,79,189
20.18 168 168,76,50,75,92,122
20.57 86 86,234,43,70
20.58 168 168,50,63,74
20.78 86 86,234,43,70
21.35 250 250,252,108,248,215,254
21.50 168 168,79,52,138,153,77
21.72 237 237,142,214,249,295,265
21.73 174 174,101,128,75,116
21.78 149 150,104
21.88 232 232,131,230,166,234,168
22.42 135 135,64,77
22.72 88 88,89,60,61,115,171
23.33 202 101,203
23.68 75 75,74,213,120,91,63
23.82 127 127,67,72,109,193,237
23.87 184 92,185
23.88 306 306,43,264,41,290
23.90 277 277,279,88,275,168
24.02 202 200,203
24.08 127 127,192,67,97,109
24.10 75 75,121,97,93,260
24.23 188 188,88,72,60,44
24.30 88 88,60,81,89,114,115
24.33 108 180,179,109,137,80
24.70 87 87,93,125,143,229
24.70 204 204,117,232,146,161
24.90 164 164,149,131,122
24.95 135 135,44,199,286,153,243
25.08 169 169,168,170,115
25.25 97 97,125,270,153
25.35 231 231,57,97,153,103
25.43 58 58,91,65,134,42
25.48 173 173,175,145,109,147
25.72 197 92,197,120,65,77
25.77 191 191,163,226,228,135,193
25.83 211 211,163,147,117,240
25.83 88 88,97,89,142,186
8270B - 23
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
Mexacarbate
4,4'-Oxydianiline
Butyl benzyl phthalate
4-Nitrobiphenyl
Phosphamidon
2-Cyclohexyl-4,6-Dinitrophenol
Methyl parathion
Carbaryl
Di methyl ami noazobenzene
Propylthiouracil
Benz(a)anthracene
Chrysene-d12 (I.S.)
3,3'-Dichlorobenzidine
Chrysene
Malathion
Kepone
Fenthion
Parathion
Anilazine
Bis(2-ethylhexyl) phthalate
3,3'-Dimethylbenzidine
Carbophenothion
5-Nitroacenaphthene
Methapyrilene
Isodrin
Captan
Chlorfenvinphos
Crotoxyphos
Phosmet
EPN
Tetrachlorvinphos
Di-n-octyl phthalate
2-Aminoanthraquinone
Barban
Aramite
Benzo(b)fluoranthene
Nitrofen
Benzo(k)fluoranthene
Chiorobenzilate
Fensulfothion
Ethion
Diethylstilbestrol
Famphur
Tri-p-tolyl phosphate"
Benzo(a)pyrene
Perylene-d12 (I.S.)
7,12-Dimethylbenz(a)anthracene
26.02 165 165,150,134,164,222
26.08 200 200,108,171,80,65
26.43 149 91,206
26.55 199 199,152,141,169,151
26.85 127 127,264,72,109,138
26.87 231 231,185,41,193,266
27.03 109 109,125,263,79,93
27.17 144 144,115,116,201
27.50 225 225,120,77,105,148,42
27.68 170 170,142,114,83
27.83 228 229,226
27.88 240 120,236
27.88 252 254,126
27.97 228 226,229
28.08 173 173,125,127,93,158
28.18 272 272,274,237,178,143,270
28.37 278 278,125,109,169,153
28.40 109 109,97,291,139,155
28.47 239 239,241,143,178,89
28.47 149 167,279
28.55 212 212,106,196,180
28.58 157 157,97,121,342,159,199
28.73 199 199,152,169,141,115
28.77 97 97,50,191,71
28.95 193 193,66,195,263,265,147
29.47 79 79,149,77,119,117
29.53 267 267,269,323,325,295
29.73 127 127,105,193,166
30.03 160 160,77,93,317,76
30.11 157 157,169,185,141,323
30.27 329 109,329,331,79,333
30.48 149 167,43
30.63 223 223,167,195
30.83 222 222,51,87,224,257,153
30.92 185 185,191,319,334,197,321
31.45 252 253,125
31.48 283 283,285,202,139,253
31.55 252 253,125
31.77 251 251,139,253,111,141
31.87 293 293,97,308,125,292
32.08 231 231,97,153,125,121
32.15 268 268,145,107,239,121,159
32.67 218 218,125,93,109,217
32.75 368 368,367,107,165,198
32.80 252 253,125
33.05 264 260,265
33.25 256 256,241,239,120
8270B - 24
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
5,5-Di phenylhydantoi n
Captafol
Dinocap
Methoxychlor
2-Acetylaminofl uorene
4,4'-Methylenebis(2-chloroaniline)
3,3'-Dimethoxybenzidine
3-Methylcholanthrene
Phosalone
Azinphos-methyl
Leptophos
Mi rex
Tris(2,3-dibromopropyl) phosphate
Dibenz(a,j)acridine
Mestranol
Coumaphos
Indeno(l,2,3-cd)pyrene
Dibenz(a,h)anthracene
Benzo(g,h,i)perylene
l,2:4,5-Dibenzopyrene
Strychnine
Piperonyl sulfoxide
Hexachlorophene
Aldrin
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
a-BHC
/3-BHC
S-BHC
7-BHC (Lindane)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
1,2-Di phenylhydrazi ne
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
33.40 180 180,104,252,223,209
33.47 79 79,77,80,107
33.47 69 69,41,39
33.55 227 227,228,152,114,274,212
33.58 181 181,180,223,152
34.38 231 231,266,268,140,195
34.47 244 244,201,229
35.07 268 268,252,253,126,134,113
35.23 182 182,184,367,121,379
35.25 160 160,132,93,104,105
35.28 171 171,377,375,77,155,379
35.43 272 272,237,274,270,239,235
35.68 201 137,201,119,217,219,199
36.40 279 279,280,277,250
36.48 277 277,310,174,147,242
37.08 362 362,226,210,364,97,109
39.52 276 138,227
39.82 278 139,279
41.43 276 138,277
41.60 302 302,151,150,300
45.15 334 334,335,333
46.43 162 162,135,105,77
47.98 196 196,198,209,211,406,408
66 263,220
222 260,292
190 224,260
190 224,260
222 256,292
292 362,326
292 362,326
360 362,394
183 181,109
181 183,109
183 181,109
183 181,109
235 237,165
246 248,176
235 237,165
79 263,279
77 105,182
195 339,341
337 339,341
272 387,422
263 82,81
67 345,250
317 67,319
8270B - 25
Revision 2
November 1990
-------
TABLE 1.
(Continued)
Retention Primary Secondary
Compound Time (min.) Ion Ion(s)
2-Fluorobiphenyl (surr.) -- 172 171
2-Fluorophenol (surr.) -- 112 64
Heptachlor -- 100 272,274
Heptachlor epoxide -- 353 355,351
Nitrobenzene-d5 (surr.) -- 82 128,54
N-Nitrosodimethylamine -- 42 74,44
Phenol-de (surr.) -- 99 42,71
Terphenyl-d14 (surr.) -- 244 122,212
2,4,6-Tribromophenol (surr.) -- 330 332,141
Toxaphene -- 159 231,233
I.S. = internal standard.
surr. = surrogate.
"Estimated retention times.
"Substitute for the non-specific mixture, tricresyl phosphate.
8270B - 26 Revision 2
November 1990
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQLs) FOR SEMIVOLATILE ORGANICS"
Estimated
Quantitation
Limits"
Ground water
Semivolatiles /ig/L
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetyl ami nof 1 uorene
l-Acetyl-2-thiourea
2-Ami noanthraqui none
Aminoazobenzene
4-Aminobiphenyl
Anilazine
o-Anisidine
Anthracene
Aramite
Azinphos-methyl
Barban
Benz(a)anthracene
Benzo (b) f 1 uoranthene
Benzo(k)fluoranthene
Benzoic acid
Benzo (g,h,i)perylene
Benzo(a)pyrene
p-Benzoquinone
Benzyl alcohol
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-bromophenyl phenyl ether
Bromoxynil
Butyl benzyl phthalate
Captafol
Captan
Carbaryl
Carbofuran
Carbophenothion
Chlorfenvinphos
4-Chloroaniline
Chi orobenzi late
5-Chloro-2-methyl aniline
4-Chl oro-3-methyl phenol
3-(Chloromethyl)pyridine hydrochloride
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Coumaphos
10
10
10
20
1000
20
10
20
100
10
10
20
100
200
10
10
10
50
10
10
10
20
10
10
10
10
10
10
20
50
10
10
10
20
20
10
10
20
100
10
10
10
10
40
Low Soil /Sediment1
M9/Kg
660
660
ND
ND
ND
ND
ND
ND
ND
ND
660
ND
ND
ND
660
660
660
3300
660
660
ND
1300
660
660
660
660
ND
660
ND
ND
ND
ND
ND
ND
1300
ND
ND
1300
ND
660
660
660
660
ND
8270B - 27 Revision 2
November 1990
-------
TABLE 2.
(Continued)
Estimated
Quantisation
Limits"
Ground water Low Soil/Sediment1
Semivolatiles
p-Cresidine
Crotoxyphos
2-Cyc1ohexyl-4,6-dinitrophenol
Demeton-o
Demeton-s
Dial late (cis or trans)
Diallate (trans or cis)
2,4-Dianrinotoluene
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Dibenzo(a,e)pyrene
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Di ethyl phthalate
Diethylstilbestrol
Di ethyl sulfate
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl ami noazobenzene
7,12-Dimethylbenz(a)anthracene
3, 3' -Dimethyl benzi dine
a, a-Dimethyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
5, 5-Diphenyl hydantoi n
Di-n-octyl phthalate
M9/L
10
20
100
10
10
10
10
20
10
10
10
10
10
NA
10
10
10
20
10
10
10
10
10
20
100
20
100
10
10
10
ND
10
10
40
20
40
50
50
10
10
100
20
20
10
M9/Kg
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
660
660
660
1300
660
ND
ND
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
3300
3300
660
660
ND
ND
ND
660
8270B - 28
Revision 2
November 1990
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits"
Ground water Low Soil/Sediment1
Semi vol allies
Disulfoton
EPN
Ethion
Ethyl carbamate
Bis(2-ethylhexyl) phthalate
Ethyl methanesulfonate
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl orophene
Hexachl oropropene
Hexamethyl phosphoramlde
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methylenebis(2-chloroanil
Methyl methanesulfonate
2-Methyl naphthal ene
Methyl parathion
2 -Methyl phenol
3-Methyl phenol
4-Methyl phenol
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
M9/L
10
10
10
50
10
20
20
40
10
20
10
10
10
10
10
10
50
10
20
ND
10
20
10
10
20
10
50
NA
20
100
10
10
ine) NA
10
10
10
10
10
10
10
20
10
40
20
MQ/Kg
ND
ND
ND
ND
660
ND
ND
ND
ND
ND
660
660
660
660
660
660
ND
ND
ND
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
ND
660
ND
660
ND
ND
ND
ND
ND
8270B - 29 Revision 2
November 1990
-------
Senrivolatiles
Naphthalene
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
4-Nitroquinol ine-1-oxide
N-Nitrosodi butyl ami ne
N-Ni trosodi ethyl ami ne
N-Nitrosodiphenylamine
N-Nitroso-di-n-propylamine
N-Nitrosopiperidine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachl orobenzene
Pentachl oron i trobenzene
Pentachl orophenol
Phenacetin
Phenanthrene
Phenobarbital
Phenol
1,4-Phenylenediamine
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
Propylthiouracil
Pyrene
TABLE 2.
(Continued)
Estimated
Quantitation
Limits"
Ground water Low Soi
M9/L
10
10
10
10
20
10
50
50
20
10
10
10
20
10
50
10
40
10
20
10
10
20
40
200
20
10
10
20
50
20
10
10
10
10
10
100
40
100
100
ND
100
10
100
10
1 /Sediment1
M9/Kg
660
ND
ND
ND
ND
ND
3300
3300
ND
ND
660
ND
ND
660
3300
ND
ND
ND
ND
660
660
ND
ND
ND
ND
ND
ND
ND
3300
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
8270B - 30
Revision 2
November 1990
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits"
Semivolatiles
Ground waterLow Soil/Sediment1
Pyridine
Resorcinol
Safrole
Strychnine
Sul fall ate
Terbufos
1,2,4, 5-Tetrachl orobenzene
2,3,4 , 6-Tetrachl orophenol
Tetrachlorvinphos
Tetraethyl pyrophosphate
Thionazine
Thiophenol (Benzenethiol)
Toluene diisocyanate
o-Toluidine
1 , 2 , 4-Tri chl orobenzene
2, 4, 5-Trichl orophenol
2, 4, 6-Tri chl orophenol
Trifluralin
2,4,5-Trimethylaniline
Trimethyl phosphate
1,3,5-Trinitrobenzene
Tris(2,3-dibromopropyl ) phosphate
Tri-p-tolyl phosphate(h)
0,0,0-Tri ethyl phosphorothioate
ND
100
10
40
10
20
10
10
20
40
20
20
100
10
10
10
10
10
10
10
10
200
10
NT
NO
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
660
660
ND
ND
ND
ND
ND
ND
ND
a EQLs listed for soil/sediment are based on wet weight. Normally data is
reported on a dry weight basis, therefore, EQLs will be higher based on the
% dry weight of each sample. This is based on a 30 g sample and gel
permeation chromatography cleanup.
b Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
ND = Not determined.
NA = Not applicable.
NT = Not tested.
Other Matrices Factor1
High-concentration soil and sludges by sonicator 7.5
Non-water miscible waste 75
1EQL = [EQL for Low Soil/Sediment (Table 2)] X [Factor].
8270B - 31
Revision 2
November 1990
-------
TABLE 3.
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA3
Mass
Ion Abundance Criteria
51
68
70
127
197
198
199
275
365
441
442
443
30-60% of mass 198
< 2% of mass 69
< 2% of mass 69
40-60% of mass 198
< 1% of mass 198
Base peak, 100% relative abundance
5-9% of mass 198
10-30% of mass 198
> 1% of mass 198
Present but less than mass 443
> 40% of mass 198
17-23% of mass 442
"See Reference 4.
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction
Acid Fraction
Acenaphthene
1,4-Di chlorobenzene
Hexachlorobutadi ene
N-Ni trosodi phenylami ne
Di-n-octyl phthalate
Fluoranthene
Benzo(a)pyrene
4-Chloro-3-methylphenol
2,4-Dichlorophenol
2-Nitrophenol
Phenol
Pentachlorophenol
2,4,6-Trichlorophenol
8270B - 32
Revision 2
November 1990
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
l,4-Dich1orobenzene-d4
Naphthalene-d8
Acenaphthene-d10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propylamine
Phenol
Phenol-d6 (surr.) ,
2-Picoline
Acetophenone
Benzoic acid
Bis(2-chloroethoxy)methane
4-Chloroaniline
4-Chloro-3-methylphenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a,a-Dimethyl-
phenethylamine
2,4-Dimethylphenol
Hexachlorobutadiene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Nitrophenol
N-Nitrosodibutyl amine
N-Nitrosopiperidine
1,2,4-Tri chlorobenzene
Acenaphthene
Acenaphthylene
1-Chloronaphthalene
2-Chloronaphthalene
4-Chlorophenyl
phenyl ether
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Fluorene
2-Fluorobiphenyl
(surr.)
Hexachlorocyclo-
pentadiene
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
4-Nitrophenol
Pentachlorobenzene
1,2,4,5-Tetra-
chlorobenzene
2,3,4,6-Tetra-
chlorophenol
2,4,6-Tribromo-
phenol (surr.)
2,4,6-Trichloro-
phenol
2,4,5-Trichloro-
phenol
(surr.) = surrogate
8270B - 33
Revision 2
November 1990
-------
TABLE 5.
(Continued)
Phenanthrene-d
10
Chrysene-d12
Perylene-d12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl ether
Di-n-butyl phthalate
4,6-Dinitro-2-methylphenol
Diphenylamine
1,2-Diphenylhydrazi ne
Fluoranthene
Hexachlorobenzene
N-Ni trosodi phenylami ne
Pentachlorophenol
Pentachloronitrobenzene
Phenacetin
Phenanthrene
Pronamide
Benzidine
Benzo(a)anthracene
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethylaminoazobenzene
Pyrene
Terphenyl-d14 (surr.)
Benzo(b)fluor-
anthene
Benzo(k)fluor-
anthene
Benzo(g,h,i)-
perylene
Benzo(a)pyrene
Dibenz(a,j)acridine
Dibenz(a,h)-
anthracene
7,12-Dimethylbenz-
(a)anthracene
Di-n-octyl phthalate
Indeno(l,2,3-cd)
pyrene
3-Methylchol-
anthrene
(surr.) = surrogate
8270B - 34
Revision 2
November 1990
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Benzo(b)fl uoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzyl butyl phthalate
0-BHC
fi-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(Z-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
D1benzo(a,h) anthracene
Di-n-butyl phthalate
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
3,3'-Dichlorobenzidine
Dieldrln
Di ethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di -n-octyl phthal ate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
27.6
40.2
39.0
32.0
27.6
38.8
32.3
39.0
58.9
23.4
31.5
21.6
55.0
34.5
46.3
41.1
23.0
13.0
33.4
48.3
31.0
32.0
61.6
70.0
16.7
30.9
41.7
32.1
71.4
30.7
26.5
23.2
21.8
29.6
31.4
16.7
32.5
32.8
20.7
37.2
54.7
24.9
26.3
Range
for x
(M9/L)(%)
60.1-132.3
53.5-126.0
7.2-152.2
43.4-118.0
41.8-133.0
42.0-140.4
25.2-145.7
31.7-148.0
D-195.0
D-139.9
41.5-130.6
D-100.0
42.9-126.0
49.2-164.7
62.8-138.6
28.9-136.8
64.9-114.4
64.5-113.5
38.4-144.7
44.1-139.9
D-134.5
19.2-119.7
D-170.6
D-199.7
8.4-111.0
48.6-112.0
16.7-153.9
37.3-105.7
8.2-212.5
44.3-119.3
D-100.0
D-100.0
47.5-126.9
68.1-136.7
18.6-131.8
D-103.5
D-188.8
42.9-121.3
71.6-108.4
D-172.2
70.9-109.4
7.8-141.5
37.8-102.2
8270B - 35
Range
P» Ps
47-145
33-145
D-166
27.133
33-143
24-159
11-162
17-163
D-219
D-152
24-149
D-110
12-158
33-184
36-166
8-158
53-127
60-118
25-158
17-168
D-145
4-136
D-203
D-227
1-118
32-129
D-172
20-124
D-262
29-136
D-114
D-112
39-139
50-158
4-146
D-107
D-209
26-137
59-121
D-192
26.155
D-152
24-116
Revision 2
November 1990
-------
Compound
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
4-Chl oro-3-methyl phenol
2-Chlorophenol
2,4-Chlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl -4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
s = Standard deviation of
x = Average recovery for
Test
cone
TABLE 6.
(Continued)
Limit
for s
Range
for x
(M9/L) (M9/L) (M9/L)(%)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
four
four
24.5
44.6
63.3
30.1
39.3
55.4
54.2
20.6
25.2
28.1
37.2
28.7
26.4
26.1
49.8
93.2
35.2
47.2
48.9
22.6
31.7
55.2-100.0
D-150.9
46.6-180.2
35.6-119.6
54.3-157.6
13.6-197.9
19.3-121.0
65.2-108.7
69.6-100.0
57.3-129.2
40.8-127.9
36.2-120.4
52.5-121.7
41.8-109.0
D-172.9
53.0-100.0
45.0-166.7
13.0-106.5
38.1-151.8
16.6-100.0
52.4-129.2
recovery measurements, in
recovery measurements, in
Range
P» Ps
40-113
D-171
21-196
21-133
35-180
D-230
D-164
54-120
52-115
44-142
22-147
23-134
39-135
32-119
D-191
D-181
29-182
D-132
14-176
5-112
37-144
M9/L.
M9/L.
p, ps = Percent recovery measured.
D = Detected; result must
a Criteria from 40 CFR
be greater than
Part
zero.
136 for Method 625. These
criteria are based
directly on the method performance data in Table 7. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 7.
8270B - 36
Revision 2
November 1990
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Compound
Accuracy, as
recovery, x'
(M9/L)
Single analyst Overall
precision, s/ precision,
S' (Mg/L)
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Chloroethane
Benzo(b)fluoranthene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzyl butyl phthalate
0-BHC
6-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo(a,h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Di ethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfar* sulfate
Endr in "aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
0.96C+0.19
0.89C+0.74
0.78C+1.66
0.80C+0.68
0.88C-0.60
0.99C-1.53
0.93C-1.80
0.87C-1.56
0.90C-0.13
0.98C-0.86
0.66C-1.68
0.87C-0.94
0.29C-1.09
0.86C-1.54
1.12C-5.04
1.03C-2.31
0.84C-1.18
0.91C-1.34
0.89C+0.01
0.91C+0.53
0.93C-1.00
0.56C-0.40
0.70C-0.54
0.79C-3.28
0.88C+4.72
0.59C+0.71
0.80C+0.28
0.86C-0.70
0.73C-1.47
1.23C-12.65
0.82C-0.16
0.43C+1.00
0.20C+1.03
0.92C-4.81
1.06C-3.60
0.76C-0.79
0.39C+0.41
0.76C-3.86
0.81C+1.10
0.90C-0.00
0.87C-2.97
0.92C-1.87
O.lBx-0.12
0.24X-1.06
0.27X-1.28
0.21X-0.32
O.lBx+0.93
0.14x-0.13
0.22X+0.43
0.19X+1.03
0.22x+0.48
0.29X+2.40
O.lSx+0.94
0.20X-0.58
0.34X+0.86
0.35X-0.99
0.16X+1.34
0.24X+0.28
0.26X+0.73
0.13X+0.66
0.07X+0.52
0.20X-0.94
0.28X+0.13
0.29X-0.32
0.26x-1.17
0.42X+0.19
0.30X+8.51
O.lSx+1.16
0.20X+0.47
0.25X+0.68
0.24X+0.23
0.28X+7.33
fl.20x-0.16
0.28X+1.44
0.54X+0.19
0.12X+1.06
0.14X+1.26
0.21X+1.19
0.12X+2.47
O.lSx+3.91
0.22X-0.73
0.12X+0.26
0.24X-0.56
0.33X-0.46
0.21X-0.67
0.26X-0.54
0.43X+1.13
0.27X-0.64
0.26X-0.21
0.17X-0.28
0.29X+0.96
0.35X+0.40
0.32X+1.35
O.Blx-0.44
0.53X+0.92
0.30X+1.94
0.93X-0.17
0.35X+0.10
0.26X+2.01
0.25X+1.04
0.36X+0.67
0.16X+0.66
0.13X+0.34
0.30X-0.46
0.33X-0.09
0.66X-0.96
0.39X-1.04
0.65X-0.58
0.59X+0.25
0.39X+0.60
0.24X+0.39
0.41X+0.11
0.29X+0.36
0.47X+3.45
0.26x-0.07
0.52X+0.22
1.05X-0.92
0.21X+1.50
0.19X+0.35
0.37X+1.19
0.63X-1.03
0.73X-0.62
0.28X-0.60
0.13X+0.61
0.50X-0.23
0.28X+0.64
8270B - 37
Revision 2
November 1990
-------
TABLE 7.
(Continued)
Compound
Accuracy, as
recovery, x'
(M9/L)
Single analyst Overall
precision, s/ precision,
(M9/L) S' (Mg/L)
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
4-Chl oro-3-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl -4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
0.74C+0.66
0.71C-1.01
0.73C-0.83
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.22
0.81C-10.86
0.87C+0.06
0.84C-0.16
0.94C-0.79
0.84C+0.35
0.78C+0.29
0.87C-0.13
0.71C+4.41
0.81C-18.04
1.04C-28.04
0.07C-1.15
0.61C-1.22
0.93C+1.99
0.43C+1.26
0.91C-0.18
0.18X-0.10
0.19X+0.92
O.Ux+0.67
0.29x+1.46
0.27X+0.77
0.21X-0.41
0.19X+0.92
0.27X+0.68
0.35X+3.61
0.12x+0.57
0.16X+0.06
O.lSx+0.85
0.23x+0.75
O.lSx+1.46
0.15X+1.25
0.16X+1.21
0.38X+2.36
O.lOx+42.29
0.16X+1.94
0.38x+2.57
0.24x+3.03
0.26X+0.73
0.16X+2.22
0.43X-0.52
0.26X+0.49
0.17X+0.80
O.BOx-0.44
0.33X+0.26
0.30X-0.68
0.27X-1-0.21
0.44X+0.47
0.43X+1.82
0.15X+0.25
O.lBx+0.31
0.21X+0.39
0.29X+1.31
0.28X+0.97
0.21X+1.28
0.22X+1.31
0.42X+26.29
0.26X+23.10
0.27X+2.60
0.44X+3.24
0.30X+4.33
0.35X+0.58
0.22X+1.81
x' = Expected recovery for one or more measurements of a sample containing a
concentration of C, in jug/L.
s/= Expected single analyst standard deviation of measurements at an average
concentration of x, in /xg/L.
S' = Expected interlaboratory_ standard deviation of measurements at an average
concentration found of x, in M9/L.
C = True value for the concentration, in M9/L.
x" = Average recovery found for measurements of samples containing a
concentration of C, in
8270B - 38
Revision 2
November 1990
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High
Surrogate Compound Water Soil/Sediment
Nitrobenzene-d5 35-114 23-120
2-Fluorobiphenyl 43-116 30-115
p-Terphenyl-d14 33-141 18-137
Phenol-d6 10-94 24-113
2-Fluorophenol 21-100 25-121
2,4,6-Tribromophenol 10-123 19-122
8270B - 39 Revision 2
November 1990
-------
TABLE 9.
METHOD PERFORMANCE DATA
PERCENT RECOVERY PERCENT RECOVERY
COMPOUND ON DAY 0 ON DAY 7
AV6. RSD AVG. RSD
3-Amino-9-ethylcarbazole 80 8 73 3
4-Chloro-l,2-phenylenediamine 91 1 108 4
4-Chloro-l,3-phenylenediamine 84 3 70 3
l,2-Dibromo-3-chloropropane 97 2 98 5
2-sec-Butyl-4,6-dinitrophenol 99 3 97 6
Ethyl parathion 100 2 103 4
4,4'-Methylenebis(N,N-dimethylaniline) 108 4 90 4
2-Methyl-5-nitroaniline 99 10 93 4
2-Methylpyridine 80 4 83 4
Tetraethyl dithiopyrophosphate 92 7 70 1
8270B - 40 Revision 2
November 1990
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METHOD 8270B
GAS CHROMATOGRAPHY/MASS SPECTROMETRY FOR SEMIVOLATILE
ORGANICS: CAPILLARY COLUMN TECHNIQUE
7 1 Prepare
sample using
Method 3540
or 3550
7 1 Prepare
sample using
Method 3510
or 3520
7.1 Prepare
sample using
Method 3540,
3550 or 3580.
7 2 Cleanup
en tract.
7.3 Set CC/MS
operating
condi tions.
Perform initial
calibration.
7 4 Perform daily
calibration Kith
SPCCs and CCCs
prior to analysis
of samples.
8270B - 42
Revision 2
November 1990
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METHOD 8270B
(Continued)
7.S.4 Diluti
e»tract
7.5 1 Screen
extract on CC/FID
or CC/PID to
eliminate samples
that are too
concentra ted.
7 5.3 Analyze
extract by CC/HS,
using appripriate
fused - si 1ica
capillary column
7 6 1 Identify
analyte by
comparing the
sample and standard
mass spectra.
762 Calculate
concentration of
each individual
analyte Report
results.
c
Stop
8270B - 43
Revision 2
November 1990
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METHOD 8275
THERMAL CHROMATOGRAPHY/MASS SPECTROMETRY FOR
SCREENING SEMIVOLATILE ORGANIC CHEMICALS
1.0 SCOPE AND APPLICATION
1.1 Method 8275 Is a screening technique that may be used for the
qualitative identification of semivolatile organic compounds in extracts prepared
from nonaqueous solid wastes and soils. Direct injection of a sample may be used
in limited applications. The following analytes can be qualitatively determined
by this method:
Compound Name CAS No."
2-Chlorophenol 95-57-8
4-Methylphenol 106-44-5
2,4-Dichlorophenol 120-83-2
Naphthalene 91-20-3
4-Chloro-3-methylphenol 59-50-7
1-Chloronaphthalene 90-13-1
2,4-Dinitrotoluene 121-14-2
Fluorene 86-73-7
Diphenylamine 122-39-4
Hexachlorobenzene 118-74-1
Dibenzothiophene 132-65-0
Phenanthrene 85-01-8
Carbazole 86-74-8
Aldrin 309-00-2
Pyrene 129-00-0
Benzo(k)fluoranthene 207-08-9
Benzo(a)pyrene 50-32-8
a Chemical Abstract Services Registry Number.
1.2 Method 8275 can be used to qualitatively identify most neutral,
acidic, and basic organic compounds that can be thermally desorbed from a sample,
and are capable of being eluted without derivatization as sharp peaks from a gas
chromatographic fused-silica capillary column coated with a slightly polar
silicone.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate
the ability to generate acceptable results with this method.
8275 - 1 Revision 0
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2.0 SUMMARY OF METHOD
2.1 A portion of the sample (0.010-0.100 g) is weighed into a sample
crucible. The crucible is placed in a pyrocell and heated. The compounds
desorbed from the sample are detected using a flame ionization detector (FID).
The FID response is used to calculate the optimal amount of sample needed for
mass spectrometry. A second sample is desorbed and the compounds are condensed
on the head of a fused silica capillary column. The column is heated using a
temperature program, and the effluent from the column is introduced into the mass
spectrometer.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever low-level samples are
analyzed after high-level samples. Whenever an unusually concentrated sample
is encountered, it should be followed by the analysis of an empty (clean)
crucible to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Thermal Chromatograph (TC) System
4.1.1 Thermal chromatograph™, Ruska Laboratories, or equivalent.
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID), 1 ^m film
thickness, silicone-coated, fused-silica capillary column (J&W Scientific
DB-5 or equivalent).
4.1.3 Flame Ionization detector (FID).
4.2 Mass Spectrometer (MS) system
4.2.1 Mass Spectrometer - Capable of scanning from 35 to 500 amu
every one second or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode.
4.2.2 TC/MS interface - Any GC-to-MS interface producing acceptable
calibration data in the concentration range of interest may be used.
4.2.3 Data System - A computer must be interfaced to the mass
spectrometer. The data system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
the duration of the chromatographic program. The computer must have
software that can search any GC/MS data file for ions of a specific mass
(or group of masses) and that can plot such ion abundances versus time or
scan number. This type of plot is defined as a reconstructed ion
chromatogram (RIC). Software must also be available that allows for
integration of the abundances in, and RIC between, specified time or scan-
number limits.
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4.3 Tools and equipment
4.3.1 Fused quartz spatula.
4.3.2 Fused quartz incinerator ladle.
4.3.3 Metal forceps for sample crucible.
4.3.4 Sample crucible storage dishes.
4.3.5 Porous fused quartz sample crucibles with lids.
4.3.6 Sample crucible cleaning incinerator.
4.3.7 Cooling rack.
4.3.8 Microbalance, 1 g capacity, 0.000001 g sensitivity,
Mettler Model M-3 or equivalent.
4.4 Vials - 10 ml, glass with Teflon lined screw-caps or crimp tops.
4.5 Volumetric flasks, Class A - 10 ml to 1000 ml.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available.
5.2 Solvents
5.2.1 Methanol, CH3OH - Pesticide grade or equivalent.
5.2.2 Acetone, CH3COCH3 - Pesticide grade or equivalent.
5.2.3 Toluene, C6H5CH3 - Pesticide grade or equivalent.
5.2.4 Methylene chloride, CH2C12 - Pesticide grade or equivalent.
5.2.5 Carbon disulfide, CS2 - Pesticide grade or equivalent.
5.2.6 Hexane, C6H14 - Pesticide grade or equivalent.
5.2.7 Other suitable solvents - Pesticide grade or equivalent.
5.3 Stock Standard solutions - Standard solutions may be prepared from
pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by weighing about 0.01 g of
pure material. Dissolve the material in pesticide quality acetone, or
other suitable solvent, and dilute to 10 ml in a volumetric flask. Larger
volumes may be used at the convenience of the analyst.
8275 - 3 Revision 0
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5.3.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp tops. Store at 4°C and protect from
light. Stock standard solutions should be checked frequently for signs
of degradation or evaporation, especially prior to use in preparation of
calibration standards.
5.3.3 Stock standard solutions must be replaced after 1 year, or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal Standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-da, acenaphthene-d10, phenanthrene-d10,
chrysene-d12, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Section 7 are met. Dissolve about 0.200 g
of each compound with a small volume of carbon disulfide. Transfer to a 50 ml
volumetric flask and dilute to volume with methylene chloride, so that the final
solvent is approximately 20/80 (V/V) carbon disulfide/methylene chloride. Most
of the compounds are also soluble in small volumes of methanol, acetone, or
toluene, except for perylene-d12. Prior to each analysis, evaporate about 10 nl
of the internal standard onto the lid of the crucible. Store internal standard
solutions at 4°C or less before, and between, use.
5.5 Calibration standards - Prepare calibration standards within the
working range of the TC/MS system. Each standard should contain each analyte
or interest (e.g. some or all of the compounds listed in Section 1.1 may be
included). Each aliquot of calibration standard should be spiked with internal
standards prior to analysis. Stock solutions should be stored at -10°C to -20°C
and should be freshly prepared once a year, or sooner if check standards indicate
a problem. The daily calibration standard should be prepared weekly, and stored
at 4°C.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Crucible Preparation
7.1.1 Turn on the incinerator and let it heat for at least 10
minutes. The bore of the incinerator should be glowing red.
7.1.2 Load the sample crucible and lid into the incinerator ladle
and insert into the incinerator bore. Leave in the incinerator for 5
minutes, then remove and place on the cooling rack.
7.1.3 Allow the crucibles and lids to cool for five minutes before
placing them in the storage dishes.
CAUTION: Do not touch the crucibles with your fingers. This can result in a
8275 - 4 Revision 0
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serious burn, as well as contamination of the crucible. Always handle
the sample crucibles and lids with forceps and tools specified.
7.1.4 All sample crucibles and lids required for the number of
analyses planned should be cleaned and placed in the storage dishes ready
for use.
7.2 Sample Preparation and Loading
7.2.1 The analyst should take care in selecting a sample for
analysis, since the sample size is generally limited to 0.100 g or less.
This implies that the sample should be mixed as thoroughly as possible
before taking an aliquot. Because the sample size is limited, the analyst
may wish to analyze several aliquots for determination.
7.2.2 The sample should be mixed or ground such that a 0.010 to
0,100 g aliquot can be removed. Remove one sample crucible from the
storage dish and place it on the microbalance. Establish the tare weight.
Remove the sample crucible from the balance with the forceps and place it
on a clean surface.
7.2.3 Load an amount of sample into the sample crucible using the
fused quartz spatula. Place the assembly on the microbalance and determine
the weight of the sample. For severely contaminated samples, less than
0.010 g will suffice, while 0.050-0.100 g is needed for low concentrations
of contaminants. Place the crucible lid on the crucible; the sample is
now ready for analysis.
7.3 FID Analysis
7.3.1 Load the sample into the TC. Hold the sample at 30°C for 2
minutes followed by linear temperature programmed heating to 260°C at
30°C/minute. Follow the temperature program with an isothermal heating
period of 10 minutes at 260°C, followed by cooling back to 30°C. The total
analysis cycle time is 24.2 minutes
7.3.2 Monitor the FID response in real time during analysis, and
note the highest response in millivolts (mv). Use this information to
determine the proper weight of sample needed for combined thermal
extraction/gas chromatography/mass spectrometry.
7.4 Thermal Extraction/GC/MS
7.4.1 Prepare a calibration curve using a clean crucible and lid
by spiking the compounds of interest at five concentrations into the
crucible and applying the internal standards to the crucible lid. Analyze
these standards and establish response factors at different concentrations.
7.4.2 Weigh out the amount of fresh sample that will provide
approximately 1000 to 3000 mv response. For example, if 0.010 g of sample
gives an FID response of 500 mv, then 0.020 to 0.060 g (0.040 g ± 50 %)
should be used. If 0.100 g gives 8000 mv, then 0.025 g ± 50 % should be
used.
8275 - 5 Revision 0
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7.4.3 After weighing out the sample into the crucible, deposit the
internal standards (10 /zL) onto the lid of the crucible. Load the
crucible into the pyrocell, using the same temperature program in Section
7.3.1. Hold the capillary at 5°C during this time to focus the released
semi-volatiles (the intermediate trap is held at 330°C to pass all
compounds onto the column). Maintain the splitter zone at 310°C, and the
GC/MS transfer line at 285°C. After the isothermal heating period is
complete, temperature program the column from 5°C to 285°C at 10°C/minute
and hold at 285°C for 5 minutes. Acquire data during the entire run time.
7.4.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the TC/MS system, a smaller sample should be
analyzed.
7.5 Data Interpretation
7.5.1 Qualitative Analysis
7.5.1.1 The qualitative identification of compounds determined
by this method is based on retention time, and on comparison of the
sample mass spectrum, after background correction, with
characteristic ions in a reference mass spectrum. The reference mass
spectrum must be generated by the laboratory using the conditions
of this method. The characteristic ions from the reference mass
spectrum are defined to be the three ions of greatest relative
intensity, or any ions over 30% relative intensity if less than three
such ions occur in the reference spectrum. Compounds should be
identified as present when the criteria below are met.
7.5.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.5.1.1.2 The RRT of the sample component is within ±
0.06 RRT units of the RRT of the standard component.
7.5.1.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions
in the reference spectrum. (Example: For an ion with an
abundance of 50% in the reference spectrum, the corresponding
abundance in a sample spectrum can range between 20% and 80%.)
7.5.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if they
have sufficiently different GC retention times. Sufficient
GC resolution is achieved if the height of the valley between
two isomer peaks is less than 25% of the sum of the two peak
heights. Otherwise, structural isomers are identified as
isomeric pairs.
8275 - 6 Revision 0
November 1990
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7.5.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributing by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background spectra
is important. Examination of extracted ion current profiles
of appropriate ions can aid in the selection of spectra, and
in qualitative identification of compounds. When analytes
coelute (i.e., only one chromatographic peak is apparent), the
identification criteria can be met, but each analyte spectrum
will contain extraneous ions contributed by the coeluting
compound.
7.5.1.2 For samples containing components not associated with
the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search routines
should not use normalization routines that would misrepresent the
library or unknown spectra when compared to each other. For example,
the RCRA permit or waste delisting requirements may require the
reporting of non-target analytes. Only after visual comparison of
sample spectra with the nearest library searches will the mass
spectral interpretation specialist assign a tentative identification.
Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in
the standard spectrum, the corresponding sample ion abundance must
be within 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from
the sample spectrum because of background contamination or coeluting.
Data system library reduction programs can sometimes create these
discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison
of sample with the nearest library searches will the mass spectral
8275 - 7 Revision 0
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interpretation specialist assign a tentative identification.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
9.0 METHOD PERFORMANCE
9.1 Table 1 presents method performance data, generated using spiked soil
samples. Method performance data in an aqueous matrix are not available.
10.0 REFERENCES
1. Zumberge, J.E., C. Sutton, R.D. Worden, T. Junk, T.R. Irvin, C.B. Henry,
V. Shirley, and E.B. Overton, "Determination of Semi-Volatile Organic
Pollutants in Soils by Thermal Chromatography-Mass Spectrometry (TC/MS):
an Assessment for Field Analysis," in preparation.
8275 - 8 Revision 0
November 1990
-------
TABLE 1
METHOD PERFORMANCE, SOIL MATRIX
Analyte
2-Chlorophenol
4-Methyl phenol
2,4-Dichlorophenol
Naphthalene
4-Chloro-3-methyl -phenol
1 -Chi oronaphthal ene
2,4-Dinitrotoluene
Fluorene
Diphenylamine
Hexachlorobenzene
Dibenzothiophene
Phenanthrene
Carbazole
Aldrin
Pyrene
Benzo(k)fluoranthene
Benzo(a)pyrene
Averaae
Clay
30
10
23
77
9
96
7
9
5
68
20
11
4
3
7
4
4
% Recovery8
Silt
22
77
20
120
12
103
10
25
6
64
35
31
8
19
19
9
8
Subsoil
2
7
26
63
9
70
10
19
6
80
50
40
9
15
20
11
11
Mean
Recovery
18
31
23
87
10
90
9
18
6
71
35
24
7
12
15
8
8
Percent theoretical recovery based upon linearity of injections deposited
on the crucible lid (slope and y-intercept). Average of 9 replicates (-10
mg soil spiked with 50 ppm of analyte); 3 different instruments at 3
different laboratories.
8275 - 9 Revision 0
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METHOD 8275
THERMAL CHROMATOGRAPHY/MASS SPECTROMETRY FOR
SCREENING SEMIVOLATILE ORGANIC CHEMICALS
Start
7.1 Prepare
crucible
7.2 Prepare
and load
sample
7.2.2
Establish
tare weight
of crucible
7.2.3 Place
sample in
crucible;
establish
weight
7.3.1 FID
Analysis using
1inear temp.
pr ogrammed
heating
7 3.2 Using
FIO response,
determine
sample weight
for TE/CC/MS
7 4.1 Prepare
cal ibralion
7.4 2 Prepare
amount of
sample for
appropriate
FID response
7.4.3 Heigh
sample into
crucible; use
temp. program
in Sec. 7 3.1
7.4.4
Is
quantitalion
ion > initial
calib. curve
range of
TC/MS
7.5.1
Qualitative
Identification
Stop
7.44 Use
smaller
sample
8275 - 10
Revision 0
November 1990
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METHOD 8290
POLYCHLORINATED DIBENZODIOXINS (PCDDs) AND POLYCHLORINATED DIBENZOFURANS (PCDFs)
BY HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-RESOLUTION
MASS SPECTROMETRY (HRGC/HRMS)
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the detection and quantitative
measurement of polychlorinated dibenzo-p-dioxins (tetra- through octachlorinated
homologues; PCDDs), and polychlorinated dibenzofurans (tetra- through
octachlorinated homologues; PCDFs) in a variety of environmental matrices and
at part-per-trillion (ppt) to part-per-quadrillion (ppq) concentrations. The
following compounds can be determined by this method:
Compound Name
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)
1,2,3,7,8-Pentachlorodibenzo-p-dioxin (PeCDD)
1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,4,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,7,8,9-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,4,6,7,8-Heptachlorodibenzo-p-dioxin (HpCDD)
2,3,7,8-Tetrachlorodibenzofuran (TCDF)
1,2,3,7,8-Pentachlorodi benzofuran (PeCDF)
2,3,4,7,8-Pentachlorodibenzofuran (PeCDF)
1,2,3,6,7,8-Hexachlorodi benzofuran (HxCDF)
1,2,3,7,8,9-Hexachlorodibenzofuran (HxCDF)
1,2,3,4,7,8-Hexachlorodi benzofuran (HxCDF)
2,3,4,6,7,8-Hexachlorodi benzofuran (HxCDF)
1,2,3,4,6,7,8-Heptachlorodi benzofuran (HpCDF)
1,2,3,4,7,8,9-Heptachlorodibenzofuran (HpCDF)
1.2 The analytical method calls for the use of high-resolution gas
chromatography and high-resolution mass spectrometry (HRGC/HRMS) on purified
sample extracts. Table 1 lists the various sample types covered by this
analytical protocol, the 2,3,7,8-TCDD-based method calibration limits (MCLs),
and other pertinent information. Samples containing concentrations of specific
congeneric analytes (PCDDs and PCDFs) considered within the scope of this method
that are greater than ten times the upper MCLs must be analyzed by a protocol
designed for such concentration levels, e.g., Method 8280. An optional method
for reporting the analytical results using a 2,3,7,8-TCDD toxicity equivalency
factor (TEF) is described.
1.3 The sensitivity of this method is dependent upon the level of inter-
ferences within a given matrix. The calibration range of the method for a 1 L
water sample is 10 to 2000 ppq for TCDD/TCDF and PeCDD/PeCDF, and 1.0 to 200 ppt
8290 - 1 Revision 0
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for a 10 g soil, sediment, fly ash, or tissue sample for the same analytes
(Table 1). Analysis of a one-tenth aliquot of the sample permits measurement
of concentrations up to 10 times the upper MCL. The actual limits of detection
and quantitation will differ from the lower MCL, depending on the complexity of
the matrix.
1.4 This method is designed for use by analysts who are experienced with
residue analysis and skilled in HRGC/HRMS.
1.5 Because of the extreme toxicity of many of these compounds, the
analyst must take the necessary precautions to prevent exposure to materials
known or believed to contain PCDDs or PCDFs. It is the responsibility of the
laboratory personnel to ensure that safe handling procedures are employed.
Section 11 of this method discusses safety procedures.
2.0 SUMMARY OF METHOD
2.1 This procedure uses matrix specific extraction, analyte specific
cleanup, and HRGC/HRMS analysis techniques.
2.2 If interferences are encountered, the method provides selected
cleanup procedures to aid the analyst in their elimination. A simplified
analysis flow chart is presented at the end of this method.
2.3 A specified amount (see Table 1) of soil, sediment, fly ash, water,
sludge (including paper pulp), still bottom, fuel oil, chemical reactor residue,
fish tissue, or human adipose tissue is spiked with a solution containing
specified amounts of each of the nine isotopically (13C12) labeled PCDDs/PCDFs
listed in Column 1 of Table 2. The sample is then extracted according to a
matrix specific extraction procedure. Aqueous samples that are judged to contain
1 percent or more solids, and solid samples that show an aqueous phase, are
filtered, the solid phase (including the filter) and the aqueous phase extracted
separately, and the extracts combined before extract cleanup. The extraction
procedures are:
a) ToluenerSoxhlet extraction for soil, sediment, fly ash and paper pulp
samples;
b) Methylene chloride:!iquid-liquid extraction for water samples;
c) Toluene:Dean-Stark extraction for fuel oil and aqueous sludge samples;
d) Toluene extraction for still bottom samples;
e) Hexane/methylene chloride:Soxhlet extraction or methylene
chloride:Soxhlet extraction for fish tissue samples; and
f) Methylene chloride extraction for human adipose tissue samples.
g) As an option, all solid samples (wet or dry) can be extracted with
toluene using a Soxhlet/Dean Stark extraction system.
8290 - 2 Revision 0
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The decision for the selection of an extraction procedure for chemical
reactor residue samples is based on the appearance (consistency, viscosity) of
the samples. Generally, they can be handled according to the procedure used
for still bottom (or chemical sludge) samples.
2.4 The extracts are submitted to an acid-base washing treatment and
dried. Following a solvent exchange step, the extracts are cleaned up by column
chromatography on alumina, silica gel, and AX-21 activated carbon on Celite 545®
(or equivalent).
2.4.1 The extracts from adipose tissue samples are treated with
silica gel impregnated with sulfuric acid before chromatography on acidic
silica gel, neutral alumina, and AX-21 on Celite 545® (or equivalent).
2.4.2 Fish tissue and paper pulp extracts are subjected to an acid
wash treatment only, prior to chromatography on alumina and
AX-21/Celite 545® (or equivalent).
2.5 The preparation of the final extract for HRGC/HRMS analysis is
accomplished by adding, to the concentrated AX-21/Celite 545® (or equivalent)
column eluate, 10 to 50 nl (depending on the matrix type) of a nonane solution
containing 50 pg/jiL of each of the two recovery standards 13C12-1,2,3,4-TCDD and
13C12-l,2,3,7,8,9-HxCDD (Table 2). The former is used to determine the percent
recoveries of tetra- and pentachlorinated PCDD/PCDF congeners, while the latter
is used to determine the percent recoveries of the hexa-, hepta- and
octachlorinated PCDD/PCDF congeners.
2.6 One to two pi of the concentrated extract are injected into an
HRGC/HRMS system capable of performing selected ion monitoring at resolving
powers of at least 10,000 (10 percent valley definition).
2.7 The identification of OCDD and nine of the fifteen 2,3,7,8-
substituted congeners (Table 3), for which a 13C-labeled standard is available
in the sample fortification and recovery standard solutions (Table 2), is based
on their elution at their exact retention time (within 0.005 retention time units
measured in the routine calibration) and the simultaneous detection of the two
most abundant ions in the molecular ion region. The remaining six 2,3,7,8-
substituted congeners (i.e., 2,3,4,7,8-PeCDF; 1,2,3,4,7,8-HxCDD; 1,2,3,6,7,8-
HxCDF; 1,2,3,7,8,9-HxCDF; 2,3,4,6,7,8-HxCDF, and 1,2,3,4,7,8,9-HpCDF), for which
no carbon-labeled internal standards are available in the sample fortification
solution, and all other identified PCDD/PCDF congeners are identified by their
relative retention times falling within their respective PCDD/PCDF retention time
windows, as established from the routine calibration data, and the simultaneous
detection of the two most abundant ions in the molecular ion region. The
identification of OCDF is based on its retention time relative to 13C12-OCDD and
the simultaneous detection of the two most abundant ions in the molecular ion
region. Confirmation is based on a comparison of the ratios of the integrated
ion abundance of the molecular ion species to their theoretical abundance ratios.
2.8 Quantitation of the individual congeners, total PCDDs and total
PCDFs is achieved in conjunction with the establishment of a multipoint (five
points) calibration curve for each homologue, during which each calibration
solution is analyzed once.
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3.0 INTERFERENCES
3.1 Solvents, reagents, glassware and other sample processing hardware
may yield discrete artifacts or elevated baselines that may cause misinter-
pretation of the chromatographic data (see references 1 and 2.) All of these
materials must be demonstrated to be free from interferants under the conditions
of analysis by performing laboratory method blanks. Analysts should avoid using
PVC gloves.
3.2 The use of high purity reagents and solvents helps minimize
interference problems. Purification of solvents by distillation in all-glass
systems may be necessary.
3.3 Interferants coextracted from the sample will vary considerably from
matrix to matrix. PCDDs and PCDFs are often associated with other interfering
chlorinated substances such as polychlorinated biphenyls (PCBs), polychlorinated
diphenyl ethers (PCDPEs), polychlorinated naphthalenes, and polychlorinated
alkyldibenzofurans that may be found at concentrations several orders of
magnitude higher than the analytes of interest. Retention times of target
analytes must be verified using reference standards. These values must
correspond to the retention time windows established in Section 8.1.1.3. While
certain cleanup techniques are provided as part of this method, unique samples
may require additional cleanup steps to achieve lower detection limits.
3.4 A high-resolution capillary column (60 m DB-5, J&W Scientific, or
equivalent) is used in this method. However, no single column is known to
resolve all isomers. The 60 m DB-5 GC column is capable of 2,3,7,8-TCDD isomer
specificity (Section 8.1.1). In order to determine the concentration of the
2,3,7,8-TCDF (if detected on the DB-5 column), the sample extract must be
reanalyzed on a column capable of 2,3,7,8-TCDF isomer specificity (e.g., DB-225,
SP-2330, SP-2331, or equivalent). When a column becomes available that resolves
all isomers, then a single analysis on this column can be used instead of
analyses on more than one column.
4.0 APPARATUS AND MATERIALS
4.1 High-Resolution Gas Chromatograph/High-Resolution Mass
Spectrometer/Data System (HRGC/HRMS/DS) - The GC must be equipped for temperature
programming, and all required accessories must be available, such as syringes,
gases, and capillary columns.
4.1.1 GC Injection Port - The GC injection port must be designed
for capillary columns. The use of splitless injection techniques is
recommended. On column 1 juL injections can be used on the 60 m DB-5
column. The use of a moving needle injection port is also acceptable.
When using the method described in this protocol, a 2 /uL injection volume
is used consistently (i.e., the injection volumes for all extracts, blanks,
calibration solutions and the performance check samples are 2 /*!_). One Mi-
injections are allowed; however, laboratories must remain consistent
throughout the analyses by using the same injection volume at all times.
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4.1.2 Gas Chromatoqraph/Mass Spectrometer (GC/MS) Interface - The
GC/MS interface components should withstand 350°C. The interface must be
designed so that the separation of 2,3,7,8-TCDD from the other TCDD isomers
achieved in the gas chromatographic column is not appreciably degraded.
Cold spots or active surfaces (adsorption sites) in the GC/MS interface
can cause peak tailing and peak broadening. It is recommended that the
GC column be fitted directly into the mass spectrometer ion source without
being exposed to the ionizing electron beam. Graphite ferrules should be
avoided in the injection port because they may adsorb the PCDDs and PCDFs.
Vespel™, or equivalent, ferrules are recommended.
4.1.3 Mass Spectrometer - The static resolving power of the
instrument must be maintained at a minimum of 10,000 (10 percent valley).
4.1.4 Data System - A dedicated data system is employed to control
the rapid multiple-ion monitoring process and to acquire the data.
Quantitation data (peak areas or peak heights) and SIM-traces (displays
of intensities of each ion signal being monitored including the lock-mass
ion as a function of time) must be acquired during the analyses and stored.
Quantitations may be reported based upon computer generated peak areas or
upon measured peak heights (chart recording). The data system must be
capable of acquiring data at a minimum of 10 ions in a single scan. It is
also recommended to have a data system capable of switching to different
sets of ions (descriptors) at specified times during an HRGC/HRMS
acquisition. The data system should be able to provide hard copies of
individual ion chromatograms for selected gas chromatographic time
intervals. It should also be able to acquire mass spectral peak profiles
(Section 8.1.2.3) and provide hard copies of peak profiles to demonstrate
the required resolving power. The data system should permit the
measurement of noise on the base line.
NOTE: The detector ADC zero setting must allow peak-to-peak measurement of the
noise on the base line of every monitored channel and allow for good
estimation of the instrument resolving power. In Figure 2, the effect of
different zero settings on the measured resolving power is shown.
4.2 GC Columns
4.2.1 In order to have an isomer specific determination for 2,3,7,8-
TCDD and to allow the detection of OCDD/OCDF within a reasonable time
interval in one HRGC/HRMS analysis, use of the 60 m DB-5 fused silica
capillary column is recommended. Minimum acceptance criteria must be
demonstrated and documented (Section 8.1.1). At the beginning of each 12
hour period (after mass resolution and GC resolution is demonstrated)
during which sample extracts or concentration calibration solutions will
be analyzed, column operating conditions must be attained for the required
separation on the column to be used for samples. Operating conditions
known to produce acceptable results with the recommended column are shown
in Section 7.6.
4.2.2 Isomer specificity for all 2,3,7,8-substituted PCDDs/PCDFs
cannot be achieved on the 60 m DB-5 GC column alone. In order to determine
the proper concentrations of the individual 2,3,7,8-substituted congeners,
the sample extract must be reanalyzed on another GC column that resolves
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the isomers. When such a column becomes available, and the isomer
specificity can be documented, the performing laboratory will be required
to use it.
4.2.3 30 m DB-225 fused silica capillary column, (J&W Scientific)
or equivalent.
4.3 Miscellaneous Equipment and Materials - The following list of items
does not necessarily constitute an exhaustive compendium of the equipment needed
for this analytical method.
4.3.1 Nitrogen evaporation apparatus with variable flow rate.
4.3.2 Balances capable of accurately weighing to 0.01 g and 0.0001 g.
4.3.3 Centrifuge.
4.3.4 Water bath, equipped with concentric ring covers and capable
of being temperature controlled within ± 2°C.
4.3.5 Stainless steel or glass container large enough to hold
contents of one pint sample containers.
4.3.6 Glove box.
4.3.7 Drying oven.
4.3.8 Stainless steel spoons and spatulas.
4.3.9 Laboratory hoods.
4.3.10 Pipets, disposable, Pasteur, 150 mm long x 5 mm ID.
4.3.11 Pipets, disposable, serological, 10 mL, for the preparation
of the carbon columns specified in Section 7.5.3.
4.3.12 Reaction vial, 2 ml, silanized amber glass (Reacti-vial, or
equivalent).
4.3.13 Stainless steel meat grinder with a 3 to 5 mm hole size inner
plate.
4.3.14 Separatory funnels, 125 ml and 2000 ml.
4.3.15 Kuderna-Danish concentrator, 500 ml, fitted with 10 ml
concentrator tube and three ball Snyder column.
4.3.16 Teflon™ or carborundum (silicon carbide) boiling chips (or
equivalent), washed with hexane before use.
NOTE: Teflon™ boiling chips may float in methylene chloride, may not work in
the presence of any water phase, and may be penetrated by nonpolar organic
compounds.
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4.3.17 Chromatographic columns, glass, 300 mm x 10.5 mm, fitted with
Teflon™ stopcock.
4.3.18 Adapters for concentrator tubes.
4.3.19 Glass fiber filters.
4.3.20 Dean-Stark trap, 5 or 10 ml, with T-joints, condenser and
125 ml flask.
4.3.21 Continuous liquid-liquid extractor.
4.3.22 All glass Soxhlet apparatus, 500 ml flask.
4.3.23 Soxhlet/Dean Stark extractor (optional), all glass, 500 ml
flask.
4.3.24 Glass funnels, sized to hold 170 ml of liquid.
4.3.25 Desiccator.
4.3.26 Solvent reservoir (125 ml_), Kontes; 12.35 cm diameter (special
order item), compatible with gravity carbon column.
4.3.27 Rotary evaporator with a temperature controlled water bath.
4.3.28 High speed tissue homogenizer, equipped with an EN-8 probe,
or equivalent.
4.3.29 Glass wool, extracted with methylene chloride, dried and
stored'in a clean glass jar.
4.3.30 Extraction jars, glass, 250 ml, with teflon lined screw cap.
4.3.31 Volumetric flasks, Class A - 10 ml to 1000 mL.
4.3.32 Glass vials, 1 dram (or metric equivalent).
NOTE: Reuse of glassware should be minimized to avoid the risk of contamination.
All glassware that is reused must be scrupulously cleaned as soon as
possible after use, according to the following procedure: Rinse glassware
with the last solvent used in it. Wash with hot detergent water, then
rinse with copious amounts of tap water and several portions of organic-
free reagent water. Rinse with high purity acetone and hexane and store
it inverted or capped with solvent rinsed aluminum foil in a clean
environment.
5.0 REAGENTS AND STANDARD SOLUTIONS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
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5.2 Column Chromatography Reagents
5.2.1 Alumina, neutral, 80/200 mesh (Super 1, Woelm®, or
equivalent). Store in a sealed container at room temperature, in a
desiccator, over self-indicating silica gel.
5.2.2 Alumina, acidic AG4, (Bio Rad Laboratories catalog #132-1240,
or equivalent). Soxhlet extract with methylene chloride for 24 hours if
blanks show contamination, and activate by heating in a foil covered glass
container for 24 hours at 190°C. Store in a glass bottle sealed with a
Teflon™ lined screw cap.
5.2.3 Silica gel, high purity grade, type 60, 70-230 mesh; Soxhlet
extract with methylene chloride for 24 hours if blanks show contamination,
and activate by heating in a foil covered glass container for 24 hours at
190°C. Store in a glass bottle sealed with a Teflon™ lined screw cap.
5.2.4 Silica gel impregnated with sodium hydroxide. Add one part
(by weight) of 1 M NaOH solution to two parts (by weight) silica gel
(extracted and activated) in a screw cap bottle and mix with a glass rod
until free of lumps. Store in a glass bottle sealed with a Teflon™ lined
screw cap.
5.2.5 Silica gel impregnated with 40 percent (by weight) sulfuric
acid. Add two parts (by weight) concentrated sulfuric acid to three parts
(by weight) silica gel (extracted and activated), mix with a glass rod
until free of lumps, and store in a screw capped glass bottle. Store in
a glass bottle sealed with a Teflon™ lined screw cap.
5.2.6 Celite 545* (Supelco), or equivalent.
5.2.7 Active carbon AX-21 (Anderson Development Co., Adrian, MI),
or equivalent, prewashed with methanol and dried in vacuo at 110°C. Store
in a glass bottle sealed with a Teflon™ lined screw cap.
5.3 Reagents
5.3.1 Sulfuric acid, H2S04, concentrated, ACS grade, specific gravity
1.84.
5.3.2 Potassium hydroxide, KOH, ACS grade, 20 percent (w/v) in
organic-free reagent water.
5.3.3 Sodium chloride, NaCl, analytical reagent, 5 percent (w/v) in
organic-free reagent water.
5.3.4 Potassium carbonate, K2C03, anhydrous, analytical reagent.
5.4 Desiccating agent
5.4.1 Sodium sulfate (powder, anhydrous), Na2S04. Purify by heating
at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
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methylene chloride, a method blank must be analyzed, demonstrating that
there is no interference from the sodium sulfate.
5.5 Solvents
5.5.1 Methylene chloride, CH2C12. High purity, distilled in glass
or highest available purity.
5.5.2 Hexane, C6H14. High purity, distilled in glass or highest
available purity.
5.5.3 Methanol, CH3OH. High purity, distilled in glass or highest
available purity.
5.5.4 Nonane, CgH^. High purity, distilled in glass or highest
available purity.
5.5.5 Toluene, C6H5CH3. High purity, distilled in glass or highest
available purity.
5.5.6 Cyclohexane, C6H12. High purity, distilled in glass or highest
available purity.
5.5.7 Acetone, CH3COCH3. High purity, distilled in glass or highest
available purity.
5.6 High-Resolution Concentration Calibration Solutions (Table 5) - Five
nonane solutions containing unlabeled (totaling 17) and carbon-labeled (totaling
11) PCDDs and PCDFs at known concentrations are used to calibrate the instrument.
The concentration ranges are homologue dependent, with the lowest values for the
tetrachlorinated dioxin and furan (1.0 pg//iL) and the highest values for the
octachlorinated congeners (1000 pg//uL).
5.6.1 Depending on the availability of materials, these high-
resolution concentration calibration solutions may be obtained from the
Environmental Monitoring Systems Laboratory, U.S. EPA, Cincinnati, Ohio.
However, additional secondary standards must be obtained from commercial
sources, and solutions must be prepared in the analyst's laboratory.
Traceability of standards must be verified against EPA-supplied standard
solutions. It is the responsibility of the laboratory to ascertain that
the calibration solutions received (or prepared) are indeed at the
appropriate concentrations before they are used to analyze samples.
5.6.2 Store the concentration calibration solutions in 1 ml minivials
at room temperature in the dark.
5.7 GC Column Performance Check Solution - This solution contains the
first and last eluting isomers for each homologous series from tetra- through
heptachlorinated congeners. The solution also contains a series of other TCDD
isomers for the purpose of documenting the chromatographic resolution. The
13C12-2,3,7,8-TCDD is also present. The laboratory is required to use nonane as
the solvent and adjust the volume so that the final concentration does not exceed
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100 pg/juL per congener. Table 7 summarizes the qualitative composition (minimum
requirement) of this performance evaluation solution.
5.8 Sample Fortification Solution - This nonane solution contains the
nine internal standards at the nominal concentrations that are listed in Table 2.
The solution contains at least one carbon-labeled standard for each homologous
series, and it is used to measure the concentrations of the native substances.
(Note that 13C12-OCDF is not present in the solution.)
5.9 Recovery Standard Solution - This nonane solution contains two
recovery standards, 13C12-1,2,3,4-TCDD and 13C12-l,2,3,7,8,9-HxCDD, at a nominal
concentration of 50 pg/juL per compound. 10 to 50 juL of this solution will be
spiked into each sample extract before the final concentration step and HRGC/HRMS
analysis.
5.10 Matrix Spike Fortification Solution - Solution used to prepare the
MS and MSD samples. It contains all unlabeled analytes listed in Table 5 at con-
centrations corresponding to the HRCC 3.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Sample Collection
6.2.1 Sample collection personnel should, to the extent possible,
homogenize samples in the field before filling the sample containers.
This should minimize or eliminate the necessity for sample homogenization
in the laboratory. The analyst should make a judgment, based on the
appearance of the sample, regarding the necessity for additional mixing.
If the sample is clearly not homogeneous, the entire contents should be
transferred to a glass or stainless steel pan for mixing with a stainless
steel spoon or spatula before removal of a sample portion for analysis.
6.2.2 Grab and composite samples must be collected in glass
containers. Conventional sampling practices must be followed. The bottle
must not be prewashed with sample before collection. Sampling equipment
must be free of potential sources of contamination.
6.3 Grinding or Blending of Fish Samples - If not otherwise specified
by the U.S. EPA, the whole fish (frozen) should be blended or ground to provide
a homogeneous sample. The use of a stainless steel meat grinder with a 3 to 5
mm hole size inner plate is recommended. In some circumstances, analysis of
fillet or specific organs of fish may be requested by the U.S. EPA. If so
requested, the above whole fish requirement is superseded.
6.4 Storage and Holding Times - All samples, except fish and adipose
tissue samples, must be stored at 4°C in the dark, extracted within 30 days and
completely analyzed within 45 days of collection. Fish and adipose tissue
samples must be stored at -20°C in the dark, extracted within 30 days and
completely analyzed within 45 days of collection. Whenever samples are analyzed
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after the holding time expiration date, the results should be considered to be
minimum concentrations and must be identified as such.
Note: The holding times listed in Section 6.4 are recommendations. PCDDs and
PCDFs are very stable in a variety of matrices, and holding times under
the conditions listed in Section 6.4 may be as high as a year for certain
matrices. Sample extracts, however, should always be analyzed within 45
days of extraction.
6.5 Phase Separation - This is a guideline for phase separation for very
wet (>25 percent water) soil, sediment and paper pulp samples. Place a 50 g
portion in a suitable centrifuge bottle and centrifuge for 30 minutes at
2,000 rpm. Remove the bottle and mark the interface level on the bottle.
Estimate the relative volume of each phase. With a disposable pipet, transfer
the liquid layer into a clean bottle. Mix the solid with a stainless steel
spatula and remove a portion to be weighed and analyzed (percent dry weight
determination, extraction). Return the remaining solid portion to the original
sample bottle (empty) or to a clean sample bottle that is properly labeled, and
store it as appropriate. Analyze the solid phase by using only the soil,
sediment and paper pulp method. Take note of, and report, the estimated volume
of liquid before disposing of the liquid as a liquid waste.
6.6 Soil. Sediment, or Paper Sludge (Pulp) Percent Dry Weight
Determination - The percent dry weight of soil, sediment or paper pulp samples
showing detectable levels (see note below) of at least one 2,3,7,8-substituted
PCDD/PCDF congener is determined according to the following procedure. Weigh
a 10 g portion of the soil or sediment sample (± 0.5 g) to three significant
figures. Dry it to constant weight at 110°C in an adequately ventilated oven.
Allow the sample to cool in a desiccator. Weigh the dried solid to three
significant figures. Calculate and report the percent dry weight. Do not use
this solid portion of the sample for extraction, but instead dispose of it as
hazardous waste.
NOTE: Until detection limits have been established (Section 1.3), the lower MCLs
(Table 1) may be used to estimate the minimum detectable levels.
% dry weight = q of dry sample x 100
g of sample
CAUTION: Finely divided soils and sediments contaminated with PCDDs/PCDFs are
hazardous because of the potential for inhalation or ingestion of
particles containing PCDDs/PCDFs (including 2,3,7,8-TCDD). Such samples
should be handled in a confined environment (i.e., a closed hood or a
glove box).
6.7 Lipid Content Determination
6.7.1 Fish Tissue - To determine the lipid content of fish tissue,
concentrate 125 mL of the fish tissue extract (Section 7.2.2), in a tared
200 mL round bottom flask, on a rotary evaporator until a constant weight
(W) is achieved.
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100 (W)
Percent lipid = -
10
Dispose of the lipid residue as a hazardous waste if the results of the
analysis indicate the presence of PCDDs or PCDFs.
6.7.2 Adipose Tissue - Details for the determination of the adipose
tissue lipid content are provided in Section 7.3.3.
7.0 PROCEDURE
7.1 Internal standard addition
7.1.1 Use a portion of 1 g to 1000 g (± 5 percent) of the sample to
be analyzed. Typical sample size requirements for different matrices are
given in Section 7.4 and in Table 1. Transfer the sample portion to a
tared flask and determine its weight.
7.1.2 Except for adipose tissue, add an appropriate quantity of the
sample fortification mixture (Section 5.8) to the sample. All samples
should be spiked with 100 /uL of the sample fortification mixture to give
internal standard concentrations as indicated in Table 1. As an example,
for 13C12-2,3,7,8-TCDD, a 10 g soil sample requires the addition of 1000 pg
of 13C12-2,3,7,8-TCDD to give the required 100 ppt fortification level. The
fish tissue sample (20 g) must be spiked with 200 /il_ of the internal
standard solution, because half of the extract will be used to determine
the lipid content (Section 6.7.1).
7.1.2.1 For the fortification of soil, sediment, fly ash,
water, fish tissue, paper pulp and wet sludge samples, mix the sample
fortification solution with 1.0 ml acetone.
7.1.2.2 Do not dilute the nonane solution for the other
matrices.
7.1.2.3 The fortification of adipose tissue is carried out
at the time of homogenization (Section 7.3.2.3).
7.2 Extraction and Purification of Fish and Paper Pulp Samples
7.2.1 Add 60 g anhydrous sodium sulfate to a 20 g portion of a
homogeneous fish sample (Section 6.3) and mix thoroughly with a stainless
steel spatula. After breaking up any lumps, place the fish/sodium sulfate
mixture in the Soxhlet apparatus on top of a glasswool plug. Add 250 ml
methylene chloride or hexane/methylene chloride (1:1) to the Soxhlet
apparatus and reflux for 16 hours. The solvent must cycle completely
through the system five times per hour. Follow the same procedure for the
partially dewatered paper pulp sample (using a 10 g sample, 30 g of
anhydrous sodium sulfate and 200 ml of toluene).
NOTE: As an option, a Soxhlet/Dean Stark extractor system may be used, with
toluene as the solvent. No sodium sulfate is added when using this option.
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7.2.2 Transfer the fish extract from Section 7.2.1 to a 250 mL
volumetric flask and fill to the mark with methylene chloride. Mix well,
then remove 125 ml for the determination of the lipid content (Section
6.7.1). Transfer the remaining 125 ml of the extract, plus two 15 ml
hexane/methylene chloride rinses of the volumetric flask, to a KD apparatus
equipped with a Snyder column. Quantitatively transfer all of the paper
pulp extract to a KD apparatus equipped with a Snyder column.
NOTE: As an option, a rotary evaporator may be used in place of the KD apparatus
for the concentration of the extracts.
7.2.3 Add a Teflon™, or equivalent, boiling chip. Concentrate the
extract in a water bath to an apparent volume of 10 mL. Remove the
apparatus from the water bath and allow to cool for 5 minutes.
7.2.4 Add 50 mL hexane and a new boiling chip to the KD flask.
Concentrate in a water bath to an apparent volume of 5 mL. Remove the
apparatus from the water bath and allow to cool for 5 minutes.
NOTE: The methylene chloride must have been completely removed before proceeding
with the next step.
7.2.5 Remove and invert the Snyder column and rinse it into the KD
apparatus with two 1 mL portions of hexane. Decant the contents of the
KD apparatus and concentrator tube into a 125 mL separatory funnel. Rinse
the KD apparatus with two additional 5 mL portions of hexane and add the
rinses to the funnel. Proceed with the cleanup according to the
instructions starting in Section 7.5.1.1, but omit the procedures described
in Sections 7.5.1.2 and 7.5.1.3.
7.3 Extraction and Purification of Human Adipose Tissue
7.3.1 Human adipose tissue samples must be stored at a temperature
of -20°C or lower from the time of collection until the time of analysis.
The use of chlorinated materials during the collection of the samples must
be avoided. Samples are handled with stainless steel forceps, spatulas,
or scissors. All sample bottles (glass) are cleaned as specified in the
note at the end of Section 4.3. Teflon lined caps should be used.
NOTE: The specified storage temperature of -20°C is the maximum storage
temperature permissible for adipose tissue samples. Lower storage
temperatures are recommended.
7.3.2 Adipose Tissue Extraction
7.3.2.1 Weigh, to the nearest 0.01 g, a 10 g portion of a
frozen adipose tissue sample into a culture tube (2.2 x 15 cm).
NOTE: The sample size may be smaller, depending on availability. In such a
situation, the analyst is required to adjust the volume of the internal
standard solution added to the sample to meet the fortification level
stipulated in Table 1.
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7.3.2.2 Allow the adipose tissue specimen to reach room
temperature (up to 2 hours).
7.3.2.3 Add 10 ml methylene chloride and 100 juL of the sample
fortification solution. Homogenize the mixture for approximately
1 minute with a tissue homogenizer.
7.3.2.4 Allow the mixture to separate, then remove the
methylene chloride extract from the residual solid material with a
disposable pi pet. Percolate the methylene chloride through a filter
funnel containing a clean glass wool plug and 10 g anhydrous sodium
sulfate. Collect the dried extract in a graduated 100 ml volumetric
flask.
7.3.2.5 Add a second 10 ml portion of methylene chloride to
the sample and homogenize for 1 minute. Decant the solvent, dry
it, and transfer it to the 100 ml volumetric flask (Section 7.3.2.4).
7.3.2.6 Rinse the culture tube with at least two additional
portions of methylene chloride (10 ml each), and transfer the entire
contents to the filter funnel containing the anhydrous sodium
sulfate. Rinse the filter funnel and the anhydrous sodium sulfate
contents with additional methylene chloride (20 to 40 ml) into the
100 ml flask. Discard the sodium sulfate.
7.3.2.7 Adjust the volume to the 100 ml mark with methylene
chloride.
7.3.3 Adipose Tissue Lipid Content Determination
7.3.3.1 Preweigh a clean 1 dram (or metric equivalent) glass
vial to the nearest 0.0001 g on an analytical balance tared to zero.
7.3.3.2 Accurately transfer 1.0 ml of the final extract
(100 ml) from Section 7.3.2.6 to the vial. Reduce the volume of the
extract on a water bath (50-60°C) by a gentle stream of purified
nitrogen until an oily residue remains. Nitrogen blowdown is
continued until a constant weight is achieved.
Note: When the sample size of the adipose tissue is smaller than 10 g, then the
analyst may use a larger portion (up to 10 percent) of the extract defined
in Section 7.3.2.7 for the lipid determination.
7.3.3.3 Accurately weigh the vial with the residue to the
nearest 0.0001 g and calculate the weight of the lipid present in
the vial based on the difference of the weights.
7.3.3.4 Calculate the percent lipid content of the original
sample to the nearest 0.1 percent as shown below:
U Y V
"Ir A * ext
Lipid content, LC (%) = x 100
W0
at
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where:
Wlr = weight of the lipid residue to the nearest 0.0001 g calculated
from Section 7.3.3.3,
Vext = total volume (100 ml) of the extract In ml from
Section 7.3.2.6,
Wat = weight of the original adipose tissue sample to the nearest
0.01 g from Section 7.3.2.1, and
Va, = volume of the aliquot of the final extract in ml used for the
quantitative measure of the lipid residue (1.0 ml).
7.3.3.5 Record the lipid residue measured in Section 7.3.3.3
and the percent lipid content from Section 7.3.3.4.
7.3.4 Adipose Tissue Extract Concentration
7.3.4.1 Quantitatively transfer the remaining extract
(99.0 ml) to a 500 ml Erlenmeyer flask. Rinse the volumetric flask
with 20 to 30 ml of additional methylene chloride to ensure
quantitative transfer.
7.3.4.2 Concentrate the extract on a rotary evaporator and
a water bath at 40°C until an oily residue remains.
7.3.5 Adipose Tissue Extract Cleanup
7.3.5.1 Add 200 ml hexane to the lipid residue in the 500 ml
Erlenmeyer flask and swirl the flask to dissolve the residue.
7.3.5.2 Slowly add, with stirring, 100 g of 40 percent (w/w)
sulfuric acid-impregnated silica* gel. Stir with a magnetic stirrer
for two hours at room temperature.
7.3.5.3 Allow the solid phase to settle, and decant the liquid
through a filter funnel containing 10 g anhydrous sodium sulfate on
a glass wool plug, into another 500 ml Erlenmeyer flask.
7.3.5.4 Rinse the solid phase with two 50 ml portions of
hexane. Stir each rinse for 15 minutes, decant, and dry as described
under Section 7.3.5.3. Combine the hexane extracts from Section
7.3.5.3 with the rinses.
7.3.5.5 Rinse the sodium sulfate in the filter funnel with
an additional 25 ml hexane and combine this rinse with the hexane
extracts from Section 7.3.5.4.
7.3.5.6 Prepare an acidic silica column as follows: Pack a
2 cm x 10 cm chromatographic column with a glass wool plug, add
approximately 20 ml hexane, add 1 g silica gel and allow to settle,
then add 4 g of 40 percent (w/w) sulfuric acid-impregnated silica
gel and allow to settle. Elute the excess hexane from the column
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until the solvent level reaches the top of the chromatographic
packing. Verify that the column does not have any air bubbles and
channels.
7.3.5.7 Quantitatively transfer the hexane extract from the
Erlenmeyer flask (Sections 7.3.5.3 through 7.3.5.5) to the silica
gel column reservoir. Allow the hexane extract to percolate through
the column and collect the eluate in a 500 mL KD apparatus.
7.3.5.8 Complete the elution by percolating 50 ml hexane
through the column into the KD apparatus. Concentrate the eluate
on a steam bath to approximately 5 mL. Use nitrogen blowdown to
bring the final volume to about 100 /iL.
NOTE: If the silica gel impregnated with 40 percent sulfuric acid is highly
discolored throughout the length of the adsorbent bed, the cleaning
procedure must be repeated beginning with Section 7.3.5.1.
7.3.5.9 The extract is ready for the column cleanups described
in Sections 7.5.2 through 7.5.3.6.
7.4 Extraction and Purification of Environmental and Waste Samples
7.4.1 Sludge/Wet Fuel Oil
7.4.1.1 Extract aqueous sludge or wet fuel oil samples by
refluxing a sample (e.g., 2 g) with 50 ml toluene in a 125 mL flask
fitted with a Dean-Stark water separator. Continue refluxing the
sample until all the water is removed.
7.4.1.2 Cool the sample, filter the toluene extract through
a glass fiber filter, or equivalent, into a 100 mL round bottom
flask.
7.4.1.3 Rinse the filter with 10 mL toluene and combine the
extract with the rinse.
7.4.1.4 Concentrate the combined solutions to near dryness
on a rotary evaporator at 50°C. Use of an inert gas to concentrate
the extract is also permitted. Proceed with Section 7.4.4.
NOTE: If the sludge or fuel oil sample dissolves in toluene, treat it according
to the instructions in Section 7.4.2 below. If the labeled sludge sample
originates from pulp (paper mills), treat it according to the instructions
starting in Section 7.2, but without the addition of sodium sulfate.
7.4.2 Still Bottom/Oil
7.4.2.1 Extract still bottom or oil samples by mixing a sample
portion (e.g., 1.0 g) with 10 mL toluene in a small beaker and
filtering the solution through a glass fiber filter (or equivalent)
into a 50 mL round bottom flask. Rinse the beaker and filter with
10 mL toluene.
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7.4.2.2 Concentrate the combined toluene solutions to near
dryness on a rotary evaporator at 50°C. Proceed with Section 7.4.4.
7.4.3 Fly Ash
Note: Because of the tendency of fly ash to "fly", all handling steps should
be performed in a hood in order to minimize contamination.
7.4.3.1 Weigh about 10 g fly ash to two decimal places and
transfer to an extraction jar. Add 100 pi sample fortification
solution (Section 5.8), diluted to 1 ml with acetone, to the sample.
Add 150 ml of 1 M HC1 to the fly ash sample. Seal the jar with the
Teflon™ lined screw cap and shake for 3 hours at room temperature.
7.4.3.2 Rinse a glass fiber filter with toluene, and filter
the sample through the filter paper, placed in a Buchner funnel, into
all flask. Wash the fly ash cake with approximately 500 ml
organic-free reagent water and dry the filter cake overnight at room
temperature in a desiccator.
7.4.3.3 Add 10 g anhydrous powdered sodium sulfate, mix
thoroughly, let sit in a closed container for one hour, mix again,
let sit for another hour, and mix again.
7.4.3.4 Place the sample and the filter paper into an
extraction thimble, and extract in a Soxhlet extraction apparatus
charged with 200 mL toluene for 16 hours using a five cycle/hour
schedule.
NOTE: As an option, a Soxhlet/Dean Stark extractor system may be used, with
toluene as the solvent. No sodium sulfate is added when using this option.
7.4.3.5 Cool and filter the toluene extract through a glass
fiber filter into a 500 ml round bottom flask. Rinse the filter
with 10 ml toluene. Add the rinse to the extract and concentrate
the combined toluene solutions to near dryness on a rotary evaporator
at 50°C. Proceed with Section 7.4.4.
7.4.4 Transfer the concentrate to a 125 ml separatory funnel using
15 ml hexane. Rinse the flask with two 5 ml portions of hexane and add
the rinses to the funnel. Shake the combined solutions in the separatory
funnel for two minutes with 50 ml of 5 percent sodium chloride solution,
discard the aqueous layer, and proceed with Section 7.5.
7.4.5 Aqueous samples
7.4.5.1 Allow the sample to come to ambient temperature, then
mark the water meniscus on the side of the 1 L sample bottle for
later determination of the exact sample volume. Add the required
acetone diluted sample fortification solution (Section 5.8).
7.4.5.2 When the sample is judged to contain 1 percent or
more solids, the sample must be filtered through a 0.45 urn glass
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fiber filter that has been rinsed with toluene. If the suspended
solids content is too great to filter through the 0.45 urn filter,
centrifuge the sample, decant, and then filter the aqueous phase.
7.4.5.3 Combine the solids from the centrifuge bottle(s) with
the particulates on the filter and with the filter itself and proceed
with the Soxhlet extraction as specified in Sections 7.4.6.1 through
7.4.6.4. Remove and invert the Snyder column and rinse it down into
the KD apparatus with two 1 ml portions of hexane.
7.4.5.4 Pour the aqueous filtrate into a 2 L separatory
funnel. Add 60 ml methylene chloride to the sample bottle, seal
and shake for 30 seconds to rinse the inner surface. Transfer the
solvent to the separatory funnel and extract the sample by shaking
the funnel for two minutes with periodic venting.
7.4.5.5 Allow the organic layer to separate from the water
phase for a minimum of 10 minutes. If the emulsion interface between
layers is more than one third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete the phase
separation (e.g., glass stirring rod).
7.4.5.6 Collect the methylene chloride into a KD apparatus
(mounted with a 10 ml concentrator tube) by passing the sample
extracts through a filter funnel packed with a glass wool plug and
5 g anhydrous sodium sulfate.
NOTE: As an option, a rotary evaporator may be used in place of the KD apparatus
for the concentration of the extracts.
7.4.5.7 Repeat the extraction twice with fresh 60 ml portions
of methylene chloride. After the third extraction, rinse the sodium
sulfate with an additional 30 ml methylene chloride to ensure quanti-
tative transfer. Combine all extracts and the rinse in the KD
apparatus.
NOTE: A continuous liquid-liquid extractor may be used in place of a separatory
funnel when experience with a sample from a given source indicates that
a serious emulsion problem will result or an emulsion is encountered when
using a separatory funnel. Add 60 ml methylene chloride to the sample
bottle, seal, and shake for 30 seconds to rinse the inner surface.
Transfer the solvent to the extractor. Repeat the rinse of the sample
bottle with an additional 50 to 100 ml portion of methylene chloride and
add the rinse to the extractor. Add 200 to 500 ml methylene chloride to
the distilling flask, add sufficient organic-free reagent water (Section
5.1) to ensure proper operation, and extract for 24 hours. Allow to cool,
then detach the distilling flask. Dry and concentrate the extract as
described in Sections 7.4.5.6 and 7.4.5.8 through 7.4.5.10. Proceed with
Section 7.4.5.11.
7.4.5.8 Attach a Snyder column and concentrate the extract
on a water bath until the apparent volume of the liquid is 5 ml.
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Remove the KD apparatus and allow It to drain and cool for at least
10 minutes.
7.4.5.9 Remove the Snyder column, add 50 ml hexane, add the
concentrate obtained from the Soxhlet extraction of the suspended
solids (Section 7.4.5.3), if applicable, re-attach the Snyder column,
and concentrate to approximately 5 ml. Add a new boiling chip to
the KD apparatus before proceeding with the second concentration
step.
7.4.5.10 Rinse the flask and the lower joint with two 5 ml
portions of hexane and combine the rinses with the extract to give
a final volume of about 15 ml.
7.4.5.11 Determine the original sample volume by filling the
sample bottle to the mark with water and transferring the water to
a 1000 ml graduated cylinder. Record the sample volume to the
nearest 5 ml. Proceed with Section 7.5.
7.4.6 Soil/Sediment
7.4.6.1 Add 10 g anhydrous powdered sodium sulfate to the
sample portion (e.g., 10 g) and mix thoroughly with a stainless
steel spatula. After breaking up any lumps, place the soil/sodium
sulfate mixture in the Soxhlet apparatus on top of a glass wool plug
(the use of an extraction thimble is optional).
NOTE: As an option, a Soxhlet/Dean Stark extractor system may be used, with
toluene as the solvent. No sodium sulfate is added when using this option.
7.4.6.2 Add 200 to 250 ml toluene to the Soxhlet apparatus
and reflux for 16 hours. The solvent must cycle completely through
the system five times per hour.
NOTE: If the dried sample is not of free flowing consistency, more sodium sulfate
must be added.
7.4.6.3 Cool and filter the extract through a glass fiber
filter into a 500 ml round bottom flask for evaporation of the
toluene. Rinse the filter with 10 ml of toluene, and concentrate
the combined fractions to near dryness on a rotary evaporator at
50°C. Remove the flask from the water bath and allow to cool for
5 minutes.
7.4.6.4 Transfer the residue to a 125 ml separatory funnel,
using 15 ml of hexane. Rinse the flask with two additional portions
of hexane, and add the rinses to the funnel. Proceed with
Section 7.5.
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7.5 Cleanup
7.5.1 Partition
7.5.1.1 Partition the hexane extract against 40 ml of
concentrated sulfuric acid. Shake for two minutes. Remove and
discard the sulfuric acid layer (bottom). Repeat the acid washing
until no color is visible in the acid layer (perform a maximum of
four acid washings).
7.5.1.2 Omit this step for the fish sample extract. Partition
the extract against 40 ml of 5 percent (w/v) aqueous sodium chloride.
Shake for two minutes. Remove and discard the aqueous layer
(bottom).
7.5.1.3 Omit this step for the fish sample extract. Partition
the extract against 40 mL of 20 percent (w/v) aqueous potassium
hydroxide (KOH). Shake for two minutes. Remove and discard the
aqueous layer (bottom). Repeat the base washing until no color is
visible in the bottom layer (perform a maximum of four base
washings). Strong base (KOH) is known to degrade certain
PCDDs/PCDFs, so contact time must be minimized.
7.5.1.4 Partition the extract against 40 mL of 5 percent
(w/v) aqueous sodium chloride. Shake for two minutes. Remove and
discard the aqueous layer (bottom). Dry the extract by pouring it
through a filter funnel containing anhydrous sodium sulfate on a
glass wool plug, and collect it in a 50 ml round bottom flask.
Rinse the funnel with the sodium sulfate with two 15 ml portions of
hexane, add the rinses to the 50 mL flask, and concentrate the hexane
solution to near dryness on a rotary evaporator (35°C water bath),
making sure all traces of toluene (when applicable) are removed.
(Use of blowdown with an inert gas to concentrate the extract is also
permitted.)
7.5.2 Silica/Alumina Column Cleanup
7.5.2.1 Pack a gravity column (glass, 30 cm x 10.5 mm), fitted
with a Teflon™ stopcock, with silica gel as follows: Insert a glass
wool plug into the bottom of the column. Place 1 g silica gel in
the column and tap the column gently to settle the silica gel. Add
2g sodium hydroxide-impregnated silica gel, 4g sulfuric acid-
impregnated silica gel, and 2 g silica gel. Tap the column gently
after each addition. A small positive pressure (5 psi) of clean
nitrogen may be used if needed. Elute with 10 mL hexane and close
the stopcock just before exposure of the top layer of silica gel to
air. Discard the eluate. Check the column for channeling. If
channeling is observed, discard the column. Do not tap the wetted
column.
7.5.2.2 Pack a gravity column (glass, 300 mm x 10.5 mm),
fitted with a Teflon™ stopcock, with alumina as follows: Insert
a glass wool plug into the bottom of the column. Add a 4 g layer
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of sodium sulfate. Add a 4 g layer of Woelm* Super 1 neutral
alumina. Tap the top of the column gently. Woelm* Super 1 neutral
alumina need not be activated or cleaned before use, but it should
be stored in a sealed desiccator. Add a 4 g layer of anhydrous
sodium sulfate to cover the alumina. Elute with 10 ml hexane and
close the stopcock just before exposure of the sodium sulfate layer
to air. Discard the eluate. Check the column for channeling. If
channeling is observed, discard the column. Do not tap a wetted
column.
NOTE: Optionally, acidic alumina (Section 5.2.2) can be used in place of neutral
alumina.
7.5.2.3 Dissolve the residue from Section 7.5.1.4 in 2 ml
hexane and apply the hexane solution to the top of the silica gel
column. Rinse the flask with enough hexane (3-4 ml) to complete
the quantitative transfer of the sample to the surface of the silica
gel.
7.5.2.4 Elute the silica gel column with 90 ml of hexane,
concentrate the eluate on a rotary evaporator (35°C water bath) to
approximately 1 ml_, and apply the concentrate to the top of the
alumina column (Section 7.5.2.2). Rinse the rotary evaporator flask
twice with 2 ml of hexane, and add the rinses to the top of the
alumina column.
7.5.2.5 Add 20 ml hexane to the alumina column and elute
until the hexane level is just below the top of the sodium sulfate.
Do not discard the eluted hexane, but collect it in a separate flask
and store it for later use, as it may be useful in determining where
the labeled analytes are being lost if recoveries are not
satisfactory.
7.5.2.6 Add 15 ml of 60 percent methylene chloride in hexane
(v/v) to the alumina column and collect the eluate in a conical
shaped (15 ml) concentration tube. With a carefully regulated stream
of nitrogen, concentrate the 60 percent methylene chloride/hexane
fraction to about 2 ml.
7.5.3 Carbon Column Cleanup
7.5.3.1 Prepare an AX-21/Celite 545® column as follows:
Thoroughly mix 5.40 g active carbon AX-21 and 62.0 g Celite 545*
to produce an 8 percent (w/w) mixture. Activate the mixture at
130°C for 6 hours and store it in a desiccator.
7.5.3.2 Cut off both ends of a 10 ml disposable serological
pipet to give a 10 cm long column. Fire polish both ends and flare,
if desired. Insert a glass wool plug at one end, then pack the
column with enough Celite 545* to form a 1 cm plug, add 1 g of the
AX-21/Celite 545* mixture, top with additional Celite 545* (enough
for a 1 cm plug), and cap the packing with another glass wool plug.
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NOTE: Each new batch of AX-21/Celite 545* must be checked as follows: Add 50 /xL
of the continuing calibration solution to 950 nl hexane. Take this
solution through the carbon column cleanup step, concentrate to 50 /xL and
analyze. If the recovery of any of the analytes is <80 percent, discard
this batch of AX-21/Celite 545*.
7.5.3.3 Rinse the AX-21/Celite 545* column with 5 ml of
toluene, followed by 2 ml of 75:20:5 (v/v) methylene
chloride/methanol/toluene, 1 ml of 1:1 (v/v) cyclohexane/methylene
chloride, and 5 ml hexane. The flow rate should be less than
0.5 mL/min. Discard the rinses. While the column is still wet with
hexane, add the sample concentrate (Section 7.5.2.6) to the top of
the column. Rinse the concentrator tube (which contained the sample
concentrate) twice with 1 ml hexane, and add the rinses to the top
of the column.
7.5.3.4 Elute the column sequentially with two 2 ml portions
of hexane, 2 ml cyclohexane/methylene chloride (50:50, v/v), and 2 ml
methylene chloride/methanol/toluene (75:20:5, v/v). Combine these
eluates; this combined fraction may be used as a check on column
efficiency.
7.5.3.5 Turn the column upside down and elute the PCDD/PCDF
fraction with 20 ml toluene. Verify that no carbon fines are present
in the eluate. If carbon fines are present in the eluate, filter
the eluate through a glass fiber filter (0.45 /im) and rinse the
filter with 2 ml toluene. Add the rinse to the eluate.
7.5.3.6 Concentrate the toluene fraction to about 1 mL on a
rotary evaporator by using a water bath at 50°C. Carefully transfer
the concentrate into a 1 ml minivial and, again at elevated
temperature (50°C), reduce the volume to about 100 pi using a stream
of nitrogen and a sand bath. Rinse the rotary evaporator flask three
times with 300 pi of a solution of 1 percent toluene in methylene
chloride, and add the rinses to the concentrate. Add 10 pi of the
nonane recovery standard solution for soil, sediment, water, fish,
paper pulp and adipose tissue samples, or 50 pi of the recovery
standard solution for sludge, still bottom and fly ash samples.
Store the sample at room temperature in the dark.
7.6 Chromatographic/Mass Spectrometric Conditions and Data Acquisition
Parameters
7.6.1 Gas Chromatograph
Column coating: DB-5
Film thickness: 0.25 Mm
Column dimension: 60 m x 0.32 mm
Injector temperature: 270°C
Splitless valve time: 45 s
Interface temperature: Function of the final temperature
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Temperature program:
Stage Init. Init. Temp. Final Final
Temp. Hold Time Ramp Temp. Hold
(°C) (min) (°C/min) (°C) Time (min)
1 200 2 5 220 16
2 5 235 7
3 5 330 5
Total time: 60 min
7.6.2 Mass Spectrometer
7.6.2.1 The mass spectrometer must be operated in a selected
ion monitoring (SIM) mode with a total cycle time (including the
voltage reset time) of one second or less (Section 7.6.3.1). At a
minimum, the ions listed in Table 6 for each of the five SIM
descriptors must be monitored. Note that with the exception of the
last descriptor (OCDD/OCDF), all descriptors contain 10 ions. The
selection (Table 6) of the molecular ions M and M+2 for 13C-HxCDF and
13C-HpCDF rather than M+2 and M+4 (for consistency) was made to
eliminate, even under high-resolution mass spectrometric conditions,
interferences occurring in these two ion channels for samples
containing high levels of native HxCDDs and HpCDDs. It is important
to maintain the same set of ions for both calibration and sample
extract analyses. The selection of the lock-mass ion is left to the
performing laboratory.
Note: At the option of the analyst, the tetra- and pentachlorinated dioxins and
furans can be combined into a single descriptor.
7.6.2.2 The recommended mass spectrometer tuning conditions
are based on the groups of monitored ions shown in Table 6. By using
a PFK molecular leak, tune the instrument to meet the minimum
required resolving power of 10,000 (10 percent valley) at m/z
304.9824 (PFK) or any other reference signal close to m/z 303.9016
(from TCDF). By using peak matching conditions and the
aforementioned PFK reference peak, verify that the exact mass of m/z
380.9760 (PFK) is within 5 ppm of the required value. Note that the
selection of the low- and high-mass ions must be such that they
provide the largest voltage jump performed in any of the five mass
descriptors (Table 6).
7.6.3 Data Acquisition
7.6.3.1 The total cycle time for data acquisition must be <
1 second. The total cycle time includes the sum of all the dwell
times and voltage reset times.
7.6.3.2 Acquire SIM data for all the ions listed in the five
descriptors of Table 6.
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7.7 Calibration
7.7.1 Initial Calibration - Initial calibration is required before
any samples are analyzed for PCDDs and PCDFs. Initial calibration is also
required if any routine calibration (Section 7.7.3) does not meet the
required criteria listed in Section 9.4.
7.7.1.1 All five high-resolution concentration calibration
solutions listed in Table 5 must be used for the initial calibration.
7.7.1.2 Tune the instrument with PFK as described in
Section 7.6.2.2.
7.7.1.3 Inject 2 pi of the GC column performance check
solution (Section 5.7) and acquire SIM mass spectral data as
described earlier in Section 8.1. The total cycle time must be < 1
second. The laboratory must not perform any further analysis until
it is demonstrated and documented that the criterion listed in
Section 8.1.2 was met.
7.7.1.4 By using the same GC (Section 7.6.1) and MS
(Section 7.6.2) conditions that produced acceptable results with
the column performance check solution, analyze a 2 /zL portion of
each of the five concentration calibration solutions once with the
following mass spectrometer operating parameters.
7.7.1.4.1 The ratio of integrated ion current for the
ions appearing in Table 8 (homologous series quantitation ions)
must be within the indicated control limits (set for each
homologous series).
7.7.1.4.2 The ratio of integrated ion current for the
ions belonging to the carbon-labeled internal and recovery
standards must be within the control limits stipulated in
Table 8.
NOTE: Sections 7.7.1.4.1 and 7.7.1.4.2 require that 17 ion ratios from Section
7.7.1.4.1 and 11 ion ratios from Section 7.7.1.4.2 be within the specified
control limits simultaneously in one run. It is the laboratory's
responsibility to take corrective action if the ion abundance ratios are
outside the limits.
7.7.1.4.3 For each SICP and for each GC signal
corresponding to the elution of a target analyte and of its
labeled standards, the signal-to-noise ratio (S/N) must be
better than or equal to 2.5. Measurement of S/N is required
for any GC peak that has an apparent S/N of less than 5:1.
The result of the calculation must appear on the SICP above
the GC peak in question.
7.7.1.4.4 Referring to Table 9, calculate the 17
relative response factors (RRF) for unlabeled target analytes
[RRF(n); n = 1 to 17] relative to their appropriate internal
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standards (Table 5) and the nine RRFs for the labeled 13C12
internal standards [RRF(m); m = 18 to 26)] relative to the two
recovery standards according to the following formulae:
RRF(n)
X Qls
RRF(m) =
A,s x Qr
Q,s
where:
A,, =
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for unlabeled
PCDDs/PCDFs,
Ais = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the labeled
internal standards,
Ars = sum of the integrated "-ion abundances of the
quantitation ions (Tables 6 and 9) for the labeled
recovery standards,
QIS = quantity of the internal standard injected (pg),
Qrs = quantity of the recovery standard injected (pg),
and
Qx = quantity of the unlabeled PCDD/PCDF analyte
injected (pg).
The RRF(n) and RRF(m) are dimensionless quantities; the units
used to express Qls, Qrs and Qx must be the same.
7.7.1.4.5 Calculate the RRF and their respective
percent relative standard deviations (%RSD) for the five
calibration solutions:
RRF(n) = 1/5 Z RRFj(n)
where n represents a particular PCDD/PCDF (2,3,7,8-substituted)
congener (n = 1 to 17; Table 9), and j is the injection number
(or calibration solution number; j = 1 to 5).
7.7.1.4.6 The relative response factors to be used for
the determination of the concentration of total isomers in a
homologous series (Table 9) are calculated as follows:
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7.7.1.4.6.1 For congeners that belong to a
homologous series containing only one Isomer (e.g.,
OCDD and OCDF) or only one 2,3,7,8-substituted Isomer
(Table 4; TCDD, PeCDD, HpCDD, and TCDF), the mean RRF
used will be the same as the mean RRF determined In
Section 7.7.1.4.5.
NOTE: The calibration solutions do not contain 13C12-OCDF as an internal standard.
This is because a minimum resolving power of 12,000 is required to resolve
the [M+6]+ ion of 13C12-OCDF from the [M+2]+ ion of OCDD (and [M+4]+ from
13C12-OCDF with [M]+ of OCDD). Therefore, the RRF for OCDF is calculated
relative to 13C12-OCDD.
7.7.1.4.6.2 For congeners that belong to a
homologous series containing more than one
2,3,7,8-substituted isomer (Table 4), the mean RRF used
for those homologous series will be the mean of the RRFs
calculated for all individual 2,3,7,8-substituted
congeners using the equation below:
1 t
RRF(k) = - Z RRFn
t n=l
where:
k = 27 to 30 (Table 9), with 27 = PeCDF; 28 =
HxCDF; 29 = HxCDD; and 30 = HpCDF,
t = total number of 2,3,7,8-substituted isomers
present in the calibration solutions (Table
5) for each homologous series (e.g., two for
PeCDF, four for HxCDF, three for HxCDD, two
for HpCDF).
NOTE: Presumably, the HRGC/HRMS response factors of different isomers within
a homologous series are different. However, this analytical protocol
will make the assumption that the HRGC/HRMS responses of all isomers in
a homologous series that do not have the 2,3,7,8-substitution pattern
are the same as the responses of one or more of the 2,3,7,8-substituted
isomer(s) in that homologous series.
7.7.1.4.7 Relative response factors [RRF(m)] to be
used for the determination of the percent recoveries for the
nine internal standards are calculated as follows:
Alsm x Qrs
RRF(m)
Q,sm x Ars
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1 5
RRF(m) = - Z RRFj(m),
5 j-1
where:
m
Ars
18 to 26 (congener type) and j = 1 to 5
(injection number),
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for a given
internal standard (m = 18 to 26),
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
appropriate recovery standard (see Table 5,
footnotes),
Qrs, Qlsm = quantities of, respectively, the recovery
standard (rs) and a particular internal
standard (is = m) injected (pg),
RRF(m) = relative response factor of a particular
internal standard (m) relative to an
appropriate recovery standard, as determined
from one injection, and
RRF(m) = calculated mean relative response factor of
a particular internal standard (m) relative
to an appropriate recovery standard, as
determined from the five initial calibration
injections (j).
7.7.2 Criteria for Acceptable Calibration - The criteria listed
below for acceptable calibration must be met before the analysis is
performed.
7.7.2.1 The percent relative standard deviations for the mean
response factors [RRF(n) and RRF(m)] from the 17 unlabeled standards
must not exceed ± 20 percent, and those for the nine labeled
reference compounds must not exceed ± 30 percent.
7.7.2.2 The S/N for the GC signals present in every SICP
(including the ones for the labeled standards) must be > 10.
7.7.2.3 The isotopic ratios (Table 8) must be within the
specified control limits.
NOTE: If the criterion for acceptable calibration listed in Section 7.7.2.1 is
met, the analyte specific RRF can then be considered independent of the
analyte quantity for the calibration concentration range. The mean RRFs
will be used for all calculations until the routine calibration criteria
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(Section 7.7.4) are no longer met. At such time, new mean RRFs will be
calculated from a new set of injections of the calibration solutions.
7.7.3 Routine Calibration (Continuing Calibration Check) - Routine
calibrations must be performed at the beginning of a 12 hour period after
successful mass resolution and GC resolution performance checks. A routine
calibration is also required at the end of a 12 hour shift.
7.7.3.1 Inject 2 pi of the concentration calibration solution
HRCC-3 standard (Table 5). By using the same HRGC/HRMS conditions
as used in Sections 7.6.1 and 7.6.2, determine and document an
acceptable calibration as provided in Section 7.7.4.
7.7.4 Criteria for Acceptable Routine Calibration - The following
criteria must be met before further analysis is performed.
7.7.4.1 The measured RRFs [RRF(n) for the unlabeled standards]
obtained during the routine calibration runs must be within + 20
percent of the mean values established during the initial calibration
(Section 7.7.1.4.5).
7.7.4.2 The measured RRFs [RRF(m) for the labeled standards]
obtained during the routine calibration runs must be within
+ 30 percent of the mean values established during the initial
calibration (Section 7.7.1.4.7).
7.7.4.3 The ion-abundance ratios (Table 8) must be within the
allowed control limits.
7.7.4.4 If either one of the criteria in Sections 7.7.4.1
and 1.1 A.I is not satisfied, repeat one more time. If these
criteria are still not satisfied, the entire routine calibration
process (Section 7.7.1) must be reviewed. It is realized that it
may not always be possible to achieve all RRF criteria. For example,
it has occurred that the RRF criteria for 13C12-HpCDD and 13C12-OCDD
were not met, however, the RRF values for the corresponding unlabeled
compounds were routinely within the criteria established in the
method. In these cases, 24 of the 26 RRF parameters have met the
QC criteria, and the data quality for the unlabeled HpCDD and OCDD
values were not compromised as a result of the calibration event.
In these situations, the analyst must assess the effect on overall
data quality as required for the data quality objectives and decide
on appropriate action. Corrective action would be in order, for
example, if the compounds for which the RRF criteria were not met
included both the unlabeled and the corresponding internal standard
compounds. If the ion-abundance ratio criterion (Section 7.7.4.3)
is not satisfied, refer to the note in Section 7.7.1.4.2 for
resolution.
NOTE: An initial calibration must be carried out whenever the HRCC-3, the sample
fortification or the recovery standard solution is replaced by a new
solution from a different lot.
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7.8 Analysis
7.8.1 Remove the sample extract (from Section 7.5.3.6) or blank from
storage. With a stream of dry, purified nitrogen, reduce the extract
volume to 10 /iL to 50 /xL.
Note: A final volume of 20 /iL or more should be used whenever possible. A 10 juL
final volume is difficult to handle, and injection of 2 nl out of 10 juL
leaves little sample for confirmations and repeat injections, and for
archiving.
7.8.2 Inject a 2 ML aliquot of the extract into the GC, operated
under the conditions that have been established to produce acceptable
results with the performance check solution (Sections 7.6.1 and 7.6.2).
7.8.3 Acquire SIM data according to Sections 7.6.2 and 7.6.3. Use
the same acquisition and mass spectrometer operating conditions previously
used to determine the relative response factors (Sections 7.7.1.4.4 through
7.7.1.4.7). Ions characteristic for polychlorinated diphenyl ethers are
included in the descriptors listed in Table 6.
NOTE: The acquisition period must at least encompass the PCDD/PCDF overall
retention time window previously determined (Section 8.1). Selected ion
current profiles (SICP) for the lock-mass ions (one per mass descriptor)
must also be recorded and included in the data package. These SICPs must
be true representations of the evolution of the lock-mass ions amplitudes
during the HRGC/HRMS run (see Section 8.2.2 for the proper level of
reference compound to be metered into the ion chamber.) The analyst may
be required to monitor a PFK ion, not as a lock mass, but as a regular ion,
in order to meet this requirement. It is recommended to examine the lock-
mass ion SICP for obvious basic sensitivity and stability changes of the
instrument during the GC/MS run that could affect the measurements [Tondeur
et al., 1984, 1987]. Report any discrepancies in the case narrative.
7.8.4 Identification Criteria - For a gas chromatographic peak to
be identified as a PCDD or PCDF, it must meet all of the following
criteria:
7.8.4.1 Retention Times
7.8.4.1.1 For 2,3,7,8-substituted congeners, which
have an isotopically labeled internal or recovery standard
present in the sample extract (this represents a total of 10
congeners including OCDD; Tables 2 and 3), the retention time
(RRT; at maximum peak height) of the sample components (i.e.,
the two ions used for quantitation purposes listed in Table
6) must be within -1 to +3 seconds of the isotopically labelled
standard.
7.8.4.1.2 For 2,3,7,8-substituted compounds that do
not have an isotopically labeled internal standard present in
the sample extract (this represents a total of six congeners;
Table 3), the retention time must fall within 0.005 retention
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time units of the relative retention times measured in the
routine calibration. Identification of OCDF is based on its
retention time relative to 13C12-OCDD as determined from the
daily routine calibration results.
7.8.4.1.3 For non-2,3,7,8-substituted compounds (tetra
through octa; totaling 119 congeners), the retention time must
be within the corresponding homologous retention time windows
established by analyzing the column performance check solution
(Section 8.1.3).
7.8.4.1.4 The ion current responses for both ions used
for quantitative purposes (e.g., for TCDDs: m/z 319.8965 and
321.8936) must reach maximum simultaneously (+ 2 seconds).
7.8.4.1.5 The ion current responses for both ions used
for the labeled standards (e.g., for 13C12-TCDD: m/z 331.9368
and m/z 333.9339) must reach maximum simultaneously (± 2
seconds).
NOTE: The analyst is required to verify the presence of 1,2,8,9-TCDD and
1,3,4,6,8-PeCDF (Section 8.1.3) in the SICPs of the daily performance
checks. Should either one compound be missing, the analyst is required
to take corrective action as it may indicate a potential problem with the
ability to detect all the PCDDs/PCDFs.
7.8.4.2 Ion Abundance Ratios
7.8.4.2.1 The integrated ion current for the two ions
used for quantitation purposes must have a ratio between the
lower and upper limits established for the homologous series
to which the peak is assigned. See Sections 7.7.1.4.1 and
7.7.1.4.2 and Table 8 for details.
7.8.4.3 Signal-to-Noise Ratio
7.8.4.3.1 All ion current intensities must be > 2.5
times noise level for positive identification of a PCDD/PCDF
compound or a group of coeluting isomers. Figure 6 describes
the procedure to be followed for the determination of the S/N.
7.8.4.4 Polychlorinated Diphenyl Ether Interferences
7.8.4.4.1 In addition to the above criteria, the
identification of a GC peak as a PCDF can only be made if no
signal having a S/N > 2.5 is detected, at the same retention
time (+ 2 seconds), in the corresponding polychlorinated
diphenyl ether (PCDPE, Table 6) channel.
7.9 Calculations
7.9.1 For gas chromatographic peaks that have met the criteria
outlined in Sections 7.8.4.1.1 through 7.8.4.3.1, calculate the concen-
tration of the PCDD or PCDF compounds using the formula:
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AX x Qls
Cx = ==—
Als x W x RRF(n)
where:
Cx = concentration of unlabeled PCDD/PCDF congeners (or group of
coeluting isomers within an homologous series) in pg/g,
A,, = sum of the integrated ion abundances of the quantitation ions
(Table 6) for unlabeled PCDDs/PCDFs,
Als = sum of the integrated ion abundances of the quantitation ions
(Table 6) for the labeled internal standards,
Qls = quantity, in pg, of the internal standard added to the sample
before extraction,
W = weight, in g, of the sample (solid or liquid), and
RRF= calculated mean relative response factor for the analyte
[RRF(n) with n = 1 to 17; Section 7.7.1.4.5].
If the analyte is identified as one of the 2,3,7,8-substituted PCDDs or
PCDFs, RRF(n) is the value calculated using the equation in
Section_7.7.1.4.5. However, if it is a non-2,3,7,8-substituted congener,
the RRF(k) value is the one calculated using the equation in
Section 7.7.1.4.6.2. [RRF(k) with k = 27 to 30].
7.9.2 Calculate the percent recovery of the nine internal standards
measured in the sample extract, using the formula:
Ais x Qrs
Internal standard percent recovery = ——x 100
Qis x Ars x RRF(m)
where:
Als = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled internal standard,
Ars = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled recovery standard; the
selection of the recovery standard depends on the type of
congeners (see Table 5, footnotes),
Qls = quantity, in pg, of the internal standard added to the
sample before extraction,
Qrs = quantity, in pg, of the recovery standard added to the
cleaned-up sample residue before HRGC/HRMS analysis, and
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RRF(m) = calculated mean relative response factor for the labeled
internal standard relative to the appropriate (see Table
5, footnotes) recovery standard._This represents the mean
obtained in Section 7.7.1.4.7 [RRF(m) with m = 18 to 26].
NOTE: For human adipose tissue, adjust the percent recoveries by adding
1 percent to the calculated value to compensate for the 1 percent of the
extract diverted for the lipid determination.
7.9.3 If the concentration in the final extract of any of the fifteen
2,3,7,8-substituted PCDD/PCDF compounds (Table 3) exceeds the upper method
calibration limits (MCL) listed in Table 1 (e.g., 200 pg//iL for TCDD in
soil), the linear range of response versus concentration may have been
exceeded, and a second analysis of the sample (using a one tenth aliquot)
should be undertaken. The volumes of the internal and recovery standard
solutions should remain the same as described for the sample preparation
(Sections 11.1 to 11.9.3). For the other congeners (including OCDD),
however, report the measured concentration and indicate that the value
exceeds the MCL.
7.9.4 The total concentration for each homologous series of PCDD
and PCDF is calculated by summing up the concentrations of all positively
identified isomers of each homologous series. Therefore, the total should
also include the 2,3,7,8-substituted congeners. The total number of GC
signals included in the homologous total concentration value must be
specified in the report.
7.9.5 Sample Specific Estimated Detection Limit - The sample specific
estimated detection limit (EDL) is the concentration of a given analyte
required to produce a signal with a peak height of at least 2.5 times the
background signal level. An EDL is calculated for each
2,3,7,8-substituted congener that is not identified, regardless of whether
or not other non-2,3,7,8-substituted isomers are present. Two methods of
calculation can be used, as follows, depending on the type of response
produced during the analysis of a particular sample.
7.9.5.1 Samples giving a response for both quantitation ions
(Tables 6 and 9) that is less than 2.5 times the background level.
7.9.5.1.1 Use the expression for EDL (specific
2,3,7,8-substituted PCDD/PCDF) below to calculate an EDL for
each absent 2,3,7,8-substituted PCDD/PCDF (i.e., S/N < 2.5).
The background level is determined by measuring the range of
the noise (peak to peak) for the two quantitation ions (Table
6) of a particular 2,3,7,8-substituted isomer within an
homologous series, in the region of the SICP trace
corresponding to the elution of the internal standard (if the
congener possesses an internal standard) or in the region of
the SICP where the congener is expected to elute by comparison
with the routine calibration data (for those congeners that
do not have a 13C-labeled standard), multiplying that noise
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height by 2.5, and relating the product to an estimated
concentration that would produce that peak height.
Use the formula:
2.5 x A, x Qls
EDL (specific 2,3,7,8-subst. PCDD/PCDF)
where:
A,s x W x RRF(n)
EDL = estimated detection limit for homologous
2,3,7,8-substituted PCDDs/PCDFs.
Ax, Als, W, RRF(n), and Qls retain the same meanings as
defined in Section 7.9.1.
7.9.5.2 Samples characterized by a response above the
background level with a S/N of at least 2.5 for both quantitation
ions (Tables 6 and 9).
7.9.5.2.1 When the response of a signal having the
same retention time as a 2,3,7,8-substituted congener has a
S/N in excess of 2.5 and does not meet any of the other
qualitative identification criteria listed in Section 7.8.4,
calculate the "Estimated Maximum Possible Concentration" (EMPC)
according to the expression shown in Section 7.9.1, except that
Ax in Section 7.9.1 should represent the sum of the area under
the smaller peak and of the other peak area calculated using
the theoretical chlorine isotope ratio.
7.9.6 The relative percent difference (RPD) is calculated as follows:
I S1 - S2 |
RPD = - x 100
S1 and S2 represent sample and duplicate sample results.
7.9.7 The 2,3,7,8-TCDD toxicity equivalents (TE) of PCDDs and PCDFs
present in the sample are calculated, if requested by the data user,
according to the method recommended by the Chlorinated Dioxins Workgroup
(CDWG) of the EPA and the Center for Disease Control (CDC). This method
assigns a 2,3,7,8-TCDD toxicity equivalency factor (TEF) to each of the
fifteen 2,3,7,8-substituted PCDDs and PCDFs (Table 3) and to OCDD and
OCDF, as shown in Table 10. The 2,3,7,8-TCDD equivalent of the PCDDs and
PCDFs present in the sample is calculated by summing the TEF times their
concentration for each of the compounds or groups of compounds listed in
Table 10. The exclusion of other homologous series such as mono-, di-,
and tri- chlorinated dibenzodioxins and dibenzofurans does not mean that
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they are non-toxic. However, their toxicity, as known at this time, is
much lower than the toxicity of the compounds listed in Table 10. The
above procedure for calculating the 2,3,7,8-TCDD toxicity equivalents is
not claimed by the CDWG to be based on a thoroughly established scientific
foundation. The procedure, rather, represents a "consensus recommendation
on science policy". Since the procedure may be changed in the future,
reporting requirements for PCDD and PCDF data would still include the
reporting of the analyte concentrations of the PCDD/PCDF congener as
calculated in Sections 7.9.1 and 7.9.4.
7.9.7.1 Two GC Column TEF Determination
7.9.7.1.1 The concentration of 2,3,7,8-TCDD (see note
below), is calculated from the analysis of the sample extract
on the 60 m DB-5 fused silica capillary column. The
experimental conditions remain the same as the conditions
described previously in Section 7.8, and the calculations are
performed as outlined in Section 7.9. The chromatographic'
separation between the 2,3,7,8-TCDD and its close eluters
(1,2,3,7/1,2,3,8-TCDD and 1,2,3,9-TCDD) must be equal or less
than 25 percent valley.
7.9.7.1.2 The concentration of the 2,3,7,8-TCDF is
obtained from the analysis of the sample extract on the 30 m
DB-225 fused silica capillary column. However, the GC/MS
conditions must be altered so that: (1) only the first three
descriptors (i.e., tetra-, penta-, and hexachlorinated
congeners) of Table 6 are used; and (2) the switching time
between descriptor 2 (pentachlorinated congeners) and
descriptor 3 (hexachlorinated congeners) takes place following
the elution of 13C12-l,2,3,7,8-PeCDD. The concentration
calculations are performed as outlined in Section 7.9. The
chromatographic separation between the 2,3,7,8-TCDF and its
close eluters (2,3,4,7-TCDF and 1,2,3,9-TCDF) must be equal
or less than 25 percent valley.
NOTE: The confirmation and quantitation of 2,3,7,8-TCDD (Section 7.9.7.1.1) may
be accomplished on the SP-2330 GC column instead of the DB-5 column,
provided the criteria listed in Section 8.1.2 are met and the requirements
described in Section 17.2.2 are followed.
7.9.7.1.3 For a gas chromatographic peak to be
identified as a 2,3,7,8-substituted PCDD/PCDF congener, it
must meet the ion abundance and signal-to-noise ratio criteria
listed in Sections 7.8.4.2 and 7.8.4.3, respectively. In
addition, the retention time identification criterion described
in Section 7.8.4.1.1 applies here for congeners for which a
carbon-labeled analogue is available in the sample extract.
However, the relative retention time (RRT) of the
2,3,7,8-substituted congeners for which no carbon-labeled
analogues are available must fall within 0.006 units of the
carbon-labeled standard RRT. Experimentally, this is
accomplished by using the attributions described in Table 11
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and the results from the routine calibration run on the SP-2330
column.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control (QC) procedures.
Quality control to validate sample extraction is covered in Method 3500. If
extract cleanup was performed, follow the QC in Method 3600 and in the specific
cleanup method.
8.2 System Performance Criteria - System performance criteria are
presented below. The laboratory may use the recommended GC column described in
Section 4.2. It must be documented that all applicable system performance
criteria (specified in Sections 8.2.1 and 8.2.2) were met before analysis of any
sample is performed. Section 7.6 provides recommended GC conditions that can
be used to satisfy the required criteria. Figure 3 provides a typical 12 hour
analysis sequence, whereby the response factors and mass spectrometer resolving
power checks must be performed at the beginning and the end of each 12 hour
period of operation. A GC column performance check is only required at the
beginning of each 12 hour period during which samples are analyzed. An HRGC/HRMS
method blank run is required between a calibration run and the first sample run.
The same method blank extract may thus be analyzed more than once if the number
of samples within a batch requires more than 12 hours of analyses.
8.2.1 GC Column Performance
8.2.1.1 Inject 2 nl (Section 4.1.1) of the column performance
check solution (Section 5.7) and acquire selected ion monitoring
(SIM) data as described in Section 7.6.2 within a total cycle time
of < 1 second (Section 7.6.3.1).
8.2.1.2 The chromatographic separation between 2,3,7,8-TCDD
and the peaks representing any other unlabeled TCDD isomers must be
resolved with a valley of < 25 percent (Figure 4), where:
Valley percent = (x/y) (100)
x = measured as in Figure 4 from the 2,3,7,8-closest TCDD
eluting isomer, and
y = the peak height of 2,3,7,8-TCDD.
It is the responsibility of the laboratory to verify the conditions
suitable for the appropriate resolution of 2,3,7,8-TCDD from all
other TCDD isomers. The GC column performance check solution also
contains the known first and last PCDD/PCDF eluters under the
conditions specified in this protocol. Their retention times are
used to determine the eight homologue retention time windows that
are used for qualitative (Section 7.8.4.1) and quantitative purposes.
All peaks (that includes 13C12-2,3,7,8-TCDD) should be labeled and
identified on the chromatograms. Furthermore, all first eluters of
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a homologous series should be labeled with the letter F, and all last
eluters of a homologous series should be labeled with the letter L
(Figure 4 shows an example of peak labeling for TCDD isomers). Any
individual selected ion current profile (SICP) (for the tetras, this
would be the SICP for m/z 322 and m/z 304) or the reconstructed
homologue ion current (for the tetras, this would correspond to m/z
320 + m/z 322 + m/z 304 + m/z 306) constitutes an acceptable form
of data presentation. An SICP for the labeled compounds (e.g., m/z
334 for labeled TCDD) is also required.
8.2.1.3 The retention times for the switching of SIM ions
characteristic of one homologous series to the next higher homologous
series must be indicated in the SICP. Accurate switching at the
appropriate times is absolutely necessary for accurate monitoring
of these compounds. Allowable tolerance on the daily verification
with the GC performance check solution should be better than 10
seconds for the absolute retention times of all the components of
the mixture. Particular caution should be exercised for the
switching time between the last tetrachlorinated congener (i.e.,
1,2,8,9-TCDD) and the first pentachlorinated congener (i.e.,
1,3,4,6,8-PeCDF), as these two compounds elute within 15 seconds of
each other on the 60 m DB-5 column. A laboratory with a GC/MS system
that is not capable of detecting both congeners (1,2,8,9-TCDD and
1,3,4,6,8-PeCDF) within one analysis must take corrective action.
If the recommended column is not used, then the first and last
eluting isomer of each homologue must be determined experimentally
on the column which is used, and the appropriate isomers must then
be used for window definition and switching times.
8.2.2 Mass Spectrometer Performance
8.2.2.1 The mass spectrometer must be operated in the electron
ionization mode. A static resolving power of at least 10,000 (10
percent valley definition) must be demonstrated at appropriate masses
before any analysis is performed (Section 7.8). Static resolving
power checks must be performed at the beginning and at the end of
each 12 hour period of operation. However, it is recommended that
a check of the static resolution be made and documented before and
after each analysis. Corrective action must be implemented whenever
the resolving power does not meet the requirement.
8.2.2.2 Chromatography time for PCDDs and PCDFs exceeds the
long term mass stability of the mass spectrometer. Because the
instrument is operated in the high-resolution mode, mass drifts of
a few ppm (e.g., 5 ppm in mass) can have serious adverse effects on
instrument performance. Therefore, a mass drift correction is
mandatory. To that effect, it is recommended to select a lock-mass
ion from the reference compound (PFK is recommended) used for tuning
the mass spectrometer. The selection of the lock-mass ion is
dependent on the masses of the ions monitored within each descriptor.
Table 6 offers some suggestions for the lock-mass ions. However,
an acceptable lock-mass ion at any mass between the lightest and
heaviest ion in each descriptor can be used to monitor and correct
mass drifts. The level of the reference compound (PFK) metered into
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the ion chamber during HRGC/HRMS analyses should be adjusted so that
the amplitude of the most intense selected lock-mass ion signal
(regardless of the descriptor number) does not exceed 10 percent of
the full scale deflection for a given set of detector parameters.
Under those conditions, sensitivity changes that might occur during
the analysis can be more effectively monitored.
NOTE: Excessive PFK (or any other reference substance) may cause noise problems
and contamination of the ion source resulting in an increase in downtime
for source cleaning.
8.2.2.3 Documentation of the instrument resolving power must
then be accomplished by recording the peak profile of the high-mass
reference signal (m/z 380.9760) obtained during the above peak
matching experiment by using the low-mass PFK ion at m/z 304.9824
as a reference. The minimum resolving power of 10,000 must be
demonstrated on the high-mass ion while it is transmitted at a lower
accelerating voltage than the low-mass reference ion, which is
transmitted at full sensitivity. The format of the peak profile
representation (Figure 5) must allow manual determination of the
resolution, i.e., the horizontal axis must be a calibrated mass scale
(amu or ppm per division). The result of the peak width measurement
(performed at 5 percent of the maximum, which corresponds to the 10
percent valley definition) must appear on the hard copy and cannot
exceed 100 ppm at m/z 380.9760 (or 0.038 amu at that particular
mass).
8.3 Quality Control Samples
8.3.1 Performance Evaluation Samples - Included among the samples
in all batches may be samples (blind or double blind) containing known
amounts of unlabeled 2,3,7,8-substituted PCDDs/PCDFs or other PCDD/PCDF
congeners.
8.3.2 Performance Check Solutions
8.3.2.1 At the beginning of each 12 hour period during which
samples are to be analyzed, an aliquot of the 1) GC column
performance check solution and 2) high-resolution concentration
calibration solution No. 3 (HRCC-3; see Table 5) shall be analyzed
to demonstrate adequate GC resolution and sensitivity, response
factor reproducibility, and mass range calibration, and to establish
the PCDD/PCDF retention time windows. A mass resolution check shall
also be performed to demonstrate adequate mass resolution using an
appropriate reference compound (PFK is recommended). If the required
criteria are not met, remedial action must be taken before
any samples are analyzed.
8.3.2.2 To validate positive sample data, the routine or
continuing calibration (HRCC-3; Table 5) and the mass resolution
check must be performed also at the end of each 12 hour period during
which samples are analyzed. Furthermore, an HRGC/HRMS method blank
run must be recorded following a calibration run and the first sample
run.
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8.3.2.2.1 If the laboratory operates only during one
period (shift) each day of 12 hours or less, the GC performance
check solution must be analyzed only once (at the beginning
of the period) to validate the data acquired during the period.
However, the mass resolution and continuing calibration checks
must be performed at the beginning as well as at the end of
the period.
8.3.2.2.2 If the laboratory operates during consecutive
12 hour periods (shifts), analysis of the GC performance check
solution must be performed at the beginning of each 12 hour
period. The mass resolution and continuing calibration checks
from the previous period can be used for the beginning of the
next period.
8.3.2.3 Results of at least one analysis of the GC column
performance check solution and of two mass resolution and continuing
calibration checks must be reported with the sample data collected
during a 12 hour period.
8.3.2.4 Deviations from criteria specified for the GC
performance check or for the mass resolution check invalidate all
positive sample data collected between analyses of the performance
check solution, and the extracts from those positive samples shall
be reanalyzed.
If the routine calibration run fails at the beginning of a 12 hour
shift, the instructions in Section 7.7.4.4 must be followed. If
the continuing calibration check performed at the end of a 12 hour
period fails by no more than 25 percent RPD for the 17 unlabelled
compounds and 35 percent RPD for the 9 labelled reference compounds,
use the mean RRFs from the two daily routine calibration runs to
compute the analyte concentrations, instead of the RRFs obtained from
the initial calibration. A new initial calibration (new RRFs) is
required immediately (within two hours) following the analysis of
the samples, whenever the RPD from the end-of-shift routine
calibration exceeds 25 percent or 35 percent, respectively. Failure
to perform a new initial calibration immediately following the
analysis of the samples will automatically require reanalysis of all
positive sample extracts analyzed before the failed end-of-shift
continuing calibration check.
8.3.3 The GC column performance check mixture, high-resolution
concentration calibration solutions, and the sample fortification solutions
may be obtained from the EMSL-CIN. However, if not available from the
EMSL-CIN, standards can be obtained from other sources, and solutions can
be prepared in the laboratory. Concentrations of all solutions containing
2,3,7,8-substituted PCDDs/PCDFs, which are not obtained from the EMSL-CIN,
must be verified by comparison with the EPA standard solutions that are
available from the EMSL-CIN.
8.3.4 Field Blanks - Each batch of samples usually contains a field
blank sample of uncontaminated soil, sediment or water that is to be
fortified before analysis according to Section 8.3.4.1. In addition to
8290 - 38 Revision 0
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this field blank, a batch of samples may include a rinsate, which is a
portion of the solvent (usually trichloroethylene) that was used to rinse
sampling equipment. The rinsate is analyzed to assure that the samples
were not contaminated by the sampling equipment.
8.3.4.1 Fortified Field Blank
8.3.4.1.1 Weigh a 10 g portion or use 1 L (for aqueous
samples) of the specified field blank sample and add 100 ML
of the solution containing the nine internal standards
(Table 2) diluted with 1.0 ml acetone (Section 7.1).
8.3.4.1.2 Extract by using the procedures beginning
in Sections 7.4.5 or 7.4.6, as applicable, add 10 ML of the
recovery standard solution (Section 7.5.3.6) and analyze a
2 ML aliquot of the concentrated extract.
8.3.4.1.3 Calculate the concentration (Section 7.9.1)
of 2,3,7,8-substituted PCDDs/PCDFs and the percent recovery
of the internal standards (Section 7.9.2).
8.3.4.1.4 Extract and analyze a ne"w simulated fortified
field blank whenever new lots of solvents or reagents are used
for sample extraction or for column chromatographic procedures.
8.3.4.2 Rinsate Sample
8.3.4.2.1 The rinsate sample must be fortified like
a regular sample.
8.3.4.2.2 Take a 100 ml (± 0.5 ml) portion of the
sampling equipment rinse solvent (rinsate sample), filter, if
necessary, and add 100 ML of the solution containing the nine
internal standards (Table 2).
8.3.4.2.3 Using a KD apparatus, concentrate to
approximately 5 ml.
NOTE: As an option, a rotary evaporator may be used in place of the KD apparatus
for the concentration of the rinsate.
8.3.4.2.4 Transfer the 5 ml concentrate from the KD
concentrator tube in 1 ml portions to a 1 ml minivial, reducing
the volume in the minivial as necessary with a gentle stream
of dry nitrogen.
8.3.4.2.5 Rinse the KD concentrator tube with two
0.5 ml portions of hexane and transfer the rinses to the 1 ml
minivial. Blow down with dry nitrogen as necessary.
8.3.4.2.6 Just before analysis, add 10 ML recovery
standard solution (Table 2) and reduce the volume to its final
8290 - 39 Revision 0
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volume, as necessary (Section 7.8.1). No column chromatography
is required.
8.3.4.2.7 Analyze an aliquot following the same
procedures used to analyze samples.
8.3.4.2.8 Report percent recovery of the internal
standard and the presence of any PCDD/PCDF compounds in /xg/L
of rinsate solvent.
8.3.5 Duplicate Analyses
8.3.5.1 In each batch of samples, locate the sample specified
for duplicate analysis, and analyze a second 10 g soil or sediment
sample portion or 1 L water sample, or an appropriate amount of the
type of matrix under consideration.
8.3.5.1.1 The results of the laboratory duplicates
(percent recovery and concentrations of 2,3,7,8-substituted
PCDD/PCDF compounds) should agree within 25 percent relative
difference (difference expressed as percentage of the mean).
Report all results.
8.3.5.1.2 Recommended actions to help locate problems:
8.3.5.1.2.1 Verify satisfactory instrument
performance (Sections 8.2 and 8.3).
8.3.5.1.2.2 If possible, verify that no error was
made while weighing the sample portions.
8.3.5.1.2.3 Review the analytical procedures with
the performing laboratory personnel.
8.3.6 Matrix Spike and Matrix Spike Duplicate
8.3.6.1 Locate the sample for the MS and MSD analyses (the
sample may be labeled "double volume").
8.3.6.2 Add an appropriate volume of the matrix spike
fortification solution (Section 5.10) and of the sample fortification
solution (Section 5.8), adjusting the fortification level as
specified in Table 1 under IS Spiking Levels.
8.3.6.3 Analyze the MS and MSD samples as described in
Section 7.
8.3.6.4 The results obtained from the MS and MSD samples
(concentrations of 2,3,7,8-substituted PCDDs/PCDFs) should agree
within 20 percent relative difference.
8.4 Percent Recovery of the Internal Standards - For each sample, method
blank and rinsate, calculate the percent recovery (Section 7.9.2). The percent
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recovery should be between 40 percent and 135 percent for all 2,3,7,8-substituted
internal standards.
NOTE: A low or high percent recovery for a blank does not require discarding
the analytical data but it may indicate a potential problem with future
analytical data.
8.5 Identification Criteria
8.5.1 If either one of the identification criteria appearing in
Sections 7.8.4.1.1 through 7.8.4.1.4 is not met for an homologous series,
it is reported that the sample does not contain unlabeled
2,3,7,8-substituted PCDD/PCDF isomers for that homologous series at the
calculated detection limit (Section 7.9.5)
8.5.2 If the first initial identification criteria (Sections
7.8.4.1.1 through 7.8.4.1.4) are met, but the criteria appearing in
Sections 7.8.4.1.5 and 7.8.4.2.1 are not met, that sample is presumed to
contain interfering contaminants. This must be noted on the analytical
report form, and the sample should be rerun or the extract reanalyzed.
8.6 Unused portions of samples and sample extracts must be preserved
for six months after sample receipt to allow further analyses.
8.7 Reuse of glassware is to be minimized to avoid the risk of
contamination.
9.0 METHOD PERFORMANCE
9.1 Data are currently not available.
10.0 REFERENCES
1. "Control of Interferences in the Analysis of Human Adipose Tissue for
2,3,7,8-Tetrachlorodibenzo-p-dioxin". D. G. Patterson, J.S. Holler, D.F.
Grote, L.R. Alexander, C.R. Lapeza, R.C. O'Connor and J.A. Liddle. Environ.
Toxicol. Chem. 5, 355-360 (1986).
2. "Method 8290: Analytical Procedures and Quality Assurance for Multimedia
Analysis of Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans by High-
Resolution Gas Chromatography/High-Resolution Mass Spectrometry". Y.
Tondeur and W.F. Beckert. U.S. Environmental Protection Agency,
Environmental Monitoring Systems Laboratory, Las Vegas, NV.
3. "Carcinogens - Working with Carcinogens", Department of Health, Education,
and Welfare, Public Health Service, Center for Disease Control. National
Institute for Occupational Safety and Health. Publication No. 77-206,
August 1977.
4. "OSHA Safety and Health Standards, General Industry", (29 CFR 1910),
Occupational Safety and Health Administration, OSHA 2206 (revised January
1976).
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5. "Safety in Academic Chemistry Laboratories", American Chemical Society
Publication, Committee on Chemical Safety (3rd Edition, 1979.)
6. "Hybrid HRGC/MS/MS Method for the Characterization of Tetrachlorinated
Dibenzo-p-dioxins in Environmental Samples." Y. Tondeur, W.J. Niederhut,
S.R. Missler, and J.E. Campana, Mass Spectrom. 14, 449-456 (1987).
11.0 SAFETY
11.1 The following safety practices are excerpts from EPA Method 613,
Section 4 (July 1982 version) and amended for use in conjunction with this
method. The 2,3,7,8-TCOD isomer has been found to be acnegenic, carcinogenic,
and teratogenic in laboratory animal studies. Other PCDOs and PCDFs containing
chlorine atoms in positions 2,3,7,8 are known to have toxicities comparable to
that of 2,3,7,8-TCDD. The analyst should note that finely divided dry soils
contaminated with PCDDs and PCDFs are particularly hazardous because of the
potential for inhalation and ingestion. It is recommended that such samples be
processed in a confined environment, such as a hood or a glove box. Laboratory
personnel handling these types of samples should wear masks fitted with charcoal
filters to prevent inhalation of dust.
11.2 The toxicity or carcinogenicity of each reagent used in this method
is not precisely defined; however, each chemical compound should be treated as
a potential health hazard. From this viewpoint, exposure to these chemicals must
be kept to a minimum. The laboratory is responsible for maintaining a current
awareness file of OSHA regulations regarding the safe handling of the chemicals
specified in this method. A reference file of material safety data sheets should
be made available to all personnel involved in the chemical analysis of samples
suspected to contain PCDDs and/or PCDFs. Additional references to laboratory
safety are given in references 3, 4 and 5.
11.3 Each laboratory must develop a strict safety program for the handling
of PCDDs and PCDFs. The laboratory practices listed below are recommended.
11.3.1 Contamination of the laboratory will be minimized by
conducting most of the manipulations in a hood.
11.3.2 The effluents of sample splitters for the gas
chromatograph and roughing pumps on the HRGC/HRMS system should pass
through either a column of activated charcoal or be bubbled through a trap
containing oil or high boiling alcohols.
11.3.3 Liquid waste should be dissolved in methanol or ethanol
and irradiated with ultraviolet light at a wavelength less than 290 nm for
several days (use F 40 BL lamps, or equivalent). Using this analytical
method, analyze the irradiated liquid wastes and dispose of the solutions
when 2,3,7,8-TCDD and -TCDF congeners can no longer be detected.
11.4 The following precautions were issued by Dow Chemical U.S.A. (revised
11/78) for safe handling of 2,3,7,8-TCDD in the laboratory and amended for use
in conjunction with this method.
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11.4.1 The following statements on safe handling are as complete
as possible on the basis of available toxicological information. The
precautions for safe handling and use are necessarily general in nature
since detailed, specific recommendations can be made only for the
particular exposure and circumstances of each individual use. Assistance
in evaluating the health hazards of particular plant conditions may be
obtained from certain consulting laboratories and from State Departments
of Health or of Labor, many of which have an industrial health service.
The 2,3,7,8-TCDD isomer is extremely toxic to certain kinds of laboratory
animals. However, it has been handled for years without injury in
analytical and biological laboratories. Many techniques used in handling
radioactive and infectious materials are applicable to 2,3,7,8-TCDD.
11.4.1.1 Protective Equipment: Throw away plastic gloves,
apron or lab coat, safety glasses and laboratory hood adequate for
radioactive work. However, PVC gloves should not be used.
11.4.1.2 Training: Workers must be trained in the proper
method of removing contaminated gloves and clothing without
contacting the exterior surfaces.
11.4.1.3 Personal Hygiene: Thorough washing of hands and
forearms after each manipulation and before breaks (coffee, lunch,
and shift).
11.4.1.4 Confinement: Isolated work area, posted with signs,
segregated glassware and tools, plastic backed absorbent paper on
benchtops.
11.4.1.5 Waste: Good technique includes minimizing
contaminated waste. Plastic bag liners should be used in waste cans.
11.4.1.6 Disposal of Hazardous Wastes: Refer to the November
7, 1986 issue of the Federal Register on Land Ban Rulings for details
concerning the handling of dioxin containing wastes.
11.4.1.7 Decontamination: Personnel - apply a mild soap with
plenty of scrubbing action. Glassware, tools and surfaces -
Chlorothene NU Solvent (Trademark of the Dow Chemical Company) is
the least toxic solvent shown to be effective. Satisfactory cleaning
may be accomplished by rinsing with Chlorothene, then washing with
a detergent and water. Dish water may be disposed to the sewer after
percolation through a charcoal bed filter. It is prudent to minimize
solvent wastes because they require special disposal through
commercial services that are expensive.
11.4.1.8 Laundry: Clothing known to be contaminated should
be disposed with the precautions described under "Disposal of
Hazardous Wastes". Laboratory coats or other clothing worn in
2,3,7,8-TCDD work area may be laundered. Clothing should be
collected in plastic bags. Persons who convey the bags and launder
the clothing should be advised of the hazard and trained in proper
handling. The clothing may be put into a washer without contact if
the launderer knows the problem. The washer should be run through
one full cycle before being used again for other clothing.
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11.4.1.9 Wipe Tests: A useful method for determining
cleanliness of work surfaces and tools is to wipe the surface with
a piece of filter paper, extract the filter paper and analyze the
extract.
NOTE: A procedure for the collection, handling, analysis, and reporting
requirements of wipe tests performed within the laboratory is described
in Attachment A. The results and decision making processes are based on
the presence of 2,3,7,8-substituted PCDDs/PCDFs.
11.4.1.10 Inhalation: Any procedure that may generate
airborne contamination must be carried out with good ventilation.
Gross losses to a ventilation system must not be allowed. Handling
of the dilute solutions normally used in analytical and animal work
presents no significant inhalation hazards except in case of an
accident.
11.4.1.11 Accidents: Remove contaminated clothing
immediately, taking precautions not to contaminate skin or other
articles. Wash exposed skin vigorously and repeatedly until medical
attention is obtained.
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Attachment A
PROCEDURES FOR THE COLLECTION, HANDLING, ANALYSIS, AND
REPORTING OF WIPE TESTS PERFORMED WITHIN THE LABORATORY
This procedure is designed for the periodic evaluation of potential contamination
by 2,3,7,8-substituted PCDD/PCDF congeners of the working areas inside the
laboratory.
A.I Perform the wipe tests on surface areas of two inches by one foot
with glass fiber paper saturated with distilled in glass acetone using a pair
of clean stainless steel forceps. Use one wiper for each of the designated
areas. Combine the wipers to one composite sample in an extraction jar containing
200 ml distilled in glass acetone. Place an equal number of unused wipers in
200 ml acetone and use this as a control. Add 100 juL of the sample
fortification solution to each jar containing used or unused wipers (Section
5.8).
A.2.1 Close the jar containing the wipers and the acetone and extract
for 20 minutes using a wrist action shaker. Transfer the extract into a
KD apparatus fitted with a concentration tube and a three ball Snyder
column. Add two Teflon™ or Carborundum™ boiling chips and concentrate
the extract to an apparent volume of 1.0 mL on a steam bath. Rinse the
Snyder column and the KD assembly with two 1 mL portions of hexane into
the concentrator tube, and concentrate its contents to near dryness with
a gentle stream of nitrogen. Add 1.0 mL hexane to the concentrator tube
and swirl the solvent on the walls.
A.2.2 Prepare a neutral alumina column as described in Section
7.5.2.2 and follow the steps outlined in Sections 7.5.2.3 through 7.5.2.5.
A.2.3 Add 10 pi of the recovery standard solution as described in
Section 7.5.3.6.
A.3 Concentrate the contents of the vial to a final volume of 10 (j.1
(either in a minivial or in a capillary tube). Inject 2 nl of each extract
(wipe and control) onto a capillary column and analyze for 2,3,7,8-substituted
PCDDs/PCDFs as specified in the analytical method in Section 7.8. Perform
calculations according to Section 7.9.
A.4 Report the presence of 2,3,7,8-substituted PCDDs and PCDFs as a
quantity (pg or ng) per wipe test experiment (WTE). Under the conditions out-
lined in this analytical protocol, a lower limit of calibration of 10 pg/WTE is
expected for 2,3,7,8-TCDD. A positive response for the blank (control) is
defined as a signal in the TCDD retention time window at any of the masses
monitored which is equivalent to or above 3 pg of 2,3,7,8-TCDD per WTE. For
other congeners, use the multiplication factors listed in Table 1, footnote (a)
(e.g., for OCDD, the lower MCL is 10 x 5 = 50 pg/WTE and the positive response
for the blank would be 3 x 5 = 15 pg). Also, report the recoveries of the
internal standards during the simplified cleanup procedure.
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A.5 At a minimum, wipe tests should be performed when there is evidence
of contamination in the method blanks.
A.6 An upper limit of 25 pg per TCDD isomer and per wipe test experiment
is allowed (use multiplication factors listed in footnote (a) from Table 1 for
other congeners). This value corresponds to 2\ times the lower calibration limit
of the analytical method. Steps to correct the contamination must be taken
whenever these levels are exceeded. To that effect, first vacuum the working
places (hoods, benches, sink) using a vacuum cleaner equipped with a high
efficiency particulate absorbent (HEPA) filter and then wash with a detergent.
A new set of wipes should be analyzed before anyone is allowed to work in the
dioxin area of the laboratory after corrective action has been taken.
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Figure 1.
8
6 u 4
Dibenzodioxin
8
Dibenzofuran
General structures of d1benzo-p-d1ox1n and dlbenzofuran.
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Figure 2.
M/AM
5,600
B
5,600
8,550
400 ppm
Peak profile displays demonstrating the effect of the detector zero on the
measured resolving power. In this example, the true resolving power is 5,600.
A) The zero was set too high; no effect 1s observed upon the measurement
of the resolving power.
B) The zero was adjusted properly.
C) The zero was set too low; this results 1n overestimating the actual
resolving power because the peak-to-peak noise cannot be measured
accurately.
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Figure 3.
8:00 AM
Mass Resolution
Mass Accuracy
Analytical Procedure
Thaw Sample Extract
1
Concentrate to 10 uL
I
9:00 AM
Initial or
Routine
Calibration
GC Column
Performance
11:00 AM
Method
Blank
8:00 PM
Mass
Resolution
Routine
Calibration
Typical 12 hour analysis sequence of events.
8290 - 49
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o
o
Figure 4.
6'8'Z'L
8'9'C'l
o
o
n
'lA
(N
0)
M
Selected ion current profile for m/z 322 (TCOOs) produced by MS analysis of the
GC performance check solution on a 60 m DB-5 fused silica capillary column under
the conditions listed in Section 7.6.
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Figure 5.
Ref. mass 304.9824 Peak top
Span. 200 ppm
System file name
Data file name
Resolution
Group number
lonization mode
Switching
Ref. masses
YVES150
A 85Z567
10000
1
EI +
VOLTAGE
304.9824
380.9260
M/AM—10.500
Channel B 380.9260 Lock mass
Span 200 ppm
Peak profiles representing two PFK reference Ions at m/z 305 and 381. The
resolution of the high-mass signal 1s 95 ppm at 5 percent of the peak height;
this corresponds to a resolving power M/ N of 10,500 (10 percent valley
definition).
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Figure 6.
20:00
22:00
24:00
26:00
28:00
30:00
Manual determination of S/N.
The peak height (S) Is measured between the mean noise (lines C and 0). These
mean signal values are obtained by tracing the line between the baseline average
noise extremes, El and E2, and between the apex average noise extremes, E3 and
E4, at the apex of the signal.
NOTE: It 1s Imperative that the Instrument Interface amplifier electronic zero
offset be set high enough so that negative going baseline noise 1s
recorded.
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Table 1.
Types of Matrices, Sample Sizes and 2,3,7,8-TCDD-Based
Method Calibration Limits (Parts per Trillion)
Lower MCL(a)
Upper MCLta)
Weight (g)
IS Spiking
Levels (ppt)
Final Extr.
Vol. (AiL)(d)
Water
0.
2
1000
1
10-50
Soil
Sediment
Paper Pulp"
01 1.0
200
10
100
10-50
Fly
Ash
1.0
200
10
100
50
Fish
Tissue
1.0
200
20
100
10-50
Human
Adipose
c Tissue
1.0
200
10
100
10-50
Sludges,
Fuel Oil
5.0
1000
2
500
50
Still-
Bottom
10
2000
1
1000
50
(a) For other congeners multiply the values by 1 for TCDF/PeCDD/PeCDF, by 2.5
for HxCDD/HxCDF/HpCDD/HpCDF, and by 5 for OCDD/OCDF.
(b) Sample dewatered according to Section 6.5.
(c) One half of the extract from the 20 g sample is used for determination of
lipid content (Section 7.2.2).
(d) See Section 7.8.1, Note.
NOTE: Chemical reactor residues are treated as still bottoms if their
appearances so suggest.
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Table 2.
Composition of the Sample Fortification
and Recovery Standard Solutions3
Analyte
Sample Fortification
Solution
Concentration
(pg/juL; Solvent:
Nonane)
Recovery Standard
Solution
Concentration
(pg//iL; Solvent:
Nonane)
13C12-2,3,7,8-TCDD 10
13C12-2,3,7,8-TCDF 10
13C12-1,2,3,4-TCDD
13C12-l,2,3,7,8-PeCDD 10
13C12-l,2,3,7,8-PeCDF 10
13C12-l,2,3,6,7,8-HxCDD 25
13C12-l,2,3,4,7,8-HxCDF 25
13C12-l,2,3,7,8,9-HxCDD
13C12-l,2,3,4,6,7,8-HpCDD 25
13C12-l,2,3,4,6,7,8-HpCDF 25
13C12-OCDD 50
50
50
(a) These solutions should be made freshly every day because of the possibility
of adsorptive losses to glassware. If these solutions are to be kept for more
than one day, then the sample fortification solution concentrations should be
increased ten fold, and the recovery standard solution concentrations should be
doubled. Corresponding adjustments of the spiking volumes must then be made.
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Table 3.
The Fifteen 2,3,7,8-Substituted PCDD and PCDF Congeners
PCDD PCDF
2,3,7,8-TCDD(*) 2,3,7,8-TCDF(*)
l,2,3,7,8-PeCDD(*) l,2,3,7,8-PeCDF(*)
l,2,3,6,7,8-HxCDD(*) 2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDF
l,2,3,7,8,9-HxCDD(+) 1,2,3,7,8,9-HxCDF
l,2,3,4,6,7,8-HpCDD(*) l,2,3,4,7,8-HxCDF(*)
2,3,4,6,7,8-HxCDF
l,2,3,4,6,7,8-HpCDF(*)
1,2,3,4,7,8,9-HpCDF
(*) The 13C-labeled analogue is used as an internal standard.
(+) The ISC-labeled analogue is used as a recovery standard.
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Table 4.
Isomers of Chlorinated Dioxins and Furans as a
Function of the Number of Chlorine Atoms
Number of
Chlorine
Atoms
1
2
3
4
5
6
7
8
Total
Number of
Dioxin
Isomers
2
10
14
22
14
10
2
1
75
Number of
2,3,7,8
Isomers
—
—
1
1
3
1
1
7
Number of
Furan
Isomers
4
16
28
38
28
16
4
1
135
Number of
2,3,7,8
Isomers
—
—
—
1
2
4
2
1
10
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Table 5.
High-Resolution Concentration Calibration Solutions
Concentration (pq/uL. in Nonane)
Compound
HRCC
Unlabeled Analytes
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
Internal Standards
13C12-2,3,7,8-TCDD
13C12-2,3,7,8-TCDF
13C12-l,2,3,7,8-PeCDD
13C12-l,2,3,7,8-PeCDF
13C12-l,2,3,6,7,8-HxCDD
13C12-l,2,3,4,7,8-HxCDF
13C12-l,2,3,4,6,7,8-HpCDD
13C12-l,2,3,4,6,7,8-HpCDF
13C12-OCDD
Recovery Standards
13C12-l,2,3,4-TCDD(a)
13C12-l,2,3,7,8,9-HxCDDtb)
200
200
500
500
500
500
500
500
500
500
500
500
500
500
500
1,000
1,000
50
50
50
50
125
125
125
125
250
50
125
50
50
125
125
125
125
125
125
125
125
125
125
125
125
125
250
250
50
50
50
50
125
125
125
125
250
50
125
10
10
25
25
25
25
25
25
25
25
25
25
25
25
25
50
50
50
50
50
50
125
125
125
125
250
50
125
2.5
2.5
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
12.5
12.5
50
50
50
50
125
125
125
125
250
50
125
1
1
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5
5
50
50
50
50
125
125
125
125
250
50
125
(a) Used for recovery determinations of TCDD, TCDF, PeCDD and PeCDF internal
standards.
(b) Used for recovery determinations of HxCDD, HxCDF, HpCDD, HpCDF and OCDD
internal standards.
8290 - 57
Revision 0
November 1990
-------
Table 6.
Ions Monitored for HRGC/HRMS Analysis of PCDDs/PCDFs
Descriptor
1
2
3
4
Accurate'8'
Mass
303.9016
305.8987
315.9419
317.9389
319.8965
321.8936
331.9368
333.9338
375.8364
[354.9792]
339.8597
341.8567
351.9000
353.8970
355.8546
357.8516
367.8949
369.8919
409.7974
[354.9792]
373.8208
375.8178
383.8639
385.8610
389.8156
391.8127
401.8559
403.8529
445.7555
[430.9728]
407.7818
409.7788
417.8250
419.8220
423.7767
425.7737
435.8169
437.8140
479.7165
[430.9728]
Ion
ID
M
M+2
M
M+2
M
M+2
M
M+2
M+2
LOCK
M+2
M+4
M+2
M+4
M+2
M+4
M+2
M+4
M+2
LOCK
M+2
M+4
M
M+2
M+2
M+4
M+2
M+4
M+4
LOCK
M+2
M+4
M
M+2
M+2
M+4
M+2
M+4
M+4
LOCK
Elemental
Composition
C12H435C140
C12H435C1337C10
13C12H435C140
13C12H,35C1337C10
Ci2H43Cl40,
C12H435C1337C102
13p M 35/»-| n
13M*35C1*37C102
C12H43SC1537C10
C9F13
C12H335C1437C10
C12H335C1337C120
C12H3 C14 CIO
13C12H35C137C120
C12H33SC1437C102
C12H335C1337C1202
13C12H335C1437C102
13C12H35C137C1202
Ci2H3 C1637C10
C9F13
C H35C1 37C10
C3i2H235Cl437cl20
C12H235C160
13C12H235C137C10
C12H3C13C102
C12H235C1437C1202
13C12H235C1537C102
13C12H/C1/C1202
C12H23§C1637C120
C9F17
C H35C1 37C10
C12H35C1537C120
13C12H35C170
13C12H35C1 37C10
C12H C163 C102
Ci2H35Cl537Cl202
13C12H35C1637C102
13C12H35C1 37C1202
C12H C173 C120
C9F17
Analyte
TCDF
TCDF
TCDF (S)
TCDF (S)
TCDD
TCDD
TCDD (S)
TCDD (S)
HxCDPE
PFK
PeCDF
PeCDF
PeCDF (S)
PeCDF (S)
PeCDD
PeCDD
PeCDD (S)
PeCDD (S)
HpCDPE
PFK
HxCDF
HxCDF
HxCDF (S)
HxCDF (S)
HxCDD
HxCDD
HxCDD (S)
HxCDD (S)
OCDPE
PFK
HpCDF
HpCDF
HpCDF (S)
HpCDF
HpCDD
HpCDD
HpCDD (S)
HpCDD (S)
NCDPE
PFK
8290 - 58
Revision 0
November 1990
-------
Table 6.
Continued
Descriptor
5
Accurate(a)
Mass
441.7428
443.7399
457.7377
459.7348
469.7780
471.7750
513.6775
[442.9278]
Ion
ID
M+2
M+4
M+2
M+4
M+2
M+4
M+4
LOCK
Elemental
Composition
C1235C1737C10
C1235C1637C120
C18"ClŁciQ,
C1235C1 37C1202
13C1235C1737C102
13C 35C1/C1202
C123§C1837C120
C-IO' 17
Analyte
OCDF
OCDF
OCDD
OCDD
OCDD (S)
OCDD (S)
DCDPE
PFK
(a)
The following nuclidic masses were used:
H = 1.007825
C =12.000000
13 C =13.003355
F =18.9984
S = internal/recovery standard
0 = 15.994915
35C1 = 34.968853
37C1 = 36.965903
8290 - 59
Revision 0
November 1990
-------
Table 7.
PCDD and PCDF Congeners Present in the GC Performance
Evaluation Solution and Used for Defining the
Homologous GC Retention Time Windows on a
60 m DB-5 Column
No. of
Chlorine
Atoms
4
-------
Table 8.
Theoretical Ion Abundance Ratios and Their Control Limits
for PCDDs and PCDFs
Number of
Chlorine
Atoms
4
5
6
6(a)
y(b)
7
8
Ion
Type
JL
M+2
M±2
M+4
M+l
M+4
M
M+2
JL
M+2
M+2
M+4
M+l
M+4
Theoretical
Ratio
0.77
1.55
1.24
0.51
0.44
1.04
0.89
Control
lower
0.65
1.32
1.05
0.43
0.37
0.88
0.76
Limits
upper
0.89
1.78
1.43
0.59
0.51
1.20
1.02
(a) Used only for 13C-HxCDF (IS).
(b) Used only for 13C-HpCDF (IS).
8290 - 61
Revision 0
November 1990
-------
Table 9.
Relative Response Factor [RRF (number)] Attributions
Number Specific Congener Name
1 2,3,7,8-TCDD (and total TCDDs)
2 2,3,7,8-TCDF (and total TCDFs)
3 1,2,3,7,8-PeCDD (and total PeCDDs)
4 1,2,3,7,8-PeCDF
5 2,3,4,7,8-PeCDF
6 1,2,3,4,7,8-HxCDD
7 1,2,3,6,7,8-HxCDD
8 1,2,3,7,8,9-HxCDD
9 1,2,3,4,7,8-HxCDF
10 1,2,3,6,7,8-HxCDF
11 1,2,3,7,8,9-HxCDF
12 2,3,4,6,7,8-HxCDF
13 1,2,3,4,6,7,8-HpCDD (and total HpCDDs)
14 1,2,3,4,6,7,8-HpCDF
15 1,2,3,4,7,8,9-HpCDF
16 OCDD
17 OCDF
18 13C12-2,3,7,8-TCDD
19 13C12-2,3,7,8-TCDF
20 13C12-l,2,3,7,8-PeCDD
21 13C12-l,2,3,7,8-PeCDF
22 13C12-l,2,3,6,7,8-HxCDD
23 13C12-l,2,3,4,7,8-HxCDF
24 13C12-l,2,3,4,6,7,8-HpCDD
25 13C12-l,2,3,4,6,7,8-HpCDF
26 13Cr,-OCDD
27 Total PeCDFs
28 Total HxCDFs
29 Total HxCDDs
30 Total HpCDFs
8290 - 62 Revision 0
November 1990
-------
Table 10.
2,3,7,8-TCDD Toxicity Equivalency Factors (TEFs) for the
Polychlorinated Dibenzodioxins and Dibenzofurans
Number
Compound(s)
TEF
1
2
3
4
5
6
7
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8,9-OCDD
1.00
0.50
0.10
0.10
0.10
0.01
0.001
8
9
10
11
12
13
14
15
16
17
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
1,2,3,4,6,7,8,9-OCDF
0.1
0.05
0.5
1
1
1
1
01
0.01
0.001
8290 - 63
Revision 0
November 1990
-------
Table 11.
Analyte Relative Retention Time Reference Attributions
Analyte Analyte RRT Reference(a)
1,2,3,4,7,8-HxCDD 13C12-l,2,3,6,7,8-HxCDD
1,2,3,6,7,8-HxCDF 13C12-l,2,3,4,7,8-HxCDF
1,2,3,7,8,9-HxCDF 13C12-l,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF 13C12-l,2,3,4,7,8-HxCDF
(a) The retention time of 2,3,4,7,8-PeCDF on the DB-5 column is measured relative
to 13C12-l,2,3,7,8-PeCDF and the retention time of 1,2,3,4,7,8,9-HpCDF relative
to 13C12-l,2,3,4,6,7,8-HpCDF.
8290 - 64 - Revision 0
November 1990
-------
METHOD 8290
POLYCHLORINATED DIBENZODIOXINS (PCDDs) AND POLYCHLORINATED DIBENZOFURANS (PCDFs)
BY HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-RESOLUTION MASS SPECTROMETRY (HRGC/HRMS)
("START)
7.1 INTERNAL STANDARD ADDITION
7.1.1 Sample size of 1 to 1000
grams, see section 7.4 & Table 1.
Determine wt. on tared flask
7.1.2 Spike samples w/100 ul
fortification mixture yielding
internal standard cones, of
Table 1, except for adipose tissue
7.1.2.1 For soil, sediment, fly
ash, water, and fish tissue, mix
1 ml acetone with 100 ul
mixture
7.1.2.2 Do not dilute for other
sample matrices
i
[7.2 SAMPLE EXTRACTION AND PURIFICATION!
±
[7.2 Fish and Paper Pulp]
7.2.1 Mix 60 gr sodium
sulfate and 20 gr sample;
place mix in Soxhlet; add
200 mis 1: 1 hexone/MeCI;
reflux 12 hours
7.2.2 Transfer extract to a
KD apparatus with a Snyder
column
7.2.3 Add Teflon boiling
chip; concentrate to 10
mis in water bath; cool for
5 minutes
7.2.4 Add new chip, 50 mis
hexane to flask; concentrate
to 5 mis; cool for 5 mins.;
assure MeCI out before next
step
7.2.5 Rinse apparatus with
hexane; transfer contents
to a separatory funnel; do
cleanup procedure
±
J7.3 Human Adipose Tissue | I 7.4 Environmental and Waste I
7.3.1 Store samples at or
below -20 C, care taken in
handling
7.3.2 Extraction
.1 Weigh out sample
.2 Let stand to room T
.3 Add MeCI, fortification
soln., homogenize
.4 Separate MeCI layer,
filter, dry, transfer to
vol. flask
.5 Redo step 3, add to
vol. flask
.6 Rinse sample train,
add to vol. flask
.7 Adjust to mark w/
MeCI
7.3.3 Determine Lioid Content
.1 Preweigh 1 dram
glass vial
.2 Transfer and reduce 1
ml. extract to viol til
weight constant
.3 Calculate weight dried
extract
.4 Calculate % lipid content
from eon.
.5 Record lipid extract wt.
and % lipid content
T
7.3.4 Extract Concentration
.1 Transfer and rinse vol.
flask contents of 7.3.2.7
to round bottom
.2 Concentrate on rotovap
at 40 C
7.3.5 Extract Cleanup
n Dissolve SecHon 4 extract
with hexane
.2 Add acid impregnated
silica, stir for 2 hours
.3 Decant and dry liquid
with sodium sulfaie
.4 Rinse silica 2x w/hexone
dry w/sodium sulfate,
combine rinses w/step 3
.5 Rinse sodium sulfate,
combine rinse w/step 4
.6 Prepare acidic silica
column
.7 Pass hexone extract
through column, collect
eluate in 500 ml. KD
assembly
.8 Rinse column w/hexane,
combine eluate w/step 7
concentrate total eluate
to 100 ul
Note: If column discolored,
repeat cleanup (7.3.5.1)
.9 Extract ready for column
cleanup
8290 - 65
Revision 0
November 1990
-------
METHOD 8290
continued
I 7.4 Environmentol ond Woste Somplesl
7.4.1 Sludge/Wet Fuel Oil
.1 Extract sample with toluene
using Dean-Stark water
separator
.2 Cool sample, filter through
glass fiber filter
.3 Rinse litter w/toluene,
combine w/extract
.4 Concentrate to near dryness
using rolovap
Note: Sample dissolves in toluene.
treat as In Section 7.4.2;
sample from pulp, treat as
In Section 7.2
L
7.4.2 Still Bottom/On
.1 Extract sample w/toluene
filter through glass fiber
filter Into round bottom
.2 Concentrate on rotovap
at 50 C
7.4.4 Transfer concentrate to sep
funnel using hexane; rinse
container, add to funnel;
add 5X NaCI soln., shake
2 minutes; discard aqueous
layer
7.4.5 Aqueous
.1 Let sample stand to room T;
mark meniscus on bottle; add
fortification soln.
.2 Filter sample: centrifuge first
if needed
.3 Combine filtered/centrifuged
solids along w/filter; do Soxhlet
extraction of Section 7.4.6.1;
rinse assembly ft combine
.4 Transfer aqueous phase to sep
funnel; rinse sample bottles
w/MeCI & transfer to funnel;
shake and extract water
.5 Let phases separate, use
mechanical means if needed
.6 Pass UeCI lover through drying
agent, collect in KD assembly
w/concentrotor tube
.7 Repeat step 4-6 2x, rinse
drying agent, combine all
in KD assembly
Note: Continous liquid-liquid
extractor may be used if
emulsion problems occur
.8 Attach Snyder column,
concentrate on water bath
til 5 mis left; remove KD
assembly, allow to drain ft
cool
.9 Remove column; add hexone,
extraction concentrate of solids,
ft new boiling chip; attach column,
concentrate to 5 mis
.10 Rinse flask ond assembly to final
volume 15 mis
.11 Determine original sample volume
by transferring meniscus volume to
graduated cylinder
7.4.3 Fly Ash
.1 Weigh sample; add
fortification soln. in acetone,
1M HCI; shake in extraction
jar for 3 hours
.2 Filter mix in Buchner funnel;
rinse filter cake w/water; dry
filter coke at room T
.3 Add sodium sulfote to cake,
mix and let stand for 1 hr.,
mix again and let stand
.4 Place sample in extraction
thimble; extract in Soxhlet
for 16 hours w/toluene
.5 Cool and filter extract; rinse
containers ft combine; rotovap
to near dryness at 50 C
7.4.6 Soil
.1 Add sodium sulfate, mix; transfer mixture to
Soxhlet assembly atop glass wool plug
.2 Add toluene, reflux for 24 hours
Note: Add more sodium sulfate if sample does not
flow freely
.3 Transfer extract to round bottom
.4 Concentrate to 10 mis on rotovap, allow to
cool
.5 Transfer concentrate and hexane rinses to KD
assembly; concentrate to tO mis, allow to
cool
.6 Rinse Snyder column into KD; transfer KD
ft concentrator tube liquids to sep funnel;
rinse KD assembly w/hexane ft add to funnel
8290 - 66
Revision 0
November 1990
-------
METHOD 8290
continued
|7.5 CLEANUP)
7.5.1 Partition
.1 Partition txlract w/concentroted
sulturic acid: shak*. discard
acid layer; repeat acid vast) til
no color present or done 4x
.2 OMIT FOR FISH SAMPLES. Partition
extract w/NoCI soln.; snake.
discard aqueous layer
.3 OMIT FOR FISH SAMPLES. Partition
extract w/KOH soln.; shake.
discard base layer; repeat base
wash til no color obtained In wash
or done 4x
.4 Partition extract w/NoCI soln.;
shake, discord oaueous layer.
Dry extract w/sodium sulfate
into round bottom flask; rinse
sodium sulfate w/hexone;
concentrate hexane soln. in
rotovap
7.5.2
eg/Alumina Column
imn
km
.1 Pock a gravity column w/silica gel;
fill w/ hexane. elute to top of bed;
check for channeling
.2 Pack a gravity column w/olumina;
fill w/hexane. elute to top of bed.
check for channeling
Note: Acidic alumina may be used Instead of
neutral alumina.
.3 Dissolve residue of Section 7.5.1.4
in hexane; transfer soln. to lop of
silica column
.4 Elute silica column w/hexane
directly onto alumina column
.5 Add hexone to alumina column;
elute to top of sodium sulfate in
collect ana save eluted hexane
.6 Add MeCI/hexane soln. to alumina
column; collect eluate in concentrator
tube
7.5.3 Carbon Column
.1 Prepare AX-21
repare AX-21/Celite 545 column;
activate mixture at 130 C for 6
hours; store in dessicator
.2 Pack a 10 ml serologicol pipet
w/prepored AX-2t/Celite 545 mix
Note: Each batch of AX-21/Celite 545
must be checked for 7. recovery
of anatytes.
.3 Concentrate MeCI/hexane fraction
of Section 7.5.2.6 to 2 mis
w/nitrogen; rinse column
w/several solns.; add sample
concentrate and rinses to top
of column
.4 Elute column sequentially
w/: cyclohexane/MeCI; MeCI/
methanol/toluene; combine eluates
.5 Turn column upside down, elute
PCDD/PCDF fraction w/loluene;
filter if carbon fines present
.6 Concentrate toluene fraction on
rotovap: further concentrate to
100 ul in miniviol using nitrogen
at 50 C; rinse tlosk 3x w/1%
toluene in MeCI; add tridecane
recovery std.; store room temp.
in the dark
8290 - 67
Revision
November
0
1990
-------
METHOD 8290
continued
7.6
1
Chromatoqraphlc. Mass Spectromttric, and
Data Acquisition Parameters
1
76
1
1 fins Chramataaranh
Select correct dimensions and parameters
of column, and set-up chromotographlc
conditions
7fi
9 Un«« ^n«-trnm«t«r
.1 Operate mass spectrometer h selected
Ion monitoring (SIM) mode; monitor Ions
of five SIM descriptors
.2 Tune mass spectrometer based on ions
of SIM descriptors
i
7fi
V IViln Ac«iiil«!Hnn
.1 Total cycle time of < or = t second
.2 Acquire SIM data for tons of 5
descriptors
771 Initial Calibration
Required before any sample analysis,
and if routine calibration does not
meet criteria
.1 All 5 calibration solns. must be
used for initial calibration
.2 Tune mass spectrometer w/PFK as
described in Section 7.7.3
.3 Inject 2 ul of CC column performance
check soln. and acquire SIM data;
assure Section 8.1.2 criterion are met
.4 Analyze each of S calibration standards
using the same conditions, with the
following MS operating parameters:
.1 Ratio of integrated ion current for
Table 8 ions within control limits
.2 Ratio of integrated ion current for
carbon labeled internal and recovery
standards within control limits
Note: Control limits must be achieved in
one run for oH Ions.
.3 Signal to noise (S/N) ratio for each
target anatyte and labeled std. selected
ion current profiles (SICP) and
CC signals > 2.S
7.7.1.4
.4 Calcute relative response factors (RRF)
for unlabeled and labeled target analytes
relative to internal stds. (Table S)
.5 Calculate average and relative standard
deviation for the 5 calibration solutions
.6 RRFs for concentration determination of
total isomers in a homologous series
are calculated as:
.1 Congeners in a homologous series w/one ,
isomer, mean RRF used is some as Section
7.7.1.4.5
Note: Calibration solns. do not contain
labeled OCOF; therefore. RRF OCDF
relative to labeled OCOD
.2 Calculation lor mean RRF for congeners
in a homologous series w/more than one
isomer
Note: Isomers in homologous series w/o
2,3,7,8 substitution pattern allotted
same response factor as other 2,3,
7,8 isomers in series
.7 Calculation of RRFs used to determine
% recoveries of nine internal standards
7.7.2
itqble Cofibrotio
ist be met befoi
listed musToe met before
analysis
.1 The X RSO for unlabeled stds. must
be within +/- 20X; for labeled.
+/- MX
.2 S/N ratio for CC signals > or = 2.5
.3 Table 8 Isotopic ratios within limits
Note: When criteria for acceptable calibration
ore met, mean RRFs used for calculations
until routine calibration criteria are not
met
771 Bnnlin« Cnlihrntion
Performed at 12 hour periods after
successful resolution checks
.1 Inject 2 ul calibration soln. HRCC-3;
use some HRCC/HRMS conditions of
Sections 7.6.1 and 7.6.2; document
an acceptable calibration
771 Triform fnr Arr^plnhl* Rnnlina (tnlihrnlipn
.1 Measured unlabeled RRFs must be w/in
+/- 20% of initial calibration values
.2 Measured labeled RRFs must be w/in
+/~ 30X of initial calibration values
.3 Table 8 ion abundance ratios must be
w/in limits
.4 Review routine calibration process if
criteria at steps I and 2 are not
satisfied
An initial calibration must be done
when new HRCC-3, sample fortification,
or recovery std. soln. from another lot
is used
Note:
8290 - 68
Revision 0
November 1990
-------
METHOD 8290
continued
_L
I 7.8 Anolvsls I
Anal
7.8.1 Reduce extract or blonk volume
to 10 or SO ul
7.8.2 Into! 2 ul aliquot of the sample
into Ih. GC
7.8.3 Acquire SIM data according to
Sections 7.6.2 and 7.6.3
'
Note: Acquisition period must at
least encompass PCOD/PCDF
overall retention time window
7.8.4 CC Identification CriUrin
.1 Relative Retention Times
.1 2.3.7.8 sub: Sample components
relative retention time (RRT) w/ln
-1 to 3 seconds of retention
time of labeled internal or
recovery std.
.2 2.3.7.8 sub: Sample RRTs
w/ln homologous retention
time windows if w/o labeled
internal std.
.3 non 2,3.7.8 sub: Retention
time w/in homologous
retention time window
.4 Ion current responses for
quantitotion must reach maximum
w/in 2 seconds
.5 Ion current responses lor labeled
stds. must reach maximum w/in
2 seconds
Note: Verify presence ol 1,2.8.9-TCUD and
1.3.4.6.8-PeCOF in SICPs
.2 Ion Abundance Ratios
.1 Ratio of integrated ion current (or
two ions used for quantification
w/in limits of homologous series
.3 Siqnol-to-Noise Ratio
.1 All ion current intensities > = 2.5
.4 Polychlorinated Diphenyl Ether
Interferences
.1 Corresponding PCDPE channel clear
of signal > = S/N 2.5 at same
retention time
_L
I 7.9 Calculation*!
J
7.9.1 Calculate concentration of PCOD
or PCOF compounds w/formulo
1
7.9.2 Calculate X recovery of nine
internal stds. using formula
Note: Add IK recovery for human
adipose tissue samples
1
7.9.3 Use smaller sample ami. if
calculated concentration exceeds
method calibration limits
1
7.9.4 Sum ol isomer concentration is
total concentration for a
homologous series
1
Lirj['»'fDl|
I
climated Detection
EDL Analyte concentration yielding
peak hi. 2.5x noise level. EOLs calculated
for non-identified 2,3.7 .8- sub congeners
Two methods of calculation:
.1 Samples w/response <2.Sx noise for
both quantification ions
.1 Use EDL expression to calculate for
absent 2,3.7.8 substituted PCDD/PCDF
.2 Samples w/response >2.Sx noise for
at least 1 quantilication ion
.1 Calculate "Estimated Maximum Possible
Concentration" (EMPC) when signal >
2.5x noise and retention time the some
1
7.9.6 Relative percent difference (RPO) formula |
*
7.9.7 Calculation ol 2.3.7 8-TCDD toxicity
eauivalenl factors (TEF) ol PCDDs and PCDFs
.1 Two CC Column TEF Determination:
Reanalyze sample extract on 60 meter
SP-2330 column
.1 Concentrations ol specified congeners
calculated from analysis done on DB-5
column
.2 Concentrations of specified congeners
calculated from analysis done on
SP-2330 column w/dilferent GC/MS
conditions
TCDD done on either column as long as
Section 8.1.2 criteria met
.3 GC peak must meet criteria of Sections
784.2 7 843 and/or 7.8.4.1.1. RRTs
of 2.3.7.8-sub congeners w/no carbon-
labeled analogues referred to w/in 0.006
RRT units of carbon-labeled std
1
fsrop^
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METHOD 8315
FORMALDEHYDE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 Method 8315 covers the determination of free formaldehyde in aqueous
samples and leachates. The following compounds can be determined by this method:
Compound Name CAS No.a
Formaldehyde 50-00-0
Acetaldehyde 75-07-0
a Chemical Abstract Services Registry Number.
1.2 Method 8315 is a high performance liquid chromatographic (HPLC) method
optimized for the determination of formaldehyde and acetaldehyde in aqueous
environmental matrices and leachates of solid samples. When this method is used
to analyze unfamiliar sample matrices, compound identification should be
supported by at least one additional qualitative technique. A gas
chromatograph/mass spectrometer (GC/MS) may be used for the qualitative
confirmation of results for the target analytes, using the extract produced by
this method.
1.3 The method detection limits (MDL) are listed in Tables 1 and 2. The
MDL for a specific sample may differ from that listed, depending upon the nature
of interferences in the sample matrix and the amount of sample used in the
procedure.
1.4 The extraction procedure for solid samples is similar to that
specified in Method 1311 (1). Thus, a single sample may be extracted to measure
the analytes included in the scope of other appropriate methods. The analyst
is allowed the flexibility to select chromatographic conditions appropriate for
the simultaneous measurement of combinations of these analytes.
1.5 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of chromatography and in the interpretation of
chromatograms. Each analyst must demonstrate the ability to generate acceptable
results with this method, using the procedure described in Section 8.2.
1.6 The toxicity or carcinogenicity of each reagent used in this method
has not been precisely defined; however, each chemical compound should be treated
as a potential health hazard. From this viewpoint, exposure to these chemicals
must be reduced to the lowest possible level by whatever means available. The
laboratory is responsible for maintaining a current awareness file of OSHA
regulations regarding the safe handling of the chemicals specified in this
method. A reference file of material safety data sheets should also be made
8315 - 1 Revision 0
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available to all personnel involved in the chemical analysis. Additional
references to laboratory safety are available.
1.7 Formaldehyde has been tentatively classified as a known or suspected,
human or mammalian carcinogen.
2.0 SUMMARY OF METHOD
2.1 For wastes comprised of solids or for aqueous wastes containing
significant amounts of solid material, the aqueous phase, if any, is separated
from the solid phase and stored for later analysis. If necessary, the particle
size of the solids in the waste is reduced. The solid phase is extracted with
an amount of extraction fluid equal to 20 times the weight of the solid phase.
The extraction fluid employed is a function of the alkalinity of the solid phase
of the waste. A special extractor vessel is used when testing for volatiles.
Following extraction, the aqueous extract is separated from the solid phase by
filtration employing 0.6 to 0.8 /im glass fiber filter.
2.2 If compatible (i.e., multiple phases will not form on combination),
the initial aqueous phase of the waste is added to the aqueous extract, and
these liquids are analyzed together. If incompatible, the liquids are analyzed
separately and the results are mathematically combined to yield a volume-weighted
average concentration.
2.3 A measured volume of aqueous sample or an appropriate amount of solids
leachate is buffered to pH 5 and derivatized with 2,4-dinitrophenylhydrazine
(DNPH), using either the solid sorbent or the methylene chloride
derivatization/extraction option. If the solid sorbent option is used, the
derivative is extracted using solid sorbent cartridges, followed by elution with
ethanol. If the methylene chloride option is used, the derivative is extracted
with methylene chloride. The methylene chloride extracts are concentrated using
the Kuderna-Danish (K-D) procedure and solvent exchanged into methanol prior to
HPLC analysis. Liquid chromatographic conditions are described which permit the
separation and measurement of formaldehyde in the extract by absorbance detection
at 360 nm.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts and/or elevated baselines in the chromatograms. All of these materials
must be routinely demonstrated to be free from interferences under the conditions
of the analysis by analyzing laboratory reagent blanks as described in
Section 8.5.
3.1.1 Glassware must be scrupulously cleaned. Clean all glassware
as soon as possible after use by rinsing with the last solvent used. This
should be followed by detergent washing with hot water, and rinses with
tap water and organic-free reagent water. It should then be drained,
dried, and heated in a laboratory oven at 130°C for several hours before
use. Solvent rinses with methanol may be substituted for the oven heating.
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After drying and cooling, glassware should be stored in a clean environment
to prevent any accumulation of dust or other contaminants.
3.1.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all
glass systems may be required.
3.2 Analysis for formaldehyde is especially complicated by its ubiquitous
occurrence in the environment.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from source to source, depending upon the nature and diversity of
the matrix being sampled. No interferences have been observed in the matrices
studied as a result of using solid sorbent extraction as opposed to liquid
extraction. If interferences occur in subsequent samples, some additional
cleanup may be necessary.
3.4 The extent of interferences that may be encountered using liquid
chromatographic techniques has not been fully assessed. Although the HPLC
conditions described allow for a resolution of the specific compounds covered
by this method, other matrix components may interfere.
4.0 APPARATUS AND MATERIALS
4.1 Reaction vessel - 250 ml Florence flask.
4.2 Separatory funnel - 250 ml, with Teflon stopcock.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Vials - 10, 25 ml, glass with Teflon lined screw caps or crimp tops.
4.5 Boiling chips - Solvent extracted with methylene chloride,
approximately 10/40 mesh (silicon carbide or equivalent).
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4.6 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.7 pH meter - Capable of measuring to the nearest 0.01 units.
4.8 High performance liquid chromatograph (modular)
4.8.1 Pumping system - Isocratic, with constant flow control capable
of 1.00 mL/min.
4.8.2 High pressure injection valve with 20 /*L loop.
4.8.3 Column - 250 mm x 4.6 mm ID, 5 /urn particle size, C18 (or
equivalent).
4.8.4 Absorbance detector - 360 rim.
4.8.5 Strip-chart recorder compatible with detector - Use of a data
system for measuring peak areas and retention times is recommended.
4.9 Glass fiber filter paper.
4.10 Solid sorbent cartridges - Packed with 500 mg C18 (Baker or
equivalent).
4.11 Vacuum manifold - Capable of simultaneous extraction of up to 12
samples (Supelco or equivalent).
4.12 Sample reservoirs - 60 ml capacity (Supelco or equivalent).
4.13 Pipet - Capable of accurately delivering 0.10 ml solution (Pipetman
or equivalent).
4.14 Water bath - Heated, with concentric ring cover, capable of
temperature control (+) 2°C). The bath should be used under a hood.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methylene chloride, CH2C12 - HPLC grade or equivalent.
5.4 Methanol, CH3OH - HPLC grade or equivalent.
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5.5 Ethanol (absolute), CH3CH2OH - HPLC grade or equivalent.
5.6 2,4-Dinitrophenylhydrazine (DNPH) (70% (W/W)), [2,4-(02N)2C6H3]NHNH2,
in organic-free reagent water.
5.7 Formalin (37.6 percent (w/w)), formaldehyde in organic-free reagent
water.
5.8 Acetic acid (glacial), CH3C02H.
5.9 Sodium hydroxide solutions, NaOH, 1.0 N and 5 N.
5.10 Sodium chloride, NaCl.
5.11 Sodium sulfite solution, Na2S03, 0.1 M.
5.12 Hydrochloric Acid, HC1, 0.1 N.
5.13 Extraction fluid - Dilute 64.3 ml of 1.0 N NaOH and 5.7 ml glacial
acetic acid to 900 ml with organic-free reagent water. Dilute to 1 liter with
organic-free reagent water. The pH should be 4.93 ± 0.02.
5.14 Stock standard solutions
5.14.1 Stock formaldehyde (approximately 1.00 mg/mL) - Prepare by
diluting 265 /zL formalin to 100 mL with organic-free reagent water.
5.14.1.1 Standardization of formaldehyde stock solution -
Transfer a 25 mL aliquot of a 0.1 M Na2S03 solution to a beaker and
record the pH. Add a 25.0 ml aliquot of the formaldehyde stock
solution (Section 5.14.1) and record the pH. Titrate this mixture
back to the original pH using 0.1 N HC1. The formaldehyde
concentration is calculated using the following equation:
Concentration (mg/mL) = 30.03 x (N HC1) x (mL HC1) 25.0
where:
N HC1 = Normality of HC1 solution used
mL HC1 = mL of standardized HC1 solution used
30.03 = MW of formaldehyde
5.14.2 Stock formaldehyde and acetaldehyde - Prepare by adding
265 juL formalin and 0.1 g acetaldehyde to 90 mL of organic-free reagent
water and dilute to 100 mL. The concentration of acetaldehyde in this
solution is 1.00 mg/mL. Calculate the concentration of formaldehyde in
this solution using the results of the assay performed in Section 5.14.1.1.
5.14.3 Stock standard solutions must be replaced after six months,
or sooner, if comparison with check standards indicates a problem.
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5.15 Reaction Solutions
5.15.1 DNPH (1.00 M9/L) - Dissolve 142.9 mg of 70% (w/w) reagent
in 100 ml absolute ethanol. Slight heating or sonication may be necessary
to effect dissolution.
5.15.2 Acetate buffer (5 N) - Prepare by neutralizing glacial
acetic acid to pH 5 with 5 N NaOH solution. Dilute to standard volume with
organic-free reagent water.
5.15.3 Sodium chloride solution (saturated) - Prepare by mixing an
excess of the reagent grade solid with organic-free reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
6.2 Samples must be refrigerated at 4°C, and must be derivatized within
5 days of sample collection and analyzed within 3 days of derivatization.
7.0 PROCEDURE
7.1 Extraction of Solid Samples
7.1.1 All solid samples should be homogeneous. When the sample is
not dry, determine the dry weight of the sample, using a representative
aliquot.
7.1.1.1 Determination of dry weight - In certain cases, sample
results are desired based on a dry weight basis. When such data is
desired, or required, a portion of sample for dry weight
determination should be weighed out at the same time as the portion
used for analytical determination.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from drying a heavily contaminated
hazardous waste sample.
7.1.1.2 Immediately after weighing the sample for extraction,
weigh 5-10 g of the sample into a tared crucible. Determine the %
dry weight of the sample by drying overnight at 105°C. Allow to cool
in a desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.1.2 Measure 25 g of solid into a 500 mL bottle with a Teflon lined
screw cap or crimp top, and add 500 mL of extraction fluid (Section 5.13).
Extract the solid by rotating the bottle at approximately 30 rpm for 18
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hours. Filter the extract through glass fiber filter paper and store in
sealed bottles at 4°C. Each ml of extract represents 0.050 g solid.
7.2 Cleanup and Separation
7.2.1 Cleanup procedures may not be necessary for a relatively clean
sample matrix. The cleanup procedures recommended in this method have been
used for the analysis of various sample types. If particular circumstances
demand the use of an alternative cleanup procedure, the analyst must
determine the elution profile and demonstrate that the recovery of
formaldehyde is no less than 85% of recoveries specified in Table 3.
Recovery may be lower for samples which form emulsions.
7.2.2 If the sample is not clean, or the complexity is unknown, the
entire sample should be centrifuged at 2500 rpm for 10 minutes. Decant
the supernatant liquid from the centrifuge bottle, and filter through glass
fiber filter paper into a container which can be tightly sealed.
7.3 Derivatization
7.3.1 For aqueous samples, measure a 50 to 100 ml aliquot of the
sample. Quantitatively transfer the sample aliquot to the reaction vessel
(Section 4.1).
7.3.2 For solid samples, 1 to 10 ml of leachate (Section 7.1) will
usually be required. The amount used for a particular sample must be
determined through preliminary experiments.
Note: For all reactions, the total volume of the aqueous layer should be
adjusted to 100 ml with water.
7.3.3 Derivatization and extraction of the derivative can be
accomplished using the solid sorbent (Section 7.3.4) or methylene chloride
option (Section 7.3.5).
7.3.4 Solid Sorbent Option
7.3.4.1 Add 4 ml of acetate buffer and adjust the pH to 5.0
± 0.1 with glacial acetic acid or 5 N NaOH. Add 6 mL of DNPH
reagent, seal the container, and place on a wrist-action shaker for
30 minutes.
7.3.4.2 Assemble the vacuum manifold and connect to a water
aspirator or vacuum pump. Assemble solid sorbent cartridges
containing a minimum of 1.5 g of CIS sorbent, using connectors
supplied by the manufacturer, and attach the sorbent train to the
vacuum manifold. Condition each cartridge by passing 10 ml dilute
acetate buffer (10 ml 5 N acetate buffer dissolved in 250 ml of
organic-free reagent water) through the sorbent cartridge train.
7.3.4.3 Remove the reaction vessel from the shaker and add
10 ml saturated NaCl solution to the vessel.
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7.3.4.4 Add the reaction solution to the sorbent train and
apply a vacuum so that the solution is drawn through the cartridges
at a rate of 3 to 5 mL/min. Release the vacuum after the solution
has passed through the sorbent.
7.3.4.5 Elute each cartridge train with approximately 9 ml
of absolute ethanol, directly into a 10 ml volumetric flask. Dilute
the solution to volume with absolute ethanol, mixed thoroughly, and
place in a tightly sealed vial until analyzed.
7.3.5 Methylene Chloride Option
7.3.5.1 Add 5 ml of acetate buffer and adjust the pH to 5.0
± 0.5 with glacial acetic acid or 5 N NaOH. Add 10 ml of DNPH
reagent, seal the container, and place on a wrist-action shaker for
1 hour.
7.3.5.2 Extract the solution with three 20 ml portions of
methylene chloride, using a 250 ml separatory funnel, and combine
the methylene chloride layers. If an emulsion forms upon extraction,
remove the entire emulsion and centrifuge at 2000 rpm for 10 minutes.
Separate the layers and proceed with the next extraction.
7.3.5.3 Assemble a Kuderna-Danish (K-D) concentrator by
attaching a 10 mL concentrator tube to a 500 ml evaporator flask.
Wash the K-D apparatus with 25 ml of extraction solvent to complete
the quantitative transfer.
7.3.5.4 Add one to two clean boiling chips to the evaporative
flask and attach a three ball Snyder column. Prewet the Snyder
column by adding about 1 ml methylene chloride to the top. Place
the K-D apparatus on a hot water bath (80-90°C) so that the
concentrator tube is partially immersed in the hot water and the
entire lower rounded surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus and the water
temperature, as required, to complete the concentration in 10-15 min.
At the proper rate of distillation the balls of the column will
actively chatter, but the chambers will not flood with condensed
solvent. When the apparent volume of liquid reaches 10 ml, remove
the K-D apparatus and allow it to drain and cool for at least 10 min.
7.3.5.5 Prior to liquid chromatographic analysis, the solvent
must be exchanged to methanol. The analyst must ensure quantitative
transfer of the extract concentrate. The exchange is performed as
follows:
7.3.5.5.1 Following K-D concentration of the methylene
chloride extract to < 10 ml using the macro Snyder column,
allow the apparatus to cool and drain for at least 10 minutes.
7.3.5.5.2 Momentarily remove the Snyder column, add 5 ml
of methanol, a new glass bead, or boiling chip, and attach
the micro Snyder column. Concentrate the extract using 1 ml
of methanol to prewet the Snyder column. Place the K-D
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apparatus on the water bath so that the concentrator tube is
partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as
required, to complete concentration. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of
liquid reaches < 5 ml, remove the K-D apparatus and allow it
to drain and cool for at least 10 minutes.
7.3.5.5.3 Remove the Snyder column and rinse the flask
and its lower joint with 1-2 ml of methanol and add to
concentrator tube. A 5 ml syringe is recommended for this
operation. Adjust the extract volume to 10 ml. Stopper the
concentrator tube and store refrigerated at 4°C if further
processing will not be performed immediately. If the extract
will be stored longer than two days, it should be transferred
to a vial with a Teflon lined screw cap or crimp top. Proceed
with liquid chromatographic analysis if further cleanup is not
required.
7.4 Chromatographic Conditions (Recommended):
Column: CIS, 250 mm x 4.6 mm ID, 5 urn particle size
Mobile Phase: methanol/water, 75:25 (v/v), isocratic
Flow Rate: 1.0 mL/min
UV Detector: 360 nm
Injection Volume: 20 ML
7.5 Calibration
7.5.1 Establish liquid chromatographic operating conditions to
produce a retention time equivalent to that indicated in Table 1 for the
solid sorbent option, or in Table 2 for methylene chloride option.
Suggested chromatographic conditions are provided in Section 7.4. Prepare
derivatized calibration standards according to the procedure in Section
7.5.1.1. Calibrate the chromatographic system using the external standard
technique (Section 7.5.1.2).
7.5.1.1 Preparation of calibration standards
7.5.1.1.1 Prepare calibration standard solutions of
formaldehyde and acetaldehyde in organic-free reagent water
from the stock standard solution (Section 5.13.2). Prepare
these solutions at the following concentrations (in M9/mL) by
serial dilution of the stock standard solution: 50, 20, 10.
Prepare additional calibration standard solutions at the
following concentrations, by dilution of the appropriate 50,
20, or 10 /ug/mL standard: 5, 0.5, 2, 0.2, 1, 0.1.
7.5.1.1.2 Process each calibration standard solution
through the derivatization option used for sample processing
(Section 7.3.4 or 7.3.5).
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7.5.1.2 External standard calibration procedure
7.5.1.2.1 Analyze each derivatized calibration standard
using the chromatographic conditions listed in Tables 1 and 2,
and tabulate peak area against concentration injected. The
results may be used to prepare calibration curves for
formaldehyde and acetaldehyde.
7.5.1.2.2 The working cal ibration curve must be veri f ied
on each working day by the measurement of one or more
calibration standards. If the response for any analyte varies
from the previously established responses by more than 10%,
the test must be repeated using a fresh calibration standard
after it is verified that the analytical system is in control.
Alternatively, a new calibration curve may be prepared for
that compound. If an autosampler is available, it is
convenient to prepare a calibration curve daily by analyzing
standards along with test samples.
7.6 Analysis
7.6.1 Analyze samples by HPLC, using conditions established in
Section 7.5.1. Tables 1 and 2 list the retention times and MDLs that were
obtained under these conditions. Other HPLC columns, chromatographic
conditions, or detectors may be used if the requirements of Section 8.2
are met, or if the data are within the limits described in Tables 1 and 2.
7.6.2 The width of the retention time window used to make
identifications should be based upon measurements of actual retention time
variations of standards over the course of a day. Three times the standard
deviation of a retention time for a compound can be used to calculate a
suggested window size; however, the experience of the analyst should weigh
heavily in the interpretation of the chromatograms.
7.6.3 If the peak area exceeds the linear range of the calibration
curve, a smaller sample volume should be used. Alternatively, the final
solution may be diluted with ethanol and reanalyzed.
7.6.4 If the peak area measurement is prevented by the presence of
observed interferences, further cleanup is required. However, none of the
3600 method series have been evaluated for this procedure.
7.7 Calculations
7.7.1 Calculate each response factor as follows (mean value based
on 5 points):
concentration of standard
RF =
area of the signal
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5
(s RFi)
1
mean RF = RF =
7.7.2 Calculate the concentration of formaldehyde and acetaldehyde
as follows:
/ig/ml = (RF) (area of signal) (concentration factor)
where:
Final volume of extract
concentration factor =
Initial sample (or leachate) volume
Note: For solid samples, a dilution factor must be included in the equation
to account for the weight of the sample used.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 The MDLs listed in Table 1 were obtained using organic-free reagent
water and solid sorbent extraction. Similar results were achieved using a final
effluent and sludge leachate. The MDLs listed in Table 2 were obtained using
organic-free reagent water and methylene chloride extraction. Similar results
were achieved using representative matrices.
9.2 This method has been tested for linearity of recovery from spiked
organic-free reagent water and has been demonstrated to be applicable over the
range from 2 x MDL to 200 x MDL.
9.3 In a single laboratory evaluation using several spiked matrices, the
average recoveries presented in Tables 3 and 4 were obtained using solid sorbent
and methylene chloride extraction, respectively. The standard deviations of the
percent recovery are also included in Tables 3 and 4.
9.4 A representative chromatogram is presented in Figure 1.
10.0 REFERENCES
1. Federal Register, 1986, 51, 40643-40652; November 7.
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TABLE 1.
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY CONDITIONS
AND METHOD DETECTION LIMITS USING SOLID
SORBENT EXTRACTION
Analyte Retention Time MDL
(minutes)
Formaldehyde 7.1 7.2
HPLC conditions: Reverse phase CIS column, 4.6 X 250 mm; isocratic elution
using methanol/water (75:25, v/v); flow rate 1.0 mL/min.; detector 360 nm.
a After correction for laboratory blank.
TABLE 2.
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY CONDITIONS
AND METHOD DETECTION LIMITS USING METHYLENE
CHLORIDE EXTRACTION
Analyte Retention Time MDL
(minutes)
Formaldehyde 7.1 7.2
Acetaldehyde 8.6 171a
HPLC conditions: Reverse phase CIS column, 4.6 X 250 mm; isocratic elution using
methanol/water (75:25, v/v); flow rate 1.0 mL/min.; detector 360 nm.
a These values include reagent blank concentrations of approximately 13
formaldehyde and 130 ng/l acetaldehyde.
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TABLE 3.
SINGLE OPERATOR ACCURACY AND PRECISION
USING SOLID SORBENT EXTRACTION
Analyte
Formaldehyde
Average
Matrix Percent
Type Recovery
Organic- free 86
Reagent Water
Final 90
Effluent
Standard
Deviation
Percent
9.4
11.0
Spike Number
Range of
(/ig/L) Analyses
15-1430 39
46.8-1430 16
Phenol
formaldehyde
Sludge
93
12.0
457-1430
15
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TABLE 4.
SINGLE OPERATOR ACCURACY AND PRECISION
USING METHYLENE CHLORIDE EXTRACTION
Analyte
Formaldehyde
Acetaldehyde
Average
Matrix Percent
Type Recovery
Organic-free
Reagent Water
Ground-
water
Liquids
Organic-free
Reagent Water
Ground-
water
Liquids
(2 types)
Solids
X
91
92.5
69.6
60.3
63.6
44.0
58.4
Standard
Deviation
Percent
P
2.5
8.2
16.3
3.2
10.9
20.2
2.7
Spike
Range
(M9/L)
50-1000
50
250
50-1000
50
250
0.10-1.08
Number
of
Analyses
9
6
12
9
12
12
12
a Spike range in units of mg/g.
x = Average recovery expected for this method
p = Average standard deviation expected for this method.
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FIGURE 1
REPRESENTATIVE CHROMATOGRAM OF A 50
SOLUTION OF FORMALDEHYDE
3*
FOR-D - Formaldehyde derivative
ACET-D - Acetaldehyde derivative
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METHOD 8315
FORMALDEHYDE BY HIGH PERFORMANCE LIQUID CHRQMATOGRAPHY [HPLCI
7.1.1 Ensure sample
homogeneity:
perform % solid
determination, if
appropriate
7.1.2 Weigh sampli
into bottle; add
extraction fluid;
extract 18 hours;
filter
7.2.1 Perform
cleanup, if
necessary
7.2.2 Centrifuge
sample, if
necessary
7.3 Derivatization:
measure aliquot for
liquid sample or
liquid extract of solid
sample; dilute to
total volume of 100
mis
solid
sorbent
7.3.4.1 Add acetate
buffer and adjust
pH with acetic acid
or sodium
hydroxide; add DNPH
reagent; seal
container; shake 30
minutes
7.3.4.2 Assemble
vacuum manifold and
connect to pump;
assemble
cartridges; attach
sorbent train to
manifold; condition
cartridge
7.3.4.3 Remove
reaction vessels
from shaker; add
sodium chloride
sol'n
7.3.4.4 Add the
reaction solution to
sorbent train; elute
under vacuum
7.3.4.5 Elute train
with ethanol
dilute to volume
w/ethonol; mix
7.3.3
Extraction:
solid sorbent or
methylene
..chloride?,,
•©•
methylene
chloride
7.3.5.1 Add acetate
buffer and adjust
pH with acetic acid
or sodium
hydroxide; add DNPH
reagent; seal
container; shake 60
minutes
7.3.5.2 Extract
sol'n with 3 20-ml
portions of
methylene chloride;
combine methylene
chloride layers
7.3.5.3 Assemble a
Kuderno-Danish
(K-D) apparatus to
concentrate extract
layers of methylene
chloride
7.3.5.4 Add boiling
chips to evaporation
flask and attach a
three-ball Snyder
column to the K-D
assembly; immerse
apparatus in hot
water bath; adjust
positioning to finish
concentration in 10-
15 minutes
7.3.5.5 Exchange
solvent to methanol
using K-D assembly
8315 - 16
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METHOD 8315
continued
7.4 Establish LC
operating parameters
7.5.1.1.1 Prepare
calibration standards
7.5.1.1.2 Derivatize
standard solutions
7.5.1.2.1 Analyze
standards and tabulate
peak area against
concentration injected
7.5.1.2.2 Verify
working calibration
curve daily with 1
or more standards
7.6.1 Analyze by
HPLC using
specified
conditions; other
conditions or
hardware may be
used if QC
requirements are met
7.6.2 Use retention
times to interpret
chromatograms
7.6.3 If peak area
exceeds linear
working range, use
a smaller sample
volume or the final
solution may be
diluted with
ethanol and
reanalyzed
7.6.4 If peak area
measurement is
prevented by
interferences,
further cleanup is
needed
7.7.1 Calculate
response factors
for analytes
7.7.2 Calculate
concentrations of
aldehydes in sample,
noting dilution factor
for solid samples
8315 - 17
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METHOD 8316
ACRYLAMIDE. ACRYLONITRILE AND ACROLEIN BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 The following compounds can be determined by this method:
Compound Name CAS No.a
Acrylamide 79-06-1
Acrylonitrile 107-13-1
Acrolein (Propenal) 107-02-8
a Chemical Abstract Services Registry Number.
1.2 The method detection limits (MDLs) for the target analytes in organic-
free reagent water are listed in Table 1. The method may be applicable to other
matrices.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Water samples are analyzed by high pressure liquid chromatography
(HPLC). A 200 /iiL aliquot is injected onto a C-18 reverse-phase column, and
compounds in the effluent are detected with an ultraviolet (UV) detector.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 One high pressure pump.
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4.1.2 Octadecyl Silane (ODS, C-18) reverse phase HPLC column,
25 cm x 4.6 mm, 10 jum, (Zorbax, or equivalent).
4.1.3 Variable wavelength UV detector.
4.1.4 Data system.
4.2 Other apparatus
4.2.1 Water degassing unit - 1 liter filter flask with stopper and
pressure tubing.
4.2.2 Analytical balance - + 0.0001 g.
4.2.3 Magnetic stirrer and magnetic stirring bar.
4.2.4 Sample filtration unit - syringe filter with 0.45 urn filter
membrane, or equivalent disposable filter unit.
4.3 Materials
4.3.1 Syringes - 10, 25, 50 and 250 /*L and 10 ml.
4.3.2 Volumetric pipettes, Class A, glass -1,5 and 10 ml.
4.3.3 Volumetric flasks - 5, 10, 50 and 100 ml.
4.3.4 Vials - 25 ml, glass with Teflon lined screw caps or crimp
tops.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Acrylamide, CH2:CHCONH2, 99+% purity, electrophoresis reagent grade.
5.3 Acrylonitrile, H2C:CHCN, 99+% purity.
5.4 Acrolein, CH2:CHCHO, 99+% purity.
5.5 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.6 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially prepared
stock standards can be used if they are verified against EPA standards. If EPA
standards are not available for verification, then standards certified by the
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manufacturer and verified against a standard made from pure material is
acceptable.
5.6.1 Acrylamide
5.6.1.1 Weigh 100 mg of aery 1 amide neat standard into a 100 ml
volumetric flask, and dilute to the mark with organic-free reagent
water. Calculate the concentration of the standard solution from the
actual weight used. When compound purity is assayed to be 96% or
greater, the weight can be used without correction to calculate the
concentration of the stock standard.
5.6.1.2 Transfer the stock solution into vials with Teflon
lined screw caps or crimp tops. Store at 4°C, protected from light.
5.6.1.3 Stock solutions must be replaced after one year, or
sooner if comparison with the check standards indicates a problem.
5.6.2 Acrylonitrile and Acrolein - Prepare separate stock solutions
for acrylonitrile and acrolein.
5.6.2.1 Place about 9.8 ml of organic-free reagent water into
a 10 ml volumetric flask before weighing the flask and stopper. Weigh
the flask and record the weight to the nearest 0.1 mg. Add two drops
of neat standard, using a 50 juL syringe, to the flask. The liquid
must fall directly into the water, without contacting the inside wall
of the flask.
CAUTION; Acrylonitrile and acrolein are toxic. Standard preparation should
be performed in an laboratory fume hood.
5.6.2.2 Stopper the flask and then reweigh. Dilute to volume
with organic-free reagent water. Calculate the concentration from
the net gain in weight. When compound purity is assayed to be 96%
or greater, the weight can be used without correction to calculate
the concentration of the stock standard.
5.6.2.3 Stock solutions must be replaced after one year, or
sooner if comparison with the check standards indicates a problem.
5.7 Calibration standards
5.7.1 Prepare calibration standards at a minimum of five
concentrations by diluting the stock solutions with organic-free reagent
water.
5.7.2 One calibration standard should be prepared at a concentration
near, but above, the method detection limit; the remaining standards should
correspond to the range of concentrations found in real samples, but should
not exceed the working range of the HPLC system (1 mg/L to 10 mg/L).
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 HPLC Conditions
Mobile Phase: Degassed organic-free reagent water
Injection Volume: 200 /xL
Flow Rate: 2.0 mL/min
Pressure: 38 atm
Temperature: 25°C
Detector UV wavelength: 195 nm
7.2 Calibration:
7.2.1 Prepare standard solutions of acrylamide as described in
Section 5.7.1. Inject 200 ^L aliquots of each solution, in triplicate,
into the chromatograph. See Method 8000 for additional guidance on
calibration by the external standard method.
7.3 Chromatographic analysis:
7.3.1 Analyze the samples using the same Chromatographic conditions
used to prepare the standard curve. Suggested Chromatographic conditions
are given in Section 7.1. Table 1 provides the retention times that were
obtained under these conditions during method development.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Before processing any samples, the analyst must demonstrate, through
the analysis of a method blank, that all glassware and reagents are interference
free.
9.0 METHOD PERFORMANCE
9.1 Method performance data are not available.
10.0 REFERENCES
1. Hayes, Samm "Acrylamide, Acrylonitrile, and Acrolein Determination in Water
by High Pressure Liquid Chromatography," USEPA.
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TABLE 1
ANALYTE RETENTION TIMES AND METHOD DETECTION LIMITS
Retention MDL
Compound Time (min) (/.iQ/L)
Acrylamide 3.5 10
Acrylonitrile 8.9 20
Acrolein (Propenal) 10.1 30
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METHOD 8316
ACRYLAMIDE. ACRYLONITRILE AND ACROLEIN BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC)
Start
7.1 Set by
HPLC
Conditions
7 .2 Calibrate
Chromatograph
7.3
Chromatographic
analysis
Stop
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METHOD 8318
N-METHYLCARBAMATES BY HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 Method 8318 is used to determine the concentration of
N-methylcarbamates in soil, water and waste matrices. The following compounds
can be determined by this method:
Compound Name CAS No.a
Aldicarb (Temik) 116-06-3
Aldicarb Sulfone 1646-88-4
Carbaryl (Sevin) 63-25-2
Carbofuran (Furadan) 1563-66-2
Dioxacarb 6988-21-2
3-Hydroxycarbofuran 16655-82-6
Methiocarb (Mesurol) 2032-65-7
Methomyl (Lannate) 16752-77-5
Promecarb 2631-37-0
Propoxur (Baygon) 114-26-1
a Chemical Abstract Services Registry Number.
1.2 The method detection limits (MDLs) of Method 8318 for determining the
target analytes in organic-free reagent water and in soil are listed in Table 1.
1.3 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of high performance liquid chromatography (HPLC)
and skilled in the interpretation of chromatograms. Each analyst must
demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 N-methylcarbamates are extracted from aqueous samples with methylene
chloride, and from soils, oily solid waste and oils with acetonitrile. The
extract solvent is exchanged to methanol/ethylene glycol, and then the extract
is cleaned up on a C-18 cartridge, filtered, and eluted on a C-18 analytical
column. After separation, the target analytes are hydrolyzed and derivatized
post-column, then quantified fluorometrically.
2.2 Due to the specific nature of this analysis, confirmation" by a
secondary method is not essential. However, fluorescence due to post-column
derivatization may be confirmed by substituting the NaOH and o-phthalaldehyde
solutions with organic-free reagent water and reanalyzing the sample. If
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fluorescence is still detected, then a positive interference is present and care
should be taken in the interpretation of the results.
2.3 The sensitivity of the method usually depends on the level of
interferences present, rather than on the instrumental conditions. Waste samples
with a high level of extractable fluorescent compounds are expected to yield
significantly higher detection limits.
3.0 INTERFERENCES
3.1 Fluorescent compounds, primarily alkyl amines and compounds which
yield primary alkyl amines on base hydrolysis, are potential sources of
interferences.
3.2 Coeluting compounds that are fluorescence quenchers may result in
negative interferences.
3.3 Impurities in solvents and reagents are additional sources of
interferences. Before processing any samples, the analyst must demonstrate
daily, through the analysis of solvent blanks, that the entire analytical system
is interference free.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 An HPLC system capable of injecting 20 n\. aliquots and
performing multilinear gradients at a constant flow. The system must also
be equipped with a data system to measure the peak areas.
4.1.2 C-18 reverse phase HPLC column, 25 cm x 4.6 mm (5 urn).
4.1.3 Post Column Reactor with two solvent delivery systems (Kratos
PCRS 520 with two Kratos Spectroflow 400 Solvent Delivery Systems, or
equivalent).
4.1.4 Fluorescence detector (Kratos Spectroflow 980, or equivalent).
4.2 Other apparatus
4.2.1 Centrifuge.
4.2.2 Analytical balance - ± 0.0001 g.
4.2.3 Top loading balance - ± 0.01 g.
4.2.4 Platform shaker.
4.2.5 Heating block, or equivalent apparatus, that can accommodate
10 mL graduated vials (Section 4.3.11).
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4.3 Materials
4.3.1 HPLC injection syringe - 50 /xL.
4.3.2 Filter paper, (Whatman #113 or #114, or equivalent).
4.3.3 Volumetric pipettes, Class A, glass, assorted sizes.
4.3.4 Reverse phase cartridges, (C-18 Sep-PakR [Waters Associates],
or equivalent).
4.3.5 Glass syringes - 5 ml.
4.3.6 Volumetric flasks, Class A - 5 ml, 10 ml, 25 ml, 50 mL,
100 mL, and 1 L.
4.3.7 Erlenmeyer flasks with teflon-lined screw caps, 250 mL.
4.3.8 Assorted glass funnels.
4.3.9 Separatory funnels, with ground glass stoppers and teflon
stopcocks - 250 ml.
4.3.10 Graduated cylinders - 100 mL.
4.3.11 Graduated glass vials - 10 mL, 20 mL.
4.3.12 Centrifuge tubes - 250 mL.
4.3.13 Vials - 25 mL, glass with Teflon lined screw caps or crimp
tops.
4.3.14 Positive displacement micro-pipettor, 3 to 25 /xL
displacement, (Gilson Microman [Rainin #M-25] with tips, [Rainin #CP-25],
or equivalent).
4.3.15 Nylon filter unit, 25 mm diameter, 0.45 pm pore size,
disposable (Alltech Associates, #2047, or equivalent).
5.0 REAGENTS
5.1 HPLC grade chemicals shall be used in all tests. It is intended that
all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society, where such specifications are
available. Other grades may be used, provided it is first ascertained that the
reagent is of sufficiently high purity to permit its use without lowering the
accuracy of the determination.
5.2 General
5.2.1 Acetonitrile, CH3CN - HPLC grade - minimum UV cutoff at 203 nm
(EM Omnisolv #AX0142-1, or equivalent).
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5.2.2 Methanol, CH3OH - HPLC grade - minimum UV cutoff at 230 nm (EM
Omni sol v #MX0488-1, or equivalent).
5.2.3 Methylene chloride, CH2C12 - HPLC grade - minimum UV cutoff
at 230 nm (EM Omni sol v #0X0831-1, or equivalent).
5.2.4 Hexane, C6H14 - pesticide grade - (EM Omnisolv #HX0298-1, or
equivalent).
5.2.5 Ethylene glycol, HOCH2CH2OH - Reagent grade - (EM Science, or
equivalent).
5.2.6 Organic-free reagent water - All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.7 Sodium hydroxide, NaOH - reagent grade - prepare 1 L of 0.05N
NaOH solution.
5.2.8 Phosphoric acid, H3P04 - reagent grade.
5.2.9 pH 10 borate buffer (J.T. Baker #5609-1, or equivalent).
5.2.10 o-Phthalaldehyde, o-C6H4(CHO)2 - reagent grade (Fisher
#0-4241, or equivalent).
5.2.11 2-Mercaptoethanol , HSCH2CH2OH - reagent grade (Fisher
#0-3446, or equivalent).
5.2.12 N-methylcarbamate neat standards (equivalence to EPA
standards must be demonstrated for purchased solutions).
5.2.13 Chloroacetic acid, C1CH2COOH, 0.1 N.
5.3 Reaction solution
5.3.1 Dissolve 0.500 g of o-phthal aldehyde in 10 ml of methanol,
in a 1 L volumetric flask. To this solution, add 900 ml of organic-free
reagent water, followed by 50 ml of the borate buffer (pH 10). After
mixing well, add 1 ml of 2-mercaptoethanol , and dilute to the mark with
organic-free reagent water. Mix the solution thoroughly. Prepare fresh
solutions on a weekly basis, as needed. Protect from light and store under
refrigeration.
5.4 Standard solutions
5.4.1 Stock standard solutions: prepare individual 1.0 mg/mL
solutions by adding 0.025 g of carbamate to a 25 ml volumetric flask, and
diluting to the mark with methanol. Store solutions, under refrigeration,
in glass vials with Teflon lined screw caps or crimp tops. Replace every
six months.
5.4.2 Intermediate standard solution: prepare a mixed 50.0
solution by adding 2.5 ml of each stock solution to a 50 ml volumetric
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flask, and diluting to the mark with methanol. Store solutions, under
refrigeration, in glass vials with Teflon lined screw caps or crimp tops.
Replace every three months.
5.4.3 Working standard solutions: prepare 0.5, 1.0, 2.0, 3.0 and
5.0 /xg/mL solutions by adding 0.25, 0.5, 1.0, 1.5 and 2.5 mL of the
intermediate mixed standard to respective 25 mL volumetric flasks, and
diluting each to the mark with methanol. Store solutions, under
refrigeration, in glass vials with Teflon lined screw caps or crimp tops.
Replace every two months, or sooner if necessary.
5.4.4 Mixed QC standard solution: prepare a 40.0 tig/ml solution
from another set of stock standard solutions, prepared similarly to those
described in Section 5.4.1. Add 2.0 ml of each stock solution to a 50 ml
volumetric flask and dilute to the mark with methanol. Store the solution,
under refrigeration, in a glass vial with a Teflon lined screw cap or crimp
top. Replace every three months.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Due to the extreme instability of N-methylcarbamates in alkaline
media, water, waste water and leachates should be preserved immediately after
collection by acidifying to pH 4-5 with 0.1 N chloroacetic acid.
6.2 Store samples at 4°C and out of direct sunlight, from the time of
collection through analysis. N-methylcarbamates are sensitive to alkaline
hydrolysis and heat.
6.3 All samples must be extracted within seven days of collection, and
analyzed within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Water, domestic wastewater, aqueous industrial wastes, and
leachates
7.1.1.1 Measure 100 mL of sample into a 250 mL separatory
funnel and extract by shaking vigorously for about 2 minutes with
30 mL of methylene chloride. Repeat the extraction two more times.
Combine all three extracts in a 100 mL volumetric flask and dilute
to volume with methylene chloride. If cleanup is required, go to
Section 7.2. If cleanup is not required, proceed directly to Section
7.3.1.
7.1.2 Soils, solids, sludges, and heavy aqueous suspensions
7.1.2.1 Determination of sample % dry weight - In certain
cases, sample results are desired based on dry-weight basis. When
such data is desired, a portion of sample for this determination
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should be weighed out at the same time as the portion used for
analytical determination.
WARNING: The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.
7.1.2.1.1 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight
at 105°C. Allow to cool in a desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.1.2.2 Extraction - Weigh out 20 ± 0.1 g of sample into a
250 ml erlenmeyer flask with a teflon-lined screw cap. Add 50 ml
of acetonitrile and shake for 2 hours on a platform shaker. Allow
the mixture to settle (5-10 min), then decant the extract into a
250 ml centrifuge tube. Repeat the extraction two more times with
20 mL of acetonitrile and 1 hour shaking each time. Decant and
combine all three extracts. Centrifuge the combined extract at
200 rpm for 10 min. Carefully decant the supernatant into a 100 ml
volumetric flask and dilute to volume with acetonitrile. (Dilution
factor = 5) Proceed to Section 7.3.2.
7.1.3 Soils heavily contaminated with non-aqueous substances, such
as oils
7.1.3.1 Determination of sample % dry weight - Follow Sections
7.1.2.1 through 7.1.2.1.1.
7.1.3.2 Extraction - Weigh out 20 + 0.1 g of sample into a
250 ml erlenmeyer flask with a teflon-lined screw cap. Add 60 ml
of hexane and shake for 1 hour on a platform shaker. Add 50 ml of
acetonitrile and shake for an additional 3 hours. Allow the mixture
to settle (5-10 min), then decant the solvent layers into a 250 ml
separatory funnel. Drain the acetonitrile (bottom layer) through
filter paper into a 100 mL volumetric flask. Add 60 mL of hexane and
50 mL of acetonitrile to the sample extraction flask and shake for
1 hour. Allow the mixture to settle, then decant the mixture into
the separatory funnel containing the hexane from the first
extraction. Shake the separatory funnel for 2 minutes, allow the
phases to separate, drain the acetonitrile layer through filter paper
into the volumetric flask, and dilute to volume with acetonitrile.
(Dilution factor = 5) Proceed to Section 7.3.2.
7.1.4 Non-aqueous liquids such as oils
7.1.4.1 Extraction - Weigh out 20 ± 0.1 g of sample into a
125 mL separatory funnel. Add 40 mL of hexane and 25 mL of
acetonitrile and vigorously shake the sample mixture for 2 minutes.
Allow the phases to separate, then drain the acetonitrile (bottom
layer) into a 100 mL volumetric flask. Add 25 mL of acetonitrile to
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the sample funnel, shake for 2 minutes, allow the phases to separate,
drain the acetonitrile layer into the volumetric flask. Repeat the
extraction with another 25 ml portion of acetonitrile, combining the
extracts. Dilute to volume with acetonitrile. (Dilution
factor = 5). Proceed to Section 7.3.2.
7.2 Cleanup - Pipet 20.0 ml of the extract into a 20 ml glass vial
containing 100 /uL of ethylene glycol. Place the vial in a heating block set
at 50° C, and gently evaporate the extract under a stream of nitrogen (in a fume
hood) until only the ethylene glycol keeper remains. Dissolve the ethylene
glycol residue in 2 ml of methanol, pass the extract through a pre-washed C-18
reverse phase cartridge, and collect the eluate in a 5 ml volumetric flask.
Elute the cartridge with methanol, and collect the eluate until the final volume
of 5.0 ml is obtained. (Dilution factor = 0.25) Using a disposable 0.45 /xm
filter, filter an aliquot of the clean extract directly into a properly labelled
autosampler vial. The extract is now ready for analysis. Proceed to
Section 7.4.
7.3 Solvent Exchange
7.3.1 Water, domestic wastewater, aqueous industrial wastes, and
leachates:
Pipet 10.0 ml of the extract into a 10 ml graduated glass vial
containing 100 /nL of ethylene glycol. Place the vial in a heating block
set at 50° C, and gently evaporate the extract under a stream of nitrogen
(in a fume hood) until only the ethylene glycol keeper remains. Add
methanol to the ethylene glycol residue, dropwise, until the total volume
is 1.0 ml. (Dilution factor = 0.1). Using a disposable 0.45 jum filter,
filter this extract directly into a properly labelled autosampler vial.
The extract is now ready for analysis. Proceed to Section 7.4.
7.3.2 Soils, solids, sludges, heavy aqueous suspensions, and non-
aqueous liquids:
Elute 15 ml of the acetonitrile extract through a C-18 reverse phase
cartridge, prewashed with 5 ml of acetonitrile. Discard the first 2 ml
of eluate and collect the remainder. Pipet 10.0 ml of the clean extract
into a 10 ml graduated glass vial containing 100 pi of ethylene glycol.
Place the vial in a heating block set at 50° C, and gently evaporate the
extract under a stream of nitrogen (in a fume hood) until only the ethylene
glycol keeper remains. Add methanol to the ethylene glycol residue,
dropwise, until the total volume is 1.0 ml. (Additional dilution
factor =0.1; overall dilution factor = 0.5). Using a disposable 0.45 /xm
filter, filter this extract directly into a properly labelled autosampler
vial. The extract is now ready for analysis. Proceed to Section 7.4.
7.4 Sample Analysis
7.4.1 Analyze the samples using the chromatographic conditions,
post-column reaction parameters and instrument parameters given in Sections
7.4.1.1, 7.4.1.2, 7.4.1.3 and 7.4.1.4. Table 2 provides the retention
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times that were obtained under these conditions during method development.
A chromatogram of the separation is shown in Figure 1.
7.4.1.1 Chromatographic Conditions
Solvent A: Organic-free reagent water, acidified with 0.4 ml
of phosphoric acid per liter of water
Solvent B: Methanol/acetonitrile (1:1, v/v)
Flow rate: 1.0 mL/min
Injection Volume: 20 /uL
Solvent delivery system program:
Time Duration
(min) Function Value (min) File
0.00 FR 1.0 0
0.00 B% 10% 0
0.02 B% 80% 20 0
20.02 B% 100% 5 0
25.02 B% 100% 5 0
30.02 B% 10% 3 0
33.02 B% 10% 7 0
36.02 ALARM 0.01 0
7.4.1.2 Post-column Hydrolysis Parameters
Solution: 0.05 N aqueous sodium hydroxide
Flow Rate: 0.7 mL/min
Temperature: 95° C
Residence Time: 35 seconds (1 mL reaction coil)
7.4.1.3 Post-column Derivatization Parameters
Solution: o-phthalaldehyde/2-mercaptoethanol (Section 5.3.1)
Flow Rate: 0.7 mL/min
Temperature: 40° C
Residence time: 25 seconds (1 mL reaction coil)
7.4.1.4 Fluorometer Parameters
Cell: 10 ML
Excitation wavelength: 340 nm
Emission wavelength: 418 nm cutoff filter
Sensitivity wavelength: 0.5 /itA
PMT voltage: -800 V
Time constant: 2 sec
7.4.2 If the peak areas of the sample signals exceed the calibration
range of the system, dilute the extract as necessary and reanalyze the
diluted extract.
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7.5 Calibration:
7.5.1 Analyze a solvent blank (20 /uL of methanol) to ensure that
the system is clean. Analyze the calibration standards (Section 5.4.3),
starting with the 0.5 jug/mL standards and ending with the 5.0 jug/mL
standard. If the percent relative standard deviation (%RSD) of the mean
response factor (RF) for each analyte does not exceed 20%, the system is
calibrated and the analysis of samples may proceed. If the %RSD for any
analyte exceeds 20%, recheck the system and/or recalibrate with freshly
prepared calibration solutions.
7.5.2 Using the established calibration mean response factors, check
the calibration of the instrument at the beginning of each day by analyzing
the 2.0 ng/ml mixed standard. If the concentration of each analyte falls
within the range of 1.70 to 2.30 Aig/mL (i.e., within + 15% of the true
value), the instrument is considered to be calibrated and the analysis of
samples may proceed. If the observed value of any analyte exceeds its true
value by more than ± 15%, the instrument must be recalibrated (Section
7.5.1).
7.5.3 After 10 sample runs, or less, the 2.0 jug/mL standards must
be analyzed to ensure that the retention times and response factors are
still within acceptable limits. Significant variations (i.e., observed
concentrations exceeding the true concentrations by more than + 15%) may
require a re-analysis of the samples.
7.6 Calculations
7.6.1 Calculate each response factor as follows (mean value based
on 5 points):
concentration of standard
area of the signal
1
mean RF = RF = —
A RF,)
[(I RF, - RF)2]172 / 4
%RSD of RF = = X 100%
RF
7.6.2 Calculate the concentration of each N-methylcarbamate as
fol1ows:
or Mg/mL = ( RF) (area of signal) (dilution factor)
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8.0 QUALITY CONTROL
8.1 Before processing any samples, the analyst must demonstrate, through
the analysis of a method blank for each matrix type, that all glassware and
reagents are interference free. Each time there is a change of reagents, a
method blank must be processed as a safeguard against laboratory contamination.
8.2 A QC check solution must be prepared and analyzed with each sample
batch that is processed. Prepare this solution, at a concentration of 2.0 ng/mi
of each analyte, from the 40.0 pg/mL mixed QC standard solution (Section 5.4.4).
The acceptable response range is 1.7 to 2.3 /ig/mL for each analyte.
8.3 Negative interference due to quenching may be examined by spiking
the extract with the appropriate standard, at an appropriate concentration, and
examining the observed response against the expected response.
8.4 Confirm any detected analytes by substituting the NaOH and OPA
reagents in the post column reaction system with deionized water, and reanalyze
the suspected extract. Continued fluorescence response will indicate that a
positive interference is present (since the fluorescence response is not due to
the post column derivatization). Exercise caution in the interpretation of the
chromatogram.
9.0 METHOD PERFORMANCE
9.1 Table 1 lists the single operator method detection limit (MDL) for
each compound in organic-free reagent water and soil. Seven/ten replicate
samples were analyzed, as indicated in the table. See reference 7 for more
details.
9.2 Tables 2, 3 and 4 list the single operator average recoveries and
standard deviations for organic-free reagent water, wastewater and soil. Ten
replicate samples were analyzed at each indicated spike concentration for each
matrix type.
9.3 The method detection limit, accuracy and precision obtained will be
determined by the sample matrix.
10.0 REFERENCES
1. California Department of Health Services, Hazardous Materials Laboratory,
"N-Methylcarbamates by HPLC", Revision No. 1.0, September 14, 1989.
2. Krause, R.T. Journal of Chromatographic Science, 1978, vol. 16, pg 281.
3. Klotter, Kevin, and Robert Cunico, "HPLC Post Column Detection of Carbamate
Pesticides", Varian Instrument Group, Walnut Creek, CA 94598.
4. USEPA, "Method 531. Measurement of N-Methylcarbomyloximes and N-
Methylcarbamates in Drinking Water by Direct Aqueous Injection HPLC with
8318 - 10 Revision 0
November 1990
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Post Column Derivatization", EPA 600/4-85-054, Environmental Monitoring
and Support Laboratory, Cincinnati, OH 45268.
5. USEPA, "Method 632. The Determination of Carbamate and Urea Pesticides in
Industrial and Municipal Wastewater", EPA 600/4-21-014, Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268.
6. Federal Register, "Appendix B to Part 136 - Definition and Procedure for
the Determination of the Method Detection Limit - Revision 1.11", Friday,
October 26, 1984, 49, No. 209, 198-199.
7. Okamoto, H.S., D. Wijekoon, C. Esperanza, J. Cheng, S. Park, J. Garcha, S.
Gill, K. Perera "Analysis for N-Methylcarbamate Pesticides by HPLC in
Environmental Samples", Proceedings of the Fifth Annual USEPA Symposium on
Waste Testing and Quality Assurance, July 24-28, 1989, Vol. II, 57-71.
8318 - 11 Revision 0
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TABLE 1
ELUTION ORDER, RETENTION TIMES8 AND
SINGLE OPERATOR METHOD DETECTION LIMITS
Method Detection Limits"
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
a-Naphthold
Methiocarb (Mesurol)
Promecarb
Retention
Time
(min)
9.59
9.59
12.70
13.50
16.05
18.06
18.28
19.13
20.30
22.56
23.02
Organic- free
Reagent Water
(M9/L)
1.9e
1.7
2.6
2.2
9.4°
2.4
2.0
1.7
-
3.1
2.5
Soil
(Mg/Kg)
44C
12
10C
>50C
12C
17
22
31
-
32
17
See Section 7.4 for chromatographic conditions
MDL for organic-free reagent water, sand, soil were determined by analyzing
10 low concentration spike replicate for each matrix type (except where
noted). See reference 7 for more details.
MDL determined by analyzing 7 spiked replicates.
Breakdown product of Carbaryl.
8318 - 12
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TABLE 2
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA8 FOR ORGANIC-FREE REAGENT WATER
Compound Recovered % Recovery SD %RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
225
244
210
241
224
232
239
242
231
227
75.0
81.3
70.0
80.3
74.7
77.3
79.6
80.7
77.0
75.7
7.28
8.34
7.85
8.53
13.5
10.6
9.23
8.56
8.09
9.43
3.24
3.42
3.74
3.54
6.03
4.57
3.86
3.54
3.50
4.1
Spike Concentration = 300 ng/l of each compound, n = 10
8318 - 13 Revision 0
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TABLE 3
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA3 FOR WASTEWATER
Compound
Recovered
% Recovery
a Spike Concentration
b No recovery
300 /ig/L of each compound, n = 10
SD
%RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
235
247
251
b
258
263
262
262
254
263
78.3
82.3
83.7
-
86.0
87.7
87.3
87.3
84.7
87.7
17.6
29.9
25.4
-
16.4
16.7
15.7
17.2
19.9
15.1
7.49
12.10
10.11
-
6.36
6.47
5.99
6.56
7.83
5.74
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TABLE 4
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA8 FOR SOIL
Compound
Recovered
Recovery
SD
Spike Concentration =2.00 mg/Kg of each compound, n = 10
%RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
1.57
1.48
1.60
1.51
1.29
1.33
1.46
1.53
1.45
1.29
78.5
74.0
80.0
75.5
64.5
66.5
73.0
76.5
72.5
64.7
0.069
0.086
0.071
0.073
0.142
0.126
0.092
0.076
0.071
0.124
4.39
5.81
4.44
4.83
11.0
9.47
6.30
4.90
4.90
9.61
8318 - 15
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FIGURE 1
100
R
E
S
P
0
N
S
E
90
10
12 IS
TIME (HIM)
1.00 ug/oL EACH OF:
1. ALDICAR3 SULFONE
2. METHOMYL
3 . 3 - HYTDROXYCARBO FURAN
-------
( START
|_7.1 Extraction
00
U>
CO
i
15
n>
to
o
7.1.1 Water, domestic wostewoter.
aqueous industrial wastes.
ond leochotes
.1 Extract 100 mis sample
w/30 mts MeCI 3x in sep.
funnel: combine extracts
in 100 ml. vol. flask ond
dilute to mark
No
7.1.2 Soils, solids, sludges.
and heavy aqueous
suspensions
Yes
7.2 Cleanup
Combine 20 mis. extract
and 100 ul ethylene glycol
in o gloss vial: blowdown
mixture w/N2 in heating
block set at 50 C: dissolve
residue in 2 mis. MeOH.
pass soln. through pre-
woshed CIS cartridge:
collect nluale in 5 ml. vol.
flask: elule cartridge
w/MeOH into vol. flask up
to mark: filter MeOH soln.
through 0.45 um litter into
outosompler vial
^T Determine % dry wt.:
.1 Weigh 5-10 gr sample
into crucible: oven dry
overnight at 105 C; cool
in dessicaton reweigh
.2 Extraction:
Weigh 20 gr. sample into
250 Erlenmeyen add 50
mis. acetonitrile. shake for
2 hrs.; decant extract into
centrifuge tube; repeat
extraction 2x w/ 20 mis.
ocetonitrile. shake 1 hr.;
combine extracts and
centrifuge 10 mkis. 200 rpm;
decant supernatant to 100 ml.
vol. flask ond dilute to mark
7.1.3 Soils heavily contaminated
with non-aqueous substances,
such as pits
etermine % dry wt.:
Follow Section 7.1.2.1
.2 Extraction:
Weigh 20 gr. sample into
250 Erienmoyen odd 60 mis.
hexone, shake 1 hr.: add 50
mis. ocetonrtrile. shake 3 hrs.;
let settle, decant extract layers
to 250 ml. sep. funnel; fitter
bottom ocetonifrite layer into
100 ml. vol. fbsk; repeat
sample fbsk extraction w/some
volumes; decant extract layers
on top of first hexone layer;
shake funnel: fitter bottom layer
into vol. fbsk; dilute to mark
7.1.4 Non-aqueous liquids
iuch os oils
-ft!
x (ruction:
Weigh 20 gr. sample into
125 ml. sap. funnel; odd
40 mis. hexone ond 25 mis.
ocelonttrite; shake, settle, and
drain bottom ocetonitrile layer
into 100 ml vol. flask: repeat
extraction 2x by adding 25 mis.
acetonitrie to initial fbsk mix;
combine ocetonitrile layers into
vol. flask; dfate to mark
o
o
a:
§
5
• o
;o
CD
• (*>
7.3 Solvent Exchange
7.3.1 Water, domestic wostewoter,
aqueous industrial wastes.
ond leachates:
Combine 10 mis extract ond
100 ul ethylene glycol in a
glass vial; blowdown mixture
w/N2 in heating block at
50 C: odd MeOH to residue
to total volume of 1 ml.; filter
MeOH soln. through 0.45 urn
filter into ouloscmpler vial
7.3 Solvent Exchange
7.3.2 Soils, solids, sludges.
heavy aqueous suspensions.
and non-aqueous liquids:
Elute 15 mis. extract through
acetonilrile prewoshed CIS
cartridge, collect tatter 13 mis.:
combine 10 mis. cleaned
extract and 100 ul ethylene
glycol in gbss viol; bbwdown
mixture w/N2 in heating block
at 50 C: odd MeOH to residue
to total volume of 1 ml.: filter
MeOH soln. through 0.45 urn
filter into outosampler vial
-------
I 7.4 Sample Analysis
7.4.1 Initialize Instrumentation:
.1 Set chromatographic parameters
.2 Set Post-column Hydrolysis parameters
.3 Set Post-column Oerivotizotion parameters
.4 Set Fkiorometer parameters
I
7.4.2 Dilute sample extract and reanalyze if
calibration range is exceeded
CD
u>
H^
CD
I
t—•
00
-M7.5 Calibration J
T
7.5.1 Analyze a solvent blank then the calibration
stds. of Section 5.4.3; ensure that %RSD of
each analyte response factor (RF) is < 20%;
recheck system and recalibrate w/fresh
solns. if %RSD > 20%
I
7.5.2 Check calibration doily w/2 ug/ml std.;
ensure that individual onolyte cones, fall
w/in +/- %15 of true value; recalibrate
if observed difference > 15%
r> m
o —»
3 3:
c-f O
— O
C CD
(A)
Q. •—
7.5.3 Check calibratbn every 10 samples or less
w/2 ug/ml std.; variations > 15% may
require re-analysis of samples
7.6 Calculations
I
7.6.1 Calculate response factors and % RSD
according to equation
7.6.2 Calculate sample onolyte cones, according
to equation
( STOP )
-------
METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH- PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TSP/MS) OR ULTRAVIOLET (UV) DETECTION
1.0 SCOPE AND APPLICATION
1.1 This method covers the use of high performance liquid chromatography
(HPLC), coupled with either thermospray-mass spectrometry (TSP-MS), and/or
ultraviolet/visible (UV/VIS), for the determination of disperse azo dyes,
organophosphoruscompounds, andTris-(2,3-dibromopropyl)phosphate, inwastewater,
ground water, sludge, and soil/sediment matrices. Additionally, it may apply
to other non-volatile compounds that are solvent extractable, are amenable to
HPLC, and are ionizable under thermospray introduction for mass spectrometric
detection. The following compounds can be determined by this method:
Compound Name
CAS No.Ł
Azo Dyes
Disperse Red 1
Disperse Red 5
Disperse Red 13
Disperse Yellow 5
Disperse Orange 3
Disperse Orange 30
Disperse Brown 1
Solvent Red 3
Solvent Red 23
Anthraquinone Dves
Disperse Blue 3
Disperse Blue 14
Disperse Red 60
Coumarin Dyes
(Fluorescent Brighteners)
Fluorescent Brightener 61
Fluorescent Brightener 236
Alkaloids
Caffeine
Strychnine
2872-
3180-
2832-
6439-
730-
5261-
17464-
6535-
85-
52-8
81-2
40-8
53-8
40-5
31-4
91-4
42-8
86-9
2475-46-9
2475-44-7
17418-58-5
8066-05-5
63590-17-0
58-08-2
57-24-9
8321 - 1
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Compound Name CAS No.a
Orqanophosphorus Compounds
Methomyl 16752-77-5
Thiofanox 39196-18-4
Famphur 52-85-7
Asulam 3337-71-1
Dichlorvos 62-73-7
Dimethoate 60-51-5
Disulfoton 298-04-4
Fensulfothion 115-90-2
Merphos 150-50-5
Methyl parathion 298-00-0
Monocrotophos 919-44-8
Naled 300-76-5
Phorate 298-02-2
Trichlorfon 52-68-6
Tris-(2,3-Dibromopropyl) phosphate, (Tris-BP) 126-72-7
a Chemical Abstract Services Registry Number.
1.2 This method may be applicable to the analysis of other non-volatile
or semivolatile compounds.
1.3 Tris-BP has been classified as a carcinogen. Purified standard
material and stock standard solutions should be handled in a hood.
1.4 The compounds were chosen for analysis by HPLC/MS because they have
been designated as problem compounds that are hard to analyze by traditional
chromatographic methods (e.g. gas chromatography). The sensitivity of this
method is dependent upon the level of interferants within a given matrix, and
varies with compound class and even with compounds within that class.
Additionally, the limit of detection (LOD) is dependent upon the mode of
operation of the mass spectrometer. For example, the LOD for caffeine in the
selected reaction monitoring (SRM) mode is 45 pg of standard injected (10 n\.
injection), while for Disperse Red 1 the LOD is 180 pg. The LOD for caffeine
under single quadrupole scanning is 84 pg and is 600 pg for Disperse
Red 1 under similar scanning conditions.
1.5 The experimentally determined method detection limits (MDL) for the
target analytes are presented in Tables 3, 10 and 13. For further compound
identification, MS/MS (CAD - collision activated dissociation) can be used as
an optional extension of this method.
1.6 This method is restricted to use by or under the supervision of
analysts experienced in the use of high performance liquid chromatographs/mass
spectrometers and skilled in the interpretation of liquid chromatograms and mass
8321 - 2 Revision 0
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spectra. Each analyst must demonstrate the ability to generate acceptable results
with this method.
2.0 SUMMARY OF METHOD
2.1 This method provides reverse phase high performance liquid
chromatographic (RP/HPLC) and thermospray (TSP) mass spectrometric (MS)
conditions for the detection of the target analytes. Quantitative analysis is
performed by TSP/MS, using an external standard approach. Sample extracts can
be analyzed by direct injection into the thermospray or onto a liquid
chromatographic-thermospray interface. A gradient elution program is used on
the chromatograph to separate the compounds. Since this method is based on an
HPLC technique, the use of ultraviolet/visible (UV/VIS) detection is optional
on routine samples.
2.2 Prior to the use of this method, appropriate sample preparation
techniques must be used. In general, water samples are extracted at a neutral
pH with methylene chloride, using a separatory funnel (Method 3510) or a
continuous liquid-liquid extractor (Method 3520). Soxhlet (Method 3540) or
ultrasonic (Method 3550) extraction using methylene chloride/acetone (1:1) is
used for solid samples. A micro-extraction technique is included for the
extraction of Tris-BP from aqueous and non-aqueous matrices.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600 and 8000.
3.2 The use of Florisil Column Cleanup (Method 3620) has been demonstrated
to yield recoveries less than 85% for some of the compounds in this method, and
is therefore not recommended for all compounds. Refer to Table 2 of Method 3620
for recoveries of organophosphorus compounds as a function of Florisil fractions.
3.3 Compounds with high proton affinity may mask some of the target
analytes. Therefore, an HPLC must be used as a chromatographic separator, for
quantitative analysis.
3.4 Analytical difficulties encountered with specific organophosphorus
compounds, as applied in this method, may include (but are not limited to) the
following:
3.4.1 Methyl parathion shows some minor degradation upon analysis.
3.4.2 Naled can undergo debromination to form dichlorvos.
3.4.3 Merphos often contains contamination from merphos oxide.
Oxidation of merphos can occur during storage, and possibly upon
introduction into the mass spectrometer.
Refer to Method 8141 for other compound problems as related to the various
extraction methods.
8321 - 3
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3.5 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts or elevated baselines, or both, causing
misinterpretation of chromatograms or spectra. All of these materials must be
demonstrated to be free from interferences under the conditions of the analysis
by running reagent blanks. Specific selection of reagents and purification of
solvents by distillation in all-glass systems may be required.
3.6 Interferants co-extracted from the sample will vary considerably from
source to source. Retention times of target analytes must be verified by using
reference standards.
3.7 The optional use of HPLC/MS/MS methods aids in the confirmation of
specific analytes. These methods are less subject to chemical noise than other
mass spectrometric methods.
4.0 APPARATUS AND MATERIALS
4.1 HPLC/MS
4.1.1 High Performance Liquid Chromatograph (HPLC) - An analytical
system with programmable solvent delivery system and all required
accessories including 10 /xL injection loop, analytical columns, purging
gases, etc. The solvent delivery system must be capable, at a minimum,
of a binary solvent system. The chromatographic system must be capable
of interfacing with a Mass Spectrometer (MS).
4.1.1.1 HPLC Post-Column Addition Pump - A pump for post
column addition should be used. Ideally, this pump should be a
syringe pump, and does not have to be capable of solvent programming.
4.1.1.2 HPLC Columns - A guard column and an analytical column
are required.
4.1.1.2.1 Guard Col umn - CIS reverse phase guard col umn,
10 mm x 2.6 mm ID, 0.5 p.m frit, or equivalent.
4.1.1.2.2 Analytical Column - CIS reverse phase column,
100 mm x 2 mm.ID, 5 pm particle size of ODS-Hypersil; or CIS
reversed phase column, 100 mm x 2 mm ID, 3 /xm particle size
of MOS2-Hypersil, or equivalent.
4.1.2 HPLC/MS interface(s)
4.1.2.1 Micromixer - 10 /xL, interfaces HPLC column system
with HPLC post-column addition solvent system.
4.1.2.2 Interface - Thermospray ionization interface and
source that will give acceptable calibration response for each
analyte of interest at the concentration required. The source must
be capable of generating both positive and negative buffer assisted
ions, and have both a repeller and a discharge electrode for
enhancement of the ion signal in both modes, respectively.
8321 - 4 Revision 0
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4.1.3 Mass spectrometer system - A single quadrupole mass
spectrometer capable of scanning from 1 to 1000 amu. The spectrometer
must also be capable of scanning from 150 to 450 amu in 1.5 sec or less,
using 70 volts (nominal) electron energy in the positive or negative
electron impact modes. In addition, the mass spectrometer must be capable
of producing a calibrated mass spectrum for PEG 400, 600, or 800 (see
Section 5.12).
4.1.3.1 Optional triple quadrupole mass spectrometer - capable
of generating daughter ion spectra with a collision gas in the second
quadrupole and operation in the single quadrupole mode.
4.1.4 Data System - A computer system that allows the continuous
acquisition and storage on machine-readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows any MS data file to be searched for ions of a specified mass, and
such ion abundances to be plotted versus time or scan number. This type
of plot is defined as an Extracted Ion Current Profile (EICP). Software
must also be available that allows integration of the abundances in any
EICP between specified time or scan-number limits. There must be computer
software available to operate the specific modes of the mass spectrometer.
4.2 HPLC with UV/VIS detector - An analytical system with solvent
programmable pumping system for at least a binary solvent system, and all
required accessories including syringes, 10 /uL injection loop, analytical
columns, purging gases, etc. An automatic injector is optional, but is useful
for multiple samples. The columns specified in Section 4.1.1.2 are also used
with this system.
4.2.1 If the UV/VIS detector is to be used in tandem with the
thermospray interface, then the detector cell must be capable of
withstanding high pressures (at least 6000 psi).
4.3 Purification Equipment for Azo Dye Standards
4.3.1 Soxhlet extraction apparatus.
4.3.2 Extraction thimbles, single thickness, 43 x 123 mm.
4.3.3 Filter paper, 9.0 cm (Whatman qualitative No. 1 or
equivalent).
4.3.4 Silica-gel column - 3 in. x 8 in., packed with Silica gel
(Type 60, EM reagent 70/230 mesh).
4.4 Kuderna-Danish (K-D) apparatus (optional).
4.4.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025
or equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
8321 - 5 Revision 0
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4.4.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.4.3 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.4.4 Springs - 1/2 in. (Kontes K-662750 or equivalent).
4.5 Disposable serological pipets - 5 ml x 1/10, 5.5 mm ID.
4.6 Collection tube - 15 ml conical, graduated (Kimble No. 45165 or
equivalent).
4.7 Vials - 5 ml conical, glass, with Teflon lined screw-caps or crimp
tops.
4.8 Glass wool - Supelco No. 2-0411 or equivalent.
4.9 Microsyringes - 100 ML, 50 ML, 10 juL (Hamilton 701 N or equivalent),
and 50 ML (Blunted, Hamilton 705SNR or equivalent).
4.10 Rotary evaporator - Equipped with 1,000 ml receiving flask.
4.11 Balances - Analytical, 0.0001 g, Top-loading, 0.01 g.
4.12 Volumetric flasks, Class A - 10 mL to 1000 mL.
4.13 Graduated cylinder - 100 ml.
4.14 Separatory funnel - 250 ml.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene
chloride,a method blank must be analyzed, demonstrating that there is no
interference from the sodium sulfate.
5.4 Ammonium acetate, NH4OOCCH3, solution (0.1 M). Filter through a 0.45
micron membrane filter (Millipore HA or equivalent).
8321 - 6 Revision 0
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5.5 Argon gas, 99+% pure.
5.6 Solvents
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.6.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.6.4 Diethyl Ether, C2H5OC2H5 - Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.6.5 Methanol, CH3OH - HPLC quality or equivalent.
5.6.6 Acetonitrile, CH3CN - HPLC quality or equivalent.
5.6.7 Ethyl acetate CH3C02C2H5 - Pesticide quality or equivalent.
5.7 Standard Materials - pure standard materials or certified solutions
of each analyte targeted for analysis. Disperse azo dyes must be purified before
use according to Section 5.8. Tris-(2,3-dibromopropyl) phosphate, 98+% pure,
may be obtained from the U.S.EPA Repository, Research Triangle Park, North
Carolina.
5.8 Disperse Azo Dye Purification
5.8.1 Two procedures are involved. The first step is the Soxhlet
extraction of the dye for 24 hours with toluene and evaporation of the
liquid extract to dryness, using a rotary evaporator. The solid is then
recrystallized from toluene, and dried in an oven at approximately 100°C.
If this step does not give the required purity, column chromatography
should be employed. Load the solid onto a 3 x 8 inch silica gel column
(Section 4.3.5), and elute with diethyl ether. Separate impurities
chromatographically, and collect the major dye fraction.
5.9 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially prepared
stock standards can be used if they are verified against EPA standards. If EPA
standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable.
5.9.1 Prepare stock standard solutions by accurately weighing
0.0100 g of pure material. Dissolve the material in methanol or other
suitable solvent (e.g. prepare Tris-BP in ethyl acetate), and dilute to
known volume in a volumetric flask. If compound purity is certified at
96% or greater, the weight can be used without correction to calculate
the concentration of the stock standard. Commercially prepared stock
standards can be used at any concentration if they are certified by the
manufacturer or by an independent source.
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5.9.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp-tops. Store at 4°C and protect from
light. Stock standard solutions should be checked frequently for signs
of degradation or evaporation, especially just prior to preparing
calibration standards.
5.10 Calibration standards - A minimum of five concentrations for each
parameter of interest should be prepared through dilution of the stock standards
with methanol (or other suitable solvent). One of these concentrations should
be near, but above, the MDL. The remaining concentrations should correspond to
the expected range of concentrations found in real samples, or should define the
working range of the HPLC-UV/VIS or HPLC-TSP/MS. Calibration standards must be
replaced after one or two months, or sooner if comparison with check standards
indicates a problem.
5.11 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, along with the
effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two surrogates (e.g. organophosphorus
compounds not expected to be present in the sample).
5.12 HPLC/MS tuning standard - Polyethylene glycol 400 (PEG-400), PEG-
600 or PEG-800. Dilute to 10 percent (v/v) in methanol. Dependent upon analyte
molecular weight range: m.w. < 500 amu, use PEG-400; m.w. > 500 amu, use PEG-600,
or PEG-800.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample preparation - Samples must be prepared by one of the following
methods prior to HPLC/MS analysis:
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3550
Waste 3540, 3550, 3580
7.1.1 Microextraction for Tris-BP:
7.1.1.1 Non-Aqueous Samples
7.1.1.1.1 Weigh a 1 gram portion of the sample into a
tared beaker. If the sample is moist, add an equivalent amount
of anhydrous sodium sulfate and mix well. Add 100 /xL of
Tris-BP (approximate concentration 1000 mg/L) to the sample
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selected for spiking; the amount added should result in a final
concentration of 100 ng//iL in the 1 ml extract.
7.1.1.1.2 Pack the sample into a disposable pipet
prepared according to Section 7.1.1.1.2.1. If packing material
has dried, rinse with methanol first, then pack sample into
the pipet.
7.1.1.1.2.1 Remove the glass wool plug. Insert
a 1 cm plug of clean si lane treated glass wool to the
bottom (narrow end) of the pipet. Pack 2 cm of sodium
sulfate (dried) onto the top of the glass wool. Wash
pipet with 3-5 ml of methanol.
7.1.1.1.3 Extract the sample with 3 ml of methanol
followed by 4 ml of 50% (v/v) methanol/methylene chloride.
Collect extract in 15 ml graduated glass tubes.
7.1.1.1.4 Evaporate the extract to 1 ml using the micro
Snyder column technique (Section 7.1.1.1.5) or nitrogen
blowdown technique (Section 7.1.1.1.6). Record the volume.
It may not be possible to evaporate some sludge samples to a
reasonable concentration.
7.1.1.1.5 Micro-Snyder Column Technique
7.1.1.1.5.1 Add the sample and one or two clean
boiling chips to the concentrator tube and attach a two
ball micro-Snyder column. Prewet the column by adding
about 0.5 ml of methylene chloride to the top of the
column. Place the K-D apparatus in a hot water bath so
that the concentrator tube is partially immersed in the
hot water. Adjust the vertical position of the apparatus
and the water temperature, as required, to complete the
concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively
chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5 ml, remove the K-
D apparatus from the water bath and allow it to drain
and cool for at least 10 minutes. Remove the Snyder
column and rinse the flask and its lower joints with
about 0.2 mL of methyl ene chloride and add to the
concentrator tube. Proceed to Section 7.1.1.1.7.
7.1.1.1.6 Nitrogen Blowdown Technique
7.1.1.1.6.1 PI ace the concentrator tube in a warm
water bath (approximately 35°C) and evaporate the solvent
volume to the required level using a gentle stream of
clean, dry nitrogen (filtered through a column of
activated carbon).
CAUTION; Do not use plasticized tubing between the carbon trap and the sample.
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7.1.1.1.6.2 The internal wall of the tube must
be rinsed down several times with methylene chloride
during the operation. During evaporation, the solvent
level in the tube must be positioned to prevent water
from condensing into the sample (i.e., the solvent level
should be below the level of the water bath). Under
normal operating conditions, the extract should not be
allowed to become dry. Proceed to Section 7.1.1.1.7.
7.1.1.1.7 Transfer the extract to a glass vial with a
Teflon lined screw-cap or crimp-top and store refrigerated at
4°C. Proceed with HPLC analysis.
7.1.1.2 Aqueous (Water and Municipal Waste Water) Samples
7.1.1.2.1 Using a 100 ml graduated cylinder, measure
100 ml of sample and transfer it to a 250 ml separatory funnel.
Add 200 pi of Tris-BP (approximate concentration 1000 mg/L)
to the sample selected for spiking; the amount added should
result in a final concentration of 200 ng//iL in the 1 ml
extract.
7.1.1.2.2 Add 10 ml of methyl ene chloride to the
separatory funnel. Seal and shake the separatory funnel three
times, approximately 30 seconds each time, with periodic
venting to release excess pressure. NOTE: Methylene chloride
creates excessive pressure rapidly; therefore, initial venting
should be done immediately after the separatory funnel has been
sealed and shaken once. Methylene chloride is a suspected
carcinogen, use necessary safety precautions.
7.1.1.2.3 Allow the organic layer to separate from the
water phase for a minimum of 10 minutes. If the emulsion
interface between layers is more than one-third the size of
the solvent layer, the analyst must employ mechanical
techniques to complete phase separation. See Section 7.5,
Method 3510.
7.1.1.2.4 Collect the extract in a 15 ml graduated
glass tube. Proceed as in Section 7.1.1.1.4.
7.2 Prior to HPLC analysis, the extraction solvent must be exchanged to
methanol. The exchange is performed during the K-D procedures listed in all of
the extraction methods.
7.3 HPLC Chromatographic Conditions:
7.3.1 Analyte-specific chromatographic conditions are shown in
Table 1. Chromatographic conditions which are not analyte-specific are
as follows:
Flow rate: 0.4 mL/min
Post-column mobile phase: 0.1 M ammonium acetate (1% methanol)
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Post-column flow rate: 0.8 mL/min
7.3.2 If there is a chromatographic problem from compound retention,
a 2% constant flow of methylene chloride may be applied as needed.
Methylene chloride/aqueous methanol solutions must be used with caution
as HPLC eluants. Acetic acid (1%), another mobile phase modifier, can be
used with compounds with acid functional groups.
7.3.3 A total flow rate of 1.0 to 1.5 mL/min is necessary to
maintain thermospray ionization.
7.3.4 Retention times for organophosphorus compounds on the
specified analytical column are presented in Table 9.
7.4 Recommended HPLC/Thermospray/MS operating conditions:
7.4.1 Positive Ionization mode
Repeller (wire or plate): 170 to 250 v (sensitivity optimized).
Mass range: 150 to 450 amu (compound dependent, expect 1 to 18 amu higher
than molecular weight of the compound).
Scan time: 1.50 sec/scan.
7.4.2 Negative Ionization mode
Discharge electrode: on
Filament: off
Mass Range: 135 to 450 amu (compound dependent, expect 1 amu lower than
molecular weight of the compound).
Scan time: 1.50 sec/scan.
7.4.3 Thermospray temperatures:
Vaporizer control » 110°C to 130°C (as necessary to achieve proper stable
tip and jet temperatures without loss of sensitivity.
See Manufacturer's recommendations).
Vaporizer tip - 200°C.
Jet « 210°C to 220°C.
Source block « 240°C to 265°C. (Some compounds may degrade in the
source block at higher temperatures, operator should
use knowledge of chemical properties to estimate
proper source temperature).
7.4.4 Sample injection volume: 20 /iL is necessary in order to
overfill the 10 nl injection loop.
7.5 Calibration:
7.5.1 Thermospray/MS system - Must be hardware-tuned, on
quadrupole 1 (and quadrupole 3 for triple quadrupoles), for accurate mass
assignment, sensitivity, and resolution. This is accomplished using
polyethylene glycol (PEG) 400, 600, or 800 (see Section 5.12) which has
average molecular weights of 400, 600, and 800, respectively. A mixture
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of these PEGs can be made such that it will approximate the expected
working mass range for the analyses. The PEG is introduced via the
thermospray interface, circumventing the HPLC.
7.5.1.1 The mass calibration parameters are as follows:
for PEG 400 and 600 for PEG 800
Mass range: 15 to 765 amu Mass range: 15 to 900 amu
Scan time: 5.00 sec/scan Scan time: 5.00 sec/scan
Approximately 100 scans should be acquired, with 2 to 3
injections made. The scan with the best fit to the accurate mass
table (see Tables 7 and 8) should be used as the calibration table.
7.5.1.2 The low mass range from 15 to 100 amu is covered by
the ions from the ammonium acetate buffer used in the thermospray
process: NH4+ (18 amu), NH4+H20 (36), CH3OHNH4+ (50) (methanol), or
CH3CNNH4+ (59) (acetonitrile), and CH3COOHNH4+ (78) (acetic acid).
The appearance of the m/z 50 or 59 ion depends upon the use of
methanol or acetonitrile as the organic modifier. The higher mass
range is covered by the ammonium ion adducts of the various ethylene
glycols (e.g. H(OCH2CH2)nOH where n=4, gives the H(OCH2CH2)4OHNH4+ ion
at m/z 212).
7.5.2 Liquid Chromatograph
7.5.2.1 Prepare calibration standards as outlined in Section
5.10.
7.5.2.2 Choose the proper ionization conditions, as outlined
in Section 7.4.1. Inject each calibration standard onto the HPLC,
using the chromatographic conditions outlined in Table 1. Calculate
the area under the curve for the mass chromatogram of each
quantitation ion. For example, Table 9 lists the retention times
and the major ions (> 5%) present in the positive ionization
thermospray single quadrupole spectra of the organophosphorus
compounds. In most cases the (M+H)+ and (M+NH4)+ adduct ions are the
only ions of significant abundance. Plot these ions as area response
versus the amount injected. The points should fall on a straight
line, with a correlation coefficient of at least 0.99.
7.5.2.3 If HPLC-UV/VIS detection is also being used, calibrate
the instrument by preparing calibration standards as outlined in
Section 5.10, and injecting each calibration standard onto the HPLC
using the chromatographic conditions outlined in Table 1. Integrate
the area under the full chromatographic peak for each concentration.
7.5.2.4 For the methods specified in Section 7.5.2.2 and
7.5.2.3, the retention time of the chromatographic peak is an
important variable in analyte identification. Therefore, the ratio
of the retention time of the sample analyte to the standard analyte
should be 1.0 ± 0.1.
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7.5.2.5 The concentration of the sample analyte will be
determined by using the calibration curves determined in Sections
7.5.2.2 and 7.5.2.3. These calibration curves must be generated on
the same day as each sample is analyzed. At least duplicate
determinations must be made for each sample extract. Concentrated
samples must be diluted by a known amount.
7.5.2.6 Refer to Method 8000 for further information on
calculations.
7.5.2.7 Precision can also be calculated from the ratio of
response (area) to the amount injected; this is defined as the
calibration factor (CF) for each standard concentration. If the
percent relative standard deviation (%RSD) of the CF is less than
20 percent over the working range, linearity through the origin can
be assumed, and the average calibration factor can be used in place
of a calibration curve. The CF and %RSD can be calculated as
follows:
CF = Total Area of Peak/Mass injected (ng)
%RSD = SD/CF x 100
where:
SD = Standard deviation between CFs
CF = Average CF
7.5.2.8 The working calibration curve, or the CF, must be
verified on each working day by the injection of one or more
calibration standards. If the response varies from the predicted
response by more than ± 20 percent, a new calibration curve must be
prepared. The % Difference is calculated as follows:
% Difference = (R, - R2)/R, x 100.
where: R., = CF first analysis.
R2 = CF from succeeding analyses.
7.6 Sample Analysis
7.6.1 Once the LC/MS system has been calibrated as outlined in
Section 7.5, then it is ready for sample analysis.
7.6.1.1 A blank 20-/uL injection (methanol) must be analyzed
after the standard(s) analyses, in order to determine any residual
contamination of the Thermospray/HPLC/MS system.
7.6.1.2 Take a 20-jiL aliquot of the sample extract from
Section 7.1. Start the HPLC gradient elution, load and inject the
sample aliquot, and start the mass spectrometer data system analysis.
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7.7 Calculations
7.7.1 Using the external standard calibration procedure (Method
8000), determine the identity and quantity of each component peak in the
sample reconstructed ion chromatogram which corresponds to the compounds
used for calibration processes. See Method 8000 for calculation equations.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Tables 4, 5, 11, and 12 indicate the single operator accuracy and
precision for this method. Compare the results obtained with the results in the
tables to determine if the data quality is acceptable.
8.3.1 If recovery is not acceptable, check the following:
8.3.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.3.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.3.1.3 If no problem is found, re-extract and re-analyze the
sample.
8.3.1.4 If, upon re-analysis, the recovery is again not within
limits, flag the data as "estimated concentration".
8.4 Instrument performance - Check the performance of the entire
analytical system daily using data gathered from analyses of blanks, standards,
and replicate samples.
8.4.1 See Section 7.4 for required HPLC/MS parameters for standard
calibration curve %RSD limits.
8.4.2 See Section 7.5.2.4 regarding retention time window QC limits.
8.4.3 If any of the chromatographic QC limits are not met, the
analyst should examine the LC system for:
o Leaks,
o Proper pressure delivery,
o A dirty guard column; may need replacing or repacking, and
o Possible partial thermospray plugging.
Any of the above items will necessitate shutting down the HPLC/TSP
system, making repairs and/or replacements, and then restarting the
analyses. The calibration standard should be reanalyzed before any sample
analyses, as described in Section 7.5.
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8.4.4 The experience of the analyst performing 1 iquid chromatography
is invaluable to the success of the method. Each day that analysis is
performed, the daily calibration standard should be evaluated to determine
if the chromatographic system is operating properly. If any changes are
made to the system (e.g. column change), the system must be recalibrated.
8.5 Optional Thermospray HPLC/MS/MS confirmation
8.5.1 With respect to this method, MS/MS shall be defined as the
daughter ion collision activated dissociation acquisition with quadrupole
one set on one mass (parent ion), quadrupole two pressurized with argon
and with a higher offset voltage than normal, and quadrupole three set to
scan desired mass range.
8.5.2 Since the thermospray process often generates only one or
two ions per compound, the use of MS/MS is a more specific mode of
operation yielding molecular structural information. In this mode, fast
screening of samples can be accomplished through direct injection of the
sample into the thermospray.
8.5.3 For MS/MS experiments, the first quadrupole should be set to
the protonated molecule or ammoniated adduct of the analyte of interest.
The third quadrupole should be set to scan from 30 amu to just above the
mass region of the protonated molecule.
8.5.4 The collision gas pressure (Ar) should be set at about
1.0 mTorr and the collision energy at 20 eV. If these parameters fail to
give considerable fragmentation, they may be raised above these settings
to create more and stronger collisions.
8.5.5 For analytical determinations, the base peak of the collision
spectrum shall be taken as the quantification ion. For extra specificity,
a second ion should be chosen as a backup quantification ion.
8.5.6 Generate a calibration curve as outlined in Section 7.5.2.
t
8.5.7 For analytical determinations, calibration blanks must be
run in the MS/MS mode to determine specific ion interferences. If no
calibration blanks are available, chromatographic separation must be
performed to assure no interferences at specific masses. For fast
screening, the MS/MS Spectra of the standard and the analyte could be
compared and the ratios of the three major (most intense) ions examined.
These ratios should be approximately the same unless there is an
interference. If an interference appears, chromatography must be utilized.
8.5.8 For unknown concentrations, the total area of the quantitation
ion(s) is calculated and the calibration curves generated as in Section
7.6 are used to attain an injected weight number.
8.5.9 MS/MS techniques can also be used to perform structural
analysis on ions represented by unassigned m/z ratios. The procedure for
compounds of unknown structures is to set up a CAD experiment on the ion
of interest. The spectrum generated from this experiment will reflect the
structure of the compound by its fragmentation pattern. A trained mass
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spectroscopist and some history of the sample are usually needed to
interpret the spectrum. (CAD experiments on actual standards of the
expected compound are necessary for confirmation or denial of that
substance.)
9.0 METHOD PERFORMANCE
9.1 Single operator accuracy and precision studies have been conducted
using spiked sediment, wastewater, sludge, and water samples. The results are
presented in Tables 4, 5, 6, 11, and 12. Tables 3, 10, and 13 list precision
and bias data that are typical with this method.
9.2 MDLs should be calculated for the known analytes, on each instrument
to be used.
9.2.1 The MDLs presented in this method were calculated by analyzing
three replicates of four standard concentrations, with the lowest
concentration being near the instrument detection limit. A linear
regression was performed on the data set to calculate the slope and
intercept. Three times the standard deviation (3a) of the lowest standard
amount, along with the calculated slope and intercept, was used to find
the MDL. The MDL was not calculated using the specifications in Chapter
One, but according to the ACS guidelines specified in Reference 4.
10.0 REFERENCES
1. Voyksner, R.D.; Haney, C.A. "Optimization and Application of Thermospray
High-Performance Liquid Chromatography/Mass Spectrometry"; Anal. Chem. 1985,
5Z, 991-996.
2. Blakley, C.R.; Vestal, M.L. "Thermospray Interface for Liquid
Chromatography/Mass Spectrometry"; Anal. Chem. 1983, 55, 750-754.
3. Taylor, V.; Hickey, D. M., Marsden, P. J. "Single Laboratory Validation of
EPA Method 8140"; EPA-600/4-87/009, U.S. Environmental Protection Agency,
Las Vegas, NV, 1987, 144 pp.
4. "Guidelines for Data Acquisition and Data Quality Evaluation in Environmental
Chemistry"; Anal. Chem. 1980, 52, 2242-2249.
5. Betowski, L. D.; Jones, T. L. "The Analysis of Organophosphorus Pesticide
Samples by HPLC/MS and HPLC/MS/MS"; Environmental Science and Technology.
1988,
8. EPA: 2nd Annual Report on Carcinogens, NTP 81-43, Dec. 1981, pp. 236-237.
9. Blum, A.; Ames, B. N. Science 195. 1977, 17.
10. Zweidinger, R. A.; Cooper, S. D.; Pellazari, E. D., Measurements of Organic
Pollutants in Water and Wastewater. ASTM 686.
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TABLE 1.
RECOMMENDED HPLC CHROMATOGRAPHIC CONDITIONS
Initial
Mobile
Phase
(%)
Analytes:
Orqanoohosphorus
Initial
Time
(min)
Compounds
Gradient
(linear)
(min)
Final
Mobile
Phase
(%)
Final
Time
(min)
50/50
(water/methanol)
10 100 5
(methanol)
Azo Dves (e.g. Disperse Red 1)
50/50 0
(water/CH3CN)
Tris-(2.3-dibromopropv1)phosphate
50/50 0
(water/methanol)
10
100 5
(CH3CN)
100 5
(methanol)
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TABLE 2.
COMPOUNDS AMENABLE TO THERMOSPRAY MASS SPECTROMETRY
Disperse Azo Dyes
Methine Dyes
Arylmethane Dyes
Coumarin Dyes
Anthraquinone Dyes
Xanthene Dyes
Flame retardants
Alkaloids
Aromatic ureas
Amides
Amines
Ami no acids
Organophosphorus Compounds
Chlorinated Phenoxyacid Herbicides
TABLE 3.
LIMITS OF DETECTION AND METHOD SENSITIVITIES
FOR DISPERSE RED 1 AND CAFFEINE
Compound
Disperse Red 1
Caffeine
Mode
SRM
Single Quad
CAD
SRM
Single Quad
CAD
LOD
pg
180
600
2,000
45
84
240
EQL(7s)
P9
420
1400
4700
115
200
560
EQL(lOs)
pg
600
2000
6700
150
280
800
EQL = Estimated Quantitation Limit
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TABLE 4.
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR ORGANIC-FREE REAGENT WATER SPIKED WITH DISPERSE RED 1
Percent Recovery
Sample HPLC/UV MS CAD SRM
Spike 1 82.2 ± 0.2 92.5 ± 3.7 87.6 ± 4.6 95.5 ± 17.1
Spike 2 87.4 ± 0.6 90.2 ± 4.7 90.4 ± 9.9 90.0 ± 5.9
RPD 6.1% 2.5% 3.2% 5.9%
TABLE 5.
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR MUNICIPAL WASTEWATER SPIKED WITH DISPERSE RED 1
Percent Recovery
Sample
Spike 1
Spike 2
RPD
HPLC/UV
93.4 ± 0.3
96.2 ± 0.1
3.0%
MS
102.0 ± 31
79.7 ± 15
25%
CAD
82.7 ± 13
83.7 ± 5.2
1.2%
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TABLE 6.
RESULTS FROM ANALYSES OF ACTIVATED SLUDGE PROCESS WASTEWATER
Sample
5 mg/L Spiking
Concentration
UF05A
UF05A-D
UF06A
UF16A
RPD
0 mg/L Spiking
Concentration
UH05A
UH05A-D
UH06A
UH16A
RPD
Recovery
HPLC/UV
0.721 ± 0.003
0.731 ± 0.021
0.279 ± 0.000
0.482 ± 0.001
1.3%
0.000
0.000
0.000
0.000
--
of Disperse Red 1
MS
0.664 ± 0.030
0.600 ± 0.068
0.253 ± 0.052
0.449 ± 0.016
10.1%
0.005 ± 0.0007
0.006 ± 0.001
0.002 ± 0.0003
0.003 ± 0.0004
18.2%
(mq/L)
CAD
0.796 ± 0.008
0.768 ± 0.093
0.301 ± 0.042
0.510 ± 0.091
3.6%
<0.001
<0.001
<0.001
<0.001
--
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TABLE 7.
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 400
Mass
18.0
35.06
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
697.44
% Relative
Abundances8
32.3
13.5
40.5
94.6
27.0
5.4
10.3
17.6
27.0
45.9
64.9
100
94.6
81.1
67.6
32.4
16.2
4.1
8.1
2.7
Intensity is normalized to mass 432.
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TABLE 8.
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 600
Mass
18.0
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
% Relative
Abundances9
4.7
11.4
64.9
17.5
9.3
43.9
56.1
22.8
28.1
38.6
54.4
64.9
86.0
100
63.2
17.5
5.6
1.8
Intensity is normalized to mass 564.
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TABLE 9.
RETENTION TIMES AND THERMOSPRAY MASS SPECTRA
OF ORGANOPHOSPHORUS COMPOUNDS
Compound
Monocrotophos
Trichlorfon
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
Retention Time
(minutes)
1:09
1:22
1:28
4:40
9:16
9:52
10:52
13:30
13:55
18:51
Mass Spectra
(% Relative Abundance)8
241 (100), 224 (14)
274 (100), 257 (19),
230 (100), 247 (20)
238 (100), 221 (40)
398 (100), 381 (23),
221 (2)
326 (10), 309 (100)
281 (100), 264 (8),
234 (48)
278 (4), 261 (100)
292 (10), 275 (100)
315 (100), 299 (15)
238 (19)
238 (5),
251 (21),
For molecules containing Cl, Br and S, only the base peak of the isotopic
cluster is listed.
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TABLE 10.
PRECISION AND LIMITS OF DETECTION FOR
ORGANOPHOSPHORUS COMPOUND STANDARDS
Compound
Dichlorvos
Dimethoate
Phorate
Disulfoton
Fensulfothion
Naled
Merphos
Methyl
parathion
Ion
238
230
261
275
309
398
299
281
Standard
Quantltation
Concentration
(ng/ML)
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
%RSD
16
13
5.7
4.2
2.2
4.2
13
7.3
0.84
14
7.1
4.0
2.2
14
6.7
3.0
4.1
9.2
9.8
2.5
9.5
9.6
5.2
6.3
5.5
17
3.9
5.3
7.1
4.8
1.5
MDL (ng)
4
2
2
1
0.4
0.2
1
30
8321 - 24
Revision 0
November 1990
-------
TABLE 11.
SINGLE OPERATOR ACCURACY AND PRECISION FOR LOW CONCENTRATION DRINKING
WATER (A), LOW CONCENTRATION SOIL (B), MEDIUM CONCENTRATION DRINKING
WATER (C), MEDIUM CONCENTRATION SEDIMENT (D)
Average
Recovery
Compound (%)
A
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
B
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
C
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
D
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
70
40
0.5
112
50
16
3.5
237
16
ND
ND
45
ND
78
36
118
52
146
4
65
85
10
2
101
74
166
ND
72
84
58
56
78
Standard
Deviation
7.7
12
1.0
3.3
28
35
8
25
4
5
15
7
19
4
29
3
7
24
15
1
13
8.5
25
8.6
9
6
5
4
Spike
Amount
ug/L
5
5
5
5
10
5
5
5
uq/Kq
50
50
50
50
100
50
50
50
uq/L
50
50
50
50
100
50
50
50
mq/Kq
2
2
2
2
3
2
2
2
Range of
Recovery
(%)
85 -
64 -
2 -
119 -
105 -
86 -
19 -
287 -
24 -
56 -
109 -
49 -
155 -
61 -
204 -
9 -
79 -
133 -
41 -
4 -
126 -
91 -
216 -
90 -
102 -
70 -
66 -
86 -
54
14
0
106
0
0
0
187
7
34
48
22
81
43
89
0
51
37
0
0
75
57
115
55
66
46
47
70
Number
of
Analyses
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
12
12
12
12
12
12
12
12
15
15
15
15
15
15
15
12
8321 - 25
Revision 0
November 1990
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TABLE 12.
SINGLE OPERATOR ACCURACY AND PRECISION FOR MUNICIPAL WASTE
WATER (A), DRINKING WATER (B), CHEMICAL SLUDGE WASTE (C)
Compound
Tris-BP (A)
(B)
(C)
Concentration
(ng/ML)
50
100
150
200
Average
Recovery
(%)
25
40
63
SINGLE
Average
Area
2675
5091
7674
8379
MDL
(ng/ML)
33
Standard
Deviation
8.0
5.0
11
TABLE
OPERATOR EQL
Standard
Deviation
782
558
2090
2030
Lower
EQL
(ng/ML)
113
Spike Range
Amount of % Number of
(ng/ML) Recovery Analyses
2 41 - 9.0 15
2 50-30 12
100 84-42 8
13.
TABLE FOR TRIS-BP
3*Std 7*Std 10*Std
Dev. Dev. Dev.
2347 5476 7823
Upper
EQL
(ng/ML)
172
EQL = Estimated Quantitation Limit
8321 - 26
Revision 0
November 1990
-------
FIGURE 1.
SCHEMATIC OF THE THERMOSPRAY PROBE AND ION SOURCE
Flange
i
Source
Mounting
Plate
Ion Sampling
Cone
Ions
Electron Vaporizer
Beam ^ Probe
\
Vapor
Temperature
T
Heater Vaporizer
Coupling
— 1C
8321 - 27
Revision 0
November 1990
-------
METHOD 8321
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/
MASS SPECTROMETRY OR UV-VIS DETECTION
C
surt
7.3 S.t HPLC
Chroutographie
condition.
7.4 S.t HPLC/
Th«rBoiprajr/MS
oonditiona
7.5
procedure
7.6 P.rforo
LC/MS
•naly»i«
8321 - 28
Revision 0
November 1990
-------
METHOD 8330
NITROAROMATICS AND NITRAMINES BY HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 Method 8330 is intended for the analysis of explosives residues.
This method is limited to use by analysts experienced in handling and analyzing
explosive materials. This method is used to determine the concentration of the
following compounds in a water, soil or sediment matrix:
Compound Abbrev CAS Noa
Octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine
Hexahydro-l,3,5-trinitro-l,3,5-triazine
1,3,5-Trinitrobenzene
1,3-Dinitrobenzene
Methyl-2,4,6-trinitrophenylnitramine
Nitrobenzene
2,4,6-Trinitrotoluene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
o-Nitrotoluene
m-Nitrotoluene
p-Nitrotoluene
HMX
RDX
TNB
DNB
Tetryl
NB
TNT
24DNT
26DNT
2NT
3NT
4NT
2691-41-0
121-82-4
99-35-4
99-65-0
479-45-8
98-95-3
118-96-7
121-14-2
606-20-2
88-72-2
99-08-1
99-99-0
a Chemical Abstracts Service Registry number
1.2 All of these compounds are either used in the manufacture of
explosives or are the degradation products of compounds used for that purpose.
When making stock solutions for calibration, treat each compound as if it were
extremely explosive.
1.3 The estimated quantitation limits (EQLs) of target analytes determined
by Method 8330 in water and soil are presented in Table 1.
1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of HPLC, skilled in the interpretation of
chromatograms, and experienced in handling explosive materials. Each analyst
must demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Aqueous samples are diluted 1/1 (v/v) with methanol, filtered,
separated on a C-18 reverse phase column, determined at 254 nm, and confirmed
on a CN reverse phase column.
8330 - 1 Revision 0
November 1990
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2.2 Soil and sediment samples are extracted using acetonitrile in an
ultrasonic bath, filtered, and chromatographed as in Section 2.1.
3.0 INTERFERENCES
3.1 2,4-DNT and 2,6-DNT elute at similar retention times (retention time
difference of 0.2 minutes). A large concentration of one isomer may mask the
response of the other isomer. If it is not apparent that both isomers are
present (or are not detected), an isomeric mixture should be reported.
3.2 Tetryl decomposes rapidly in methanol/water solutions, as well as
with heat. All aqueous samples expected to contain tetryl should be diluted
with acetonitrile prior to filtration. All samples expected to contain tetryl
should not be exposed to temperatures above room temperature.
3.3 Degradation products of tetryl appear as a shoulder on the TNT peak.
Peak heights rather than peak areas should be used when tetryl is present in
concentrations that are significant relative to the concentration of TNT.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC - equipped with a pump capable of achieving 4000 psi,
a 100 M! loop injector and a 254 nm UV detector (Perkin Elmer Series 3,
or equivalent).
4.1.2 Columns:
4.1.2.1 Primary column: C-18 Reverse phase HPLC column, 25 cm
x 4.6 mm (5 /nm), (Supelco LC-18, or equivalent).
4.1.2.2 Secondary column: CN Reverse phase HPLC column, 25 cm
x 4.6 mm (5 p.m) , (Supelco LC-CN, or equivalent).
4.1.3 Strip chart recorder.
4.1.4 Digital integrator (optional).
4.1.5 Autosampler (optional).
4.2 Other Equipment
4.2.1 Temperature controlled ultrasonic bath.
4.2.2 Vortex mixer.
4.2.3 Balance ± 0.0001 g.
8330 - 2 Revision 0
November 1990
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4.3 Materials
4.3.1 Injection syringe.
4.3.2 Filters - 0.5 pm Millex-SR, disposable, or equivalent.
4.3.3 Pipettes, volumetric, Class A, glass - 50 ml, 10 ml, 5 ml,
4 ml, 2 ml, 1 ml.
4.3.4 Vials, 20 ml, glass.
4.3.5 Vials, 15 ml, glass, Teflon lined screw cap or crimp top.
4.3.6 Syringes - 3 ml and 10 ml.
4.3.7 Volumetric flasks, Class A - 10 ml, 20 ml, 50 ml, 100 ml,
200 ml, 250 ml.
4.3.8 Mortar and pestle.
4.4 Preparation
4.4.1 Prepare all materials to be used as described in Chapter 4
for volatile organics.
5.0 REAGENTS
5.1 HPLC grade chemicals shall be used in all tests. It is intended that
all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society, where such specifications are
available. Other grades may be used, provided it is first ascertained that the
reagent is of sufficiently high purity to permit its use without lowering the
accuracy of the determination.
5.1.1 Acetonitrile, CH3CN - HPLC grade.
5.1.2 Methanol, CH3OH - HPLC grade.
5.1.3 Calcium Chloride, CaCl2 - Reagent grade. Prepare an aqueous
solution of 5 g/L.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock Standard Solutions
5.3.1 Analyte Standards
5.3.1.1 HMX - Standard Analytical Reference Material.
5.3.1.2 RDX - Standard Analytical Reference Material.
8330 - 3 Revision 0
November 1990
-------
5.3.1.3 DNB - Standard Analytical Reference Material.
5.3.1.4 Tetryl - Standard Analytical Reference Material.
5.3.1.5 TNT - Standard Analytical Reference Material.
5.3.1.6 2,4-DNT - Standard Analytical Reference Material.
5.3.1.7 2,6-DNT - Standard Analytical Reference Material.
5.3.1.8 1,3,5-TNB - Standard Analytical Reference Material.
5.3.1.9 NB - Standard Analytical Reference Material.
5.3.1.10 2-NT - Reagent grade.
5.3.1.11 3-NT - Reagent grade.
5.3.1.12 4-NT - Reagent grade.
5.3.2 Dry each analyte standard to constant weight in a vacuum
desiccator in the dark. Place about 0.100 g (weighed to 0.0001 g) of a
single analyte into a 100 mL volumetric flask and dilute to volume with
acetonitrile. Invert flask several times until dissolved. Store in
refrigerator at 4°C in the dark. Calculate the concentration of the stock
solution from the actual weight used (nominal concentration = 1,000 mg/L).
Stock solutions may be used for up to one year.
5.4 Intermediate Standards Solutions
5.4.1 If both 2,4-DNT and 2,6-DNT are to be determined, prepare
two separate intermediate stock solutions containing (1) HMX, RDX, 1,3,5-
TNB, 1,3-DNB, NB, TNT, and 2,4-DNT and (2) Tetryl, 2,6-DNT, 2-NT, 3-NT,
and 4-NT. Intermediate stock standard solutions should be prepared at
1,000 Mg/L, in acetonitrile when analyzing soil samples, and in methanol
when analyzing aqueous samples.
5.4.2 Dilute the two concentrated intermediate stock solutions,
with the appropriate solvent, to prepare intermediate standard solutions
that cover the range of 2.5 - 1,000 p.g/1. These solutions should be
refrigerated on preparation, and may be used for 30 days.
5.5 Working standards
5.5.1 Prepare working standards by diluting intermediate standards
solutions by 50% (v/v) with (1) organic-free reagent water, when analyzing
aqueous solutions, or (2) 5 g/L calcium chloride solution (Section 5.1.3),
when analyzing soil and sediment samples. These solutions must be
refrigerated, and may be used for 28 days after preparation.
5.6 Eluent
5.6.1 To prepare 1 liter of eluent, add 500 ml of methanol to 500 ml
8330 - 4 Revision 0
November 1990
-------
of organic-free reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Grab samples must be collected and stored in glass containers. Follow
conventional sampling procedures.
6.2 Samples must be kept below 4°C from the time of collection through
analysis.
6.3 Soil and sediment samples should be air dried to constant weight at
room temperature or colder after collection. While it is possible to analyze
wet soil samples, it is much more difficult to obtain a homogeneous subsample
on a wet sample. If wet soil samples are to be analyzed, a moisture
determination must be made on a separate subsample.
7.0 PROCEDURE
7.1 Sample Preparation
7.1.1 Aqueous Samples
7.1.1.1 Sample Filtration: Place a 5 mL portion of each
water sample in a scintillation vial, add 5 mL of methanol, shake
thoroughly, and filter through a 0.5 /xm filter. Discard the first
3 mL of filtrate, and retain the remainder for analysis.
7.1.2 Soil and Sediment Samples
7.1.2.1 Sample homogenization: Dry soil samples in air at
room temperature or colder, being careful not to expose the samples
to direct sunlight. Grind sample thoroughly in an acetonitrile
rinsed mortar.
7.1.2.2 Sample extraction
7.1.2.2.1 Place a 2.0 g subsample of each soil sample
in a 15 mL glass vial. Add 10.0 mL of acetonitrile, cap with
teflon lined cap, vortex swirl for one minute, and place in
ultrasonic bath for 18 hours. If tetryl is being analyzed,
keep ultrasonic bath at room temperature or below.
7.1.2.2.2 After sonication, allow sample to settle for
30 minutes. Remove 5.0 mL of supernatant, and combine with
5.0 mL of calcium chloride solution (Section 5.1.3) in a 20 mL
vial. Shake, and let stand for 15 minutes.
7.1.2.2.3 Place supernatant in syringe and filter
through a 0.5 /zm filter. Discard first 2 to 3 mL and retain
remainder for analysis.
8330 - 5 Revision 0
November 1990
-------
7.2 Chromatographic Conditions
Mobile Phase: 50/50 (v/v) methanol/organic-free reagent water
Flow rate: 1.5 mL/min
Injection volume: 100 jil
UV Detector: 254 nm
7.3 Calibration of HPLC
7.3.1 Analyze working standards in triplicate, using the
chromatographic conditions given in Section 7.2. Prepare calibration
curve using peak heights or peak areas, as appropriate. The calibration
curve should be linear with zero intercept.
7.3.2 At the beginning of each analysis day, after the midpoint of
a sample run, and after the last sample of the day, inject midpoint
calibration standards. Compare mean peak heights obtained during the day
with the peak heights obtained in the morning. If these values do not
agree within 20%, reinject all solutions in triplicate and recalculate
calibration curve.
7.4 Sample Analysis
7.4.1 Analyze the samples using the chromatographic conditions
given in Section 7.2. Confirm each measurement by injecting onto the CN
column.
7.4.2 Table 2 presents the retention times for the analytes on both
the C18 and CN columns. Figure 1 presents typical chromatograms.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Prior to preparation of stock solutions, acetonitrile, methanol, and
water blanks should be run to determine possible interferences with analyte
peaks. If the acetonitrile, methanol, or water blanks show contamination, a
different batch should be used.
8.3 Method Blanks
8.3.1 Method blanks for the analysis of aqueous samples should be
organic-free reagent water carried through all sample storage and handling
procedures.
8.3.2 Method blanks for the analysis of soil samples should be
uncontaminated soil carried through all sample storage, extraction, and
handling procedures.
8330 - 6 Revision 0
November 1990
-------
9.0 METHOD PERFORMANCE
9.1 Method 8330 was tested by six laboratories. The results of this testing
indicate that the results presented in Tables 3 through 5 are to be expected.
10.0 REFERENCES
1. Bauer, C.F., S.M. Koza, and T.F. Jenkins, "Collaborative Test Results for
a Liquid Chromatographic Method for the Determination of Explosives Residues
in Soil," manuscript submitted to the Journal of the AOAC, April 1989.
2. Department of the Army, "Reversed-Phase HPLC Method for the Determination
of Explosive Residues in Soil," Appendix B, provided by Dennis J. Wynne,
Chief, Technology Division, U.S. Army Toxic and Hazardous Materials Agency,
Aberdeen Proving Ground, Maryland 21010-5401.
3. Department of the Army, "An Improved RP-HPLC Method for the Determination
of Nitroaromatics and Nitramines in Water" Appendix B, provided by Dennis
J. Wynne, Chief, Technology Division, U.S. Army Toxic and Hazardous
Materials Agency, Aberdeen Proving Ground, Maryland 21010-5401.
11.0 SAFETY
11.1 Standard precautionary measures used for handling other organic
compounds should be sufficient for safe handling of the analytes targeted by
Method 8330.
8330 - 7 Revision 0
November 1990
-------
O r-
Figure 1
Absorbance
HMX
CJl
-r
(0
o O
ro
O
TNB
DNB
2-Am-DNT
2,6-DNT
2,4 DNT
8330 - 8
Revision 0
November 1990
-------
TABLE 1
ESTIMATED QUANTITATION LIMITS
Compound
Water Soil
Abbrev (M9/L) (M9/9)
Octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine
Hexahydro-l,3,5-trinitro-l,3,5-triazine
1, 3, 5-Tri nitrobenzene
1,3-Dinitrobenzene
Methy1-2,4,6-trinitrophenylnitramine
Nitrobenzene
2,4,6-Trinitrotoluene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
o-Nitrotoluene
m-Nitrotoluene
p-Nitrotoluene
HMX
RDX
TNB
DNB
Tetryl
NB
TNT
24DNT
26DNT
2NT
3NT
4NT
13.0
14.0
7.3
4.0
44.0
NA
6.9
5.7
9.4
12.0
7.9
8.5
2.2
1.0
0.25
0.25
0.65
0.26
0.25
0.25
0.26
0.25
0.25
0.25
NA Not available
TABLE 2
RETENTION TIMES FOR ANALYTES ON C-18 AND CN COLUMNS
C-18
Analyte
HMX
RDX
TNB
DNB
Tetryl
NB
TNT
26DNT
24DNT
2NT
4NT
3NT
Retention
Time (min)
2.4
3.7
5.1
6.2
6.9
7.2
8.4
9.8
10.1
12.3
13.3
14.2
Analyte
NB
TNB
DNB
2NT
4NT
3NT
26DNT
24DNT
TNT
RDX
Tetryl
HMX
CN
Retention
Time (min)
3.8
4.1
4.2
4.4
4.4
4.5
4.6
4.9
5.0
6.2
7.4
8.4
8330 - 9
Revision 0
November 1990
-------
TABLE 3
INTRALABORATORY PRECISION OF METHOD FOR SOIL SAMPLES
Spiked soils
Mean
Concentration
(M9/9)
%rsd
Field-contaminated soils
Mean
Concentration
(M9/9) SD %rsd
HMX
RDX
TNB
DNB
tetryl
TNT
24DNT
46
60
8.6
46
3.5
17
40
5.0
1.7
1.4
0.4
1.9
0.14
3.1
1.4
0.17
3.7
2.3
4.6
4.1
4.0
17.9
3.5
3.4
14
153
104
877
2.8
72
1.1
2.3
7.0
669
1.0
1.8
21.6
12
29.6
0.2
6.0
0.11
0.41
0.61
55
0.44
12.8
14.1
11.5
3.4
7.1
8.3
9.8
18.0
9.0
8.2
42.3
8330 - 10
Revision 0
November 1990
-------
TABLE 4
INTERLABORATORY ERROR OF METHOD FOR SOIL SAMPLES
Spi
ked soils
Mean
Concentration
(M/9) SD
HMX 46
RDX 60
TNB 8.6
46
DNB 3.5
tetryl 17
TNT 40
24DNT 5.0
2.6
2.6
0.61
2.97
0.24
5.22
1.88
0.22
INTERLABORATORY ERROR
HMX
RDX
TNB
TNT
Sample 1
mean
cone.
(M9/L)
nd
431
74.3
10635
Field contaminated
%rsd
5.7
4.4
7.1
6.5
6.9
30.7
4.7
4.4
Mean
Concentration
(M9/9)
14
153
104
877
2.8
72
1.1
2.3
7.0
669
1.0
SD
3.7
37.3
17.4
67.3
0.23
8.8
0.16
0.49
1.27
63.4
0.74
soils
%rsd
26.0
24.0
17.0
7.7
8.2
12.2
14.5
21.3
18.0
9.5
74.0
TABLE 5
OF METHOD FOR WATER SAMPLES8
%rsd
22.9
3.2
59.4
Samole 2
mean
cone.
(M9/L)
184"
2117
27. 6C
1746
%rsd
8.4
29.5
4.2
26.8
10 replicate determinations, except where noted
6 replicate determinations
7 replicate determinations
8330 - 11
Revision 0
November 1990
-------
Aqueous Somple
7.1.1.1 Somole Filtration:
Sample Filtral
Place 5 mis.
Place 5 mis. sample in
scintillation vial. Add
5 mis. methoncJ; shake;
filter through 0.5 um
filter. Discard first 3 mis.
retain remainder for use.
oo
OJ
o
7.1 Is sample in
an aqueous or
soil/sediment
matrix?
Soil and Sediment
Samples
7.1.2.1 Sample Homoaenization
Air dry sample at room T
or below. Avoid exposure
to direct sunlight. Grind
sample in an acetonitrile
rinsed mortar.
7.3 Calibration of HPLC
7.3.1 Run working stds. in triplicate.
Plot [ ] vs. peak area or ht.
Curve should be linear with
zero intercept.
( 7.1.2.2 Sample Extraction
i
7.1.2.2.1 Place 2 grs. soil
subsomple, 10 mis.
ocetonitrile in 15 ml.
glass vial. Cap, vortex
swirl, place in room T
or below ultrasonic bath
for 18 hrs.
7.3.2 Analyze midrange calibration
std. at beginning, middle,
and end of sample analyses.
Redo Section 7.3.1 if peak
areas or hts. do not agree
to w/in +/- 20X of initial
calibration values.
1
7.1.2.2.2 Let sdn. settle. Add 5
mis. supernatant to 5
mis. calcium chloride
soln. in 20 ml. vial.
Shake, let stand 15 mins.
| 7.4 Sample Analysis
7.4.1 Analyze samples. Confirm
measurement w/injection
onto CN column.
7..1.2.2.3 Filter supernatant through
0.5 um filter. Discard
initial 3 mis., retain
remainder for analysis.
7.2 Set Chromatographic Conditions
7.4.2 Refer to Table 2 for typical
anolyte retention times.
•30
i>
—I
r>
oo:
IO
o
i 00
I U>
JO U>
o
oo
Stop
-------
METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PRESSURE LIQUID CHRQMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 This method is intended for the analysis of tetrazene, an explosive
residue, in soil and water. This method is limited to use by analysts
experienced in handling and analyzing explosive materials. The following
compounds can be determined by this method:
Compound CAS Noa
Tetrazene 31330-63-9
a Chemical Abstracts Service Registry number
1.2 Tetrazene degrades rapidly in water and methanol at room temperature.
Special care must be taken to refrigerate or cool all solutions throughout the
analytical process.
1.3 Tetrazene, in its dry form, is extremely explosive. Caution must be
taken during preparation of standards.
1.4 The estimated quantitation limit (EQL) of Method 8331 for determining
the concentration of tetrazene is approximately 7 ng/l in water and
approximately 1 ing/Kg in soil.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of HPLC, skilled in the interpretation of
chromatograms, and experienced in handling explosive materials. Each analyst
must demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A 10 ml water sample is filtered, eluted on a C-18 column using ion
pairing reverse phase HPLC, and quantitated at 280 nm.
2.2 2 g of soil are extracted with 55:45 v/v methanol-water and
1-decanesulfonic acid on a platform shaker, filtered, and eluted on a C-18 column
using ion pairing reverse phase HPLC, and quantitated at 280 nm.
8331 - 1 Revision 0
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3.0 INTERFERENCES
3.1 No interferences are known. Tetrazene elutes early, however, and if
a computing integrator is used for peak quantification, the baseline setting may
have to be set to exclude baseline aberrations. Baseline setting is particularly
important at low concentrations of analyte.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC - Pump capable of achieving 4000 psi.
4.1.2 100 /xL loop injector.
4.1.3 Variable or fixed wavelength detector capable of reading
280 nm.
4.1.4 C-18 reverse phase HPLC column, 25 cm x 4.6 mm (5 nm)
(Supelco LC-18, or equivalent).
4.1.5 Digital integrator - HP 3390A (or equivalent)
4.1.6 Strip chart recorder.
4.2 Other apparatus
4.2.1 Platform orbital shaker.
4.2.2 Analytical balance - + 0.0001 g.
4.2.3 Desiccator.
4.3 Materials
4.3.1 Injection syringe - 500 /xL-
4.3.2 Filters - 0.5 p.m Millex-SR and 0.5 p.m Millex-HV, disposable,
or equivalent.
4.3.3 Pipets - volumetric, glass, Class A.
4.3.4 Scintillation vials - 20 mL, glass.
4.3.5 Syringes - 10 mL.
4.3.6 Volumetric flasks, Class A - 100 mL, 200 mL.
4.3.7 Erlenmeyer flasks with ground glass stoppers - 125 mL.
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4.4 Preparation
4.4.1 Prepare all materials as described in Chapter 4 for volatile
organics.
5.0 REAGENTS
5.1 HPLC grade chemicals shall be used in all tests. It is intended that
all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society, where such specifications are
available. Other grades may be used, provided it is first ascertained that the
reagent is of sufficiently high purity to permit its use without lowering the
accuracy of the determination.
5.2 General
5.2.1 Methanol, CH3OH - HPLC grade.
5.2.2 Organic-free reagent water - All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.3 1-Decanesulfonic acid, sodium salt, C10H21S03Na - HPLC grade.
5.2.4 Acetic acid (glacial), CH3COOH - reagent grade.
5.3 Standard Solutions
5.3.1 Tetrazene - Standard Analytical Reference Material.
5.3.2 Stock standard solution - Dry tetrazene to constant weight
in a vacuum desiccator in the dark. (Tetrazene is extremely explosive in
the dry state. Do not dry more reagent than is necessary to prepare stock
solutions.) Place about 0.0010 g (weighed to 0.0001 g) into a 100-ml
volumetric flask and dilute to volume with methanol. Invert flask several
times until tetrazene is dissolved. Store in freezer at -10°C. Stock
solution is about 100 mg/L. Replace stock standard solution every week.
5.3.3 Intermediate standard solutions
5.3.3.1 Prepare a 4 mg/L standard by diluting the stock
solution 1/25 v/v with methanol.
5.3.3.2 Pipet 0.5, 1.0, 2.0, 5.0, 10.0, and 20.0 mL of the
4 mg/L standard solution into 6 separate 100 mL volumetric flasks,
and make up to volume with methanol. Pipet 25.0 mL of the 4 mg/L
standard solution into a 50 mL volumetric flask, and make up to
volume with methanol. This results in intermediate standards of
about 0.02, 0.04, 0.08, 0.2, 0.4, 0.8, 2 and 4 mg/L.
5.3.3.3 Cool immediately on preparation in refrigerator or
ice bath.
8331 - 3 Revision 0
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5.3.4 Working standard solutions
5.3.4.1 Inject 4 ml of each of the intermediate standard
solutions into 6.0 mL of water. This results in concentrations of
about 0.008, 0.016, 0.032, 0.08, 0.16, 0.3, 0.8 and 1.6 mg/L.
5.3.4.2 Cool immediately on preparation in refrigerator or
ice bath.
5.5 QC spike concentrate solution
5.5.1 Dry tetrazene to constant weight in a vacuum desiccator in
the dark. (Tetrazene is extremely explosive in the dry state. Do not
dry any more than necessary to prepare standards.) Place about 0.0011 g
(weighed to 0.0001 g) into a 200-ml volumetric flask and dilute to volume
with methanol. Invert flask several times until tetrazene is dissolved.
Store in freezer at -10eC. QC spike concentrate solution is about 55 mg/L.
Replace stock standard solution every week.
5.5.2 Prepare spiking solutions, at concentrations appropriate to
the concentration range of the samples being analyzed, by diluting the QC
spike concentrate solution with methanol. Cool on preparation in
refrigerator or ice bath.
5.6 Eluent
5.6.1 To make about 1 liter of eluent, add 2.44 g of
1-decanesulfonic acid, sodium salt to 400/600 v/v methanol/water, and add
2.0 ml of glacial acetic acid.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
6.2 Samples must be collected and stored in glass containers. Follow
conventional sampling procedures.
6.3 Samples must be kept below 4°C from the time of collection through
analysis.
7.0 PROCEDURE
7.1 Sample Preparation
7.1.1 Filtration of Water Samples
7.1.1.1 Place a 10 mL portion of each water sample in a
syringe and filter through a 0.5 /urn Millex-HV filter unit. Discard
first 5 mL of filtrate, and retain 5 mL for analysis.
8331 - 4 Revision 0
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7.1.2 Extraction and Filtration of Soil Samples
7.1.2.1 Determination of sample % dry weight - In certain
cases, sample results are desired based on dry-weight basis. When
such data is desired, a portion of sample for this determination
should be weighed out at the same time as the portion used for
analytical determination.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.
7.1.2.1.1 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight
at 105°C. Allow to cool in a desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.1.2.2 Weigh 2 g soil subsamples into 125 ml Erlenmeyer
flasks with ground glass stoppers.
7.1.2.3 Add 50 mL of 55/45 v/v methanol-water with
1-decanesulfonic acid, sodium salt added to make a 0.1 M solution.
7.1.2.4 Vortex for 15 seconds.
7.1.2.5 Shake for 5 hr at 2000 rpm on platform shaker.
7.1.2.6 Place a 10 mL portion of each soil sample extract in
a syringe and filter through a 0.5 /urn Millex-SR filter unit.
Discard first 5 ml of filtrate, and retain 5 ml for analysis.
7.2 Sample Analysis
7.2.1 Analyze the samples using the chromatographic conditions
given in Section 7.2.1.1. Under these conditions, the retention time of
tetrazene is 2.8 min. A sample chromatogram, including other compounds
likely to be present in samples containing tetrazene, is shown in Figure 1.
7.2.1.1 Chromatographic Conditions
Solvent: 0.01 M 1-decanesulfonic acid, in acidic
methanol/water (Section 5.5)
Flow rate: 1.5 mL/min
Injection volume: 100 nl
UV Detector: 280 nm
7.3 Calibration of HPLC
7.3.1 Initial Calibration - Analyze the working standards (Section
5.3.4), starting with the 0.008 mg/L standards and ending with the
8331 - 5 Revision 0
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0.30 mg/L standard. If the percent relative standard deviation (%RSD) of
the mean response factor (RF) for each analyte does not exceed 20%, the
system is calibrated and the analysis of samples may proceed. If the %RSD
for any analyte exceeds 20%, recheck the system and/or recalibrate with
freshly prepared calibration solutions.
7.3.2 Continuing Calibration - On a daily basis, inject 250 /uL of
stock standard into 20 ml water. Keep solution in refrigerator until
analysis. Analyze in triplicate (by overfilling loop) at the beginning
of the day, singly after each five samples, and singly after the' last
sample of the day. Compare response factors from the mean peak area or
peak height obtained over the day with the response factor at initial
calibration. If these values do not agree within 10%, recalibrate.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Prior to preparation of stock solutions, methanol should be analyzed
to determine possible interferences with the tetrazene peak. If the methanol
shows contamination, a different batch of methanol should be used.
8.3 Method Blanks
8.3.1 Method blanks for the analysis of water samples should be
organic-free reagent water carried through all sample storage and handling
procedures.
8.3.2 Method blanks for the analysis of soil samples should be
uncontaminated soil carried through all sample storage, extraction, and
handling procedures.
9.0 METHOD PERFORMANCE
9.1 Method 8331 was tested in a laboratory over a period of four days.
Spiked organic-free reagent water and standard soil were analyzed in duplicate
each day for four days. The H.PLC was calibrated daily according to the
procedures given in Section 7.1. Method performance data are presented in Tables
1 and 2.
10.0 REFERENCES
1. Walsh, M.E., and T.F. Jenkins, "Analytical Method for Determining Tetrazene
in Water," U.S. Army Corps of Engineers, Cold Regions Research & Engineering
Laboratory, Special Report 87-25, 1987.
2. Walsh, M.E., and T.F. Jenkins, "Analytical Method for Determining Tetrazene
in Soil," U.S. Army Corps of Engineers, Cold Regions Research & Engineering
Laboratory, Special Report 88-15, 1988.
8331 - 6 Revision 0
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11.0 SAFETY
11.1 Standard precautionary measures used for handling other organic
compounds should be sufficient for safe handling of the analytes targeted by
Method 8331.
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I6f
12
6
8
FIGURE 1
TNT
0.064
Absorbonct Units
8331 - 8
Revision 0
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TABLE 1.
METHOD PERFORMANCE, WATER MATRIX
Spike
Cone.
(M9/L)
0.00
7.25
14.5
29
72.5
145
290
725
OVERALL
Ava % Recovery
Replicate
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Day 1
0.0
NA
0.0
NA
8.9
122
6.6
91
14.6
101
14.8
102
31.8
110
29.5
102
71.1
98
71.2
98
140.6
97
138.5
96
289.4
100
282.0
97
737.6
102
700.2
97
Day 2
0.0
NA
0.0
NA
7.8
108
9.9
137
14.6
101
14.1
97
30.0
103
29.7
102
73.6
102
71.3
98
143.8
99
140.8
97
288.5
99
284.2
98
707.2
98
695.8
96
Day 3
0.0
NA
0.0
NA
7.4
102
8.5
117
13.8
95
14.1
98
30.8
106
30.4
105
75.7
104
70.7
98
144.7
100
140.9
97
291.0
100
281.9
97
714.3
99
714.2
99
Day 4
0.0
NA
0.0
NA
9.4
130
6.7
92
14.6
101
15.2
105
28.7
99
30.7
106
73.9
102
71.6
99
142.1
98
136.9
94
289.8
100
282.5
97
722.0
100
716.3
99
Average
% Recovery
NA
. NA
116
109
99
100
105
104
101
98
98
96
100
97
99
97
102
8331 - 9
Revision 0
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TABLE 2
METHOD PERFORMANCE, SOIL MATRIX
Spike
Cone.
(M9/L)
0.00
1.28
2.56
5.12
12.8
25.6
OVERALL
Ava % Recovery
Replicate
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Day 1
0.0
NA
0.0
NA
0.6
49
1.2
92
1.4
56
1.5
59
2.9
57
3.0
58
7.8
61
8.0
62
17.2
67
16.7
65
Day 2
0.0
NA
0.0
NA
0.9
73
0.7
56
1.5
58
2.0
79
3.0
58
3.0
59
7.6
59
8.4
66
16.7
65
16.8
66
Day 3
0.0
NA
0.0
NA
0.6
48
0.8
63
1.6
61
1.4
56
2.9
56
3.5
69
7.8
61
7.7
60
17.4
68
17.6
69
Day 4
0.0
NA
0.0
NA
1.0
74
0.7
56
1.6
61
1.3
50
2.9
56
3.1
60
8.1
63
8.2
64
17.3
68
17.2
67
Average
% Recovery
NA
NA
61
67
59
61
57
61
61
63
67
67
62
8331 - 10
Revision 0
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METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC1
Start
7.1.1 Filter 10
ml water
sample; discard
first 5 ml;
analyze last 5
7.1.2.1
Determine X
dry weight
7.1.2.2-7.1.2.5
Extract 2g soil
with 50 ml
solvent
7.1.2.6 Filter
10 ml extract;
discard 5 ml;
analyze last 5
ml
7 . 2 Analyze
samples using
chromatographic
conditions in
Section 7.2.1.1
7.3.1 Initial
Calibration:
Analyze working
standards
(Section 5.3.3)
7.3.1 Is XRSD
of mean RF
>20%?
7.3.1 Recheck
system/
recalibrate
with new cali-
bration solu.
No
7.3.2
Continuing
Calibration
Stop
8331 - 11
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METHOD 8410
GAS CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED
(GC/FT-IR) SPECTROMETRY FOR SEMIVOLATILE ORGANICS:
CAPILLARY COLUMN
1.0 SCOPE AND APPLICATION
1.1 This method covers the automated Identification, or compound class
assignment of unidentifiable compounds, of solvent extractable semivolatile
organic compounds which are amenable to gas chromatography, by GC/FT-IR. GC/FT-
IR can be a useful complement to GC/MS analysis (Method 8270). It is
particularly well suited for the identification of specific isomers that are
not differentiated using GC/MS. Compound class assignments are made using
infrared group absorption frequencies. The presence of an infrared band in the
appropriate group frequency region may be taken as evidence of the possible
presence of a particular compound class, while its absence may be construed as
evidence that the compound class in question is not present. This evidence will
be further strengthened by the presence of confirmatory group frequency bands.
Identification limits of the following compounds have been demonstrated by this
method.
Compound Name
8410 - 1
CAS No.'
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzo(b)pyrene
Benzoic acid
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
4-Chloroaniline
4-Chloro-3-methyl phenol
2-Chl oronaphthal ene
2-Chlorophenol
4-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
2,4-Dichlorophenol
Dimethyl phthalate
83-32-9
208-96-8
120-12-7
56-55-3
50-32-8
65-85-0
111-91-1
111-44-4
39638-32-9
117-81-7
101-55-3
85-68-7
106-47-8
59-50-7
91-58-7
95-57-8
106-48-9
7005-72-3
218-01-9
132-64-9
84-74-2
95-50-1
541-73-1
106-46-7
120-83-2
131-11-3
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November 1990
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Compound Name CAS No.8
Diethyl phthalate 84-66-2
4,6-Dinitro-2-methylphenol 534-52-1
2,4-Dinitrophenol 51-28-5
2,4-Dinitrotoluene 121-14-2
2,6-Dinitrotoluene 606-20-2
Di-n-octyl phthalate 117-84-0
Di-n-propyl phthalate 131-16-8
Fluoranthene 206-44-0
Fluorene 86-73-7
Hexachlorobenzene 118-74-1
1,3-Hexachlorobutadiene 87-68-3
Hexachlorocyclopentadi ene 77-47-4
Hexachloroethane 67-72-1
Isophorone 78-59-1
2-Methylnaphthalene 91-57-6
2-Methylphenol 95-48-7
4-Methylphenol 106-44-5
Naphthalene 91-20-3
2-Nitroaniline 88-74-4
3-Nitroaniline 99-09-2
4-Nitroaniline 100-01-6
Nitrobenzene 98-95-3
2-Nitrophenol 88-75-5
4-Nitrophenol 100-02-7
N-Nitrosodimethylamine 62-75-9
N-Nitrosodiphenylamine 86-30-9
N-Nitroso-di-n-propylamine 621-64-7
Pentachlorophenol 87-86-5
Phenanthrene 85-01-8
Phenol 108-95-2
Pyrene 129-00-0
1,2,4-Trichlorobenzene 120-82-1
2,4,5-Trichlorophenol 95-95-4
2,4,6-Trichlorophenol 88-06-2
a Chemical Abstract Services Registry Number.
1.2 This method is applicable to the determination of most extractable,
semivolatile-organic compounds in wastewater, soils and sediments, and solid
wastes. Benzidine can be subject to losses during solvent concentration and GC
analysis; o-BHC, 6-BHC, endosulfan I and II, and endrin are subject to
decomposition under the alkaline conditions of the extraction step; endrin is
subject to decomposition during GC analysis; and hexachlorocyclopentadiene and
N-nitrosodiphenylamine may decompose during extraction and GC analysis. Other
extraction and/or instrumentation procedures should be considered for unstable
analytes.
8410 - 2 Revision 0
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1.3 The identification limit of this method may depend strongly upon the
level and type of gas chromatographable (GC) semi volatile extractants. The
values listed in Tables 1 and 2 represent the minimum quantities of semivolatile
organic compounds which have been identified by the specified GC/FT-IR system,
using this method and under routine environmental analysis conditions. Capillary
GC/FT-IR wastewater identification limits of 25 /ug/L may be achieved for weak
infrared absorbers with this method, while the corresponding identification
limits for strong infrared absorbers is 2 p.g/1. Identification limits for other
sample matrices can be calculated from the wastewater values after choice of the
proper sample workup procedure (see Section 7.1).
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract and uses FT-IR for detection and
quantitation of the target analytes.
3.0 INTERFERENCES
3.1 Glassware and other sample processing hardware must be thoroughly
cleaned to prevent contamination and misinterpretation. All of these materials
must be demonstrated to be free from interferences under the conditions of the
analysis by running method blanks. Specific selection of reagents or
purification of solvents by distillation in all-glass systems may be required.
3.2 Matrix interference will vary considerably from source to source,
depending upon the diversity of the residual waste being sampled. While general
cleanup techniques are provided as part of this method, unique samples may
require additional cleanup to isolate the analytes of interest from interferences
in order to achieve maximum sensitivity.
3.3 4-Chlorophenol and 2-nitrophenol are subject to interference from
co-eluting compounds.
3.4 Clean all glassware as soon as possible after use by rinsing with
the last solvent used. Glassware should be sealed/stored in a clean environment
immediately after drying to prevent any accumulation of dust or other
contaminants.
4.0 APPARATUS AND MATERIALS
4.1 Gas Chromatographic/Fourier Transform Infrared Spectrometric Equipment
4.1.1 Fourier Transform-Infrared Spectrometer - A spectrometer
capable of collecting at least one scan set per second at 8 cm"1 resolution
is required. In general, a spectrometer purchased after 1985, or
retrofitted to meet post-1985 FT-IR improvements, will be necessary to meet
the detection limits of this protocol. A state-of-the-art A/D converter
8410 - 3 Revision 0
November 1990
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is required, since it has been shown that the signal-to-noise ratio of
single beam GC/FT-IR systems is A/D converter limited.
4.1.2 GC/FT-IR Interface - The interface should be lightpipe volume-
optimized for the selected chromatographic conditions (lightpipe volume
of 100-200 juL for capillary columns). The shortest possible inert
transfer line (preferably fused silica) should be used to interface the
end of the chromatographic column to the lightpipe. If fused silica
capillary columns are employed, the end of the GC column can serve as the
transfer line if it is adequately heated. It has been demonstrated that
the optimum lightpipe volume is equal to the full width at half height of
the GC eluate peak.
4.1.3 Capillary Column - A fused silica DB-5 30 m x 0.32 mm
capillary column with 1.0 /xm film thickness (or equivalent).
4.1.4 Data Acquisition - A computer system dedicated to the GC/FT-
IR system to allow the continuous acquisition of scan sets for a full
chromatographic run. Peripheral data storage systems should be available
(magnetic tape and/or disk) for the storage of all acquired data. Software
should be available to allow the acquisition and storage of every scan set
to locate the file numbers and transform high S/N scan sets, and to provide
a real time reconstructed chromatogram.
4.1.5 Detector - A cryoscopic, medium-band HgCdTe (MCT) detector
with the smallest practical focal area. Typical narrow-band MCT detectors
operate from 3800-800 cm"1 but medium-band MCT detectors can reach 650 cm"1.
A 750 cm'1 cutoff (or lower) is desirable since it allows the detection of
typical carbon-chlorine stretch and aromatic out-of-plane carbon-hydrogen
vibrations of environmentally important organo-chlorine and polynuclear
aromatic compounds. The MCT detector sensitivity (D)" should be > 1 x 1010
cm.
4.1.6 Lightpipe - Constructed of inert materials, gold coated, and
volume-optimized for the desired chromatographic conditions (see Section
7.3).
4.1.7 Gas Chromatograph - The FT-IR spectrometer should be
interfaced to a temperature programmable gas Chromatograph equipped with
a Grob-type (or equivalent) purged splitless injection system suitable for
capillary glass columns or an on-column injector system.
A short, inert transfer line should interface the gas Chromatograph
to the FT-IR lightpipe and, if applicable, to the GC detector. Fused
silica GC columns may be directly interfaced to the lightpipe inlet and
outlet.
4.2 Dry Purge Gas - If the spectrometer is the purge-type, provisions
should be made to provide a suitable continuous source of dry purge-gas to the
FT-IR spectrometer.
4.3 Dry Carrier Gas - The carrier gas should be passed through an
efficient cartridge-type drier.
8410 - 4 Revision 0
November 1990
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4.4 Syringes - 1-juL, 10-^L.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Acetone, CH3COCH3 - Pesticide quality, or equivalent.
5.3.2 Methylene chloride, CH2C12 - Pesticide quality, or equivalent.
5.4 Stock Standard Solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as a certified solution.
5.4.1 Prepare stock standard solutions by accurately weighing 0.1000
± 0.0010 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 100 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96 percent or greater, the
weight may be used without correction to calculate the concentration of
the stock standard. Commercially prepared stock standards may be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.4.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps. Store at 4°C and protect from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.4.3 Stock standard solutions must be replaced after 6 months or
sooner if comparison with quality control reference samples indicates a
problem.
5.5 Calibration Standards and Internal Standards - For use in situations
where GC/FT-IR will be used for primary quantitation of analytes rather than
confirmation of GC/MS identification.
5.5.1 Prepare calibration standards that contain the compounds of
interest, either singly or mixed together. The standards should be
prepared at concentrations that will completely bracket the working range
of the chromatographic system (at least one order of magnitude is
suggested).
8410 - 5 Revision 0
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5.5.2 Prepare Internal standard solutions. Suggested Internal
standards are 1-fluoronaphthalene, terphenyl, 2-chlorophenol, phenol,
bis(2-chloroethoxy)methane, 2,4-dichlorophenol, phenanthrene, anthracene,
and butyl benzyl phthalate. Determine the internal standard concentration
levels from the minimum identifiable quantities.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample Preparation - Samples must be prepared by one of the following
methods prior to GC/FT-IR analysis.
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3550
Waste 3540, 3550, 3580
7.2 Extracts may be cleaned up by Method 3640, Gel-Permeation Cleanup.
7.3 Initial Calibration - Recommended GC/FT-IR conditions:
Scan time: At least 2 scan/sec.
Initial column temperature and hold time: 40°C for 1 minute.
Column temperature program: 40-280°C at 10°C/min.
Final column temperature hold: 280°C.
Injector temperature: 280-300°C.
Transfer line temperature: 270°C.
Lightpipe: 280°C.
Injector: Grob-type, splitless or on-column.
Sample volume: 2-3 pi.
Carrier gas: Dry helium at about 1 mL/min.
7.4 With an oscilloscope, check the detector centerburst intensity versus
the manufacturer's specifications. Increase the source voltage, if necessary,
to meet these specifications. For reference purposes, laboratories should
prepare a plot of time versus detector voltage over at least a 5 day period.
7.5 Capillary Column Interface Sensitivity Test - Install a 30 m x 0.32 mm
fused silica capillary column coated with 1.0 jum of DB-5 (or equivalent). Set
the lightpipe and transfer lines at 280°C, the injector at 225°C and the GC
detector at 280°C (if used). Under splitless Grob-type or on-column injection
conditions, inject 25 ng of nitrobenzene, dissolved in 1 juL of methylene
chloride. The nitrobenzene should be identified by the on-line library software
search within the first five hits (nitrobenzene should be contained within the
search library).
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7.6 Interferometer - If the interferometer is air-driven, adjust the
interferometer drive air pressure to manufacturer's specifications.
7.7 MCT Detector Check - If the centerburst intensity is 75 percent or
less of the mean intensity of the plot maximum obtained by the procedure of
Section 7.4, install a new source and check the MCT centerburst with an
oscilloscope versus the manufacturer's specifications (if available). Allow at
least five hours of new source operation before data acquisition.
7.8 Frequency Calibration - At the present time, no consensus exists
within the spectroscopic community on a suitable frequency reference standard
for vapor-phase FT-IR. One reviewer has suggested the use of indene as an on-
the-fly standard.
7.9 Minimum Identifiable Quantities - Using the GC/FT-IR operating
parameters specified in Section 7.3, determine the minimum identifiable
quantities for the compounds of interest.
7.9.1 Prepare a plot of lightpipe temperature versus MCT centerburst
intensity (in volts or other vertical height units). This plot should span
the temperature range between ambient and the lightpipe thermal limit in
increments of about 20°C. Use this plot for daily QA/QC (see Section 8.4).
Note that modern GC/FT-IR interfaces (1985 and later) may have eliminated
most of this temperature effect.
7.10 GC/FT-IR Extract Analysis
7.10.1 Analysis - Analyze the dried methylene chloride extract using
the chromatographic conditions specified in Section 7.3 for capillary
column interfaces.
7.10.2 GC/FT-IR Identification - Visually compare the analyte
infrared (IR) spectrum versus the search library spectrum of the most
promising on-line library search hits. Report, as identified, those
analytes with IR frequencies for the five (maximum number) most intense
IR bands (S/N > 5) which are within ± 5.0 cm"1 of the corresponding bands
in the library spectrum. Choose IR bands which are sharp and well
resolved. The software used to locate spectral peaks should employ the
peak "center of gravity" technique. In addition, the IR frequencies of
the analyte and library spectra should be determined with the same computer
software.
7.10.3 Retention Time Confirmation - After visual comparison of the
analyte and library spectrum as described in Section 7.10.2, compare the
relative retention times (RRT) of the analyte and an authentic standard
of the most promising library search hit. The standard and analyte RRT
should agree within + 0.01 RRT units when both are determined at the same
chromatographic conditions.
7.10.4 Compound Class or Functionality Assignment - If the analyte
cannot be unequivocally identified, report its compound class or
functionality. See Table 3 for gas-phase group frequencies to be used as
an aid for compound class assignment. It should be noted that FT-IR gas-
8410 - 7 Revision 0
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phase group stretching frequencies are 0-30 cm"1 higher in frequency than
those of the condensed phase.
7.10.5 Quantitation - Although this protocol can be used to confirm
GC/MS identifications, with subsequent quantisation, FT-IR quantitation
guidelines are also provided.
7.10.6 Integrated Absorbance Technique - After analyte
identification, construct a standard calibration curve of concentration
versus integrated infrared absorbance. For this purpose, choose for
integration only those FT-IR scans which are at or above the peak half-
height. The calibration curve should span at least one order of magnitude
and the working range should bracket the analyte concentration.
7.10.7 Maximum Absorbance Infrared Band Technique - Following
analyte identification, construct a standard calibration curve of
concentration versus maximum infrared band intensity. For this purpose,
choose an intense, symmetrical and well resolved IR absorbance band.
(Note that IR transmission is not proportional to concentration).
Select the FT-IR scan with the highest absorbance to plot against
concentration. The calibration curve should span at least one order of
magnitude and the working range should bracket the analyte concentration.
This method is most practical for repetitive, target compound analyses.
It is more sensitive than the integrated absorbance technique.
7.10.8 Supplemental GC Detector Technique - If a GC detector is used
in tandem with the FT-IR detector, the following technique may be used:
following analyte identification, construct a standard calibration curve
of concentration versus integrated peak area. The calibration curve should
span at least one order of magnitude and the working range should bracket
the analyte concentration. This method is most practical for repetitive,
target compound analyses.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Quality
control to validate sample extraction is covered in Method 3500 and in the
extraction method utilized. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8.2 One Hundred Percent Line Test - Set the GC/FT-IR operating conditions
to those employed for the Sensitivity Test (see Section 7.5). Collect 16 scans
over the entire detector spectral range. Plot the test and measure the peak-
to-peak noise between 1800 and 2000 cm'1. This noise should be < 0.15%. Store
this plot for future reference.
8.3 Single Beam Test - With the GC/FT-IR at analysis conditions, collect
16 scans in the single beam mode. Plot the co-added file and compare with a
subsequent file acquired in the same fashion several minutes later. Note if the
spectrometer is at purge equilibrium. Also check the plot for signs of
deterioration of the lightpipe potassium bromide windows. Store this plot for
future reference.
8410 - 8 Revision 0
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deterioration of the lightpipe potassium bromide windows. Store this plot for
future reference.
8.4 Align Test - With the lightpipe and MCT detector at thermal
equilibrium, check the intensity of the centerburst versus the signal temperature
calibration curve. Signal intensity deviation from the predicted intensity may
mean thermal equilibrium has not yet been achieved, loss of detector coolant,
decrease in source output, or a loss in signal throughput resulting from
lightpipe deterioration.
8.5 Mirror Alignment - Adjust the interferometer mirrors to attain the
most intense signal. Data collection should not be initiated until the
interferogram is stable. If necessary, align the mirrors prior to each GC/FT-
IR run.
8.6 Lightpipe - The lightpipe and lightpipe windows should be protected
from moisture and other corrosive substances at all times. For this purpose,
maintain the lightpipe temperature above the maximum GC program temperature but
below its thermal degradation limit. When not in use, maintain the lightpipe
temperature slightly above ambient. At all times maintain a flow of dry, inert,
carrier gas through the lightpipe.
8.7 Beamsplitter - If the spectrometer is thermostated, maintain the
beamsplitter at a temperature slightly above ambient at all times. If the
spectrometer is not thermostated, minimize exposure of the beamsplitter to
atmospheric water vapor.
9.0 METHOD PERFORMANCE
9.1 Method 8410 has been in use at the U.S. Environmental Protection
Agency Environmental Monitoring Systems Laboratory for more than two years.
Portions of it have been reviewed by key members of the FT-IR spectroscopic
community (9). Side by side comparisons with GC/MS sample analyses indicate
similar demands upon analytical personnel for the two techniques. Extracts
previously subjected to GC/MS analysis are generally compatible with GC/FT-IR.
However, it should be kept in mind that lightpipe windows are typically water
soluble. Thus, extracts must be vigorously dried prior to analysis.
10.0 REFERENCES
1. Handbook for Analytical Quality Control in Water and Wastewater
Laboratories; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory. ORD Publication Offices of Center for
Environmental Research Information: Cincinnati, OH, March 1979; Section 4,
EPA-600/4-79-019.
2. Freeman, R.R. Hewlett Packard Application Note: Quantitative Analysis
Using a Purged Solitless In.iection Technique; ANGC 7-76.
3. Cole, R.H. Tables of Wavenumbers for the Calibration of Infrared
Spectrometers; Pergamon: New York, 1977.
8410 - 9 Revision 0
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4. Grasselli, J.G.; Griffiths, P.R.; Hannah, R.W. "Criteria for Presentation
of Spectra from Computerized IR Instruments"; ADD!. Spectrosc. 1982, 36>
87.
5. Nyquist, R.A. The Interpretation of Vaoor-Phase Infrared Spectra. Group
Frequency Data; Volume I. Sadtler Laboratories: Philadelphia, PA, 1984.
6. Socrates, G. Infrared Characteristic Group Frequencies; John Wiley and
Sons: New York, NY, 1980.
7. Bellamy, L.J. The Infrared Spectra of Complex Organic Molecules; 2nd ed.;
John Wiley and Sons: New York, NY, 1958.
8. Szymanski, H.A. Infrared Band Handbook. Volumes I and II; Plenum: New
York, NY, 1965.
9. Gurka, D.F. "Interim Protocol for the Automated Analysis of Semivolatile
Organic Compounds by Gas Chromatography/Fourier Transform- Infrared
Spectrometry"; APP!. Spectrosc. 1985, 39, 826.
10. Griffiths, P.R.; de Haseth, J.A.; Azarraga, L.V. "Capillary GC/FT-IR";
Anal. Chem. 1983, 55, 1361A.
11. Griffiths, P.R.; de Haseth, J.A. Fourier Transform-Infrared Spectrometrv;
Wiley-Interscience: New York, NY, 1986.
12. Gurka, D. F.; Farnham, I.; Potter, B. B.; Pyle, S.; Titus, R. and Duncan,
W. "Quantitation Capability of a Directly Linked Gas Chromatography/Fourier
Transform Infrared/Mass Spectrometry System"; Anal. Chem.. 1989, 61, 1584.
8410 - 10 Revision 0
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TABLE 1.
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED IDENTIFICATION LIMITS FOR BASE/NEUTRAL EXTRACTABLES
Compound
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzo(a)pyrene
Bis(2-chloroethyl) ether
Bi s (2-chl oroethoxy)methane
Bis (2-chl oroisopropyl) ether
Butyl benzyl phthalate
4-Bromophenyl phenyl ether
2-Chl oronaphthal ene
4-Chloroaniline
4-Chlorophenyl phenyl ether
Chrysene
Di-n-butyl phthalate
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
Di-n-propyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Bis-(2-ethylhexyl) phthalate
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachlorocyclopentadiene
Hexachloroethane
1 , 3-Hexachl orobutadi ene
Isophorone
2-Methyl naphthal ene
Naphthalene
Nitrobenzene
N-Nitrosodimethylamine
N-Nitrosodi-n-propylamine
N-Nitrosodiphenylaminee
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
Identification
ng injected8
40(25)
50(50)
40(50)
(50)
(100)
70(10)
50(10)
50(10)
25(10)
40(5)
110
40
20(5)
(100)
20(5)
40
20(5)
20(5)d
25(10)
25(5)
50
50
50
20
20
25(10)
100(50)
40(50)
40
120
50
120
40
110
40(25)
25
20(5)
50(5)
40
40
40
40
50(50)
100(50)
50(25)
Limit
M9/Lb
20(12.5)
25(25)
20(25)
(25)
(50)
35(5)
25(5)
25(5)
12.5(5)
20(2.5)
55
20
10(2.5)
(50)
10(2.5)
20
10(2.5)
10(2.5)
12.5(5)
12.5(2.5)
25
25
25
10
10
12.5(5)
50(25)
20(25)
20
60
25
60
20
55
20(12.5)
12.5
10(2.5)
25(2.5)
20
20
20
20
25(25)
50(25)
25(12.5)
i/max, cm"1
799
799
874
745
756
1115
1084
1088
1748
1238
851
1543
1242
757
1748
1192
1748
1751
1748
1748
1458
779
1474
1547
1551
1748
773
737
1346
814
783
853
1690
3069
779
1539
1483
1485
1501
1564
1583
1362
729
820
750
8410 - 11
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TABLE 1.
(Continued)
Determined using on-column injection and the conditions of Section 7.3. A
medium band HgCdTe detector [3800-700cm~1; D'value (Apeak 1000 Hz, 1) 4.5 x
1010 cm Hz1/2W"1] type with a 0.25 mm2 focal chip was used. The GC/FT-IR system
is a 1976 retrofitted model.
Based on a 2 /xL injection of a one liter sample that has been extracted and
concentrated to a volume of 1.0 ml_.
Most intense IR peak and suggested quantitation peak.
Values in parentheses were determined with a new (1986) GC/FT-IR system. A
narrow band HgCdTe detector [3800-750cm'1; D'value (Apeak 1000 Hz, 1) 4 x 1010
cm Hz1/2W"1] was used. Chromatographic conditions are those of Section 7.3.
Detected as diphenylamine.
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TABLE 2.
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED ON-LINE AUTOMATED IDENTIFICATION LIMITS FOR ACIDIC EXTRACTABLES
Identification
Compound
ng injected8
»/niax, cm
.-1 c
Benzoic acid
2-Chlorophenol
4-Chlorophenold
4-Chl oro-3-methyl phenol
2-Methyl phenol
4-Methyl phenol
2,4-Dichlorophenol
2,4-Dinitrophenol
4, 6-Dinitro-2-methyl phenol
2-Nitrophenold
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
70
50
100
25
50
50
50
60
60
40
50
50
70
120
120
35
25
50
12.5
25
25
25
30
30
20
25
25
35
60
60
1751
1485
1500
1177
748
1177
1481
1346
1346
1335
1350
1381
1184
1470
1458
a Operating conditions are the same as those cited in Section 7.3.
b Based on a 2 /iL injection of a one liter sample that has been extracted and
concentrated to a volume of 1.0 mL.
0 Most intense IR peak and suggested quantitation peak.
d Subject to interference from co-eluting compounds.
8410 - 13
Revision 0
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TABLE 3.
GAS-PHASE GROUP FREQUENCIES
Number of
Functionality Class Compounds
Ether
Ester
Nitro
Nitrile
Ketone
Amide
Al kyne
Acid
Phenol
Aryl, Alkyl
Benzyl, Alkyl
Diaryl
Dialkyl
Alkyl, Vinyl
Unsubstituted Aliphatic
Aromatic
Monosubstituted Acetate
Aliphatic
Aromatic
Aliphatic
Aromatic
Aliphatic (acyclic)
(a, 6 unsaturated)
Aromatic
Substituted Acetamides
Aliphatic
Aliphatic
Dimerized-Aliphatic
Aromatic
1,4-Disubstituted
1,3-Disubstituted
14
3
5
12
3
29
11
34
5
18
9
9
13
2
16
8
8
24
22
2
10
10
15
15
15
10
10
10
Frequency
Range, i/cm"1
1215-1275
1103-1117
1238-1250
1084-1130
1204-1207
1128-1142
1748-1761
1703-1759
1753-1788
1566-1594
1548-1589
1377-1408
1327-1381
1535-1566
1335-1358
2240-2265
2234-2245
1726-1732
1638-1699
1701-1722
1710-1724
3323-3329
3574-3580
1770-1782
3586-3595
3574-3586
1757-1774
3645-3657
1233-1269
1171-1190
3643-3655
1256-1315
1157-1198
8410 - 14
Revision 0
November 1990
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TABLE 3.
(Continued)
Functionality
Phenol (continued)
Alcohol
Amine
Al kane
Aldehyde
Benzene
Class
1,2-Disubstituted
Primary Aliphatic
Secondary Aliphatic
Tertiary Aliphatic
Primary Aromatic
Secondary Aromatic
Aliphatic
Aromatic
Aliphatic
Monosubstituted
Number of
Compounds
6
20
11
16
17
10
10
6
15
5
10
14
12
12
12
6
6
6
7
24
24
11
23
25
Frequency
Range, i/cm"1
3582-3595
1255-1274
3630-3680
1206-1270
1026-1094
3604-3665
1231-1270
3640-3670
1213-1245
3480-3532
3387-3480
760- 785
2930-2970
2851-2884
1450-1475
1355-1389
1703-1749
2820-2866
2720-2760
1742-1744
2802-2877
2698-2712
1707-1737
1582-1630
1470-1510
831- 893
735- 790
675- 698
8410 - 15
Revision 0
November 1990
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TABLE 4. FUSED SILICA CAPILLARY COLUMN GC/FT-IR QUANTITATION RESULTS
Compound
Concentration
Range, and
Identification
Limit, nga
Maximum
Absorbance"
Correlation
Coefficient11
Integrated
Absorbancec
Correlation
Coefficient*
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzoic acid
Benzo(a)pyrene
Bi s (2-chl oroethoxyjmethane
Bis(2-chloroethyl) ether
Bis (2-chl oroisopropyl) ether
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
4-Chloroaniline
4-Chl oro-3-methyl phenol
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenole
4-Chlorophenyl phenyl ether
Chrysene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
2,4-Dichlorophenol
Dimethyl phthalate
Dimethyl phthalate
Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Bis(2-ethylhexyl) phthalate
Fluoranthene
Fluorene
Hexachlorobenzene
1,3-Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Isophorone
2-Methyl naphthalene
25-250
25-250
50-250
50-250
50-250
100-250
25-250
25-250
50-250
25-250
25-250
25-250
25-250
100-250
25-250
25-250
100-250
25-250
25-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
50-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
50-250
100-250
25-250
25-250
50-250
0.9995
0.9959
0.9969
0.9918
0.9864
0.9966
0.9992
0.9955
0.9981
0.9995
0.9999
0.9991
0.9975
0.9897
0.9976
0.9999
0.9985
0.9697
0.9998
0.9937
0.9985
0.9994
0.9964
0.9998
0.9998
0.9936
0.9920
0.9966
0.9947
0.9983
0.9991
0.9983
0.9987
0.9981
0.9960
0.9862
0.9986
0.9984
0.9981
0.9985
0.9985
0.9971
0.9921
0.9892
0.9074
0.9991
0.9992
0.9998
0.9996
0.9994
0.9965
0.9946
0.9988
0.9965
0.9997
0.9984
0.8579
0.9996
0.9947
0.9950
0.9994
0.9969
0.9996
0.9997
0.9967
0.9916
0.9928
0.9966
0.9991
0.9993
0.9966
0.9989
0.9995
0.9979
0.9845
0.9992
0.9990
0.9950
(continued)
8410 - 16
Revision 0
November 1990
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TABLE 4. (Continued)
Compound
Concentration
Range, and
Identification
Limit, nga
Maximum
Absorbance"
Correlation
Coefficient11
Integrated
Absorbance0
Correlation
Coefficient"
2-Methyl phenol
4-Methyl phenol
Naphthalene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenole
4-Nitrophenol
N-Ni trosodimethyl ami ne
N-Ni trosodi phenyl ami ne
N-Ni trosodi -n-propyl amine
Pentachlorophenol
Phenanthrene
Phenol
Pyrene
1,2,4-Trichlorobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
25-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
25-250
25-250
25-250
50-250
25-250
25-250
50-250
50-250
25-250
25-250
0.9972
0.9972
0.9956
0.9996
0.9985
0.9936
0.9997
0.9951
0.9982
0.9994
0.9991
0.9859
0.9941
0.9978
0.9971
0.9969
0.9952
0.9969
0.9964
0.9959
0.9954
0.9994
0.9990
0.9992
0.9979
0.9953
0.9993
0.9971
0.9995
0.9883
0.9989
0.9966
0.9977
0.991
0.9966
0.9965
Lower end of range is at or near the identification limit.
FT-IR scan with highest absorbance plotted against concentration.
Integrated absorbance of combined FT-IR scans which occur at or above the
chromatogram peak half-height.
Regression analysis carried out at four concentration levels. Each level
analyzed in duplicate chromatographic conditions are stated in Section 7.3.
Subject to interference from co-eluting compounds.
8410 - 17
Revision 0
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METHOD 8410
GAS CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED (GC/FT-IR)
SPECTROMETRY FOR SEMIVOLATILE ORGANICS: CAPILLARY COLUMN
Start
7 7 Replace
Source
7 . 8 Frequency
No
7.10.7 Standard
calibration
curve of cone .
v* . max . IR
band int«n*i ty
8410 - 18
Revision 0
November 1990
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METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE
1.0 SCOPE AND APPLICATION
1.1 Method 1312 is designed to determine the mobility of both organic
and inorganic analytes present in samples of soils, wastes, and wastewaters.
1.2 If a total analysis of the soil, waste, or wastewater demonstrates
that individual analytes are not present, or that they are present but at such
low concentrations that the appropriate regulatory levels could not possibly be
exceeded, Method 1312 need not be run.
1.3 If an analysis of any one of the liquid fractions of the 1312
extract indicates that a regulated compound is present at such high
concentrations that, even after accounting for dilution from the other fractions
of the extract, the concentration would be above the regulatory level for that
compound, then the waste is hazardous and it is not necessary to analyze the
remaining fractions of the extract.
1.4 If an analysis of extract obtained using a bottle extractor shows
that the concentration of any regulated volatile analyte exceeds the regulatory
level for that compound, then the waste is hazardous and extraction using the ZHE
is not necessary. However, extract from a bottle extractor cannot be used to
demonstrate that the concentration of volatile compounds is below the regulatory
level.
2.0 SUMMARY OF METHOD
2.1 For liquid samples (i.e.. those containing less than 0.5 percent
dry solid material), the sample, after filtration through a 0.6 to 0.8 p.m glass
fiber filter, is defined as the 1312 extract.
2.2 For samples containing greater than 0.5 percent solids, the liquid
phase, if any, is separated from -the solid phase and stored for later analysis;
the particle size of the solid phase is reduced, if necessary. The solid phase
is extracted with an amount of extraction fluid equal to 20 times the weight of
the solid phase. The extraction fluid employed is a function of the region of
the country where the sample site is located if the sample is a soil. If the
sample is a waste or wastewater, the extraction fluid employed is a pH 4.2
solution. A special extractor vessel is used when testing for volatile analytes
(see Table 1 for a list of volatile compounds). Following extraction, the liquid
extract is separated from the sample by 0.6 to 0.8 /Ltm glass fiber filter.
2.3 If compatible (i.e., multiple phases will not form on combination),
the initial liquid phase of the waste is added to the liquid extract, and these
are analyzed together. If incompatible, the liquids are analyzed separately and
the results are mathematically combined to yield a volume-weighted average
concentration.
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3.0 INTERFERENCES
3.1 Potential interferences that may be encountered during analysis are
discussed in the individual analytical methods.
4.0 APPARATUS AND MATERIALS
4.1 Agitation apparatus: The agitation apparatus must be capable of
rotating the extraction vessel in an end-over-end fashion (see Figure 1) at 30
+ 2 rpm. Suitable devices known to EPA are identified in Table 2.
4.2 Extraction Vessels
4.2.1 Zero Headspace Extraction Vessel (ZHE). This device is for
use only when the sample is being tested for the mobility of volatile
analytes (i.e., those listed in Table 1). The ZHE (depicted in Figure 2)
allows for liquid/solid separation within the device and effectively
precludes headspace. This type of vessel allows for initial liquid/solid
separation, extraction, and final extract filtration without opening the
vessel (see Step 4.3.1). These vessels shall have an internal volume of
500-600 ml and be equipped to accommodate a 90-110 mm filter. The devices
contain VITON*1 0-rings which should be replaced frequently. Suitable ZHE
devices known to EPA are identified in Table 3.
For the ZHE to be acceptable for use, the piston within the ZHE
should be able to be moved with approximately 15 psi or less. If it takes
more pressure to move the piston, the 0-rings in the device should be
replaced. If this does not solve the problem, the ZHE is unacceptable for
1312 analyses and the manufacturer should be contacted.
The ZHE should be checked for leaks after every extraction. If the
device contains a built-in pressure gauge, pressurize the device to 50
psi, allow it to stand unattended for 1 hour, and recheck the pressure.
If the device does not have a built-in pressure gauge, pressurize the
device to 50 psi, submerge it in water, and check for the presence of air
bubbles escaping from any of the fittings. If pressure is lost, check all
fittings and inspect and replace 0-rings, if necessary. Retest the
device. If leakage problems cannot be solved, the manufacturer should be
contacted.
Some ZHEs use gas pressure to actuate the ZHE piston, while others
use mechanical pressure (see Table 3). Whereas the volatiles procedure
(see Step 7.3) refers to pounds-per-square-inch (psi), for the
mechanically actuated piston, the pressure applied is measured in torque-
inch-pounds. Refer to the manufacturer's instructions as to the proper
conversion.
4.2.2 Bottle Extraction Vessel. When the sample is being
evaluated using the nonvolatile extraction, a jar with sufficient capacity
to hold the sample and the extraction fluid is needed. Headspace is
allowed in this vessel.
VlTON® is a trademark of Du Pont.
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The extraction bottles may be constructed from various materials,
depending on the analytes to be analyzed and the nature of the waste (see
Step 4.3.3). It is recommended that borosilicate glass bottles be used
instead of other types of glass, especially when inorganics are of
concern. Plastic bottles, other than polytetrafluoroethylene, shall not
be used if organics are to be investigated. Bottles are available from a
number of laboratory suppliers. When this type of extraction vessel is
used, the filtration device discussed in Step 4.3.2 is used for initial
liquid/solid separation and final extract filtration.
4.3 Filtration Devices: It is recommended that all filtrations be
performed in a hood.
i
4.3.1 Zero-Headspace Extraction Vessel (ZHE): When the sample
is evaluated for volatiles, the zero-headspace extraction vessel described
in Step 4.2.1 is used for filtration. The device shall be capable of
supporting and keeping in place the glass fiber filter and be able to
withstand the pressure needed to accomplish separation (50 psi).
NOTE: When it is suspected that the glass fiber filter has been ruptured, an
in-line glass fiber filter may be used to filter the material within the
ZHE.
4.3.2 Filter Holder: When the sample is evaluated for other than
volatile analytes, a filter holder capable of supporting a glass fiber
filter and able to withstand the pressure needed to accomplish separation
may be used. Suitable filter holders range from simple vacuum units to
relatively complex systems capable of exerting pressures of up to 50 psi
or more. The type of filter holder used depends op the properties of the
material to be filtered (see Step 4.3.3). These devices shall have a
minimum internal volume of 300 mL and be equipped to accommodate a minimum
filter size of 47 mm (filter hol'ders having an internal capacity of 1.5 L
or greater, and equipped to accommodate a 142 mm diameter filter, are
recommended). Vacuum filtration can only be used for wastes with low
solids content (<10 percent) and for highly granular, liquid-containing
wastes. All other types of wastes should be filtered using positive
pressure filtration. Suitable filter holders known to EPA are shown in
Table 4.
4.3.3 Materials of Construction: Extraction vessels and
filtration devices shall be made of inert materials which will not leach
or absorb sample components. Glass, polytetrafluoroethylene (PTFE), or
type 316 stainless steel equipment may be used when evaluating the
mobility of both organic and inorganic components. Devices made of high-
density polyethylene (HOPE), polypropylene (PP), or polyvinyl chloride
(PVC) may be used only when evaluating the mobility of metals.
Borosilicate glass bottles are recommended for use over other types of
glass bottles, especially when inorganics are analytes of concern.
4.4 Filters: Filters shall be made of borosilicate glass fiber, shall
contain no binder materials, and shall have an effective pore size of 0.6 to
0.8-/xm or equivalent. Filters known to EPA which meet these specifications are
identified in Table 5. Pre-filters must not be used. When evaluating the
mobility of metals, filters shall be acid-washed prior to use by rinsing with IN
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nitric acid followed by three consecutive rinses with deionized distilled water
(a minimum of 1-L per rinse is recommended). Glass fiber filters are fragile and
should be handled with care.
4.5 pH Meters: The meter should be accurate to + 0.05 units at 25°C.
4.6 ZHE Extract Collection Devices: TEDLAR*2 bags or glass, stainless
steel or PTFE gas-tight syringes are used to collect the initial liquid phase and
the final extract when using the ZHE device. These devices listed are
recommended for use under the following conditions:
4.6.1 If a waste contains an aqueous liquid phase or if a waste
does not contain a significant amount of nonaqueous liquid (i.e.. <1
percent of total waste), the TEDLAR* bag or a 600 ml syringe should be used
to collect and combine the initial liquid and solid extract.
4.6.2 If a waste contains a significant amount of nonaqueous
liquid in the initial^liquid phase (i.e., >1 percent of total waste), the
syringe or the TEDLAR* bag may be used for both the initial solid/liquid
separation and the final extract filtration. However, analysts should use
one or the other, not both.
4.6.3 If the waste contains no initial liquid phase (is 100
percent solid) or^has no significant solid phase (is 100 percent liquid),
either the TEDLAR* bag or the syringe may be used. If the syringe is used,
discard the first 5 mL of liquid expressed from the device. The remaining
aliquots are used for analysis.
4.7 ZHE Extraction Fluid Transfer Devices: Any device capable of
transferring the extraction fluid into the ZHE without changing the nature of the
extraction fluid is acceptable (e.g., a positive displacement or peristaltic
pump, a gas-tight syringe, pressure filtration unit (see Step 4.3.2), or other
ZHE device).
4.8 Laboratory Balance: Any laboratory balance accurate to within ±
0.01 grams may be used (all weight measurements are to be within ±0.1 grams).
4.9 Beaker or Erlenmeyer flask, glass, 500 mL.
4.10 Watchglass, appropriate diameter to cover beaker or Erlenmeyer
flask.
4.11 Magnetic stirrer.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
TEDLAR* is a registered trademark of Du Pont.
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provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to reagent water in this method
refer to one of the following, as appropriate.
5.2.1 Inorganic Analytes: Water which is generated by any method
which would achieve the performance standards for ASTM Type II water. The
analyte(s) of concern must be no higher than the highest of either (1.) the
detection limit, or (2) five percent of the regulatory level for that
analyte, or (3) five percent of the measured concentration in the sample.
5.2.2 Volatile Analytes: Water in which an interferant is not
observed at the method detection limit of the compounds of interest.
Organic-free water can be generated by passing tap water through a carbon
filter bed containing about 1 Ib. of activated carbon. A water
purification system may be used to generate organic-free deionized water.
Organic-free water may also be prepared by boiling water for 15 minutes.
Subsequently, while maintaining the temperature at 90°C, bubble a
contaminant-free inert gas through the water for 1 hour. The analyte(s)
of concern must be no higher than the highest of either (1) the detection
limit, or (2) five percent of the regulatory level for that analyte, or
(3) five percent of the measured concentration in the sample.
5.2.3 Semivolatile Analytes: Water in which an interferant is
not observed at the method detection limit of the compounds of interest.
Organic-free water can be generated by passing tap water through a carbon
filter bed containing about 1 Ib. of activated carbon. A water
purification system may be used to generate organic-free deionized water.
The analyte(s) of concern must be no higher than the highest of either (1)
the detection limit, or (2) five percent of the regulatory level for that
analyte, or (3) five percent of the measured concentration in the sample.
5.3 Sulfuric acid/nitric acid (60/40 weight percent mixture) H2S04/HN03.
Cautiously mix 60 g of concentrated sulfuric acid with 40 g of concentrated
nitric acid.
5.4 Extraction fluids.
5.4.1 Extraction fluid #1: This fluid is made by adding the
60/40 weight percent mixture of sulfuric and nitric acids to reagent water
(Step 5.2) until the pH is 4.20 ± 0.05. The fluid is used to determine
the Teachability of soil from a site that is east of the Mississippi
River, and the Teachability of wastes and wastewaters.
NOTE: Solutions are unbuffered and exact pH may not be attained.
5.4.2 Extraction fluid #2: This fluid is made by adding the
60/40 weight percent mixture of sulfuric and nitric acids to reagent water
(Step 5.2) until the pH is 5.00 ± 0.05. The fluid is used to determine
the Teachability of soil from a site that is west of the Mississippi
River.
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5.4.3 Extraction fluid #3: This fluid is reagent water (Step
5.2) and is used to determine cyanide and volatiles Teachability.
NOTE: These extraction fluids should be monitored frequently for impurities.
The pH should be checked prior to use to ensure that these fluids are made
up accurately. If impurities are found or the pH is not within the above
specifications, the fluid shall be discarded and fresh extraction fluid
prepared.
5.5 Analytical standards shall be prepared according to the appropriate
analytical method.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples shall be collected using an appropriate sampling plan.
6.2 There may be requirements on the minimal size of the field sample
depending upon the physical state or states of the waste and the analytes of
concern. An aliquot is needed for the preliminary evaluations of the percent
solids and the particle size. An aliquot may be needed to conduct the
nonvolatile analyte extraction procedure (see Step 1.4 concerning the use of this
extract for volatile organics). If volatile organics are of concern, another
aliquot may be needed. Quality control measures may require additional aliquots.
Further, it is always wise to collect more sample just in case something goes
wrong with the initial attempt to conduct the test.
6.3 Preservatives shall not be added to samples before extraction.
6.4 Samples may be refrigerated unless refrigeration results in
irreversible physical change to the waste. If precipitation occurs, the entire
sample (including precipitate) should be extracted.
6.5 When the sample is to be evaluated for volatile analytes, care
shall be taken to minimize the loss of volatiles. Samples shall be collected and
stored in a manner intended to prevent the loss of volatile analytes (e.g..
samples should be collected in Teflon-lined septum capped vials and stored at
4°C. Samples should be opened only immediately prior to extraction).
6.6 1312 extracts should be prepared for analysis and analyzed as soon
as possible following extraction. Extracts or portions of extracts for metallic
analyte determinations must be acidified with nitric acid to a pH < 2, unless
precipitation occurs (see Step 7.2.14 if precipitation occurs). Extracts should
be preserved for other analytes according to the guidance given in the individual
analysis methods. Extracts or portions of extracts for organic analyte
determinations shall not be allowed to come into contact with the atmosphere
(i.e.. no headspace) to prevent losses. See Section 8.0 (Quality Control) for
acceptable sample and extract holding times.
7.0 PROCEDURE
7.1 Preliminary Evaluations
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Perform preliminary 1312 evaluations on a minimum 100 gram aliquot of
sample. This aliquot may not actually undergo 1312 extraction. These
preliminary evaluations include: (1) determination of the percent solids (Step
7.1.1); (2) determination of whether the waste contains insignificant solids and
is, therefore, its own extract after filtration (Step 7.1.2); and (3)
determination of whether the solid portion of the waste requires particle size
reduction (Section 7.1.3).
7.1.1 Preliminary determination of percent solids: Percent
solids is defined as that fraction of a waste sample (as a percentage of
the total sample) from which no liquid may be forced out by an applied
pressure, as described below.
7.1.1.1 If the sample will obviously yield no free
liquid when subjected to pressure filtration (i.e., is 100%
solids), weigh out a representative subsample (100 g minimum) and
proceed to Step 7.1.3.
7.1.1.2 If the sample is liquid or multiphasic,
liquid/solid separation to make a preliminary determination of
percent solids is required. This involves the filtration device
discussed in Step 4.3.2, and is outlined in Steps 7.1.1.3 through
7.1.1.9.
7.1.1.3 Pre-weigh the filter and the container that will
receive the filtrate.
7.1.1.4 Assemble filter holder and filter following the
manufacturer's instructions. Place the filter on the support
screen and secure.
7.1.1.5 Weigh out a subsample of the waste (100 gram
minimum) and record the weight.
7.1.1.6 Allow slurries to stand to permit the solid phase
to settle. Samples that settle slowly may be centrifuged prior to
filtration. Centri.fugation is to be used only as an aid to
filtration. If used, the liquid should be decanted and filtered
followed by filtration of the solid portion of the waste through
the same filtration system.
7.1.1.7 Quantitatively transfer the sample to the filter
holder (liquid and solid phases). Spread the sample evenly over
the surface of the filter. If filtration of the waste at 4°C
reduces the amount of expressed liquid over what would be expressed
at room temperature, then allow the sample to warm up to room
temperature in the device before filtering.
NOTE: If sample material (>1 percent of original sample weight) has obviously
adhered to the container used to transfer the sample to the filtration
apparatus, determine the weight of this residue and subtract it from the
sample weight determined in Step 7.1.1.5 to determine the weight of the
sample that will be filtered.
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Gradually apply vacuum or gentle pressure of 1-10 psi, until air
or pressurizing gas moves through the filter. If this point is not
reached under 10 psi, and if no additional liquid has passed through the
filter in any 2-minute interval, slowly increase the pressure in 10 psi
increments to a maximum of 50 psi. After each incremental increase of 10
psi, if the pressurizing gas has not moved through the filter, and if no
additional liquid has passed through the filter in any 2-minute interval,
proceed to the next 10-psi increment. When the pressurizing gas begins to
move through the filter, or when liquid flow has ceased at 50 psi (i.e.,
filtration does not result in any additional filtrate within any 2-minute
period), stop the filtration.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.1.1.8 The material in the filter holder is defined as
the solid phase of the sample, and the filtrate is defined as the
liquid phase.
NOTE: Some samples, such as oily wastes and some paint wastes, will obviously
contain some material that appears to be a liquid, but even after applying
vacuum or pressure filtration, as outlined in Step 7.1.1.7, this material
may not filter. If this is the case, the material within the filtration
device is defined as a solid. Do not replace the original filter with a
fresh filter under any circumstances. Use only one filter.
7.1.1.9 Determine the weight of the liquid phase by
subtracting the weight of the filtrate container (see Step 7.1.1.3)
from the total weight of the filtrate-filled container. Determine
the weight of the solid phase of the sample by subtracting the
weight of the liquid phase from the weight of the total sample, as
determined in Step 7.1.1.5 or 7.1.1.7.
Record the weight of the liquid and solid phases.
Calculate the percent solids as follows:
Weight of solid (Step 7.1.1.9)
Percent solids = x 100
Total weight of waste (Step 7.1.1.5 or 7.1.1.7)
7.1.2 If the percent solids determined in Step 7.1.1.9 is equal
to or greater than 0.5%, then proceed either to Step 7.1.3 to determine
whether the solid material requires particle size reduction or to Step
7.1.2.1 if it is noticed that a small amount of the filtrate is entrained
in wetting of the filter. If the percent solids determined in Step
7.1.1.9 is less than 0.5%, then proceed to Step 7.2.9 if the nonvolatile
1312 analysis is to be performed, and to Section 7.3 with a fresh portion
of the waste if the volatile 1312 analysis is to be performed.
7.1.2.1 Remove the solid phase and filter from the
filtration apparatus.
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7.1.2.2 Dry the filter and solid phase at 100 ± 20°C
until two successive weighings yield the same value within + 1
percent. Record the final weight.
Note: Caution should be taken to ensure that the subject solid will not flash
upon heating. It is recommended that the drying oven be vented to a hood
or other appropriate device.
7.1.2.3 Calculate the percent dry solids as follows:
Percent (Weight of dry sample + filter) - tared weight of filter
dry solids = x 100
Initial weight of sample (Step 7.1.1.5 or 7.1.1.7)
7.1.2.4 If the percent dry solids is less than 0.5%,
then proceed to Step 7.2.9 if the nonvolatile 1312 analysis is to
be performed, and to Step 7.3 if the volatile 1312 analysis is to
be performed. If the percent dry solids is greater than or equal
to 0.5%, and if the nonvolatile 1312 analysis is to be performed,
return to the beginning of this Section (7.1) and, with a fresh
portion of sample, determine whether particle size reduction is
necessary (Step 7.1.3).
7.1.3 Determination of whether the sample requires particle-size
reduction (particle-size is reduced during this step): Using the solid
portion of the sample, evaluate the solid for particle size. Particle-
size reduction is required, unless the solid has a surface area per gram
of material equal to or greater than 3.1 cm , or is smaller than 1 cm in
its narrowest dimension (i.e., is capable of passing through a 9.5 mm
(0.375 inch) standard sieve). If the surface area is smaller or the
particle size larger than described above, prepare the solid portion of
the sample for extraction by crushing, cutting, or grinding the waste to
a surface area or particle size as described above. If the solids are
prepared for organic volatiles extraction, special precautions must be
taken (see Step 7.3.6).
Note: Surface area criteria are meant for filamentous (e.g., paper, cloth, and
similar) waste materials. Actual measurement of surface area is not
required, nor is it recommended. For materials that do not obviously meet
the criteria, sample-specific methods would need to be developed and
employed to measure the surface area. Such methodology is currently not
available.
7.1.4 Determination of appropriate extraction fluid:
7.1.4.1 For soils, if the sample is from a site that is
east of the Mississippi River, extraction fluid #1 should be used.
If the sample is from a site that is west of the Mississippi River,
extraction fluid #2 should be used.
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7.1.4.2 For wastes and wastewater, extraction fluid #1
should be used.
7.1.4.3 For cyanide-containing wastes and/or soils,
extraction fluid #3 (reagent water) must be used because leaching
of cyanide- containing samples under acidic conditions may result
in the formation of hydrogen cyanide gas.
7.1.5 If the aliquot of the sample used for the preliminary
evaluation (Steps 7.1.1 - 7.1.4) was determined to be 100% solid at Step
7.1.1.1, then it can be used for the Section 7.2 extraction (assuming at
least 100 grams remain), and the Section 7.3 extraction (assuming at least
25 grams remain). If the aliquot was subjected to the procedure in Step
7.1.1.7, then another aliquot shall be used for the volatile extraction
procedure in Section 7.3. The aliquot of the waste subjected to the
procedure in Step 7.1.1.7 might be appropriate for use for the Section 7.2
extraction if an adequate amount of solid (as determined by Step 7.1.1.9)
was obtained. The amount of solid necessary is dependent upon whether a
sufficient amount of extract will be produced to support the analyses. If
an adequate amount of solid remains, proceed to Step 7.2.10 of the
nonvolatile 1312 extraction.
7.2 Procedure when Volatiles are not Involved
A minimum sample size of 100 grams (solid and liquid phases) is
recommended. In some cases, a larger sample size may be appropriate, depending
on the solids content of the waste sample (percent solids, See Step 7.1.1),
whether the initial liquid phase of the waste will be miscible with the aqueous
extract of the solid, and whether inorganics, semivolatile organics, pesticides,
and herbicides are all analytes of concern. Enough solids should be generated
for extraction such that the volume of 1312 extract will be sufficient to support
all of the analyses required. If the amount of extract generated by a single
1312 extraction will not be sufficient to perform all of the analyses, more than
one extraction may be performed and the extracts from each combined and aliquoted
for analysis.
7.2.1 If the sample will obviously yield no liquid when subjected
to pressure filtration (i.e., is 100 percent solid, see Step 7.1.1), weigh
out a subsample of the sample (100 gram minimum) and proceed to Step
7.2.9.
7.2.2 If the sample is liquid or multiphasic, liquid/solid
separation is required. This involves the filtration device described in
Step 4.3.2 and is outlined in Steps 7.2.3 to 7.2.8.
7.2.3 Pre-weigh the container that will receive the filtrate.
7.2.4 Assemble the filter holder and filter following the
manufacturer's instructions. Place the filter on the support screen and
secure. Acid wash the filter if evaluating the mobility of metals (see
Step 4.4).
Note: Acid washed filters may be used for all nonvolatile extractions even when
metals are not of concern.
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7.2.5 Weigh out a subsample of the sample (100 gram minimum) and
record the weight. If the waste contains <0.5 percent dry solids (Step
7.1.2), the liquid portion of the waste, after filtration, is defined as
the 1312 extract. Therefore, enough of the sample should be filtered so
that the amount of filtered liquid will support all of the analyses
required of the 1312 extract. For wastes containing >0.5 percent dry
solids (Steps 7.1.1 or 7.1.2), use the percent solids information obtained
in Step 7.1.1 to determine the optimum sample size (100 gram minimum) for
filtration. Enough solids should be generated by filtration to support
the analyses to be performed on the 1312 extract.
7.2.6 Allow slurries to stand to permit the solid phase to settle.
Samples that settle slowly may be centrifuged prior to filtration. Use
centrifugation only as an aid to filtration. If the sample is
centrifuged, the liquid should be decanted and filtered followed by
filtration of the solid portion of the waste through the same filtration
system.
7.2.7 Quantitatively transfer the sample (liquid and solid phases)
to the filter holder (see Step 4.3.2). Spread the waste sample evenly
over the surface of the filter. If filtration of the waste at 4°C reduces
the amount of expressed liquid over what would be expressed at room
temperature, then allow the sample to warm up to room temperature in the
device before filtering.
NOTE: If waste material (>1 percent of the original sample weight) has obviously
adhered to the container used to transfer the sample to the filtration
apparatus, determine the weight of this residue and subtract it from the
sample weight determined in Step 7.2.5, to determine the weight of the
waste sample that will be filtered.
Gradually apply vacuum or gentle pressure of 1-10 psi, until air
or pressurizing gas moves through the filter. If this point if not
reached under 10 psi, and if no additional liquid has passed through the
filter in any 2-minute interval, slowly increase the pressure in 10-psi
increments to maximum of 50 psi. After each incremental increase of 10
psi, if the pressurizing gas has not moved through the filter, and if no
additional liquid has passed through the filter in any 2-minute interval,
proceed to the next 10-psi increment. When the pressurizing gas begins to
move through the filter, or when the liquid flow has ceased at 50 psi
(i.e.. filtration does not result in any additional filtrate within a
2-minute period), stop the filtration.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.2.8 The material in the filter holder is defined as the solid
phase of the sample, and the filtrate is defined as the liquid phase.
Weigh the filtrate. The liquid phase may now be either analyzed (see
Steps 7.2.12) or stored at 4°C until time of analysis.
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NOTE: Some wastes, such as oily wastes and some paint wastes, will obviously
contain some material which appears to be a liquid. Even after applying
vacuum or pressure filtration, as outlined in Step 7.2.7, this material
may not filter. If this is the case, the material within the filtration
device is defined as a solid, and is carried through the extraction as a
solid. Do not replace the original filter with a fresh filter under any
circumstances. Use only one filter.
7.2.9 If the sample contains <0.5% dry solids (see Step 7.1.2),
proceed to Step 7.2.13. If the sample contains >0.5 percent dry solids
(see Step 7.1.1 or 7.1.2), and if particle-size reduction of the solid was
needed in Step 7.1.3, proceed to Step 7.2.10. If the sample as received
passes a 9.5 mm sieve, quantitatively transfer the solid material into the
extractor bottle along with the filter used to separate the initial liquid
from the solid phase, and proceed to Step 7.2.11.
7.2.10 Prepare the solid portion of the sample for extraction by
crushing, cutting, or grinding the waste to a surface area or particle-
size as described in Step 7.1.3. When the surface area or particle-size
has been appropriately altered, quantitatively transfer the solid material
into an extractor bottle. Include the filter used to separate the initial
liquid from the solid phase.
NOTE: Sieving of the waste is not normally required. Surface area requirements
are meant for filamentous (e.g., paper, cloth) and similar waste
materials. Actual measurement of surface area is not recommended. If
sieving is necessary, a Teflon-coated sieve should be used to avoid
contamination of the sample.
7.2.11 Determine the amount of extraction fluid to add to the
extractor vessel as follows:
20 x % solids (Step 7.1.1) x weight of waste
filtered (Step 7.2.5 or 7.2.7)
Weight of =
extraction fluid
100
Slowly add this amount of appropriate extraction fluid (see Step
7.1.4) to the extractor vessel. Close the extractor bottle tightly (it is
recommended that Teflon tape be used to ensure a tight seal), secure in
rotary extractor device, and rotate at 30 ± 2 rpm for 18 + 2 hours.
Ambient temperature (i.e.. temperature of room in which extraction takes
place) shall be maintained at 23 + 2°C during the extraction period.
NOTE: As agitation continues, pressure may build up within the extractor bottle
for some types of sample (e.g.. limed or calcium carbonate-containing
sample may evolve gases such as carbon dioxide). To relieve excess
pressure, the extractor bottle may be periodically opened (e.g.. after 15
minutes, 30 minutes, and 1 hour) and vented into a hood.
7.2.12 Following the 18 + 2 hour extraction, separate the material
in the extractor vessel into its component liquid and solid phases by
filtering through a new glass fiber filter, as outlined in Step 7.2.7.
1312 - 12 Revision 0
November 1990
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For final filtration of the 1312 extract, the glass fiber filter may be
changed, if necessary, to facilitate filtration. Filter(s) shall be
acid-washed (see Step 4.4) if evaluating the mobility of metals.
7.2.13 Prepare the 1312 extract as follows:
7.2.13.1 If the sample contained no initial liquid phase,
the filtered liquid material obtained from Step 7.2.12 is defined
as the 1312 extract. Proceed to Step 7.2.14.
7.2.13.2 If compatible (e.g.. multiple phases will not
result on combination), combine the filtered liquid resulting from
Step 7.2.12 with the initial liquid phase of the sample obtained
in Step 7.2.7. This combined liquid is defined as the 1312
extract. Proceed to Step 7.2.14.
7.2.13.3 If the initial liquid phase of the waste, as
obtained from Step 7.2.7, is not or may not be compatible with the
filtered liquid resulting from Step 7.2.12, do not combine these
liquids. Analyze these liquids, collectively defined as the 1312
extract, and combine the results mathematically, as described in
Step 7.2.14.
7.2.14 Following collection of the 1312 extract, the pH of the
extract should be recorded. Immediately aliquot and preserve the extract
for analysis. Metals aliquots must be acidified with nitric acid to pH <
2. If precipitation is observed upon addition of nitric acid to a small
aliquot of the extract, then the remaining portion of the extract for
metals analyses shall not be acidified and the extract shall be analyzed
as soon as possible. All other aliquots must be stored under
refrigeration (4°C) until analyzed. The 1312 extract shall be prepared
and analyzed according to appropriate analytical methods. 1312 extracts
to be analyzed for metals shall be acid digested except in those instances
where digestion causes loss of metallic analytes. If an analysis of the
undigested extract shows that the concentration of any regulated metallic
analyte exceeds the regulatory level, then the waste is hazardous and
digestion of the extract is not necessary. However, data on undigested
extracts alone cannot be used to demonstrate that the waste is not
hazardous. If the individual phases are to be analyzed separately,
determine the volume of the individual phases (to + 0.5 percent), conduct
the appropriate analyses, and combine the results mathematically by using
a simple volume-weighted average:
(V,) (C,) + (V2) (C2)
Final Analyte Concentration =
V, + V2
where:
V, = The volume of the first phase (L).
C, = The concentration of the analyte of concern in the first phase (mg/L).
V2 = The volume of the second phase (L).
C2 = The concentration of the analyte of concern in the second phase
(mg/L).
1312 - 13 . Revision 0
November 1990
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7.2.15 Compare the analyte concentrations in the 1312 extract with
the levels identified in the appropriate regulations. Refer to Section
8.0 for quality assurance requirements.
7.3 Procedure when Volatiles are Involved
Use the ZHE device to obtain 1312 extract for analysis of volatile
compounds only. Extract resulting from the use of the ZHE shall not be used to
evaluate the mobility of non-volatile analytes (e.g., metals, pesticides, etc.).
The ZHE device has approximately a 500 ml internal capacity. The ZHE can
thus accommodate a maximum of 25 grams of solid (defined as that fraction of a
sample from which no additional liquid may be forced out by an applied pressure
of 50 psi), due to the need to add an amount of extraction fluid equal to 20
times the weight of the solid phase.
Charge the ZHE with sample only once and do not open the device until the
final extract (of the solid) has been collected. Repeated filling of the ZHE to
obtain 25 grams of solid is not permitted.
Do not allow the sample, the initial liquid phase, or the extract to be
exposed to the atmosphere for any more time than is absolutely necessary. Any
manipulation of these materials should be done when cold (4°C) to minimize loss
of volatiles.
7.3.1 Pre-weigh the (evacuated) filtrate collection container
(see Step 4.6) and set aside. If using a TEDLAR* bag, express all liquid
from the ZHE device into the bag, whether for the initial or final
liquid/solid separation, and take an aliquot from the liquid in the bag
for analysis. The containers listed in Step 4.6 are recommended for use
under the conditions stated in Steps 4.6.1-4.6.3.
7.3.2 Place the ZHE piston within the body of the ZHE (it may be
helpful first to moisten the piston 0-rings slightly with extraction
fluid). Adjust the piston within the ZHE body to a height that will
minimize the distance the piston will have to move once the ZHE is charged
with sample (based upon sample size requirements determined from Step 7.3,
Step 7.1.1 and/or 7.1.2). Secure the gas inlet/outlet flange (bottom
flange) onto the ZHE body in accordance with the manufacturer's
instructions. Secure the glass fiber filter between the support screens
and set aside. Set liquid inlet/outlet flange (top flange) aside.
7.3.3 If the sample is 100% solid (see Step 7.1.1), weigh out
a subsample (25 gram maximum) of the waste, record weight, and proceed to
Step 7.3.5.
7.3.4 If the sample contains <0.5% dry solids (Step 7.1.2), the
liquid portion of waste, after filtration, is defined as the 1312 extract.
Filter enough of the sample so that the amount of filtered liquid will
support all of the volatile analyses required. For samples containing
>0.5% dry solids (Steps 7.1.1 and/or 7.1.2), use the percent solids
information obtained in Step 7.1.1 to determine the optimum sample size to
charge into the ZHE. The recommended sample size is as follows:
1312 - 14 Revision 0
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7.3.4.1 For samples containing <5% solids (see Step
7.1.1), weigh out a 500 gram subsample of waste and record the
weight.
7.3.4.2 For wastes containing >5% solids (see Step
7.1.1), determine the amount of waste to charge into the ZHE as
fol1ows:
25
Weight of waste to charge ZHE = x 100
percent solids (Step 7.1.1)
Weigh out a subsample of the waste of the appropriate size and
record the weight.
7.3.5 If particle-size reduction of the solid portion of the
sample was required in Step 7.1.3, proceed to Step 7.3.6. If particle-
size reduction was not required in Step 7.1.3, proceed to Step 7.3.7.
7.3.6 Prepare the sample for extraction by crushing, cutting, or
grinding the solid portion of the waste to a surface area or particle size
as described in Step 7.1.3.1. Wastes and appropriate reduction equipment
should be refrigerated, if possible, to 4°C prior to particle-size
reduction. The means used to effect particle-size reduction must not
generate heat in and of itself. If reduction of the solid phase of the
waste is necessary, exposure of the waste to the atmosphere should be
avoided to the extent possible.
NOTE: Sieving of the waste is not recommended due to the possibility that
volatiles may be lost. The use of an appropriately graduated ruler is
recommended as an acceptable alternative. Surface area requirements are
meant for filamentous (e.g.. paper, cloth) and similar waste materials.
Actual measurement of surface area is not recommended.
When the surface area or particle-size has been appropriately
altered, proceed to Step 7.3.7.
7.3.7 Waste slurries need not be allowed to stand to permit the
solid phase to settle. Do not centrifuge samples prior to filtration.
7.3.8 Quantitatively transfer the entire sample (liquid and solid
phases) quickly to the ZHE. Secure the filter and support screens into
the top flange of the device and secure the top flange to the ZHE body in
accordance with the manufacturer's instructions. Tighten all ZHE fittings
and place the device in the vertical position (gas inlet/outlet flange on
the bottom). Do not attach the extraction collection device to the top
plate.
Note: If sample material (>1% of original sample weight) has obviously adhered
to the container used to transfer the sample to the ZHE, determine the
weight of this residue and subtract it from the sample weight determined
in Step 7.3.4 to determine the weight of the waste sample that will be
filtered.
1312 - 15 Revision 0
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Attach a gas line to the gas inlet/outlet valve (bottom flange)
and, with the liquid inlet/outlet valve (top flange) open, begin applying
gentle pressure of 1-10 psi (or more if necessary) to force all headspace
slowly out of the ZHE device into a hood. At the first appearance of
liquid from the liquid inlet/outlet valve, quickly close the valve and
discontinue pressure. If filtration of the waste at 4°C reduces the
amount of expressed liquid over what would be expressed at room
temperature, then allow the sample to warm up to room temperature in the
device before filtering. If the waste is 100 percent solid (see Step
7.1.1), slowly increase the pressure to a maximum of 50 psi to force most
of the headspace out of the device and proceed to Step 7.3.12.
7.3.9 Attach the evacuated pre-weighed filtrate collection
container to the liquid inlet/outlet valve and open the valve. Begin
applying gentle pressure of 1-10 psi to force the liquid phase of the
sample into the filtrate collection container. If no additional liquid
has passed through the filter in any 2-minute interval, slowly increase
the pressure in 10-psi increments to a maximum of 50 psi. After each
incremental increase of 10 psi, if no additional liquid has passed through
the filter in any 2-minute interval, proceed to the next 10-psi increment.
When liquid flow has ceased such that continued pressure filtration at 50
psi does not result in any additional filtrate within a 2-minute period,
stop the filtration. Close the liquid inlet/outlet valve, discontinue
pressure to the piston, and disconnect and weigh the filtrate collection
container.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.3.10 The material in the ZHE is defined as the solid phase of
the sample and the filtrate is defined as the liquid phase.
NOTE: Some samples, such as oily wastes and some paint wastes, will obviously
contain some material which appears to be a liquid. Even after applying
pressure filtration, this material will not filter. If this is the case,
the material within the filtration device is defined as a solid, and is
carried through the 1312 extraction as a solid.
If the original waste contained <0.5 percent dry solids (see Step
7.1.2), this filtrate is defined as the 1312 extract and is analyzed
directly. Proceed to Step 7.3.15.
7.3.11 The liquid phase may now be either analyzed immediately
(see Steps 7.3.13 through 7.3.15) or stored at 4°C under minimal headspace
conditions until time of analysis. Determine the weight of extraction
fluid #3 to add to the ZHE as follows:
20 x % solids (Step 7.1.1) x weight
of waste filtered (Step 7.3.4 or 7.3.8)
Weight of extraction fluid =
100
1312 - 16 Revision 0
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7.3.12 The following steps detail how to add the appropriate
amount of extraction fluid to the solid material within the ZHE and
agitation of the ZHE vessel. Extraction fluid #3 is used in all cases
(see Step 5.7).
7.3.12.1 With the ZHE in the vertical position, attach a
line from the extraction fluid reservoir to the liquid inlet/outlet
valve. The line used shall contain fresh extraction fluid and
should be preflushed with fluid to eliminate any air pockets in the
line. Release gas pressure on the ZHE piston (from the gas
inlet/outlet valve), open the liquid inlet/outlet valve, and begin
transferring extraction fluid (by pumping or similar means) into
the ZHE. Continue pumping extraction fluid into the ZHE until the
appropriate amount of fluid has been introduced into the device.
7.3.12.2 After the extraction fluid has been added,
immediately close the liquid inlet/outlet valve and disconnect the
extraction fluid line. Check the ZHE to ensure that all valves are
in their closed positions. Manually rotate the device in an
end-over-end fashion 2 or 3 times. Reposition the ZHE in the
vertical position with the liquid inlet/outlet valve on top.
Pressurize the ZHE to 5-10 psi (if necessary) and slowly open the
liquid inlet/outlet valve to bleed out any headspace (into a hood)
that may have been introduced due to the addition of extraction
fluid. This bleeding shall be done quickly and shall be stopped
at the first appearance of liquid from the valve. Re-pressurize
the ZHE with 5-10 psi and check all ZHE fittings to ensure that
they are closed.
7.3.12.3 Place the ZHE in the rotary extractor apparatus
(if it is not already there) and rotate at 30 ± 2 rpm for 18+2
hours. Ambient temperature (i .e.. temperature of room in which
extraction occurs) shall be maintained at 23 ± 2°C during
agitation.
7.3.13 Following the 18+2 hour agitation period, check the
pressure behind the ZHE piston by quickly opening and closing the gas
inlet/outlet valve and noting the escape of gas. If the pressure has not
been maintained (i.e., no gas release observed), the ZHE is leaking.
Check the ZHE for leaking as specified in Step 4.2.1, and perform the
extraction again with a new sample of waste. If the pressure within the
device has been maintained, the material in the extractor vessel is once
again separated into its component liquid and solid phases. If the waste
contained an initial liquid phase, the liquid may be filtered directly
into the same filtrate collection container (i.e., TEDLAR* bag) holding the
initial liquid phase of the waste. A separate filtrate collection
container must be used if combining would create multiple phases, or there
is not enough volume left within the filtrate collection container.
Filter through the glass fiber filter, using the ZHE device as discussed
in Step 7.3.9. All extracts shall be filtered and collected if the TEDLAR*
bag is used, if the extract is multiphasic, or if the waste contained an
initial liquid phase (see Steps 4.6 and 7.3.1).
1312 - 17 Revision 0
November 1990
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NOTE: An in-line glass fiber filter may be used to filter the material within
the ZHE if it is suspected that the glass fiber filter has been ruptured
7.3.14 If the original sample contained no initial liquid phase,
the filtered liquid material obtained from Step 7.3.13 is defined as the
1312 extract. If the sample contained an initial liquid phase, the
filtered liquid material obtained from Step 7.3.13 and the initial liquid
phase (Step 7.3.9) are collectively defined as the 1312 extract.
7.3.15 Following collection of the 1312 extract, immediately
prepare the extract for analysis and store with minimal headspace at 4°C
until analyzed. Analyze the 1312 extract according to the appropriate
analytical methods. If the individual phases are to be analyzed
separately (i.e.. are not miscible), determine the volume of the
individual phases (to 0.5%), conduct the appropriate analyses, and combine
the results mathematically by using a simple volume- weighted average:
(V,) (C,) + (V2) (C2)
Final Analyte =
Concentration
V2
where:
V1 = The volume of the first phases (L).
C1 = The concentration of the analyte of concern in the first phase (mg/L).
V2 = The volume of the second phase (L).
C2 = The concentration of the analyte of concern in the second phase
(mg/L).
7.3.16 Compare the analyte concentrations in the 1312 extract with
the levels identified in the appropriate regulations. Refer to Section
8.0 for quality assurance requirements.
8.0 QUALITY CONTROL
8.1 A minimum of one blank (using the same extraction fluid as used for
the samples) for every 20 extractions that have been conducted in an extraction
vessel.
8.2 A matrix spike shall be performed for each waste type (e.g..
wastewater treatment sludge, contaminated soil, etc.) unless the result exceeds
the regulatory level and the data is being used solely to demonstrate that the
waste property exceeds the regulatory level. A minimum of one matrix spike must
be analyzed for each analytical batch. The bias determined from the matrix spike
determination shall be used to correct the measured values. (See Steps 8.2.4 and
8.2.5) As a minimum, follow the matrix spike addition guidance provided in each
analytical method.
8.2.1 Matrix spikes are to be added after filtration of the 1312
extract and before preservation. Matrix spikes should not be added prior
to 1312 extraction of the sample.
1312 - 18 Revision 0
November 1990
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8.2.2 In most cases, matrix spike levels should be added at a
concentration equivalent to the corresponding regulatory level. If the
analyte concentration is less than one half the regulatory level, the
spike concentration may be as low as one half of the analyte
concentration, but may not be less than five times the method detection
limit. In order to avoid differences in matrix effects, the matrix spikes
must be added to the same nominal volume of 1312 extract as that which was
analyzed for the unspiked sample.
8.2.3 The purpose of the matrix spike is to monitor the
performance of the analytical methods used, and to determine whether
matrix interferences exist. Use of other internal calibration methods,
modification of the analytical methods, or use of alternate analytical
methods may be needed to accurately measure the analyte concentration in
the 1312 extract when the recovery of the matrix spike is below the
expected analytical method performance.
8.2.4 Matrix spike recoveries are calculated by the following
formula:
%R (% Recovery) = 100 (Xs - Xu) / K
where:
Xs = measured value for the spiked sample
Xu = measured value for the unspiked sample, and
K = known value of the spike in the sample.
8.2.5 Measured values are corrected for analytical bias using the
following formula:
Xc = 100 (Xu / %R)
where:
Xc = corrected value, and
Xu = measured value of the unspiked sample.
8.3 All quality control measures described in the appropriate analytical
methods shall be followed.
8.4 Samples must undergo 1312 extraction within the following time
periods:
1312 - 19 Revision 0
November 1990
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SAMPLE MAXIMUM HOLDING TIMES (davs)
Volatiles
Semi -
volatiles
Mercury
Metals,
except
mercury
From: Field
Collec-
tion
To: 1312
extrac-
tion
14
14
28
180
From: 1312
extrac-
tion
To: Prepara-
tive
extrac-
tion
NA
7
NA
NA
From: Prepara-
tive
extrac-
tion
To: determi-
native
analysis
14
40
28
180
Total
Elapsed
Time
28
61
56
360
NA = Not Applicable
If sample holding times are exceeded, the values obtained will be considered
minimal concentrations. Exceeding the holding time is not acceptable in
establishing that a waste does not exceed the regulatory level. Exceeding the
holding time will not invalidate characterization if the waste exceeds the
regulatory level.
9.0 METHOD PERFORMANCE
9.1 Precision results for semi-volatiles and metals: An eastern soil
with high organic content and a western soil with low organic content were used
for the semi-volatile and metal leaching experiments. Both types of soil were
analyzed prior to contaminant spiking. The results are shown in Table 6. The
concentrations of contaminants leached from the soils were consistently
reproducible, as shown by the low relative standard deviations (RSDs) of the
recoveries (generally less than 10 % for most of the compounds).
9.2 Precision results for volatiles: Four different soils were spiked
and tested for the extraction of volatiles. Soils One and Two were from western
and eastern Superfund sites. Soils Three and Four were mixtures of a western
soil with low organic content and two different municipal sludges. The results
are shown in Table 7. Extract concentrations of volatile organics from the
eastern soil were lower than from the western soil. Replicate Teachings of Soils
Three and Four showed lower precision than the leachates from the Superfund
soils.
1312 - 20
Revision 0
November 1990
-------
10.0 REFERENCES
1.0 Environmental Monitoring Systems Laboratory, "QA Support for RCRA Testing:
Annual Report". EPA Contract 68-03-3249, January 1989.
2.0 Research Triangle Institute, "Interlaboratory Comparison of Methods 1310,
1311, and 1312 for Lead in Soil". U.S. EPA Contract 68-01-7075, November
1988.
1312 - 21 Revision 0
November 1990
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Table 1. Volatile Analytes1
Compound CAS No.
Acetone 67-64-1
Benzene 71-43-2
n-Butyl alcohol 71-36-3
Carbon disulfide 75-15-0
Carbon tetrachloride 56-23-5
Chlorobenzene 108-90-7
Chloroform 67-66-3
1,2-Dichloroethane 107-06-2
1,1-Dichloroethylene 75-35-4
Ethyl acetate 141-78-6
Ethyl benzene 100-41-4
Ethyl ether 60-29-7
Isobutanol 78-83-1
Methanol 67-56-1
Methylene chloride 75-09-2
Methyl ethyl ketone 78-93-3
Methyl isobutyl ketone 108-10-1
Tetrachloroethylene 127-18-4
Toluene 108-88-3
1,1,1,-Trichloroethane 71-55-6
Trichloroethylene 79-01-6
Trichlorofluoromethane 75-69-4
l,l,2-Trichloro-l,2,2-trifluoroethane 76-13-1
Vinyl chloride 75-01-4
Xylene 1330-20-7
When testing for any or all of these analytes, the zero-headspace extractor
vessel shall be used instead of the bottle extractor.
1312 - 22 Revision 0
November 1990
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Table 2. Suitable Rotary Agitation Apparatus1
Company
Location
Model No.
Analytical Testing and
Consulting Services,
Inc.
Associated Design and
Manufacturing Company
Environmental Machine and
Design, Inc.
IRA Machine Shop and
Laboratory
Lars Lande Manufacturing
Millipore Corp.
Warrington, PA
(215) 343-4490
Alexandria, VA
(703) 549-5999
Lynchburg, VA
(804) 845-6424
Santurce, PR
(809) 752-4004
4-vessel extractor (DC20S);
8-vessel extractor (DC20);
12-vessel extractor (DC20B)
2-vessel
4-vessel
6-vessel
8-vessel
12-vessel
24-vessel
(3740-2);
(3740-4);
(3740-6);
(3740-8);
(3740-12);
(3740-24)
8-vessel (08-00-00)
4-vessel (04-00-00)
8-vessel (011001)
Whitmore Lake, MI 10-vessel (10VRE)
(313) 449-4116 5-vessel (5VRE)
Bedford, MA
(800) 225-3384
4-ZHE or
4 l-liter
bottle extractor
(YT300RAHW)
1 Any device that rotates the extraction vessel in an end-over-end fashion at 30
±2 rpm is acceptable.
1312 - 23
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Table 3. Suitable Zero-Headspace Extractor Vessels1
Company
Location
Model No.
Analytical Testing &
Consulting Services, Inc.
Associated Design and
Manufacturing Company
Lars Lande Manufacturing2
Millipore Corporation
Environmental Machine
and Design, Inc.
Warrington, PA
(215) 343-4490
Alexandria, VA
(703) 549-5999
Whitmore Lake, MI
(313) 449-4116
Bedford, MA
(800) 225-3384
Lynchburg, VA
(804) 845-6424
C102, Mechanical
Pressure Device
3745-ZHE, Gas
Pressure Device
ZHE-11, Gas
Pressure Device
YT30090HW, Gas
Pressure Device
VOLA-TOX1, Gas
Pressure Device
1 Any device that meets the specifications listed in Step 4.2.1 of the method is
suitable.
2 This device uses a 110 mm filter.
1312 - 24
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November 1990
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Table 4. Suitable Filter Holders1
Company
Nucleopore Corporation
Micro Filtration
Systems
Millipore Corporation
Location
Pleasanton, CA
(800) 882-7711
Dublin, CA
(800) 334-7132
(415) 828-6010
Bedford, MA
(800) 225-3384
Model/
Catalogue #
425910
410400
302400
311400
YT30142HW
XX1004700
Size
142 mm
47 mm
142 mm
47 mm
142 mm
47 mm
1 Any device capable of separating the liquid from the solid phase of the waste
is suitable, providing that it is chemically compatible with the waste and the
constituents to be analyzed. Plastic devices (not listed above) may be used when
only inorganic analytes are of concern. The 142 mm size filter holder is
recommended.
Table 5. Suitable Filter Media1
Company
Millipore Corporation
Nucleopore Corporation
Whatman Laboratory
Products, Inc.
Micro Filtration
Systems
Location Model
Bedford, MA AP40
(800) 225-3384
Pleasanton, CA 211625
(415) 463-2530
Clifton, NJ GFF
(201) 773-5800
Dublin, CA GF75
(800) 334-7132
(415) 828-6010
Pore
Size
(Mm)
0.7
0.7
0.7
0.7
1 Any filter that meets the specifications in Step 4.4 of the Method is suitable.
1312 - 25 Revision 0
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TABLE 6 - METHOD 1312 PRECISION RESULTS FOR SEMI-VOLATILES AND METALS
Eastern Soil (t>H 4.
FORTIFIED ANALYTES
bis(2-chloroethyl) -
ether
2-Chlorophenol
1,4-Dichlorobenzene
1 , 2 -Dichlorobenzene
2 -Methylphenol
Nitrobenzene
2 , 4 - D ime thy Ipheno 1
Hexachlorobutadiene
Acenaphthene
2,4-Dinitrophenol
2 , 4-Dinitrotoluene
Hexachlorobenzene
gamma BHC (Lindane)
beta BHC
METALS
Lead
Cadmium
Amount
Spiked
(Mg)
1040
1620
2000
8920
3940
1010
1460
6300
3640
1300
1900
1840
7440
640
5000
1000
* •= Triplicate analyses.
** = Duplicate analyses: one v
Amount
Recovered*
(Mg)
834
1010
344
1010
1860
812
200
95
210
896**
1150
3.7
230
35
70
387
alue was rejc
% RSD
12.5
6.8
12.3
8.0
7.7
10.0
18.4
12.9
8.1
6.1
5.4
12.0
16.3
13.3
4.3
2.3
jcted as
,2) Western Soil (vti 5.0)
Amount
Recovered*
(Mg)
616
525
272
1520
1130
457
18
280
310**
23**
585
10
1240
65.3
10
91
an outlier at the
% RSD
14.2
54.9
34.6
28.4
32.6
21.3
87.6
22.8
7.7
15.7
54.4
173.2
55.2
51.7
51.7
71.3
90%
confidence level using the Dixon Q test.
1312 - 26
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November 1990
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TABLE 7 - METHOD 1312 PRECISION RESULTS FOR VOLATILES
Soil
No. 1
(Western)
ound Name
:one
•lonitrile
ene
ityl Alcohol
-Butanol)
ion disulfide
ion tetrachloride
irobenzene
iroform
Dichloroe thane
Dichloroethane
•1 acetate
'Ibenzene
•1 ether
iutanol (4 -Methyl
.-propanol)
tylene chloride
iyl ethyl ketone
:-Butanone)
iyl isobutyl
:tone
1,2-Tetrachloro-
:hane
2,2-Tetrachloro-
:hane
•achloroethene
tene
1-Trichloro-
:hane
2-Trichloro-
:hane
ihloroethene
i_ i _ ~_
inloro-
.uororae thane
2-Trichloro-
•ifluoroe thane
rl chloride
Avg.
%Rec.
44.0
52.5
47.8
55.5
21.4
40.6
64.4
61.3
73.4
31.4
76.4
56.2
48.0
0.0
47.5
56.7
81.1
69.0
85.3
45.1
59.2
47.2
76.2
54.5
20.7
18.1
10.2
* %RSD
12.4
68.4
8.29
2.91
16.4
18.6
6.76
8.04
4.59
14.5
9.65
9.22
16.4
ND
30.3
5.94
10.3
6.73
7.04
12.7
8.06
16.0
5.72
11.1
24.5
26.7
20.3
Soil
No. 2
(Eastern)
Avg.
%Rec.
43.8
50.5
34.8
49.2
12.9
22.3
41.5
54.8
68.7
22.9
75.4
23.2
55.1
0.0
42.2
61.9
88.9
41.1
58.9
15.2
49.3
33.8
67.3
39.4
12.6
6.95
7.17
* %RSD
2.25
70.0
16.3
14.6
49.5
29.1
13.1
16.4
11.3
39.3
4.02
11.5
9.72
ND
42.9
3.94
2.99
11.3
4.15
17.4
10.5
22.8
8.43
19.5
60.1
58.0
72.8
Soil No
(Western
Sludge)
Avg.
%Rec . **
116.0
49.3
49.8
65.5
36.5
36.2
44.2
61.8
58.3
32.0
23.0
37.5
37.3
61.8
52.0
73.7
58.3
50.8
64.0
26.2
45.7
40.7
61.7
38.8
28.5
21.5
25.0
. 3
and
%RSD
11.5
44.9
36.7
37.2
51.5
41.4
32.0
29.1
33.3
54.4
119.8
36.1
31.2
37.7
37.4
31.3
32.6
31.5
25.7
44.0
35.2
40.6
28.0
40.9
34.0
67.8
61.0
Soil No. 4
(Western and
Sludge)
Avg.
%Rec.*** %RSD
21.3 71.4
51.8 4.6
33.4 41.1
73.0 13.9
21.3 31.5
24.0 34.0
33.0 24.9
45.8 38.6
41.2 37.8
16.8 26.4
11.0 115.5
27.2 28.6
42.0 17.6
76.0 12.2
37.3 16.6
40.6 39.0
39.8 40.3
36.8 23.8
53.6 15.8
18.6 24.2
31.4 37.2
26.2 38.8
46.4 25.4
25.6 34.1
19.8 33.9
15.3 24.8
11.8 25.4
* Triplicate analyses
** Six replicate analyses
*** Five replicate analyses
1312 - 27
Revision 0
November 1990
-------
Motor
(30 ± 2 rpm
Extraction Vessel Holder
UUU
Figure 1. Rotary Agitation Apparatus
1312 - 28
Revision 0
November 1990
-------
Liquid Inlet/Outlet Valve
Top Flange
^•^MMM
Support Screen S*
Filter//
Support Screen
Viton O-Rings
Bottom Flange
Pressurized Gas
Inlet/Outlet Valve
Pressure
Gauge
Figure 2. Zero-Headspace Extractor (ZHE)
1312 - 29
Revision 0
November 1990
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METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE
Separate
liquids from
solids with 0.6
- 0.8 urn glass
fiber filter
Discard
solids
Separate
liquids from
solids with 0.6
- 0.8 urn glass
fiber filter
Solid
Yes
Extract »/
appropriate fluid
1) Bottle extractor
for non-volatile!
2) ZHE device for
volatile!
Reduce
particle size
to <9.5 mm
1312 - 30
Revision 0
November 1990
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METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
Discard
solids
Solid
Store
at
1 iquid
4 C
Separate
extract from
iclida u/ 0.6 -
0.8 urn glass
fiber filter
I
liquid
compatible \ No
with the
extract?
Measure amount of
liquid and analyze
(mathematically
combine result w/
result of extract
analysis)
Combine
extract «/
liquid phase
of waste
Analyze
liquid
STOP
1312 - 31
Revision 0
November 1990
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METHOD 9040A
pH ELECTROMETRIC MEASUREMENT
1.0 SCOPE AND APPLICATION
1.1 Method 9040 is used to measure the pH of aqueous wastes and those
multiphase wastes where the aqueous phase constitutes at least 20% of the total
volume of the waste.
1.2 The corrosivity of concentrated acids and bases, or of concentrated
acids and bases mixed with inert substances, cannot be measured. The pH
measurement requires some water content.
2.0 SUMMARY
2.1 The pH of the sample is determined electrometrically using either
a glass electrode in combination with a reference potential or a combination
electrode. The measuring device is calibrated using a series of standard
solutions of known pH.
3.0 INTERFERENCES
i
3.1 The glass electrode, in general, is not subject to solution
interferences from color, turbidity, colloidal matter, oxidants, reductants, or
high salinity.
3.2 Sodium error at pH levels >10 can be reduced or eliminated by using
a low-sodium-error electrode.
3.3 Coatings of oily material or particulate matter can impair
electrode response. These coatings can usually be removed by gentle wiping or
detergent washing, followed by rinsing with distilled water. An additional
treatment with hydrochloric acid (1:10) may be necessary to remove any remaining
film.
3.4 Temperature effects on the electrometric determination of pH arise
from two sources. The first is caused by the change in electrode output at
various temperatures. This interference can be controlled with instruments
having temperature compensation or by calibrating the electrode-instrument system
at the temperature of the samples. The second source of temperature effects is
the change of pH due to changes in the sample as the temperature changes. This
error is sample-dependent and cannot be controlled. It should, therefore, be
noted by reporting both the pH and temperature at the time of analysis.
4.0 APPARATUS AND MATERIALS
4.1 pH meter: Laboratory or field model. Many instruments are commer-
cially available with various specifications and optional equipment.
4.2 Glass electrode.
9040A - 1 Revision 1
November 1990
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4.3 Reference electrode: A silver-silver chloride or other reference
electrode of constant potential may be used.
NOTE: Combination electrodes incorporating both measuring and referenced
functions are convenient to use and are available with solid, gel-type
filling materials that require minimal maintenance.
4.4 Magnetic stirrer and Teflon-coated stirring bar.
4.5 Thermometer or temperature sensor for automatic compensation.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Primary standard buffer salts are available from the National
Institute of Standards and Technology (Special Publication 260) and should be
used in situations where extreme accuracy is necessary. Preparation of reference
solutions from these salts requires some special precautions and handling, such
as low-conductivity dilution water, drying ovens, and carbon-dioxide-free purge
gas. These solutions should be replaced at least once each month.
5.3 Secondary standard buffers may be prepared from NBS salts or
purchased as solutions from commercial vendors. These commercially available
solutions have been validated by comparison with NIST standards and are
recommended for routine use.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be analyzed as soon as possible.
7.0 PROCEDURE
7.1 Calibration:
7.1.1 Because of the wide variety of pH meters and accessories,
detailed operating procedures cannot be incorporated into this method.
Each analyst must be acquainted with the operation of each system and
familiar with all instrument functions. Special attention to care of the
electrodes is recommended.
7.1.2 Each instrument/electrode system must be calibrated at a
minimum of two points that bracket the expected pH of the samples and are
approximately three pH units or more apart. (For corrosivity characteri-
zation, the calibration of the pH meter should include a buffer of pH 2
for acidic wastes and a pH 12 buffer for caustic wastes.) Various
9040A - 2 Revision 1
November 1990
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instrument designs may involve use of a dial (to "balance" or
"standardize") or a slope adjustment, as outlined in the manufacturer's
instructions. Repeat adjustments on successive portions of the two buffer
solutions until readings are within 0.05 pH units of the buffer solution
value.
7.2 Place the sample or buffer solution in a clean glass beaker using
a sufficient volume to cover the sensing elements of the electrodes and to give
adequate clearance for the magnetic stirring bar. If field measurements are
being made, the electrodes may be immersed directly into the sample stream to an
adequate depth and moved in a manner to ensure sufficient sample movement across
the electrode-sensing element as indicated by drift-free readings (<0.1 pH).
7.3 If the sample temperature differs by more than 2°C from the buffer
solution, the measured pH values must be corrected. Instruments are equipped
with automatic or manual compensators that electronically adjust for temperature
differences. Refer to manufacturer's instructions.
7.4 Thoroughly rinse and gently wipe the electrodes prior to measuring
pH of samples. Immerse the electrodes into the sample beaker or sample stream
and gently stir at a constant rate to provide homogeneity and suspension of
solids. Note and record sample pH and temperature. Repeat measurement on
successive volumes of sample until values differ by <0.1 pH units. Two or three
volume changes are usually sufficient.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Electrodes must be thoroughly rinsed between samples.
9.0 METHOD PERFORMANCE
9.1 Forty-four analysts in twenty laboratories analyzed six synthetic
water samples containing exact increments of hydrogen-hydroxyl ions, with the
following results:
Accuracy as
pH Units
3.5
3.5
7.1
7.2
8.0
8.0
Standard Deviation
oH Units
0.10
0.11
0.20
0.18
0.13
0.12
Bias
-0.29
-0.00
+1.01
-0.03
-0.12
+0.16
Bias
oH Units
-0.01
+0.07
-0.002
-0.01
+0.01
10.0 REFERENCES
1. National Institute of Standards and Technology,
Material Catalog 1986-87, Special Publication 260.
Standard Reference
9040A - 3
Revision 1
November 1990
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METHOD 9040A
pH ELECTROMETRIC MEASUREMENT
Start
7.1 Calibrate pH
meter
7.2 Place sample or
buffer solution in
glass beaker
7.3 Does
temperature
differ by mor
than 2C from
buffer?
7.3 Correct
measured pH values
7.4 Immerse
electrodes and
measure pH of
sample
7.4 Note and record
pH and temperature;
repeat 2 or 3 times
with different
volumes
Stop
9040A - 4
Revision 1
November 1990
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METHOD 9045A
SOIL AND HASTE oH
1.0 SCOPE AND APPLICATION
1.1 Method 9045 is an electrometric procedure which has been approved
for measuring pH in calcareous and non-calcareous soils and waste samples.
2.0 SUMMARY OF METHOD
2.1 The soil sample is mixed either with reagent water or with a
calcium chloride solution (see Section 5.0), depending on whether the soil is
calcareous or non-calcareous, and the pH of the resulting solution is measured.
The waste sample is mixed with reagent water, and the pH of the resulting
solution is measured.
3.0 INTERFERENCES
3.1 Samples with very low or very high pH may give incorrect readings
on the meter. For samples with a true pH of >10, the measured pH may be
incorrectly low. This error can be minimized by using a low-sodium-error
electrode. Strong acid solutions, with a true pH of <1, may give incorrectly
high pH measurements.
3.2 Temperature fluctuations will cause measurement errors.
3.3 Errors will occur when the electrodes become coated. If an
electrode becomes coated with an oily material that will not rinse free, the
electrode can either (1) be cleaned with an ultrasonic bath, or (2) be washed
with detergent, rinsed several times with water, placed in 1:10 HC1 so that the
lower third of the electrode is submerged, and then thoroughly rinsed with water.
4.0 APPARATUS AND MATERIALS
4.1 pH Meter with means for temperature compensation.
4.2 Electrodes:
4.2.1 Calomel electrode.
4.2.2 Glass electrode.
4.2.3 A combination electrode can be employed instead of calomel
or glass.
4.3 Beaker: 50-mL.
4.4 Class A volumetric flasks: 1 L and 2 L.
4.5 Analytical balance: capable of weighing 0.1 g.
4.6 Aluminum foil.
9045A - 1 Revision 1
November 1990
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5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Primary standard buffer salts are available from the National
Institute of Standards and Technology (NIST) and should be used in situations
where extreme accuracy is necessary. Preparation of reference solutions from
these salts requires some special precautions and handling, such as low-
conductivity dilution water, drying ovens, and carbon-dioxide-free purge gas.
These solutions should be replaced at least once each month.
5.4 Secondary standard buffers may be prepared from NIST salts or
purchased as solutions from commercial vendors. These commercially available
solutions, which have been validated by comparison with NIST standards, are
recommended for routine use.
5.5 Stock calcium chloride solution (CaCl2), 3.6 M: Dissolve 1059 g of
CaCl? • 2H20 in reagent water in a 2-liter Class A volumetric flask. Cool the
solution, dilute it to volume with reagent water, and mix it well. Dilute 20 ml
of this solution to 1 liter with reagent water in a Class A volumetric flask and
standardize it by titrating a 25-mL aliquot of the diluted solution with standard
0.1 N AgN03, using 1 ml of 5% K2Cr04 as the indicator.
5.6 Calcium chloride (CaCl,), 0.01 M: Dilute 5 ml of stock 3.6 M CaCl,
to 1.8 liters with reagent water. If the pH of this solution is not between 5
and 6.5, adjust the pH by adding a little Ca(OH)2 or HC1. As a check on the
preparation of this solution, measure its electrical conductivity. The specific
conductivity should be 2.32 ± 0.08 umho per cm at 25°C.
5.7 Hydrochloric acid (HC1): 1:3 mixture with reagent water.
5.8 Silver nitrate (AgN03), 0.1N: volumetric standard.
5.9 Potassium chromate (K2Cr04), 5%.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be analyzed as soon as possible.
7.0 PROCEDURE
7.1 Calibration:
9045A - 2 Revision 1
November 1990
-------
7.1.1 Because of the wide variety of pH meters and accessories,
detailed operating procedures cannot be incorporated into this method.
Each analyst must be acquainted with the operation of each system and
familiar with all instrument functions. Special attention to care of the
electrodes is recommended.
7.1.2 Each instrument/electrode system must be calibrated at a
minimum of two points that bracket the expected pH of the samples and are
approximately three pH units or more apart. Repeat adjustments on
successive portions of the two buffer solutions until readings are within
0.05 pH units of the buffer solution value.
7.2 Determination of calcareous vs. non-calcareous soils:
7.2.1 Place approximately 0.5 g of sample (less than 60 mesh) on
a piece of aluminum foil.
7.2.2 Add one or two drops of 1:3 HC1 to the sample. The
presence of CaC03 is indicated by a bubbling or audible fizz.
7.2.3 If the sample produces bubbling or fizzing, it is a
calcareous soil. If no bubbling or fizzing occurs, the sample is a non-
calcareous soil.
7.3 Sample preparation and pH measurement of non-calcareous soils:
7.3.1 To 20 g of soil in a 50-mL beaker, add 20 mL of reagent
water and stir the suspension several times during the next 30 min.
7.3.2 Let the soil suspension stand for about 1 hr to allow most
of the suspended clay to settle out from the suspension.
7.3.3 Adjust the electrodes in the clamps of the electrode holder
so that, upon lowering the electrodes into the beaker, the glass electrode
will be immersed just deep enough into the clear supernatant solution to
establish a good electrical contact through the ground-glass joint or the
fiber-capillary hole. Insert the electrodes into the sample solution in
this manner. For combination electrodes, immerse just below the
suspension.
7.3.4 If the sample temperature differs by more than 2°C from the
buffer solution, the measured pH values must be corrected.
7.3.5 Report the results as "soil pH measured in water."
7.4 Sample preparation and pH measurement of calcareous soils:
7.4.1 To 10 g of soil in a 50-mL beaker, add 20 mL of 0.01 M
CaCl2 (Step 5.6) solution and stir the suspension several times during the
next 30 min.
7.4.2 Let the soil suspension stand for about 30 min to allow
most of the suspended clay to settle out from the suspension.
9045A - 3 Revision 1
November 1990
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7.4.3 Adjust the electrodes in the clamps of the electrode holder
so that, upon lowering the electrodes into the beaker, the glass electrode
will be immersed well into the partly settled suspension and the calomel
electrode will be immersed just deep enough into the clear supernatant
solution to establish a good electrical contact through the ground-glass
joint or the fiber-capillary hole. Insert the electrode into the sample
solution in this manner.
7.4.4 If the sample temperature differs by more than 2°C from the
buffer solution, the measured pH values must be corrected.
7.4.5 Report the results as "soil pH measured in 0.01 M CaCl2".
7.5 Sample preparation and pH measurement of waste materials:
7.5.1 To 20 g of waste sample in a 50-mL beaker add 20 ml reagent
water and stir the suspension several times during the next 30 min.
7.5.2 Let the waste suspension stand for about 15 min to allow
most of the suspended waste to settle out from the suspension.
NOTE: If the waste is hydroscopic and absorbs all the reagent water,
begin the experiment again using 20 g of waste and 40 ml of reagent
water.
NOTE: If the supernatant is multiphasic, decant the oily phase and
measure the pH of the aqueous phase. The electrode may need to be
cleaned (Step 3.3) if it becomes coated with an oily material.
7.5.3 Adjust the electrodes in the clamps of the electrode holder
so that, upon lowering the electrodes into the beaker, the glass electrode
will be immersed just deep enough into the clear supernatant to establish
a good electrical contact through the ground-glass joint or the fiber-
capillary hole. Insert the electrodes into the sample solution in this
manner. For combination electrodes, immerse just below the suspension.
7.5.4 If the sample temperature differs by more than 2°C from the
buffer solution, the measured pH values must be corrected.
7.5.5 Report the results as "waste pH measured in water."
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Electrodes must be thoroughly rinsed between samples.
9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
1.0 Black, Charles Allen; Methods of Soil Analysis; American Society of
Agronomy: Madison, WI, 1973.
9045A - 4 Revision 1
November 1990
-------
METHOD 9045A
SOIL AND WASTE pH
START
7 1 Calibrate
each
ins t rument/
elect rode
ays tern
7.2.1 Place
0 5 g sample
on aluminum
foil
7.S.I Add
water to 20 g
was te; stir
7 5 2 Let
was te
suspension
stand for 15
minu tea
7 4.1 Add
calcium
chloride
solution to lOg
soil; stir
7 4 2 Let
soi 1
suspension
-stand for 30
minu tes
7.3.1 Add
water to 20 g
soil; stir
7 3 2 Let
soil
suspension
stand for 1
hour
Is
supernatan t
u 1 tiphasic?
Repeat
experiment
with 20 g
waste and 40
mL water
Decant oily
phase.
measure pH of
aqueous phase
9045A - 5
Revision 1
November 1990
-------
METHOD 9045A
SOIL AND WASTE pH
(CONTINUED)
Insert
eleclrodej
into sample
solution
Correct
measured pH
values
9045A - 6
Revision 1
November 1990
-------
METHOD 9056
AN ION CHROMATOGRAPHY METHOD
1.0 SCOPE AND APPLICATION
1.1 This method addresses the sequential determination of the anions
chloride, fluoride, bromide, nitrate, nitrite, phosphate, and sulfate in the
collection solutions from the bomb combustion of solid waste samples, as well as
all water samples.
1.2 The minimum detection limit (MDL), the minimum concentration of a
substance that can be measured and reported with 99% confidence that the value
is above zero, varies for anions as a function of sample size and the
conductivity scale used. Generally, minimum detectable concentrations are in the
range of 0.05 mg/L for F- and 0.1 mg/L for Br", Cl", N03", N02", P04 , and SO,
with a 100-/zL sample loop and a 10-umho full-scale setting on the conductivity
detector. Similar values may be achieved by using a higher scale setting and an
electronic integrator. Idealized detection limits of an order of magnitude lower
have been determined in reagent water by using a 1 umho full-scale setting (Table
1).
The upper limit of the method is dependent on total anion concentration and
may be determined experimentally. These limits may be extended by appropriate
dilution.
2.0 SUMMARY OF METHOD
A small volume of combustate collection solution or other water sample,
typically 2 to 3 ml, is injected into an ion chromatograph to flush and fill a
constant volume sample loop. The sample is then injected into a stream of
carbonate-bicarbonate eluent of the same strength as the collection solution or
water sample.
The sample is pumped through three different ion exchange columns and into
a conductivity detector. The first two columns, a precolumn or guard column and
a separator column, are packed with low-capacity, strongly basic anion exchanger.
Ions are separated into discrete bands based on their affinity for the exchange
sites of the resin. The last column is a suppressor column that reduces the
background conductivity of the eluent to a low or negligible level and converts
the anions in the sample to their corresponding acids. The separated anions in
their acid form are measured using an electrical-conductivity cell. Anions are
identified based on their retention times compared to known standards.
Quantitation is accomplished by measuring the peak height or area and comparing
it to a calibration curve generated from known standards.
3.0 INTERFERENCES
3.1 Any species with a retention time similar to that of the desired ion
will interfere. Large quantities of ions eluting close to the ion of interest
will also result in an interference. Separation can be improved by adjusting the
eluent concentration and/or flow rate.
9056 - 1 Revision 0
November 1990
-------
Sample dilution and/or the use of the method of standard additions can also
be used.
For example, high levels of organic acids may be present in industrial
wastes, which may interfere with inorganic anion analysis. Two common species,
formate and acetate, elute between fluoride and chloride.
3.2 Because bromide and nitrate elute very close together, they are
potential interferents for each other. It is advisable not to have Br"/N03"
ratios higher than 1:10 or 10:1 if both anions are to be quantified. If nitrate
is observed to be an interference with bromide, use of an alternate detector
(e.g.. electrochemical detector) is recommended.
3.3 Method interferences may be caused by contaminants in the reagent
water, reagents, glassware, and other sample processing apparatus that lead to
discrete artifacts or elevated baseline in ion chromatograms.
3.4 Samples that contain particles larger than 0.45 urn and reagent
solutions that contain particles larger than 0.20 jum require filtration to
prevent damage to instrument columns and flow systems.
3.5 If a packed bed suppressor column is used, it will be slowly consumed
during analysis and, therefore, will need to be regenerated. Use of either an
anion fiber suppressor or an anion micromembrane suppressor eliminates the time-
consuming regeneration step through the use of a continuous flow of regenerant.
4.0 APPARATUS AND MATERIALS
4.1 Ion chromatograph, capable of delivering 2 to 5 ml of eluent per
minute at a pressure of 200 to 700 psi (1.3 to 4.8 MPa). The chromatograph shall
be equipped with an injection valve, a 100-juL sample loop, and set up with the
following components, as schematically illustrated in Figure 1.
4.1.1 Precolumn, a guard column placed before the separator column
to protect the separator column from being fouled by particulates or
certain organic constituents (4 x 50 mm, Dionex P/N 030825 [normal run],
or P/N 030830 [fast run], or equivalent).
4.1.2 Separator column, a column packed with low-capacity
pellicular anion exchange resin that is styrene divinylbenzene-based has
been found to be suitable for resolving F", CT, N02", P04 , Br", N03", and
SO/2 (see Figure 2) (4 x 250 mm, Dionex P/N 03827 [normal run], or P/N
030831 [fast run], or equivalent).
4.1.3 Suppressor column, a column that is capable of converting
the eluent and separated anions to their respective acid forms (fiber,
Dionex P/N 35350, micromembrane, Dionex P/N 38019 or equivalent).
4.1.4 Detector, a low-volume, flowthrough, temperature-
compensated, electrical conductivity cell (approximately 6 pi volume,
Dionex, or equivalent) equipped with a meter capable of reading from 0 to
1,000 iuseconds/cm on a linear scale.
9056 - 2 Revision 0
November 1990
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4.1.5 Pump, capable of delivering a constant flow of approximately
2 to 5 mL/min throughout the test and tolerating a pressure of 200 to 700
psi (1.3 to 4.8 MPa).
4.2 Recorder, compatible with the detector output with a full-scale
response time in 2 seconds or less.
4.3 Syringe, minimum capacity of 2 ml and equipped with a male pressure
fitting.
4.4 Eluent and regenerant reservoirs, suitable containers for storing
eluents and regenerant. For example, 4 L collapsible bags can be used.
4.5 Integrator, to integrate the area under the chromatogram. Different
integrators can perform this task when compatible with the electronics of the
detector meter or recorder. If an integrator is used, the maximum area
measurement must be within the linear range of the integrator.
4.6 Analytical balance, capable of weighing to the nearest 0.0001 g.
4.7 Pipets, Class A volumetric flasks, beakers: assorted sizes.
5.0 REAGENTS
5.1 Purity of reagents. Reagent-grade chemicals shall be used in all
tests. Unless otherwise indicated, it is intended that all reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One. Column life may be extended by passing
reagent water through a 0.22-/am filter prior to use.
5.3 Eluent, 0.003M NaHC03/0.0024M Na2C03. Dissolve 1.0080 g of sodium
bicarbonate (0.003M NaHC03) and 1.0176 g of sodium carbonate (0.0024M Na2C03) in
reagent water and dilute to 4 L with reagent water.
5.4 Suppressor regenerant solution. Add 100 ml of IN H?S04 to 3 L of
reagent water in a collapsible bag and dilute to 4 L with reagent water.
5.5 Stock solutions (1,000 mg/L).
5.5.1 Bromide stock solution (1.00 ml = 1.00 mg Br"). Dry
approximately 2 g of sodium bromide (NaBr) for 6 hours at 150°C, and cool
in a desiccator. Dissolve 1.2877 g of the dried salt in reagent water,
and dilute to 1 L with reagent water.
5.5.2 Chloride stock solution (1.00 ml = 1.00 mg CT). Dry sodium
chloride (NaCl) for 1 hour at 600°C, and cool in a desiccator. Dissolve
1.6484 g of the dry salt in reagent water, and dilute to 1 L with reagent
water.
9056 - 3 Revision 0
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5.5.3 Fluoride stock solution (1.00 ml = 1.00 mg F"). Dissolve
2.2100 g of sodium fluoride (NaF) in reagent water, and dilute to 1 L with
reagent water. Store in chemical-resistant glass or polyethylene.
5.5.4 Nitrate stock solution (1.00 ml = 1.00 mg NO,"). Dry
approximately 2 g of sodium nitrate (NaNO,) at 105°C for 24 hours.
Dissolve exactly 1.3707 g of the dried salt in reagent water, and dilute
to 1 L with reagent water.
5.5.5 Nitrite stock solution (1.00 ml = 1.00 mg N02"). Place
approximately 2 g of sodium nitrate (NaN02) in a 125 ml beaker and dry to
constant weight (about 24 hours) in a desiccator containing concentrated
H2SO,. Dissolve 1.4998 g of the dried salt in reagent water, and dilute
to 1 L with reagent water. Store in a sterilized glass bottle.
Refrigerate and prepare monthly.
NOTE: Nitrite is easily oxidized, especially in the presence of moisture,
and only fresh reagents are to be used.
NOTE: Prepare sterile bottles for storing nitrite solutions by heating for
1 hour at 170°C in an air oven.
5.5.6 Phosphate stock solution (1.00 ml = 1.00 mg P043"). Dissolve
1.4330 g of potassium dihydrogen phosphate (KH2P04) in reagent water, and
dilute to 1 L with reagent water. Dry sodium sulfate (Na2S04) for 1 hour
at 105°C and cool in a desiccator.
5.5.7 Sulfate stock solution (1.00 ml = 1.00 mg S042"). Dissolve
1.4790 g of the dried salt in reagent water, and dilute to 1 L with
reagent water.
5.6 Anion working solutions. Prepare a blank and at least three
different working solutions containing the following combinations of anions. The
combination anion solutions must be prepared in Class A volumetric flasks. See
Table 2.
5.6.1 Prepare a high-range standard solution by diluting the
volumes of each anion specified in Table 2 together to 1 L with reagent
water.
5.6.2 Prepare the intermediate-range standard solution by diluting
10.0 ml of the high-range standard solution (see Table 2) to 100 mL with
reagent water.
5.6.3 Prepare the low-range standard solution by diluting 20.0 ml
of the intermediate-range standard solution (see Table 2) to 100 ml with
reagent water.
5.7 Stability of standards. Stock standards are stable for at least 1
month when stored at 4°C. Dilute working standards should be prepared weekly,
except those that contain nitrite and phosphate, which should be prepared fresh
daily.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Analyze the samples as soon as possible after collection. Preserve
by refrigeration at 4°C.
7.0 PROCEDURE
7.1 Calibration
7.1.1 Establish ion chromatographic operating parameters
equivalent to those indicated in Table 1.
7.1.2 For each analyte of interest, prepare calibration standards
at a minimum of three concentration levels and a blank by adding
accurately measured volumes of one or more stock standards to a Class A
volumetric flask and diluting to volume with reagent water. If the
working range exceeds the linear range of the system, a sufficient number
of standards must be analyzed to allow an accurate calibration curve to be
established. One of the standards should be representative of a concen-
tration near, but above, the method detection limit if the system is
operated on an applicable attenuator range. The other standards should
correspond to the range of concentrations expected in the sample or should
define the working range of the detector. Unless the attenuator range
settings are proven to be linear, each setting must be calibrated
individually.
7.1.3 Using injections of 0.1 to 1.0 mL (determined by injection
loop volume) of each calibration standard, tabulate peak height or area
responses against the concentration. The results are used to prepare a
calibration curve for each analyte. During this procedure, retention
times must be recorded. The retention time is inversely proportional to
the concentration.
7.1.4 The working calibration curve must be verified on each
working day, or whenever the anion eluent is changed, and for every batch
of samples. If the response or retention time for any analyte varies from
the expected values by more than ± 10%, the test must be repeated, using
fresh calibration standards. If the results are still more than ± 10%, an
entirely new calibration curve must be prepared for that analyte.
7.1.5 Nonlinear response can result when the separator column
capacity is exceeded (overloading). Maximum column loading (all anions)
should not exceed about 400 ppm.
7.2 Analyses
7.2.1 Sample preparation. When aqueous samples are injected, the
water passes rapidly through the columns, and a negative "water dip" is
observed that may interfere with the early-eluting fluoride and/or
chloride ions. The water dip should not be observed in the combustate
samples; the collecting solution is a concentrated eluent solution that
9056 - 5 Revision 0
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will "match" the eluent strength when diluted to 100-mL with reagent water
according to the bomb combustion procedure. Any dilutions required in
analyzing other water samples should be made with the eluent solution.
The water dip, if present, may be removed by adding concentrated eluent to
all samples and standards. When a manual system is used, it is necessary
to micropipet concentrated buffer into each sample. The recommended
procedures follow:
(1) Prepare a 100-mL stock of eluent 100 times normal concentration by
dissolving 2.5202 g NaHCO, and 2.5438 g Na2C03 in 100-mL reagent
water. Protect the volumetric flask from air.
(2) Pipet 5 mL of each sample into a clean polystyrene micro-beaker.
Micropipet 50 nl of the concentrated buffer into the beaker and stir
well.
Dilute the samples with eluent, if necessary, to concentrations within the
linear range of the calibration.
7.2.2 Sample analysis.
7.2.2.1 Start the flow of regenerant through the
supressor column.
7.2.2.2 Set up the recorder range for maximum sensitivity
and any additional ranges needed.
7.2.2.3 Begin to pump the eluent through the columns.
After a stable baseline is obtained, inject a midrange standard. If
the peak height deviates by more than 10% from that of the previous
run, prepare fresh standards.
7.2.2.4 Begin to inject standards starting with the
highest concentration standard and decreasing in concentration. The
first sample should be a quality control reference sample to check
the calibration.
7.2.2.5 Using the procedures described in Step 7.2.1,
calculate the regression parameters for the initial standard curve.
Compare these values with those obtained in the past. If they
exceed the control limits, stop the analysis and look for the
problem.
7.2.2.6 Inject a quality control reference sample. A
spiked sample or a sample of known content must be analyzed with
each batch of samples. Calculate the concentration from the
calibration curve and compare the known value. If the control
limits are exceeded, stop the analysis until the problem is found.
Recalibration is necessary.
7.2.2.7 When an acceptable value has been obtained for
the quality control sample, begin to inject the samples.
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7.2.2.8 Load and Inject a fixed amount of well-mixed
sample. Flush injection loop thoroughly, using each new sample.
Use the same size loop for standards and samples. Record the
resulting peak size in area or peak height units. An automated
constant volume injection system may also be used.
7.2.2.9 The width of the retention time window used to
make identifications should be based on measurements of actual
retention time variations of standards over the course of a day.
Three times the standard deviation of a retention time can be used
to calculate a suggested window size for a compound. However, the
experience of the analyst should weigh heavily in the interpretation
of chromatograms.
7.2.2.10 If the response for the peak exceeds the working
range of the system, dilute the sample with an appropriate amount of
reagent water and reanalyze.
7.2.2.11 If the resulting chromatogram fails to produce
adequate resolution, or if identification of specific anions is
questionable, spike the sample with an appropriate amount of
standard and reanalyze.
NOTE: Retention time is inversely proportional to concentration. Nitrate
and sulfate exhibit the greatest amount of change, although all
anions are affected to some degree. In some cases, this peak
migration can produce poor resolution or misidentification.
7.3 Calculation
7.3.1 Prepare separate calibration curves for each anion of
interest by plotting peak size in area, or peak height units of standards
against concentration values. Compute sample concentration by comparing
sample peak response with the standard curve.
7.3.2 Enter the calibration standard concentrations and peak
heights from the integrator or recorder into a calculator with linear
least squares capabilities.
7.3.3 Calculate the following parameters: slope (s), intercept
(I), and correlation coefficient (r). The slope and intercept define a
relationship between the concentration and instrument response of the
form:
y, = s, x, + I (1)
where: y, = predicted instrument response
s,- = response slope
xf = concentration of standard i
I = intercept
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Rearrangement of the above equation yields the concentration corresponding
to an instrumental measurement:
Xj = (Yj - D/SJ (2)
where:
Xj = calculated concentration for a sample
yj = actual instrument response for a sample
Sj and I are calculated slope and intercept from calibration above.
7.3.4 Enter the sample peak height into the calculator, and
calculate the sample concentration in milligrams per liter.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 After every 10 injections, analyze a midrange calibration standard.
If the instrument response has changed by more than 5%, recalibrate. Verify
continuing calibration by analyzing a midrange standard with every sample batch.
8.3 Analyze all samples in duplicate. Take the duplicate sample through
the entire sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Single-operator accuracy and precision for reagent, drinking and
surface water, and mixed domestic and industrial wastewater are listed in Table
3.
9.2 Combustate samples. These data are based on 41 data points obtained
by six laboratories who each analyzed four used crankcase oils and three fuel oil
blends with crankcase in duplicate. The oil samples were combusted using Method
5050. A data point represents one duplicate analysis of a sample. One data
point was judged to be an outlier and was not included in the results.
9.2.1 Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the sample operator with the same apparatus under constant operating
conditions on identical test material would exceed, in the long run, in
the normal and correct operation of the test method, the following values
only in 1 case in 20 (see Table 4):
Repeatability = 20.9 /x*
*where x is the average of two results in M9/9-
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Reproducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Reproducibility = 42.1
*where x is the average value of two results in M9/9-
9.2.2 Bias. The bias of this method varies with concentration,
as shown in Table 5:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Environmental Protection Agency. Test Method for the Determination of
Inorganic Anions in Water by Ion Chromatography. EPA Method 300.0. EPA-600/4-
84-017. 1984.
2. Annual Book of ASTM Standards, Volume 11.01 Water D4327, Standard Test
Method for Anions in Water by Ion Chromatography, pp. 696-703. 1988.
3. Standard Methods for the Examination of Water and Wastewater, Method 429,
"Determination of Anions by Ion Chromatography with Conductivity Measurement,"
16th Edition of Standard Methods.
4. Dionex, 1C 16 Operation and Maintenance Manual, PN 30579, Dionex Corp.,
Sunnyvale, CA 94086.
5. Method detection limit (MDL) as described in "Trace Analyses for
Wastewater," J. Glaser, D. Foerst, G. McKee, S. Quave, W. Budde, Environmental
Science and Technology, Vol. 15, Number 12, p. 1426, December 1981.
6. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency Office of Solid Waste. EPA Contract No. 68-
01-7075, WA 80. July 1988.
9056 - 9 Revision 0
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION
LIMITS IN REAGENT WATER
Analyte
Fluoride
Chlorine
Nitrite-N
0-Phosphate-P
Nitrate-N
Sulfate
Retention8
time
min
1.2
3.4
4.5
9.0
11.3
21.4
Relative
retention
time
1.0
2.8
3.8
7.5
9.4
17.8
Method6
detection limit,
mg/L
0.005
0.015
0.004
0.061
0.013
0.206
Standard conditions:
Columns - As specified in 4.1.4
Detector - As specified in 4.1.4
Eluent - As specified in 5.3
Concentrations of mixed standard (mg/L):
Fluoride 3.0
Chloride 4.0
Nitrite-N 10.0
Sample loop - 100 ;uL
Pump volume - 2.30 mL/min
0-Phosphate-P 9.0
Nitrate-N 30.0
Sulfate 50.0
calculated from data obtained using an attentuator setting of 1 umho full
scale. Other settings would produce an MDL proportional to their value.
9056 - 10
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TABLE 2.
PREPARATION OF STANDARD SOLUTIONS FOR INSTRUMENT CALIBRATION
Hiqh-ranae standard (see 5.6.1)
Fluoride (F")
Chloride (CV)
Nitrite (N02~)
Phosphate (P043~)
Bromide (Br~)
Nitrate (N03")
Sulfate (S042')
Milliliters
of each
stock solution
(1.00 mL =
1.00 mg)
diluted to
1,000 ml
10
10
20
50
10
30
100
An ion
concentration
mg/L
10
10
20
50
10
30
100
Intermediate-
range standard,
mg/L
(see 5.6.2)
1.0
1.0
2.0
5.0
1.0
3.0
10.0
Low-range
standard,
mg/L (see
5.6.3)
0.2
0.2
0.4
1.0
0.2
0.6
2.0
9056 - 11
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TABLE 3.
SINGLE-OPERATOR ACCURACY AND PRECISION
Sampl e
Analyte type
Chloride
Fluoride
Nitrate-N
Nitrite-N
0-Phosphate-P
Sulfate
RW
DM
SW
WW
RW
DW
SW
WW
RW
DW
SW
WW
RW
DW
SW
WW
RW
DE
SW
WW
RW
DW
SW
WW
Spike
mg/L
0.050
10.0
1.0
7.5
0.24
9.3
0.50
1.0
0.10
31.0
0.50
4.0
0.10
19.6
0.51
0.52
0.50
45.7
0.51
4.0
1.02
98.5
10.0
12.5
Number
of
replicates
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Mean
recovery,
%
97.7
98.2
105.0
82.7
103.1
87.7
74.0
92.0
100.9
100.7
100.0
94.3
97.7
103.3
88.2
100.0
100.4
102.5
94.1
97.3
102.1
104.3
111.6
134.9
Standard
deviation,
mg/L
0.0047
0.289
0.139
0.445
0.0009
0.075
0.0038
0.011
0.0041
0.356
0.0058
0.058
0.0014
0.150
0.0053
0.018
0.019
0.386
0.020
0.04
0.066
1.475
0.709
0.466
RW = Reagent water.
DW = Drinking water.
SW = Surface water.
WW = Wastewater.
9056 - 12
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TABLE 4.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN
USED OILS BY BOMB OXIDATION AND ION CHROMATOGRAPHY
Average value, Repeatability, Reproducibility,
500
1,000
1,500
2,000
2,500
3,000
467
661
809
935
1,045
1,145
941
1,331
1,631
1,883
2,105
2,306
TABLE 5.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY
BOMB OXIDATION AND ION CHROMATOGRAPHY
Amount
Expected
Mg/g
320
480
920
1,498
1,527
3,029
3,045
Amount
found
Mg/g
567
773
1,050
1,694
1,772
3,026
2,745
Bias,
Mg/g
247
293
130
196
245
-3
-300
Percent,
bias
+77
+61
+14
+13
+16
0
-10
9056 - 13 Revision 0
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FIGURE 1
SCHEMATIC OF ION CHROMATOGRAPH
WASTE
(1) Eluent reservoir
(2) Pump
(3) Precolumn
(4) Separator column
(5) Suppressor column
(6) Detector
(7) Recorder or integrator, or both
9056 - 14
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FIGURE 2
TYPICAL ANION PROFILE
• 12
MINUTIS
I
20
9056 - 15
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METHOD 9056
AN ION CHROMATOGRAPHY METHOD
START
chroma tographic
operating
parameters
7.1.2 Prepare
cal ibration
minimum of three
concent ration
levels and a blank
1
7.1.3 Prepare
calibration curves
1
7.1.4 Verify the
cal ibration curves
each working day or
whenever the anion
eluent is changed,
and for every batch
of samples
-
/ X. 7.2.1 If a dilution
S 7.2.1 Are X. Yes is necessary the
( samples ) » dilution should be
>w aqueous? / made with eluent
N. / solution
No
7.2.2 Analyze
s tandards beginning
with the highest
concentration and
decreasing in
concentration
7.2.1 Add
concentrated eluent
<— to all sarnies and
standards to remove
water dip
1
•+
7.2.2.5 Compare
results to
cal ibration curve;
if results exceed
control 1 imi ts ,
identify problem
before proceeding
1
7.2.2.6 Inject a
spiked sample of
known cone . ;
calculate the cone..
from the cal ibration
curve ; if resul t
exceeds cont rol
limits, find problem
before proceeding
1
722.7 Begin
sample analysis
1
7 2.2.8 Analyze all
samples in same
manner
1
/I 2 .2 .10 >v
/ Doe« response N. Yes 7 2.2.10 Dilute
( for peak exceed J > sample with reagent
N. norking / water
N. range? /
No
7.3 1 Prepare
sample calibration
curves for each
anion of interest
and compute sample
concentration
concentrations from
•* instrumental
response
{ STOP )
9056 - 16
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METHOD 9070A
TOTAL RECOVERABLE OIL AND GREASE
(GRAVIMETRIC. SEPARATORY FUNNEL EXTRACTION)
1.0 SCOPE AND APPLICATION
1.1 This method measures the fluorocarbon-113 extractable matter from
surface and saline waters and industrial, domestic, and aqueous wastes. It is
applicable to the determination of relatively nonvolatile hydrocarbons, vegetable
oils, animal fats, waxes, soaps, greases, and related matter.
1.2 The method is not applicable to measurement of light hydrocarbons
that volatilize at temperatures below 70°C. Petroleum fuels, from gasoline
through No. 2 fuel oils, are completely or partially lost in the solvent removal
operation.
1.3 Some crude oils and heavy fuel oils contain a significant
percentage of residue-type materials that are not soluble in fluorocarbon-113.
Accordingly, recoveries of these materials will be low.
1.4 The method covers the range from 5 to 1,000 mg/L of extractable
material.
1.5 When determining the level of oil and grease in sludge samples,
Method 9071 is to be employed.
2.0 SUMMARY OF METHOD
2.1 The 1-liter sample is acidified to a low pH (2) and serially
extracted with fluorocarbon-113 in a separatory funnel. The solvent is
evaporated from the extract and the residue is weighed.
3.0 INTERFERENCES
3.1 Matrix interferences will likely be coextracted from the sample.
The extent of these interferences will vary from waste to waste, depending on the
nature and diversity of the waste being analyzed.
4.0 APPARATUS AND MATERIALS
4.1 Separatory funnel: 2,000-mL, with Teflon stopcock.
4.2 Vacuum pump, or other source of vacuum.
4.3 Flask: Boiling, 125-mL (Corning No. 4100 or equivalent).
4.4 Distilling head: Claisen or equivalent.
4.5 Filter paper: Whatman No. 40, 11 cm.
Replacement solvent will be specified in a forthcoming regulation.
9070A - 1 Revision 1
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5.0 REAGENTS
5.1 Reagent grade chemicals shall be used In all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Hydrochloric acid, 1:1: Mix equal volumes of concentrated HC1 and
reagent water.
5.4 Fluorocarbon-1132(l,l,2-trichloro-l,2,2-trifluoroethane): Boiling
point, 48eC.
5.5 Sodium sulfate: Anhydrous crystal.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 A representative sample should be collected in a 1-liter glass
bottle. If analysis is to be delayed for more than a few hours, the sample must
be preserved by the addition of 5 mL HC1 (Step 5.3) at the time of collection and
refrigerated at 4°C.
6.2 Collect a representative sample in a wide-mouth glass bottle that
has been rinsed with the solvent to remove any detergent film and acidify in the
sample bottle.
6.3 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.4 Because losses of grease will occur on sampling equipment, the
collection of a composite sample is impractical. Individual portions collected
at prescribed time intervals must be analyzed separately to obtain the average
concentration over an extended period.
7.0 PROCEDURE
7.1 Mark the sample bottle at the water meniscus for later determina-
tion of sample volume. If the sample was not acidified at time of collection,
add 5 mL HC1 (Step 5.3) to the sample bottle. After mixing the sample, check the
pH by touching pH-sensitive paper to the cap to ensure that the pH is 2 or lower.
Add more acid if necessary.
7.2 Pour the sample into a separatory funnel.
7.3 Tare a boiling flask (pre-dried in an oven at 103°C and stored in
a desiccator). Use gloves when handling flask to avoid adding fingerprints.
Replacement solvent will be specified in a forthcoming regulation.
9070A - 2 Revision 1
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7.4 Add 30 ml fluorocarbon-113 (Step 5.3) to the sample bottle and
rotate the bottle to rinse the sides. Transfer the solvent into the separatory
funnel. Extract by shaking vigorously for 2 min. Allow the layers to separate
and filter the solvent layer through a funnel containing solvent-moistened filter
paper.
NOTE: An emulsion that fails to dissipate can be broken by pouring about
1 g sodium sulfate (Step 5.4) into the filter paper cone and slowly
draining the emulsion through the salt. Additional 1-g portions
can be added to the cone as required.
7.5 Repeat Step 7.4 twice more, with additional portions of fresh
solvent, combining all solvent in the boiling flask.
7.6 Rinse the tip of the separatory funnel, the filter paper, and then
the funnel with a total of 10-20 ml solvent and collect the rinsings in the
flask.
7.7 Connect the boiling flask to the distilling head and evaporate the
solvent by immersing the lower half of the flask in water at 70°C. Collect the
solvent for reuse. A solvent blank should accompany each set of samples.
7.8 When the temperature in the distilling head reaches 50°C or the
flask appears dry, remove the distilling head. To remove solvent vapor, sweep
out the flask for 15 sec with air by inserting a glass tube that is connected to
a vacuum source. Immediately remove the flask from heat source and wipe the
outside to remove excess moisture and fingerprints.
7.9 Cool the boiling flask in a desiccator for 30 min. and weigh.
7.10 Calculation:
mg/L total oil and grease
R - B
where:
R = residue, gross weight of extraction flask minus
the tare weight (mg);
B = blank determination, residue of equivalent
volume of extraction solvent, mg; and
V = volume of sample in liters, determined by
refilling sample bottle to calibration line and
correcting for acid addition, if necessary.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Comprehensive quality control procedures are specified for each
target compound in the referring analytical method.
9070A - 3
Revision 1
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8.3 The matrix duplicate and matrix spike samples are brought through
the whole sample preparation and analytical process.
8.4 The use of corn oil is recommended as a reference sample solution.
9.0 METHOD PERFORMANCE
9.1 Two oil and grease methods (Methods 9070 and 9071) were tested on
sewage by a single laboratory. This method determined the oil and grease level
in the sewage to be 12.6 mg/L. When 1-liter portions of the sewage were dosed
with 14.0 mg of a mixture of No. 2 fuel oil and Wesson oil, the recovery was 93%,
with a standard deviation of + 0.9 mg/L.
10.0 REFERENCES
1. Blum, K.A., and M.J. Taras, "Determination of Emulsifying Oil in Industrial
Wastewater," JWPCF Research Suppl., 40, R404 (1968).
2. Standard Methods for the Examination of Water and Wastewater, 14th ed.,
p. 515.
9070A - 4 Revision 1
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METHOD 9070A
TOTAL RECOVERABLE OIL AND GREASE (GRAVIMETRIC, SEPARATORY FUNNEL EXTRACTION)
7.2 Pour
sample into
separatory
funnal
7.3 Tare
boiling flask
7.4 Add
f luorocarbon-
113; extract;
filtar
solvent layer
7.S Repeat
twice adding
froth solvent
7 . 5 Combine
solvent in —
boiling flask
7.7 Cvaporate
solvent;
collect for
reuse
7 . 8 Remove
solvent vapor
7.9 Cool
flask and
weigh
7.10
Calculate
total amount
of oil and
grease
Stop
9070A - 5
Revision 1
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METHOD 9075
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED
PETROLEUM PRODUCTS BY X-RAY FLUORESCENCE SPECTROMETRY (XRR
1.0 SCOPE AND APPLICATION
1.1 This test method covers the determination of total chlorine in new
and used oils, fuels, and related materials, including crankcase, hydraulic,
diesel, lubricating and fuel oils, and kerosene. The chlorine content of
petroleum products is often required prior to their use as a fuel.
1.2 The applicable range of this method is from 200 ng/g to percent
levels.
2.0 SUMMARY OF METHOD
2.1 A well-mixed sample, contained in a disposable plastic sample cup,
is loaded into an X-ray fluorescence (XRF) spectrometer. The intensities of the
chlorine K alpha and sulfur K alpha lines are measured, as are the intensities
of appropriate background lines. After background correction, the net inten-
sities are used with a calibration equation to determine the chlorine content.
The sulfur intensity is used to correct for absorption by sulfur.
3.0 INTERFERENCES
3.1 Possible interferents include metals, water, and sediment in the oil.
Results of spike recovery measurements and measurements on diluted samples can
be used to check for interferences.
Each sample, or one sample from a group of closely related samples, should
be spiked to confirm that matrix effects are not significant. Dilution of
samples that may contain water or sediment can product incorrect results, so
dilution should be undertaken with caution and checked by spiking. Sulfur
interferes with the chlorine determination, but a correction is made.
Spike recovery measurements of used crankcase oil showed that diluting
samples five to one allowed accurate measurements on approximately 80% of the
samples. The other 20% of the samples were not accurately analyzed by XRF.
3.2 Water in samples absorbs X-rays due to chlorine. For this inter-
ference, using as short an X-ray counting time as possible is beneficial. This
appears to be related to stratification of samples into aqueous and nonaqueous
layers while in the analyzer.
Although a correction for water may be possible, none is currently
available. In general, the presence of any free water as a separate phase or a
water content greater than 25% will reduce the chlorine signal by 50 to 90%.
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4.0 APPARATUS AND MATERIALS
4.1 XRF spectrometer, either energy dispersive or wavelength dispersive.
The instrument must be able to accurately resolve and measure the intensity of
the chlorine and sulfur lines with acceptable precision.
4.2 Disposable sample cups with suitable plastic film such as Mylar*.
5.0 REAGENTS
5.1 Purity of reagents. Reagent-grade chemicals shall be used in all
tests. Unless otherwise indicated, it is intended that all reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Mineral oil, mineral spirits or paraffin oil, sulfur and chlorine
free, for preparing standards and dilutions.
5.3 1-Chlorodecane (Aldrich Chemical Co.), 20.1% chlorine, or similar
chlorine compound.
5.4 Di-n-butyl sulfide (Aldrich Chemical Co.), 21.9% sulfur by weight.
5.5 Quality control standards such as the standard reference materials
NBS 1620, 1621, 1622, 1623, and 1624, sulfur in oil standards, and NBS 1818,
chlorine in oil standards.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 The collected sample should be kept headspace free prior to prepara-
tion and analysis to minimize volatilization losses of organic halogens. Because
waste oils may contain toxic and/or carcinogenic substances, appropriate field
and laboratory safety procedures should be followed.
6.3 Laboratory sampling of the sample should be performed on a well-mixed
sample of oil. The mixing should be kept to a minimum and carried out as nearly
headspace free as possible to minimize volatilization losses of organic halogens.
6.4 Free water, as a separate phase, should be removed and cannot be
analyzed by this method.
7.0 PROCEDURE
7.1 Calibration and standardization.
7.1.1 Prepare primary calibration standards by diluting the
chlorodecane and n-butyl sulfide with mineral spirits or similar material.
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7.1.2 Prepare working calibration standards that contain sulfur,
chlorine, or both according to the following table:
Cl:
S:
1.
2.
3.
4.
500, 1,000, 2,000, 4,000, and 6,000 M9/9
0.5, 1.0, and 1.5% sulfur
0.5% S, 1,000 M9/9 Cl
0.5% S, 4,000 M9/9 Cl
1.0% S, 500 M9/9 Cl
1.0% S, 2,000 M9/9 Cl
5. 1.0% S, 6,000 M9/9 Cl
6. 1.5% S, 1,000 M9/9 Cl
7. 1.5% S, 4,000 M9/9 Cl
8. 1.5% S, 6,000 M9/9 Cl
Once the correction factor for sulfur interference with chlorine is
determined, fewer standards may be required.
7.1.3 Measure the intensity of the chlorine K alpha line and the
sulfur K alpha line as well as the intensity of a suitably chosen
background. Based on counting statistics, the relative standard deviation
of each peak measurement should be 1% or less.
7.1.4 Determine the net chlorine and sulfur intensities by
correcting each peak for background. Do this for all of the calibration
standards as well as for a paraffin blank.
7.1.5 Obtain a linear calibration curve for sulfur by performing
a least squares fit of the net sulfur intensity to the standard concentra-
tions, including the blank. The chlorine content of a standard should
have little effect on the net sulfur intensity.
7.1.6 The calibration equation for chlorine must include a
correction term for the sulfur concentration. A suitable equation
follows:
where:
Cl = (ml + b) (1 + k*S)
(1)
I = net chlorine intensity
m, b, k* = adjustable parameters.
f
Using a least squares procedure, the above equation or a suitable
substitute should be fitted to the data. Many XRF instruments are
equipped with suitable computer programs to perform this fit. In any
case, the resulting equation should be shown to be accurate by analysis of
suitable standard materials.
7.2 Analysis.
7.2.1 Prepare a calibration curve as described in Step 7.1. By
periodically measuring a very stable sample containing both sulfur and
chlorine, it may be possible to use the calibration equations for more
than 1 day. During each day, the suitability of the calibration curve
should be checked by analyzing standards.
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7.2.2 Determine the net chlorine and sulfur intensities for a
sample in the same manner as was done for the standards.
7.2.3 Determine the chlorine and sulfur concentrations of the
samples from the calibration equations. If the sample concentration for
either element is beyond the range of the standards, the sample should be
diluted with mineral oil and reanalyzed.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 One sample in ten should be analyzed in triplicate and the relative
standard deviation reported. For each triplicate, a separate preparation should
be made, starting from the original sample.
8.3 Each sample, or one sample in ten from a group of similar samples,
should be spiked with the elements of interest by adding a known amount of
chlorine or sulfur to the sample. The spiked amount should be between 50% and
200% of the sample concentration, but the minimum addition should be at least
five times the limit of detection. The percent recovery should be reported and
should be between 80% and 120%. Any sample suspected of containing >25% water
should also be spiked with organic chlorine.
8.4 Quality control standard check samples should be analyzed every day
and should agree within 10% of the expected value of the standard.
9.0 METHOD PERFORMANCE
These data are based on 47 data points obtained by seven laboratories who
each analyzed four used crankcase oils and three fuel oil blends with crankcase
in duplicate. A data point represents one duplicate analysis of a sample. Two
data points were determined to be outliers and are not included in these results.
9.1 Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method, the following values only in 1 case in 20 (see
Table 1):
Repeatability =5.72
*where x is the average of two results in
Reoroducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
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Reproducibility = 9.83
*where x is the average value of two results in /xg/g.
9.2 Bias. The bias of this test method varies with concentration, as
shown in Table 2:
Bias = Amount found - Amount expected.
10.0 REFERENCE
1. Gaskill, A.; Estes, E.D.; Hardison, D.L.; and Myers, I.E. Validation of
Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract
No. 68-01-7075, WA 80. July 1988.
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TABLE 1. REPEATABILITY AND REPRODUCIBILITY
FOR CHLORINE IN USED OILS BY
X-RAY FLUORESCENCE SPECTROMETRY
Average value, Repeatability, Reproducibility,
M9/9 M9/9 M9/9
500 128 220
1,000 181 311
1,500 222 381
2,000 256 440
2,500 286 492
3,000 313 538
TABLE 2. RECOVERY AND BIAS DATA FOR CHLORINE IN
USED OILS BY X-RAY FLUORESCENCE SPECTROMETRY
Amount
expected,
M9/9
320
480
920
1,498
1,527
3,029
3,045
Amount
found,
M9/9
278
461
879
1,414
1,299
2,806
2,811
Bias,
M9/9
-42
-19
-41
-84
-228
-223
-234
Percent
bias
-13
-4
-4
-6
-15
-7
-8
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METHOD 9075
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED
PETROLEUM PRODUCTS BY X-RAY FLUORESCENCE SPECTROMETRY (XRF)
START
7.1.1 - 7.1.2
Prepare calibration
3 tandards
71.3 Measure
intensity of
3 tandards and
background
7,1.4 Determine net
intensity for
standards and a
paraffin blank
7.1.5 - 7.1.6
Construct
calibration curves
for sulfur and
chl o r me
7.2.1 Check
calibration curves
perlodica11y
throughout the day
7.2.2 Determine net
chlorine and sulfur
intensi ties
7.2.3 Determine
chlorine and sulfur
concentrations from
calibration curves
7.2.3
Is sample
concentration
beyond range of
standards?
7.2.3 Dilute sample
with mineral oil
9075 - 7
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METHOD 9076
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS BY OXIDATIVE COMBUSTION AND MICROCOULOMETRY
1.0 SCOPE AND APPLICATION
1.1 This test method covers the determination of total chlorine in new
and used oils, fuels and related materials, including crankcase, hydraulic,
diesel, lubricating and fuel oils, and kerosene by oxidative combustion and
microcoulometry. The chlorine content of petroleum products is often required
prior to their use as a fuel.
1.2 The applicable range of this method is from 10 to 10,000 ng/g
chlorine.
2.0 SUMMARY OF METHOD
2.1 The sample is placed in a quartz boat at the inlet of a high-
temperature quartz combustion tube. An inert carrier gas such as argon, carbon
dioxide, or nitrogen sweeps across^the inlet while oxygen flows into the center
of the combustion tube. The boat and sample are advanced into a vaporization
zone of approximately 300°C to volatilize the light ends. Then the boat is
advanced to the center of the combustion tube, which is at 1,000°C. The oxygen
is diverted to pass directly over the sample to oxidize any remaining refractory
material. All during this complete combustion cycle, the chlorine is converted
to chloride and oxychlorides, which then flow into an attached titration cell
where they quantitatively react with silver ions. The silver ions thus consumed
are coulometrically replaced. The total current required to replace the silver
ions is a measure of the chlorine present in the injected samples.
2.2 The reaction occurring in the titration cell as chloride enters is:
Cl" + Ag+ > AgCl (1)
The silver ion consumed in the above reaction is generated coulometrically
thus:
Ag° > Ag+ + e" y (2)
2.3 These microequivalents of silver are equal to the number of micro-
equivalents of titratable sample ion entering the titration cell.
3.0 INTERFERENCES
3.1 Other titratable halides will also give a positive response. These
titratable halides include HBr and HI (HOBr + HOI do not precipitate silver).
Because these oxyhalides do not react in the titration cell, approximately 50%
microequivalent response is detected from bromine and iodine.
3.2 Fluorine as fluoride does not precipitate silver, so it is not an
interferant nor is it detected.
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3.3 This test method is applicable in the presence of total sulfur
concentrations of up to 10,000 times the chlorine level.
4.0 APPARATUS AND MATERIALS1
4.1 Combustion furnace. The sample should be oxidized in an electric
furnace capable of maintaining a temperature of 1,000'C to oxidize the organic
matrix.
4.2 Combustion tube, fabricated from quartz and constructed so that a
sample, which is vaporized completely in the inlet section, is swept into the
oxidation zone by an inert gas where it mixes with oxygen and is burned. The
inlet end of the tube connects to a boat insertion device where the sample can
be placed on a quartz boat by syringe, micropipet, or by being weighed
externally. Two gas ports are provided, one for an inert gas to flow across the
boat and one for oxygen to enter the combustion tube.
4.3 Microcoulometer, Stroehlein Coulomat 702 CL or equivalent, having
variable gain and bias control, and capable of measuring the potential of the
sensing-reference electrode pair, and comparing this potential with a bias
potential, and applying the amplified difference to the working-auxiliary
electrode pair so as to generate a titrant. The microcoulometer output signal
shall be proportional to the generating current. The microcoulometer may have
a digital meter and circuitry to convert this output signal directly to nanograms
or micrograms of chlorine or micrograms per gram chlorine.
4.4 Titration cell. Two different configurations have been applied to
coulometrically titrate chlorine for this method.
4.4.1 Type I uses a sensor-reference pair of electrodes to detect
changes in silver ion concentration and a generator anode-cathode pair of
electrodes to maintain constant silver ion concentration and an inlet for
a gaseous sample from the pyrolysis tube. The sensor, reference, and
anode electrodes are silver electrodes. The cathode electrode is a
platinum wire. The reference electrode resides in a saturated silver
acetate half-cell. The electrolyte contains 70% acetic acid in water.
4.4.2 Type II uses a sensor-reference pair of electrodes to
detect changes in silver ion concentration and a generator anode-cathode
pair of electrodes to maintain constant silver ion concentration, an inlet
for a gaseous sample that passes through a 95% sulfuric acid dehydrating
tube from the pyrolysis tube, and a sealed two-piece titration cell with
an exhaust tube to vent fumes to an external exhaust. All electrodes can
be removed and replaced independently without reconstructing the cell
assembly. The anode electrode is constructed of silver. The cathode
electrode is constructed of platinum. The anode is separated from the
cathode by a 10% KNO, agar bridge, and continuity is maintained through an
aqueous 10% KN03 salt bridge. The sensor electrode is constructed of
1Three commercial analyzers fulfill the requirements for apparatus Steps
4.1 through 4.4 and have been found satisfactory for this method. They are
the two Dohrmann Models DX-20B and MCTS-20 and Mitsubishi Model TSX-10
available from Cosa Instrument.
9076 - 2 Revision 0
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silver. The reference electrode is a silver/silver chloride ground glass
sleeve, double-junction electrode with aqueous 1M KN03 in the outer chamber
and aqueous 1M KC1 in the inner chamber.
4.5 Sampling syringe, a microliter syringe of 10 /iL capacity capable of
accurately delivering 2 to 5 /LiL of a viscous sample into the sample boat.
4.6 Micropipet, a positive displacement micropipet capable of accurately
delivering 2 to 5 nl of a viscous sample into the sample boat.
4.7 Analytical balance. When used to weigh a sample of 2 to 5 mg onto
the boat, the balance shall be accurate to + 0.01 mg. When used to determine the
density of the sample, typically 8 g per 10 ml, the balance shall be accurate to
± 0.1 g.
4.8 Class A volumetric flasks: 100 ml.
5.0 REAGENTS
5.1 Purity of Reagents. Reagent-grade chemicals shall be used in all
tests. Unless otherwise indicated, it is intended that all reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Acetic acid, CH3C02H. Glacial.
5.4 Isooctane, (CH3)2CHCH2C(CH3)3 (2,2,4-Trimethylpentane).
5.5 Chlorobenzene, C6H5C1.
5.6 Chlorine, standard stock solution - 10,000 ng Cl//iL, weigh
accurately 3.174 g of chlorobenzene into 100-mL Class A volumetric flask. Dilute
to the mark with isooctane.
5.7 Chlorine, standard solution. 1,000 ng Cl/^L, pipet 10.0 mL of
chlorine stock solution (Step 5.6) into a 100-mL volumetric flask and dilute to
volume with isooctane.
5.8 Argon, helium, nitrogen, or carbon dioxide, high-purity grade (HP)
used as the carrier gas. High-purity grade gas has a minimum purity of 99.995%.
5.9 Oxygen, high-purity grade (HP), used as the reactant gas.
5.10 Gas regulators. Two-stage regulator must be used on the reactant and
carrier gas.
5.11 Cell Type 1.
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5.11.1 Cell electrolyte solution. 70% acetic acid: combine 300
ml reagent water with 700 mL acetic acid (Step 5.3) and mix well.
5.11.2 Silver acetate, CH3C02Ag. Powder purified for saturated
reference electrode.
5.12 Cell Type 2.
5.12.1 Sodium acetate, CH3C02Na.
5.12.2 Potassium nitrate, KN03.
5.12.3 Potassium chloride, KC1.
5.12.4 Sulfuric acid (concentrated), H2S04.
5.12.5 Agar, (jelly strength 450 to 600 g/cm2).
5.12.6 Cell electrolyte solution - 85% acetic acid: combine 150
ml reagent water with 1.35 g sodium acetate (Step 5.12.1) and mix well;
add 850 ml acetic acid (Step 5.3) and mix well.
5.12.7 Dehydrating solution - Combine 95 mL sulfuric acid (Step
5.12.4) with 5 mL reagent water and mix well.
CAUTION: This is an exothermic reaction and may proceed with bumping unless
controlled by the addition of sulfuric acid. Slowly add sulfuric
acid to reagent water. Do not add water to sulfuric acid.
5.12.8 Potassium nitrate (10%), KN03. Add 10 g potassium nitrate
(Step 5.12.2) to 100 mL reagent water and mix well.
5.12.9 Potassium nitrate (1M), KN03. Add 10.11 g potassium
nitrate (Step 5.12.2) to 100 mL reagent water and mix well.
5.12.10 Potassium chloride (1M), KC1. Add 7.46 g potassium
chloride (Step 5.12.3) to 100 mL reagent water and mix well.
5.12.11 Agar bridge solution - Mix 0.7 g agar (Step 5.12.5), 2.5g
potassium nitrate (Step 5.12.2), and 25 mL reagent water and heat to
boiling.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Because the collected sample will be analyzed for total halogens, it
should be kept headspace free and refrigerated prior to preparation and analysis
to minimize volatilization losses of organic halogens. Because waste oils may
contain toxic and/or carcinogenic substances, appropriate field and laboratory
safety procedures should be followed.
9076 - 4 Revision 0
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6.3 Laboratory subsampling of the sample should be performed on a well-
mixed sample of oil.
7.0 PROCEDURES
7.1 Preparation of apparatus.
7.1.1 Set up the analyzer as per the equipment manufacturer's
instructions.
7.1.2 Typical operating conditions: Type 1.
Furnace temperature 1,OOO'C
Carrier gas flow 43 cm /min
Oxygen gas flow 160 cm /min
Coulometer
Bias 250 mV
Gain 25%
7.1.3 Typical operating conditions: Type 2.
Furnace temperature H-l 850°C
H-2 1,QOO°C
Carrier gas flow 250 cm /min
Oxygen gas flow 250 cm /min
Coulometer
End point potential (bias) 300 mV
Gain G-l 1.5 coulombs/A mV
G-2 3.0 coulombs/A mV
G-3 3.0 coulombs/A mV
ES-1 (range 1) 25 mV
ES-2 (range 2) 30 mV
NOTE: Other conditions may be appropriate. Refer to the instrumentation manual.
7.2 Sample introduction.
7.2.1 Carefully fill a 10-jiL syringe with 2 to 5 juL of sample
depending on the expected concentration of total chlorine. Inject the
sample through the septum onto the cool boat, being certain to touch the
boat with the needle tip to displace the last droplet.
7.2.2 For viscous samples that cannot be drawn into the syringe
barrel, a positive displacement micropipet may be used. Here, the 2-5 juL
of sample is placed on the boat from the micropipet through the opened
hatch port. The same technique as with the syringe is used to displace
the last droplet into the boat. A tuft of quartz wool in the boat can aid
in completely transferring the sample from the micropipet into the boat.
NOTE: Dilution of samples to reduce viscosity is not recommended due to
uncertainty about the solubility of the sample and its chlorinated
constituents. If a positive displacement micropipet is not available,
dilution may be attempted to enable injection of viscous samples.
9076 - 5 Revision 0
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7.2.3 Alternatively, the sample boat may be removed from the
instrument and tared on an analytical balance. A sample of 2-5 mg is
accurately weighed directly into the boat and the boat and sample returned
to the inlet of the instrument.
2-5 /iL = 2-5 mg
NOTE: Sample dilution may be required to ensure that the titration system is not
overloaded with chlorine. This will be somewhat system dependent and
should be determined before analysis is attempted. For example, the MCTS-
20 can titrate up to 10,000 ng chlorine in a single injection or weighed
sample, while the DX-20B has an upper limit of 50,000 ng chlorine. For 2
to 5 juL sample sizes, these correspond to nominal concentrations in the
sample of 800 to 2,000 jzg/g and 4,000 to 10,000 M9/g» respectively. If
the system is overloaded, especially with inorganic chloride, residual
chloride may persist in the system and affect results of subsequent
samples. In general, the analyst should ensure that the baseline returns
to normal before running the next sample. To speed baseline recovery, the
electrolyte can be drained from the cell and replaced with fresh
electrolyte.
NOTE: To determine total chlorine, do not extract the sample either with reagent
water or with an organic solvent such as toluene or isooctane. This may
lower the inorganic chlorine content as well as result in losses of
volatile solvents.
7.2.4 Follow the manufacturer's recommended procedure for moving
the sample and boat into the combustion tube.
7.3 Calibration and standardization.
7.3.1 System recovery - The fraction of chlorine in a standard
that is titrated should be verified every 4 hours by analyzing the
standard solution (Step 5.7). System recovery is typically 85% or better.
The pyrolysis tube should be replaced whenever system recovery drops below
75%.
NOTE: The 1,000 /xg/g system recovery sample is suitable for all systems except
the MCTS-20 for which a 100 jug/g sample should be used.
7.3.2 Repeat the measurement of this standard at least three
times.
7.3.3 System blank - The blank should be checked daily with
isooctane. It is typically less than 1 /xg/g chlorine. The system blank
should be subtracted from both samples and standards.
7.4 Calculations.
7.4.1 For systems that read directly in mass units of chloride,
the following equations apply:
Chlorine, /*g/g (wt/wt) = Disp1ays - B (3)
(V.) (D8) (RF)
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or
Chlorine, M9/9 (wt/wt) = Disp1ays - B (4)
(M) (RF)
where:
Display = Integrated value in nanograms (when the integrated values are
displayed in micrograms, they must be multiplied by 10 )
DisplayB = blank measurement Displays = sample measurement
V = Volume of sample injected in microliters
VB = blank volume Vs = sample volume
D = Density of sample, grams per cubic centimeters
DB = blank density Ds = sample density
RF = Recovery factor = ratio of chlorine = Found - Blank
determined in standard minus the system Known
blank, divided by known standard content
B = System blank, jug/g chlorine = Display.
(VB) (DB)
M = Mass of sample, mg
7.4.2 Other systems internally compensate for recovery factor,
volume, density, or mass and blank, and thus read out directly in parts
per million chlorine units. Refer to instrumentation manual.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Each sample should be analyzed twice. If the results do not agree
to within 10%, expressed as the relative percent difference of the results,
repeat the analysis.
8.3 Analyze matrix spike and matrix spike duplicates - spike samples with
a chlorinated organic at a level of total chlorine commensurate with the levels
being determined. The spike recovery should be reported and should be between
80 and 120% of the expected value. Any sample suspected of containing >25% water
should also be spiked with organic chlorine.
9.0 METHOD PERFORMANCE
These data are based on 66 data points obtained by 10 laboratories who each
analyzed four used crankcase oils and three fuel oil blends with crankcase in
duplicate. A data point represents one duplicate analysis of a sample. One
laboratory and four additional data points were determined to be outliers and are
not included in these results.
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9.1 Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method the following values only in 1 case in 20 (see Table
Repeatability = 0.137
*where x is the average of two results in M9/g«
Reproducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Repioducibility = 0.455
-------
TABLE 1.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN
USED OILS BY MICROCOULOMETRIC TITRATION
Average value Repeatability, Reproducibility,
M9/9 M9/9 M9/9
500 69 228
1,000 137 455
1,500 206 683
2,000 274 910
2,500 343 1,138
3,000 411 1,365
TABLE 2.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS
BY MICROCOULOMETRIC TITRATION
Amount
expected,
M9/9
320
480
920
1,498
1,527
3,029
3,045
Amount
found
M9/9
312
443
841
1,483
1,446
3,016
2,916
Bias,
M9/9
-8
-37
-79
-15
-81
-13
-129
Percent
bias
-3
-8
-9
-1
-5
0
-4
9076 - 9 Revision 0
November 1990
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METHOD 9076
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS BY OXIDATIVE COMBUSTION AND MICROCOULOMETRY
7.2.2 Inject
sample into
cool boat
Kith
micropipet
7.2.4 Move
sample and
boat into
combus tion
tube
7.3.1 Verify
ays tern .
recovery
every 4 hours
7.2.1 Inject
sample into
cool boat
with syringe
7.32 Repeat
s tandard
measurement
at least
three times
73.3 Check
system blank
daily with
is ooctane
7.4 Calculate
chlo rine
concent ration
STOP
9076 - 10
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METHOD 9077
TEST METHODS FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS (FIELD TEST KIT METHODS)
1.0 SCOPE AND APPLICATION
1.1 The method may be used to determine if a new or used petroleum
product meets or exceeds requirements for total halogen measured as chloride.
An analysis of the chlorine content of petroleum products is often required prior
to their use as a fuel. The method is specifically designed for used oils
permitting onsite testing at remote locations by nontechnical personnel to avoid
the delays for laboratory testing.
1.2 In these field test methods, the entire analytical sequence,
including sampling, sample pretreatment, chemical reactions, extraction, and
quantification, are combined in a single kit using predispensed and encapsulated
reagents. The overall objective is to provide a simple, easy to use procedure,
permitting nontechnical personnel to perform a test with analytical accuracy
outside of a laboratory environment in under 10 minutes. One of the kits is
preset at 1,000 ^9/9 total chlorine to meet regulatory requirements for used
oils. The other kits provide quantitative results over a range of 750 to 7,000
/ng/g and 300 to 4,000 jug/g.
2.0 SUMMARY OF METHOD
2.1 The oil sample (around 0.4 g by volume) is dispersed in a solvent
and reacted with a mixture of metallic sodium catalyzed with naphthalene and
diglyme at ambient temperature. This process converts all organic halogens to
their respective sodium halides. All halides in the treated mixture, including
those present prior to the reaction, are then extracted into an aqueous buffer,
which is then titrated with mercuric nitrate using diphenyl carbazone as the
indicator. The end point of the titration is the formation of the blue-violet
mercury diphenylcarbazone complex. Bromide and iodide are titrated and reported
as chloride.
2.2 Reagent quantities are preset in the fixed end point kit (Method
A) so that the color of the solution at the end of the titration indicates
whether the sample is above 1,000 p,g/g chlorine (yellow) or below 1,000 ng/g
chlorine (blue).
2.3 The first quantitative kit (Method B) involves a reverse titration
of a fixed volume of mercuric nitrate with the extracted sample such that the end
point is denoted by a change from blue to yellow in the titration vessel over the
range of the kit (750 to 7,000 p.g/g). The final calculation is based on the
assumption that the oil has a specific gravity of 0.9.
2.4 The second quantitative kit (Method C) involves a titration of the
extracted sample with mercuric nitrate by means of a 1-mL microburette such that
the end point is denoted by a change from pale yellow to red-violet over the
range of the kit (300 to 4,000 ng/g). The concentration of chlorine in the
original oil is then read from a scale on the microburette.
9077 - 1 Revision 0
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NOTE: Warning--All reagents are encapsulated or contained within
ampoules. Strict adherence to the operational procedures included
with the kits as well as accepted safety procedures (safety glasses
and gloves) should be observed.
NOTE: Warning—When crushing the glass ampoules, press firmly in the
center of the ampoule once. Never attempt to recrush broken glass
because the glass may come through the plastic and cut fingers.
NOTE: Warning—In case of accidental breakage onto skin or clothing, wash
with large amounts of water. All the ampoules are poisonous and
should not be taken internally.
NOTE: Warning—The gray ampoules contain metallic sodium. Metallic
sodium is a flammable water-reactive solid.
NOTE: Warning—Do not ship kits on passenger aircraft. Dispose of used
kits properly.
NOTE: Caution—When the sodium ampoule in either kit is crushed, oils
that contain more than 25% water will cause the sample to turn
clear to light gray. Under these circumstances, the results may
be biased excessively low and should be disregarded.
3.0 INTERFERENCES
3.1 Free water, as a second phase, should be removed. However, this
second phase can be analyzed separately for chloride content if desired.
9077 - 2 Revision 0
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METHOD A
FIXED END POINT TEST KIT METHOD
4.0A APPARATUS AND MATERIALS
4.1A The CLOR-D-TECT 10001 is a complete self-contained kit. It
includes: a sampling tube to withdraw a fixed sample volume for analysis; a
polyethylene test tube #1 into which the sample is introduced for dilution and
reaction with metallic sodium; and a polyethylene tube #2 containing a buffered
aqueous extractant, the mercuric nitrate titrant, and diphenyl carbazone
indicator. Included are instructions to conduct the test and a color chart to
aid in determining the end point.
5.0A REAGENTS
5.1A Purity of reagents. Reagent-grade chemicals shall be used in all
tests. Unless otherwise indicated, it is intended that all reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the determina-
tion.
5.2A All necessary reagents are contained within the kit.
5.3A The kit should be examined upon opening to see that all of the
components are present and that all the ampoules (4) are in place and not
leaking. The liquid in Tube #2 (yellow cap) should be approximately 1/2 in.
above the 5-mL line and the tube should not be leaking. The ampoules are not
supposed to be completely full.
6.0A SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1A All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2A Because the collected sample will be analyzed for total halogens,
it should be kept headspace free and refrigerated prior to preparation and
analysis to minimize volatilization losses of organic halogens. Because waste
oils may contain toxic and/or carcinogenic substances, appropriate field and
laboratory safety procedures should be followed.
7.0A PROCEDURE
7.1A Preparation. Open analysis carton, remove contents, mount plastic
test tubes in the provided holder. Remove syringe and glass sampling capillary
from foil pouch.
Available from Dexsil Corporation, One Hamden Park Drive, Hamden, CT 06517.
9077 - 3 Revision 0
November 1990
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NOTE: Perform the test in a warm, dry area with adequate light. In cold
weather, a truck cab is sufficient. If a warm area is not
available, Step 7.3 should be performed while warming Tube #1 in
palm of hand.
7.2A Sample introduction. Remove white cap from Tube #1. Using the
plastic syringe, slowly draw the oil up the capillary tube until it reaches the
flexible adapter tube. Wipe excess oil from the tube with the provided tissue,
keeping capillary vertical. Position capillary tube into Tube #1, and detach
adapter tubing, allowing capillary to drop to the bottom of the tube. Replace
white cap on tube. Crush the capillary by squeezing the test tube several times,
being careful not to break the glass reagent ampoules.
7.3A Reaction. Break the lower (colorless) capsule containing the clear
diluent solvent by squeezing the sides of the test tube. Mix thoroughly by
shaking the tube vigorously for 30 seconds. Crush the upper grey ampoule
containing metallic sodium, again by squeezing the sides of the test tube. Shake
vigorously for 20 seconds. Allow reaction to proceed for 60 seconds, shaking
intermittently several times while timing with a watch.
NOTE: Caution—Always crush the clear ampoule in each tube first.
Otherwise, stop the test and start over using another complete kit.
False (low) results may occur and allow a contaminated sample to
pass without detection if clear ampoule is not crushed first.
7.4A Extraction. Remove caps from both tubes. Pour the clear buffered
extraction solution from Tube #2 into Tube #1. Replace the white cap on Tube #1,
and shake vigorously for 10 seconds. Vent tube by partially unscrewing the
dispenser cap. Close cap securely, and shake for an additional 10 seconds. Vent
again, tighten cap, and stand tube upside down on white cap. Allow phases to
separate for 2 minutes.
7.5A Analysis. Put filtration funnel into Tube #2. Position Tube #1
over funnel and open nozzle on dispenser cap. Squeeze the sides of Tube #1 to
dispense the clear aqueous lower phase through the filter into Tube #2 to the 5-
ml_ line on Tube #2. Remove the filter funnel. Replace the yellow cap on Tube
#2 and close the nozzle on the dispenser cap. Break the colorless lower capsule
containing mercuric nitrate solution by squeezing the sides of the tube, and
shake for 10 seconds. Then break the upper colored ampoule containing the
diphenylcarbazone indicator, and shake for 10 seconds. Observe color
immediately.
7.6A Interpretation of results
7.6.1A Because all reagent levels are preset, calculations are not
required. A blue solution in Tube #2 indicates a chlorine content in the
original oil of less than 1,000 /xg/g, and a yellow color indicates that
the chlorine concentration is greater than 1,000 ng/g. Refer to the color
chart enclosed with the kit in interpreting the titration end point.
7.6.2A Report the results as < or > 1,000 ng/g chlorine in the oil
sample.
9077 - 4 Revision 0
November 1990
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8.0A QUALITY CONTROL
8.1A Refer to Chapter One for specific quality control procedures.
8.2A Each sample should be tested two times. If the results do not
agree, then a third test must be performed. Report the results of the two that
agree.
9.0A METHOD PERFORMANCE
9.1A No formal statement is made about either the precision or bias of
the overall test kit method for determining chlorine in used oil because the
result merely states whether there is conformance to the criteria for success
specified in the procedure, (i.e.. a blue or yellow color in the final solution).
In a collaborative study, eight laboratories analyzed four used crankcase oils
and three fuel oil blends with crankcase in duplicate using the test kit. Of the
resulting 56 data points, 3 resulted in incorrect classification of the oil's
chlorine content (Table 1). A data point represents one duplicate analysis of
a sample. There were no disagreements within a laboratory on any duplicate
determinations.
9077 - 5 Revision 0
November 1990
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TABLE 1.
PRECISION AND BIAS INFORMATION FOR METHOD A-
FIXED END POINT TEST KIT METHOD
Expected
concentration,
Percent agreement*3
Expected results, Percent
correct8 Within Between
320
480
920
1,498
1,527
3,029
3,045
< 1,000
< 1,000
< 1,000
> 1,000
> 1,000
> 1,000
> 1,000
100
100
100
87
75
100
100
100
100
100
100
100
100
100
100
100
100
87
75
100
100
aPercent correct—percent correctly identified as above or below
1,000
bPercent agreement --percent agreement within or between laboratories,
9077 - 6
Revision 0
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START
METHOD 9077 A
FIXED END POINT TEST KIT METHOD
7 1A Open test kit
7 . 2A Draw oil into
capillary tuba;
remove excess oil;
drop capillary tube
into Tube tl and
cap Tube /I: crush
capillary tube
7.3A Break
colorless capsule;
mix; crush grey
capsule; mix; allow
reaction to proceed
for 60 sec.
7.1A Pour Tube t2
solution into Tube
t\; mix; vent;
allow phases to
separate
7 5A Filter aqueous
lower phase in Tube
fi into Tube #2;
remove filter
funnel; break
colorless capsule;
mix; break upper
colored capsule;
mix; observe color
7 6.1 Chlorine
content is > 1000
ug/g
7.6.1 Chlorine
content is < 1000
ug/g
7.6.2 Report
results
STOP
9077 - 7
Revision 0
November 1990
-------
METHOD B
REVERSE TITRATION QUANTITATIVE END POINT TEST KIT METHOD
4.OB APPARATUS AND MATERIALS
4.IB QuantiClor2 kit components (see Figure 1).
4.1.IB Plastic reaction bottle: 1 oz, with flip-top dropper cap
and a crushable glass ampoule containing sodium.
4.1.2B Plastic buffer bottle: contains 9.5 ml of aqueous buffer
solution.
4.1.3B Titration vial: contains buffer bottle and indicator-
impregnated paper.
4.1.4B Glass vial: contains 2.0 ml of solvents.
4.1.5B Micropipet and plunger, 0.25 ml_.
4.1.6B Activated carbon filtering column.
4.1.7B Titret and valve assembly.
4.2B The reagents needed for the test are packaged in disposable
containers.
4.3B The procedure utilizes a Titret. Titrets* are hand-held,
disposable cells for titrimetric analysis. A Titret is an evacuated glass
ampoule (13 mm diameter) that contains an exact amount of a standardized liquid
titrant^ A flexible valve assembly is attached to the tip of the ampoule.
Titrets* employ the principle of reverse titration; that is, small doses of
sample are added to the titrant to the appearance of the end point color. The
color change indicates that the equivalency point has been reached. The flow of
the sample into the Titret may be controlled by using an accessory called a
Titrettor™.
5.OB REAGENTS
5.IB The crushable glass ampoule, which is inside the reaction bottle,
contains 85 mg of metallic sodium in a light oil dispersion.
5.2B The buffer bottle contains 0.44 g of NaH2P04 • 2H20 and 0.32 ml of
HN03 in distilled water.
5.3B The glass vial contains 770 mg Stoddard Solvent (CAS No. 8052-41-
3), 260 mg toluene, 260 mg butyl ether, 260 mg diglyme, 130 mg naphthalene, and
70 mg demulsifier.
2Quanti-Chlor Kit, Titrets*, and Titrettor™ are manufactured by Chemetrics,
Inc., Calverton, VA 22016. U.S. Patent No. 4,332,769.
9077 - 8 Revision 0
November 1990
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5.4B The Titret contains 1.12 mg mercuric nitrate in distilled water.
5.5B The indicator-impregnated paper contains approximately 0.3 mg of
diphenylcarbazone and 0.2 mg of brilliant yellow.
6.OB SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See Section 6.0A of Method A.
7.OB PROCEDURE
7.IB Shake the glass vial and pour its contents into the reaction
bottle.
7.2B Fill the micropipet with a well-shaken oil sample by pulling the
plunger until its top edge is even with the top edge of the micropipet. Wipe off
the excess oil and transfer the sample into the reaction bottle (see Figure 2.1).
7.3B Gently squeeze most of the air out of the reaction bottle (see
Figure 2.2). Cap the bottle securely, and shake vigorously for 30 seconds.
7.4B Crush the sodium ampoule by pressing against the outside wall of
the reaction bottle (see Figure 2.3).
NOTE: Caution—Samples containing a high percentage of water will
generate heat and gas, causing the reaction bottle walls to
expand. To release the gas, briefly loosen the cap.
7.5B Shake the reaction bottle vigorously for 30 seconds.
7.6B Wait 1 minute. Shake the reaction bottle occasionally during this
time.
7.7B Remove the buffer bottle from the titration vial, and slowly pour
its contents into the reaction bottle (see Figure 2.4).
7.8B Cap the reaction bottle and shake gently for a few seconds. As
soon as the foam subsides, release the gas by loosening the cap. Tighten the
cap, and shake vigorously for 30 seconds. As before, release any gas that has
formed, then turn the reaction bottle upside down (see Figure 2.5).
7.9B Wait 1 minute.
7.10B While holding the filtering column in a vertical position, remove
the plug. Gently tap the column to settle the carbon particles.
7.11B Keeping the reaction bottle upside down, insert the flip top into
the end of the filtering column and position the column over the titration vial
(see Figure 2.6). Slowly squeeze the lower aqueous layer out of the reaction
bottle and into the filtering column. Keep squeezing until the first drop of oil
is squeezed out.
9077 - 9 Revision 0
November 1990
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NOTE: Caution--The aqueous layer should flow through the filtering
column into the titration vial in about 1 minute. In rare cases,
it may be necessary to gently tap the column to begin the flow.
The indicator paper should remain in the titration vial.
7.12B Cap the titration vial and shake it vigorously for 10 seconds.
7.13B Slide the flexible end of the valve assembly over the tapered tip
of the Titret so that it fits snugly (see Figure 3.1).
7.14B Lift (see Figure 3.2) the control bar and insert the assembled
Titret into the Titrettor™.
7.15B Hold the Titrettor™ with the sample pipe in the sample, and press
the control bar to snap the pre-scored tip of the Titret (see Figure 3.3).
NOTE: Caution—Because the Titret is sealed under vacuum, the fluid
inside may be agitated when the tip snaps.
7.16B With the tip of the sample pipe in the sample, briefly press the
control bar to pull in a SMALL amount of sample (see Figure 3.3). The contents
of the Titret will turn purple.
NOTE: Caution—During the titration, there will be some undissolved
powder inside the Titret. This does not interfere with the
accuracy of the test.
7.17B Wait 30 seconds.
7.18B Gently press the control bar again to allow another SMALL amount
of the sample to be drawn into the Titret.
NOTE: Caution--Do not press the control bar unless the sample pipe is
immersed in the sample. This prevents air from being drawn into
the Titret.
7.19B After each addition, rock the entire assembly to mix the contents
of the Titret. Watch for a color change from purple to very pale yellow.
7.20B Repeat Steps 13.18 and 13.19 until the color change occurs.
NOTE: Caution—The end point color change (from purple to pale yellow)
actually goes through an intermediate gray color. During this
intermediate stage, extra caution should be taken to bring in
SMALL amounts of sample and to mix the Titret contents well.
7.21B When the color of the liquid in the Titret changes to PALE YELLOW,
remove the Titret from the Titrettor™. Hold the Titret in a vertical position
and carefully read the test result on the scale opposite the liquid level.
9077 - 10 Revision 0
November 1990
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7.22B Calculation
7.22.IB To obtain results in micrograms per gram total
chlorine, multiply scale units on the Titret by 1.3 and then
subtract 200.
8.OB QUALITY CONTROL
8.IB Refer to Chapter One for specific quality control procedures.
8.2B Each sample should be tested two times. If the results do not
agree to within 10%, expressed as the relative percent difference of the results,
a third test must be performed. Report the results of the two that agree.
9.OB METHOD PERFORMANCE
9.IB These data are based on 49 data points obtained by seven
laboratories who each analyzed four used crankcase oils and three fuel oil blends
with crankcase in duplicate. A data point represents one duplicate analysis of
a sample. There were no outlier data points or laboratories.
9.2B Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method, the following values only in 1 case in 20 (see
Table 2):
Repea tabi lity = 0.31
-------
TABLE 2.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY THE QUANTITATIVE END POINT TEST KIT METHOD
Average value, Repeatability, Reproducibility,
1,000
1,500
2,000
2,500
3,000
310
465
620
775
930
600
900
1,200
1,500
1,800
TABLE 3.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY THE
QUANTITATIVE END POINT TEST KIT METHOD
Amount
expected,
M9/9
320 (< 750)a
480 (< 750)a
920
1,498
1,527
3,029
3,045
Amount
found,
M9/g
776
782
1,020
1,129
1,434
1,853
2,380
Bias,
Mg/g
+16
+32
+100
-369
-93
-1,176
-665
Percent
bias
+3
+4
+11
-25
-6
-39
-22
The lower limit of the kit is 750 p.g/g.
9077 - 12 Revision 0
November 1990
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Reaction bottle
Titration via
Filtering
Column
assembly
6
*—'r
Buffer
bottle
Micro pipet
Figure 1. Components of CHEMetrics Total Chlorine in Waste Oil Test Kit
(Cat. No. K2610).
9077 - 13
Revision 0
November 1990
-------
Push plunger
down to
transfer
sample
Figure 2.1
Figure 2.2
* Crush
Figure 2.3
Buffer Bottle
Figure 2.4
Reaction bottle
upsidedown in
component tray
Figure 2.5
Aqueous
Layer
Filtering Column
Figure 2.6
Titration Vial
Figure 2. Reaction-Extraction Procedure.
9077 - 14
Revision 0
November 1990
-------
Attaching
the Valve
Assembly
Figure 3.1
Valve
Assembly
Titret
\
/\
Snapping
the Tip
Figure 3.2
Lift control bar
Performing the
Analysis
Figure 3.3
Watch for
color change
here
Press control bar
Sample pipe
Sample
Reading
the Result
Figure 3.4
Read
scale units
when color
changes
permanently
Figure 3. Titration Procedure
9077 - 15
Revision 0
November 1990
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METHOD 9077 B
REVERSE TITRATION QUANTITATIVE END POINT TEST KIT METHOD
START
...
7. IB Shake glass
vial, pour into
reaction bottle
1
7-2B Fill
micropipet with
oil; remove excess
oil; t rans f er oil
to reaction bottle
7 . 3B Squeeze air
from reaction
bottle; cap ; mix
i
7.4B Crush sodium
ampoule
.
7 5B - 7 .68 Shake
reaction bottle for
30 seconds ; wai t
one minute
7 . 7B Pour buffer
into reaction
bottle
•»
7 SB - 7.9B Shake
gent 1 y ; rel ease
gas; shake; release
gas; turn bottle
upside down; wait
one minute
7 .10B Prepare
filtering column
1
7 .118 Filter lower
aqueous layer
through filtering
column into
ti t ra t i on vial
.
7.12B Shake vial
1
7.13B Assemble
valve assembly over
Titret
..
7.14B Insert Titret
into Titrettor
7.158 Snap tip of
Titret
7.16B - 7.20B Pull
3ma 11 amount of
sample into Titret;
mix; wai t 30
seconds; repea t
process until color
changes from purple
to pale yellow
7.21B When color
changes to pale
yel1ow, remove
Titret; record test
result from Titret
7.22B Calculate
concent ration of
chlorine in ug/g
STOP
9077 - 16
Revision 0
November 1990
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METHOD C
DIRECT TITRATION QUANTITAVE END POINT TEST KIT METHOD
4.0C APPARATUS AND MATERIALS
4.1C The CHLOR-D-TECT Q40003 is a complete self-contained kit. It
includes: a sampling syringe to withdraw a fixed sample volume for analysis; a
polyethylene test tube #1 into which the sample is introduced for dilution and
reaction with metallic sodium; a polyethylene tube #2 containing a buffered
aqueous extractant and the diphenylcarbazone indicator; a microburette containing
the mercuric nitrate titrant; and a plastic filtration funnel. Also included are
instructions to conduct the test.
5.0C REAGENTS
5.1C All necessary reagents are contained within the kit. The diluent
solvent containing the catalyst, the metallic sodium, and the diphenylcarbazone
are separately glass-encapsulated in the precise quantity required for analysis.
A predispensed volume of buffer is contained in the second polyethylene tube.
Mercuric nitrate titrant is also supplied in a sealed titration burette.
5.2C The kit should be examined upon opening to see that all of the
components are present and that all ampoules (3) are in place and not leaking.
The liquid in Tube #2 (clear cap) should be approximately 1/2 in. above the 5-mL
line and the tube should not be leaking. The ampoules are not supposed to be
completely full.
6.0C SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1C See Section 6.0A of Method A.
7.0C PROCEDURE
7.1C Preparation. Open analysis carton, remove contents, mount plastic
test tubes in the provided holder.
NOTE: Perform the test in a warm, dry area with adequate light. In cold
weather, a truck cab is sufficient. If a warm area is not
available, Step 19.3 should be performed while warming Tube #1 in
palm of hand.
7.2C Sample introduction. Unscrew the white dispenser cap from Tube #1.
Slide the plunger in the empty syringe a few times to make certain that it slides
easily. Place the top of the syringe in the oil sample to be tested, and pull
back on the plunger until it reaches the stop and cannot be pulled further.
Remove the syringe from the sample container, and wipe any excess oil from the
outside of the syringe with the enclosed tissue. Place the tip of the syringe
in Tube #1, and dispense the oil sample by depressing the plunger. Replace the
white cap on the tube.
Available from Dexsil Corporation, One Hamden Park Drive, Hamden, CT 06517.
9077 - 17 Revision 0
November 1990
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7.3C Reaction. Break the lower (colorless) capsule containing the clear
diluent solvent by squeezing the sides of the test tube. Mix thoroughly by
shaking the tube vigorously for 30 seconds. Crush the upper grey ampoule
containing metallic sodium, again by squeezing the sides of the test tube. Shake
vigorously for 20 seconds. Allow reaction to proceed for 60 seconds, shaking
intermittently several times while timing with a watch.
NOTE: Caution--Always crush the clear ampoule in each tube first.
Otherwise, stop the test and start over using another complete kit.
False (low) results may occur and allow a contaminated sample to
pass without detection if clear ampoule is not crushed first.
7.4C Extraction. Remove caps from both tubes. Pour the clear buffered
extraction solution from Tube #2 into Tube #1. Replace the white cap on Tube #1,
and shake vigorously for 10 seconds. Vent tube by partially unscrewing the
dispenser cap. Close cap securely, and shake for an additional 10 seconds. Vent
again, tighten cap, and stand tube upside down on white cap. Allow phases to
separate for 2 minutes.
NOTE: Tip Tube #2 to an angle of only about 45°. This will prevent the
holder from sliding out.
7.5C Analysis. Put filtration funnel into Tube #2. Position Tube #1
over funnel and open nozzle on dispenser cap. Squeeze the sides of Tube #1 to
dispense the clear aqueous lower phase through the filter into Tube #2 to the 5-
mL line on Tube #2. Remove the filter funnel, and close the nozzle on the
dispenser cap. Place the plunger rod in the titration burette and press until
it clicks into place. Break off (do not pull off) the tip on the titration
burette. Insert the burette into Tube #2, and tighten the cap. Break the
colored ampoule, and shake gently for 10 seconds. Dispense titrant dropwise by
pushing down on burette rod in small increments. Shake the tube gently to mix
titrant with solution in Tube #2 after each increment. Continue adding titrant
until solution turns from yellow to red-violet. An intermediate pink color may
develop in the solution, but should be disregarded. Continue titrating until a
true red-violet color is realized. The chlorine concentration of the original
oil sample is read directly off the titrating burette at the tip of the black
plunger. Record this result immediatley as the red-violet color will fade with
time.
8.0C QUALITY CONTROL
8.1C Refer to Chapter One for specific quality control procedures.
8.2C Each sample should be tested two times. If the results do not
agree to within 10%, expressed as the relative percent difference of the results,
a third test must be performed. Report the results of the two that agree.
9.0C METHOD PERFORMANCE
9.1C These data are based on 96 data points obtained by 12 laboratories
who each analyzed six used crankcase oils and two fuel oil blends with crankcase
in duplicate. A data point represents one duplicate analysis of a sample.
9077 - 18 Revision 0
November 1990
-------
9.2C Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method, the following values only in 1 case in 20 (see
Table 4):
Repeatability = 0.175
*where x is the average of two results in
Reproducibilitv - The difference between two single and independent results
obtained by different operators working in different laboratories on identical
test material would exceed, in the long run, the following values only in 1 case
in 20:
Reproducibility = 0.331
*where x is the average value of two results in pg/g.
9.3C Bias. The bias of this test method varies with concentration, as
shown in Table 5:
Bias = Amount found - Amount expected
10.0 REFERENCE
1. Gaskill, A.; Estes, E.D.; Hardison, D.L.; and Myers, L.E. Validation of
Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract
No. 68-01-7075, wA 80. July 1988.
9077 - 19 Revision 0
November 1990
-------
TABLE 4.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY THE QUANTITATIVE END POINT TEST KIT METHOD
Average value, Repeatability, Reproducibility,
M9/9 M9/9 M9/9
500
1,000
1,500
2,000
2,500
3,000
4,000
88
175
263
350
438
525
700
166
331
497
662
828
993
1,324
TABLE 5.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY THE
QUANTITATIVE END POINT TEST KIT METHOD
Amount
expected,
M9/9
664
964
1,230
1,445
2,014
2,913
3,812
4,190
Amount
found,
M9/9
695
906
1,116
1,255
1,618
2,119
2,776
3,211
Bias,
M9/9
31
-58
-114
-190
-396
-794
-1,036
-979
Percent
bias
+5
-6
-9
-13
-20
-27
-27
-23
9077 - 20 Revision 0
November 1990
-------
METHOD 9077 C
DIRECT TITRATION QUANTITAVE END POINT TEST KIT METHOD
START
7.1C Open test kit
7.2C Draw oil into
syringe; remove
excess oil;
dispense oil into
Tube ti
7.3C Break
colorless capsule;
mix; crush grey
capsule; mix; allow
reaction to proceed
for 60 seconds
7.4C Pour Tube H
solution into Tube
#1; mix; vent;
allow phases to
separate
7.SC Filter aqueous
lower phase in Tube
fl into Tube t2,
remove filter
funnel
7.SC Place plunger
in titraton
burette; press;
break off burette
tip; insert burette
in Tube #2; break
colored ampoule;
shake
7.SC Dispense
titrant; shake;
repeat process
until solution
turns from yellow
to red*violet
7.SC Record level
from titrating
buret te
STOP
9077 - 21
Revision 0
November 1990
-------
METHOD 9200A
NITRATE
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the analysis of groundwater, drinking,
surface, and saline waters, and domestic and industrial wastes. Modifications
can be made to remove or correct for turbidity, color, salinity, or dissolved
organic compounds in the sample.
1.2 The applicable range of concentration is 0.1 to 2 mg N03-N per liter
of sample.
2.0 SUMMARY OF METHOD
2.1 This method is based upon the reaction of the nitrate ion with
brucine sulfate in a 13 N H2S04 solution at a temperature of 100°C. The color
of the resulting complex is measured at 410 nm. Temperature control of the color
reaction is extremely critical.
3.0 INTERFERENCES
3.1 Dissolved organic matter will cause an off color in 13 N H2S04 and
must be compensated for by additions of all reagents except the brucine-
sulfanilic acid reagent. This also applies to natural color, not due to
dissolved organics, that is present.
3.2 If the sample is colored or if the conditions of the test cause
extraneous coloration, this interference should be corrected by running a
concurrent sample under the same conditions but in the absence of the brucine-
sulfanilic acid reagent.
3.3 Strong oxidizing or reducing agents cause interference. The
presence of oxidizing agents may be determined by a residual chlorine test;
reducing agents may be detected with potassium permanganate.
3.3.1 Oxidizing agents' interference is eliminated by the
addition of sodium arsenite.
3.3.2 Reducing agents may be oxidized by addition of H202.
3.4 Ferrous and ferric ion and quadrivalent manganese give slight
positive interferences, but in concentrations less than 1 mg/L these are
negligible.
3.5 Uneven heating of the samples and standards during the reaction
time will result in erratic values. The necessity for absolute control of
temperature during the critical color development period cannot be too strongly
emphasized.
9200A - 1 Revision 1
November 1990
-------
4.0 APPARATUS AND MATERIALS
4.1 Spectrophotometer or filter photometer suitable for measuring
absorbance at 410 nm.
4.2 Sufficient number of 40- to 50-mL glass sample tubes for reagent
blanks, standards, and samples.
4.3 Neoprene-coated wire racks to hold sample tubes.
4.4 Water bath suitable for use at 100°C. This bath should contain a
stirring mechanism so that all tubes are at the same temperature and should be
of sufficient capacity to accept the required number of tubes without a
significant drop in temperature when the tubes are immersed.
4.5 Water bath suitable for use at 10-15°C.
4.6 Analytical balance: capable of weighing to 0.0001 g.
4.7 Class A volumetric flasks: 1 L.
4.8 pH Indicator paper.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Sodium chloride solution (30%): Dissolve 300 g NaCl in reagent
water and dilute to 1 liter with reagent water.
5.4 Sulfuric acid solution: Carefully add 500 ml concentrated H2S04 to
125 ml reagent water. Cool and keep tightly stoppered to prevent absorption of
atmospheric moisture.
5.5 Brucine-sulfanilic acid reagent: Dissolve 1 g brucine sulfate --
(C23H26N204)2 . H2S04 . 7H20 -- and 0.1 g sulfanilic acid (NH2C6H4S03H • H20) in
70 ml not reagent water. Add 3 ml concentrated HC1, cool, mix, and dilute to 100
ml with reagent water. Store in a dark bottle at 5°C. This solution is stable
for several months; the pink color that develops slowly does not affect its
usefulness. Mark bottle with warning. "CAUTION: Brucine Sulfate is toxic; do
not ingest."
5.6 Potassium nitrate stock solution (1.0 ml = 0.1 mg N03-N): Dissolve
0.7218 g anhydrous potassium nitrate (KN03) in reagent water and dilute to 1
liter in a Class A volumetric flask. Preserve with 2 ml chloroform per liter.
This solution is stable for at least 6 months.
9200A - 2 Revision 1
November 1990
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5.7 Potassium nitrate standard solution (1.0 mi = 0.001 mg N03-N):
Dilute 10.0 mL of the stock solution (Step 5.6) to 1 liter in a Class A
volumetric flask. This standard solution should be prepared fresh weekly.
5.8 Acetic acid (1+3): Dilute 1 volume glacial acetic acid (CH3COOH)
with 3 volumes of reagent water.
5.9 Sodium hydroxide (1 N): Dissolve 40 g of NaOH in reagent water.
Cool and dilute to 1 liter with reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Analysis should be performed within 48 hours. If analysis can be
done within 24 hours, the sample should be preserved by refrigeration at 4°C.
When samples must be stored for more than 24 hours, they should be preserved with
sulfuric acid (2 mL/L concentrated H2S04) and refrigerated.
7.0 PROCEDURE
7.1 Adjust the pH of the samples to approximately 7 with acetic acid
(Step 5.8) or sodium hydroxide (Step 5.9). If necessary, filter to remove
turbidity. Sulfuric acid can be used in place of acetic acid, if preferred.
7.2 Set up the required number of sample tubes in the rack to handle
the reagent blank, standards, and samples. Space tubes evenly throughout the
rack to allow for even flow of bath water between the tubes. This should assist
in achieving uniform heating of all tubes.
7.3 If it is necessary to correct for color or dissolved organic matter
which will cause color on heating, run a set of duplicate samples with all of the
reagents, except the brucine-sulfanilic acid.
7.4 Pipet 10.0 mL of standards and samples or an aliquot of the samples
diluted to 10.0 mL into the sample tubes.
7.5 If the samples are saline, add 2 mL of the 30% sodium chloride
solution (Step 5.3) to the reagent blank, standards, and samples. For freshwater
samples, sodium chloride solution may be omitted. Mix contents of tubes by
swirling; place rack in cold-water bath (0-10°C).
7.6 Pipet 10.0 mL of sulfuric acid solution (Step 5.4) into each tube
and mix by swirling. Allow tubes to come to thermal equilibrium in the cold
bath. Be sure that temperatures have equilibrated in all tubes before
continuing.
7.6.1 Add 0.5 mL brucine-sulfanilic acid reagent (Step 5.5) to
each tube (except the interference control tubes) and carefully mix by
swirling; place the rack of tubes in the 100°C water bath for exactly 25
minutes.
9200A - 3 Revision 1
November 1990
-------
CAUTION: Immersion of the tube rack into the bath should not decrease the
temperature of the bath by more than 1-2°C. In order to keep this
temperature decrease to an absolute minimum, flow of bath water
between the tubes should not be restricted by crowding too many
tubes into the rack. If color development in the standards reveals
discrepancies in the procedure, the operator should repeat the
procedure after reviewing the temperature control steps.
7.7 Remove rack of tubes from the hot-water bath, immerse in the cold-
water bath, and allow to reach thermal equilibrium (20-25°C).
7.8 Read absorbance against the reagent blank at 410 nm using a 1-cm
or longer cell.
7.9 Calculation:
7.9.1 Obtain a standard curve by plotting the absorbance of
standards run by the above procedure against mg/L N03-N. (The color
reaction does not always follow Beer's law.)
7.9.2 Subtract the absorbance of the sample without the brucine-
sulfanilic reagent from the absorbance of the sample containing brucine-
sulfanilic acid and determine mg/L N03-N. Multiply by an appropriate
dilution factor if less than 10 ml of sample is taken.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Linear calibration curves must be composed of a minimum of a blank
and five standards. A set of standards must be included with each batch of
samples.
8.3 Dilute samples if they are more concentrated than the highest
standard or if they fall on the plateau of a calibration curve.
8.4 After calibrating, verify calibration with an independently
prepared check standard.
8.5 Matrix spikes and matrix spike duplicates are brought through the
whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Twenty-seven analysts in fifteen laboratories analyzed natural-
water samples containing exact increments of inorganic nitrate, with the
following results:
9200A - 4 Revision 1
November 1990
-------
Increment as Precision as Accuracy as
Nitrogen, Nitrate Standard Deviation Bias Bias
(mg/L N) (mg/L N) (%) (mg/L N)
0.16
0.19
1.08
1.24
0.092
0.083
0.245
0.214
-6.79
+8.30
+4.12
+2.82
-0.01
+0.02
+0.04
+0.04
10.0 REFERENCES
1. Annual Book of ASTM Standards, Part 31, "Water," Standard D992-71, p. 363
(1976).
2. Jenkins, D. and L. Medsken, "A Brucine Method for the Determination of
Nitrate in Ocean, Estuarine, and Fresh Water," Anal.Chem., 36, p. 610 (1964).
3. Standard Methods for the Examination of Water and Wastewater, 14th ed., p.
427, Method 419D (1975).
9200A - 5 Revision 1
November 1990
-------
METHOD 9200A
NITRATE
START
7.3 Run
duplicates with
all reagenti
except brucine
sulfanilic acid
7.5 Add 30S
sodium chloride
solution; mm;
place in cold
water bath
-
7.1 Adjust pH
of samples to
7; filter if
necessary
7.2 Set up
sample tubes
in rack
7 4 Pipette
standards and
samples into
sample tubes
7.6 Pipette
sulfuric acid
solution into
each tube;
mix
761 Add
brucine
sulfanilie
acid reagent
to each tube
7.6 1 Bathe
rack of tubes
in 100C water
for 25 min
7 . 7 Immerse
tubes in cold
water; allow to
reach thermal
equilibr ium
78 Read
absorbance
against
reagent blank
at 410 nm
791 Obtain a
std absorbance
curve and
calculate mg/L
nitrate
STOP
9200A - 6
Revision 1
November 1990
-------
METHOD 9252
CHLORIDE (TITRIMETRIC. MERCURIC NITRATE)
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to ground water, drinking, surface, and
saline waters, and domestic and industrial wastes.
1.2 The method is suitable for all concentration ranges of chloride
content; however, in order to avoid large titration volume, a sample aliquot
containing not more than 10 to 20 mg Cl" per 50 ml is used.
1.3 Automated titration may be used.
2.0 SUMMARY OF METHOD
2.1 An acidified sample is titrated with mercuric nitrate in the
presence of mixed diphenylcarbazone-bromophenol blue indicator. The end point
of the titration is the formation of the blue-violet mercury diphenylcarbazone
complex.
3.0 INTERFERENCES
3.1 Anions and cations at concentrations normally found in surface
waters do not interfere. However, at the higher concentration often found in
certain wastes, problems may occur.
3.2 Sulfite interference can be eliminated by oxidizing the 50 ml of
sample solution with 0.5-1 ml of H202.
4.0 APPARATUS AND MATERIALS
4.1 Standard laboratory titrimetric equipment, including 1 mL or 5 mL
microburet with 0.01 mL gradations.
4.2 Class A volumetric flasks: 1 L and 100 mL.
4.3 pH Indicator paper.
4.4 Analytical balance: capable of weighing to 0.0001 g.
5.0 REAGENTS
5.1 Reagent-grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
9252 - 1 Revision 1
November 1990
-------
5.3 Standard sodium chloride solution, 0.025 N: Dissolve 1.4613 g ±
0.0002 g of sodium chloride (dried at 600°C for 1 hr) in chloride-free water in
a 1 liter Class A volumetric flask and dilute to the mark with reagent water.
5.4 Nitric acid (HNO,) solution: Add 3.0 ml concentrated nitric acid
to 997 ml of reagent water ("3 + 997" solution).
5.5 Sodium hydroxide (NaOH) solution (10 g/L): Dissolve approximately
10 g of NaOH in reagent water and dilute to 1 L with reagent water.
5.6 Hydrogen peroxide (H202): 30%.
5.7 Hydroquinone solution (10 g/L): Dissolve 1 g of purified
hydroquinone in reagent water in a 100 ml Class A volumetric flask and dilute to
the mark.
5.8 Mercuric nitrate titrant (0.141 N): Dissolve 24.2 g Hg(N03)2 • H20
in 900 mL of reagent water acidified with 5.0 ml concentrated HN03 in a 1 liter
volumetric flask and dilute to the mark with reagent water. Filter, if
necessary. Standardize against standard sodium chloride solution (Step 5.3)
using the procedures outlined in Section 7.0. Adjust to exactly 0.141 N and
check. Store in a dark bottle. A 1.00 mL aliquot is equivalent to 5.00 mg of
chloride.
5.9 Mercuric nitrate titrant (0.025 N): Dissolve 4.2830 g Hg(N03)2 •
H20 in 50 mL of reagent water acidified with 0.05 mL of concentrated
HN03 (sp. gr. 1.42) in a 1 liter volumetric flask and dilute to the mark with
reagent water. Filter, if necessary. Standardize against standard sodium
chloride solution (Step 5.3) using the procedures outlined in Section 7.0.
Adjust to exactly 0.025 N and check. Store in a dark bottle.
5.10 Mercuric nitrate titrant (0.0141 N): Dissolve 2.4200 g Hg(N03)2 •
H20 in 25 mL of reagent water acidified with 0.25 mL of concentrated HN03 (sp.
gr. 1.42) in a 1 liter Class A volumetric flask and dilute to the mark with
reagent water. Filter, if necessary. Standardize against standard sodium
chloride solution (Step 5.3) using the procedures outlined in Section 7.0.
Adjust to exactly 0.0141 N and check. Store in a dark bottle. A 1 mL aliquot
is equivalent to 500 jug of chloride.
5.11 Mixed indicator reagent: Dissolve 0.5 g crystalline diphenylcar-
bazone and 0.05 g bromophenol blue powder in 75 mL 95% ethanol in a 100 mL Class
A volumetric flask and dilute to the mark with 95% ethanol. Store in brown
bottle and discard after 6 mo.
5.12 Alphazurine indicator solution: Dissolve 0.005 g of alphazurine
blue-green dye in 95% ethanol or isopropanol in 100 mL Class A volumetric flask
and dilute to the mark with 95% ethanol orv isopropanol.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual:
6.2 There are no special requirements for preservation.
9252 - 2 Revision 1
November 1990
-------
7.0 PROCEDURE
7.1 Place 50 ml of sample in a vessel for titration. If the concentra-
tion is greater than 20 mg/L chloride, use 0.141 N mercuric nitrate titrant (Step
5.8) in Step 7.6, or dilute sample with reagent water. If the concentration is
less than 2.5 mg/L of chloride, use 0.0141 N mercuric nitrate titrant (Step 5.10)
in Step 7.6. Using a 1 mL or 5 ml microburet, determine an indicator blank on
50 ml chloride-free water using Step 7.6. If the concentration is less than 0.1
mg/L of chloride, concentrate an appropriate volume to 50 ml.
7.2 Add 5 to 10 drops of mixed indicator reagent (Step 5.11); shake or
swirl solution.
7.3 If a blue-violet or red color appears, add HN03 solution (Step 5.4)
dropwise until the color changes to yellow. Proceed to Step 7.5.
7.4 If a yellow or orange color forms immediately on addition of the
mixed indicator, add NaOH solution (Step 5.5) dropwise until the color changes
to blue-violet; then add HN03 solution (Step 5.4) dropwise until the color
changes to yellow.
7.5 Add 1 ml excess HN03 solution (Step 5.4).
7.6 Titrate with 0.025 N mercuric nitrate titrant (Step 5.9) until a
blue-violet color persists throughout the solution. If volume of titrant exceeds
10 ml or is less than 1 mL, use the 0.141 N or 0.0141 N mercuric nitrate
solutions, respectively. If necessary, take a small sample aliquot. Alphazurine
indicator solution (Step 5.12) may be added with the indicator to sharpen the end
point. This will change color shades. Practice runs should be made.
Note: The use of indicator modifications and the presence of heavy metal
ions can change solution colors without affecting the accuracy of
the determination. For example, solutions containing alphazurine
may be bright blue when neutral, grayish purple when basic, blue-
green when acidic, and blue-violet at the chloride end point.
Solutions containing about 100 mg/L nickel ion and normal mixed
indicator are purple when neutral, green when acidic, and gray at
the chloride end point. When applying this method to samples that
contain colored ions or that require modified indicator, it is
recommended that the operator become familiar with the specific
color changes involved by experimenting with solutions prepared as
standards for comparison of color effects.
7.6.1 If chromate is present at <100 mg/L and iron is not
present, add 5-10 drops of alphazurine indicator solution (Step 5.12) and
acidify to a pH of 3 (indicating paper). End point will then be an olive-
purple color.
7.6.2 If chromate is present at >100 mg/L and iron is not
present, add 2 mL of fresh hydroquinone solution (Step 5.7).
7.6.3 If ferric ion is present use a volume containing no more
than 2.5 mg of ferric ion or ferric ion plus chromate ion. Add 2 mL fresh
hydroquinone solution (Step 5.7).
9252 - 3 Revision 1
November 1990
-------
7.6.4 If sulfite ion is present, add 0.5 ml of H202 solution
(Step 5.6) to a 50 ml sample and mix for 1 min.
7.7 Calculation:
(A - B)N x 35,450
mg chloride/liter =
ml of sample
where:
A = mi titrant for sample;
B = ml titrant for blank; and
N = normality of mercuric nitrate titrant.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 A matrix duplicate and matrix spike sample are brought through the
whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Water samples—A total of 42 analysts in 18 laboratories analyzed
synthetic water samples containing exact increments of chloride, with the results
shown in Table 1.
In a single laboratory, using surface water samples at an average
concentration of 34 mg CT/L, the standard deviation was +1.0.
A synthetic unknown sample containing 241 mg/L chloride, 108 mg/L Ca, 82
mg/L Mg, 3.1 mg/L K, 19.9 mg/L Na, 1.1 mg/L nitrate N, 0.25 mg/L nitrate N, 259
mg/L sulfate and 42.5 mg/L total alkalinity (contributed by NaHCOJ in reagent
water was analyzed in 10 laboratories by the mercurimetric method, with a
relative standard deviation of 3.3% and a relative error of 2.9%.
9.2 Oil combustates--These data are based on 34 data points obtained by
five laboratories who each analyzed four used crankcase oils and three fuel oil
blends with crankcase oil in duplicate. The samples were combusted using Method
5050. A data point represents one duplicate analysis of a sample. One data
point was judged to be an outlier and was not included in these results.
9.2.1 Precision and bias.
9.2.1.1 Precision. The precision of the method as determined
by the statistical examination of interlaboratory test results is as
follows:
9252 - 4 Revision 1
November 1990
-------
Repeatability - The difference between successive results
obtained by the same operator with the same apparatus under constant
operating conditions on identical test material would exceed, in the
long run, in the normal and correct operation of the test method, the
following values only in 1 case in 20 (see Table 2):
Repeatability = 7.61 Jx*
*where x is the average of two results in /ng/g.
Reproducibilitv - The difference between two single and
independent results obtained by different operators working in
different laboratories on identical test material would exceed, in
the long run, the following values only in 1 case in 20:
Reproducibility = 20.02
*where x is the average value of two results in M9/9-
9.2.1.2 Bias. The bias of this method varies with
concentration, as shown in Table 3:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Annual Book of ASTM Standards, Part 31, "Water," Standard D512-67, Method
A, p. 270 (1976).
2. Standard Methods for the Examination of Water and Wastewater, 15th ed.,
(1980).
3. U.S. Environmental Protection Agency, Methods for Chemical Analysis of
Water and Wastes, EPA 600/4-79-020 (1983), Method 325.3.
9252 - 5 Revision 1
November 1990
-------
TABLE 1. ANALYSES OF SYNTHETIC WATER SAMPLES
FOR CHLORIDE BY MERCURIC NITRATE METHOD
Increment as Precision as Accuracy as
Chloride Standard Deviation Bias Bias
(mg/L) (mg/L) (%) (mg/L)
17
18
91
97
382
398
1.54
1.32
2.92
3.16
11.70
11.80
+2.16
+3.50
+0.11
-0.51
-0.61
-1.19
+0.4
+0.6
+0.1
-0.5
-2.3
-4.7
TABLE 2. REPEATABILITY AND REPRODUCIBILITY
FOR CHLORINE IN USED OILS BY BOMB
OXIDATION AND MERCURIC NITRATE TITRATION
Average value, Repeatability, Reproducibility,
500
1,000
1,500
2,000
2,500
3,000
170
241
295
340
381
417
448
633
775
895
1,001
1,097
9252 - 6 Revision 1
November 1990
-------
TABLE 3. RECOVERY AND BIAS DATA FOR CHLORINE IN
USED OILS BY BOMB OXIDATION AND
MERCURIC NITRATE TITRATION
Amount Amount
expected, found, Bias, Percent
M9/9 M9/9 M9/9 bias
320 460 140 +44
480 578 98 +20
920 968 48 +5
1,498 1,664 166 +11
1,527 1,515 - 12 - 1
3,029 2,809 -220 - 7
3,045 2,710 -325 -11
9252 - 7 Revision 1
November 1990
-------
METHOD 9252
CHLORIDE (TITRIMETRIC, MERCURIC NITRATE)
START
7.1 Place 50 mL
sample in titration
vessel; determine
concentration of
mercuric nitrate
titrant to use in
Step 7.6; determine
an indicator blank
7.2 Add indicator
to sample; shake
7.3 Is sample
blue-violet or
red?
7.4 Is sample
yellow or
orange?
7.4 Add sodium
hydroxide unti1
sample is
blue-violet; add
nitric acid until
sample is yellow
7.3 Add nitric acid
until sample is
yellow
7.5 Add 1 mL nitric
acid
7.6 Titrate with
mercuric nitrate
until blue-violet
color persists
7.7 Calculate
concentration of
chloride in sampla
9252 - 8
STOP
Revision 1
November 1990
-------
METHOD 9253
CHLORIDE (TITRIMETRIC. SILVER NITRATE)
1.0 SCOPE AND APPLICATION
1.1 This method is intended primarily for oxygen bomb combustates or
other waters where the chloride content is 5 mg/L or more and where interferences
such as color or high concentrations of heavy metal ions render Method 9252
impracticable.
2.0 SUMMARY OF METHOD
2.1 Water adjusted to pH 8.3 is titrated with silver nitrate solution
in the presence of potassium chromate indicator. The end point is indicated by
persistence of the orange-silver chromate color.
3.0 INTERFERENCES
3.1 Bromide, iodide, and sulfide are titrated along with the chloride.
Orthophosphate and polyphosphate interfere if present in concentrations greater
than 250 and 25 mg/L, respectively. Sulfite and objectionable color or turbidity
must be eliminated. Compounds that precipitate at pH 8.3 (certain hydroxides)
may cause error by occlusion.
3.2 Residual sodium carbonate from the bomb combustion may react with
silver nitrate to produce the precipitate, silver carbonate. This competitive
reaction may interfere with the visual detection of the end point. To remove
carbonate from the test solution, add small quantities of sulfuric acid followed
by agitation.
4.0 APPARATUS AND MATERIALS
4.1 Standard laboratory titrimetric equipment, including 1 mL or 5 mL
microburet with 0.01 mL gradations, and 25 mL buret.
4.2 Analytical balance: capable of weighing to 0.0001 g.
4.3 Class A volumetric flask: 1 L.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Hydrogen peroxide (30%), H202.
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5.4 Phenolphthalein indicator solution (10 g/L).
5.5 Potassium chromate indicator solution. Dissolve 50 g of potassium
chromate (K2CrOJ in 100 ml of reagent water and add silver nitrate (AgN03) until
a slightly red precipitate is produced. Allow the solution to stand, protected
from light, for at least 24 hours after the addition of AgN03. Then filter the
solution to remove the precipitate and dilute to 1 L with reagent water.
5.6 Silver nitrate solution, standard (0.025N). Crush approximately
5 g of silver nitrate (AgNO,) crystals and dry to constant weight at 40°C.
Dissolve 4.2473 ± 0.0002 g of the crushed, dried crystals in reagent water and
dilute to 1 L with reagent water. Standardize against the standard NaCl
solution, using the procedure given in Section 7.0.
5.7 Sodium chloride solution, standard (0.025N). Dissolve 1.4613 g +
0.0002 g of sodium chloride (dried at 600°C for 1 hr) in chloride-free water in
a 1 liter Class A volumetric flask and dilute to the mark with reagent water.
5.8 Sodium hydroxide solution (0.25'N). Dissolve approximately 10 g of
NaOH in reagent water and dilute to 1 L with reagent water.
5.9 Sulfuric acid (1:19), H2S04. Carefully add 1 volume of concentrated
sulfuric acid to 19 volumes of reagent water, while mixing.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 There are no special requirements for preservation.
7.0 PROCEDURE
7.1 Pour 50 mL or less of the sample, containing between 0.25 mg and
20 mg of chloride ion, into a white porcelain container. Dilute to approximately
50 mL with reagent water, if necessary. Adjust the pH to the phenolphthalein end
point (pH 8.3) using H2S04 (Step 5.9) or NaOH solution (Step 5.8).
7.2 Add approximately 1.0 mL of K2Cr04 indicator solution and mix. Add
standard AgN03 solution dropwise from a 25 mL buret until the orange color
persists throughout the sample when illuminated with a yellow light or viewed
with yellow goggles.
7.3 Repeat the procedure described in Steps 7.1 and 7.2 using exactly
one-half as much original sample, diluted to 50 mL with halide-free water.
7.4 If sulfite ion is present, add 0.5 mL of H202 to the samples
described in Steps 7.2 and 7.3 and mix for 1 minute. Adjust the pH, then proceed
as described in Steps 7.2 and 7.3.
7.5 Calculation
7.5.1 Calculate the chloride ion concentration in the original
sample, in milligrams per liter, as follows:
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Chloride (mg/L) = [(V, - V2) x N x 71,000] / S
where:
V1 = Milliliters of standard AgNO, solution added in titrating
the sample prepared in Step 7.1.
V2 = Milliliters of standard AgNO, solution added in titrating
the sample prepared in Step 7.3.
N = Normality of standard AgN03 solution.
S = Milliliters of original sample in the 50 ml test sample
prepared in Step 7.1.
71,000 = 2 x 35,500 mg CT/equivalent, since V, - 2V2.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 A matrix duplicate and matrix spike sample are brought through the
whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 These data are based on 32 data points obtained by five
laboratories who each analyzed four used crankcase oils and three fuel oil blends
with crankcase in duplicate. The samples were combusted using Method 5050. A
data point represents one duplicate analysis of a sample. Three data points were
judged to be outliers and were not included in these results.
9.1.1 Precision. The precision of the method as determined by
the statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the same operator with the same apparatus under- constant operating
conditions on identical test material would exceed, in the long run, in
the normal and correct operation of the test method, the following values
only in 1 case in 20 (see Table 1):
Repeatability =0.36
*where x is the average of two results in M9/9-
Reoroducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
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Reproducibility =0.71 Jx*
where x is the average of two results in M9/9-
9.1.2 Bias. The bias of this method varies with concentration,
as shown in Table 2:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Rohrbough, W.G.; et al. Reagent Chemicals. American Chemical Society
Specifications. 7th ed.; American Chemical Society: Washington, DC, 1986.
2. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification for
Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
3. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. "Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels," Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract No.
68-01-7075, WA 80. July 1988.
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TABLE 1.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY BOMB OXIDATION AND SILVER NITRATE TITRATION
Average value
(M9/9)
Repeatability
(M9/9)
Reproducibility
(M9/9)
500
1,000
1,500
2,000
2,500
3,000
180
360
540
720
900
1,080
355
710
1,065
1,420
1,775
2,130
TABLE 2.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY
BOMB OXIDATION AND SILVER NITRATE TITRATION
Amount
expected
(M9/9)
320
480
920
1,498
1,527
3,029
3,045
Amount
found
(M9/9)
645
665
855
1,515
1,369
2,570
2,683
Bias,
(M9/9)
325
185
-65
17
-158
-460
-362
Percent
bias
+102
+39
-7
+1
-10
-15
-12
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METHOD 9253
CHLORIDE (TITRIMETRIC, SILVER NITRATE)
START
7.1 Place 50 ml
sample in porcelain
container
7.4 Add hydrogen
peroxide; mix for 1
minute
Yea
7.4 Is
sulfite ion
present in
sample?
No
7.1 Adjust pH to
8.3
72 Add 1.0 mL
potassium chromate;
stir; add silver
nitrate until
orange color
persis ts
7 . 3 Repeat steps
7.1 and 7.2 with
1/2 as much sample
diluted to 50 mL
75 Calculate
concentration of
chloride in sample
STOP
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1} U.a GOVERNMENT PRNTWQ OFFICE: 1981-281-724/28489
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