PB34-224674
Fused Silica Capillary Column GC/MS Quality
Control Protocol for the Determination of
Semivolatile Priority Pollutants
Acurex Corp.
Mountain View, CA
Prepared for
Environmental Monitoring Systems Lab.
Las Vegas, NV
Jul 84
{
of Commerce
Information Service
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EPA-600/4-84-068
July 1984
FUSEO SILICA CAPILLARY COLUMN
GC/MS QUALITY CONTROL PROTOCOL
FOR THE DETERMINATION OF
SEMIVOLATILE PRIORITY POLLUTANTS
by
Acurex Corporation
Energy I Environmental Division
555 Clyde Avenue
Mountain View, California 94039
Contract Number 68-03-3100
Prepared for
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89114
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
LAS VEGAS, NEVADA 89114
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TECHNICAL REPORT DATA
(Pteast rtad Instructions on the rtvcne be/on computing)
1. REPORT NO. 2.
EPA-600/4-84-068
3. RECIBJENT'S ACCESSION NO.
res*. aza
?lK£tf ^TLW'J^ILLARY COLUMN GC/MS QUALITY CONTROL
PROTOCOL FOR THE DETERMINATION OF SEMIVOLATILE
PRIORITY POLLUTANTS
S. REPORT DATE * '
July 1984
6. PERFORMING ORGANIZATION CODE
7. AUTHORIS)
Acurex Corporation
Energy & Environmental Division
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND AOORESS
Acurex Corporation
Energy and Environmental Division
555 Clyde Avenue
Mountain View, California 94039
10. PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
Contract Number 68-03-3100
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency--Las Vegas, NV
Office of Research and Development
Environmental Monitoring Systems Laboratory
.as Vegas, Nevada 89114
13. TYPE OF REPORT AND PERIOD COVERED
Resnonse P.eDort 1.2/82-5/84
14. SPONSORING AGENCY CODE
EPA/600/07
,18. SUPPLEMENTARY NOTES
I
16. ABSTRACT
This quality control protocol 1s intended to serve as a guide to those laboratories
employing fused silica capillary column (FSCC) gas chromatographlc/mass spectrometry
(6C/MS) techniques for the analysis of the extractable semivolatlle priority pollu-
tants. The document presents apparatus requirements and gives guidance for standards
preparation, extract mixing, system setup and calibration. Qualitative and quanti-
tative data Including internal standard assignment, quantitation mass, relative
retention times (RRT) values, Interlaboratory and Intralaboratory response factors
(RF) values are also presented. This document provides a schedule for GC/MS system
calibration requirements including: ion abundance calibration, column performance
testing, sensitivity verification, system linearity, and injection technique repro-
ducibility.
This protocol does not provide guidance regarding sample extraction and workup;
therefore, any quality control (QC) parameters related to this aspect c.f priority
pollutant analysis are not addressed 1n this protocol.
17. KEY V¥OROS AND DOCUMENT ANALYSIS
1. DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS
c. COSATl Field/Croup
10. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
IS. SECURITY CLASS (This Report)
UNCLASSIFIED
21. NO. OF t>AdE4
66
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
CPA Pom 2220-1 («•». 4.77)
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NOTICE
This report has been reviewed 1n accordance with the U.S. Environmental
Protection Agency's peer and administrative review policies and approved for
presentation and publication. Mention of trade names or commercial products
does not constitute endorsement or recommentiatlon for use.
11
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ACKNOWLEDGMENT
Acurex Corporation and the project team members express their appreciation
to the following Individuals who have reviewed this document and provided
helpful recommendations:
Mrs. Ann A1ford-Stevens, U.S. EPA, Environmental Monitoring Systems
Laboratory, Cincinnati, Ohio 45268
Dr. Bruce N. Colby, Systems, Science and Software, P.O. Box 1620,
La Jolla, California 92038
Dr. Randy Jenkins, J&W Scientific, Inc., 3871 Security Parx Drlva,
Rancho Cordova, California 95670
Dr. Bob Melton, U.S. EPA, Environmental Monitoring Systems Laboratory,
Cincinnati, Ohio 45268
Dr. Walt Shackelford, U.S. EPA, Environmental Research Laboratory,
Athens, Georgia 30613
111
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ABSTRACT
The purpose of this document 1s to establish standard conditions for the
fused silica capillary column (FSCC) gas chromatographlc/mass spectrometric
(GC/MS) analysis of the semlvolatlle priority pollutants (SVPP). Apparatus
requirements, column Installation instructions, and operating conditions for
spllt/splltless and on-column injection techniques are presented. Guidance for
riant^ds preparation, extract mixing, and sample Injection 1s also presented.
Qualitative and quantitative data are presented for the SVPP Including: Internal
standard assignments, quantitation 1ons, relative retention time (RRT) values,
Interlaboratory response factor (RF) values and RF relative standard deviations
(RSD's), and Intralaboratory RF data acquired with the two Injection techniques.
This quality control protocol provides an explicit schedule for fused
silica capillary column GC/MS standardization Including: 1on abundance
calibration, column performance testing, sensitivity verification, and ongoing
quality control during sample analysis. Examples of data acquired using this
protocol are also presented, as are forms required for compliance with the
documentation requirements.
This protocol does not provide guidance regarding sample "workup." The QC
parameters related to this aspect of priority pollutant analysis are not
addressed 1n this protocol.
1 v
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CONTENTS
Notice 11
Acknowledgment 111
Abstract . 1v
Figures v1
Tables v11
1 Scope and Application 1
2 Summary of Quality Control Requirements 3
2.1 Initialization Process 3
2.2 Sample Analysis 5
2.3 Ongoing Calibration 5
3 Apparatus 6
3.1 Gas Chromatograph 6
3.2 Mass Spectrometer .............. 7
3.3 Data System 7
4 Standards and Sample Extracts 9
4.1 Stock Solutions 9
4.2 Preparation of Stock Solutions 9
4.3 Internal Standards 12
4.4 Preparation of Calibration Standards 12
4.5 System Performance Standard 13
4.6 Mixing of Extracts 13
4.7 Safety Considerations 14
5 Qualitative and Quantitative Aspects of GC/MS Analysis ... 15
5.1 Determination of Relative Retention Times . . 15
5.2 Determination of Response Factors 15
5.3 Mass Spectral Criteria 22
6 Initialization 23
6.1 System Setup 23
6.2 Calibration 27
7 Ongoing QC Activities 37
7.1 Dally Calibration Check 37
7.2 System Recallbratlon 38
7.3 Internal Standards Verification 38
7.4 Saturation Effects 40
8 Documentation 41
9 References 51
Appendices
A. Labeled/Unlabeled — Compound Pairs 52
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FIGURES
Number Page
1 Flowchart summarizing QC requirements for FSCC GC/MS
analysis ........ 4
2 Chromatography/sensitivity check 28
3 Reconstructed ion chromatogram of column performance
test mixture 32
4 Reconstructed 1on chromatogram of composite priority
polutant standard (splltless Injection) ......... 35
6 Reconstructed 1on chromatogram of composite priority
pollutant standard (on-column Injection) .... 36
6 Area counts for the Internal standards versus time 39
v1
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TABLES
Number Page
1 TraceabiHty of Stock Solutions for Semi volatile Priority
Pollutants 10
la Traceabllity of Stock Solutions for Pesticides 11
2 Internal Standards Recommended for GC/MS Analysis 12
3 Interlaboratory Response Factors of Semivolatlle Priority
Pollutants 16
3a Interlaboratroy Response Factors of Pesticides 18
4 Intralaboratory Response Factors of "Semivolatlle Priority
Pollutants 19
4a Intralaboratory Response Factors of Pesticides 21
5 Gas Chromatograph1c/Mass Spectrometrlc Operating Conditions ... 25
6 DFTPP Ion Abundance Criteria 29
7 Composition of Capillary Column Performance Test Mixtures .... 31
8 DFTPP Ion Abundance Verification 42
9 FSCC GC/MS Initial QC Calibration Data -- Semivolatlle Priority
Pollutants 43
9a FSCC GC/MS Initial QC Calibration Data -- Pesticides 45
10 FSCC GC/MS Ongoing QC Data -- Semivolatlle Priority Pollutants. . 46
10a FSCC GC/MS Ongoing QC Data -- Pesticides 48
11 FSCC GC/MS Ongoing QC Data -- Absolute Area Counts for the
Internal Standards. 50
v11
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SECTION 1
SCOPE AND APPLICATION
This quality control protocol 1s Intended to serve as a guide to those
laboratories employing gas chromatographic mass/spectrometr1c (GC/MS)
techniques for the analysis of the semlvolatile priority pollutants (SVPP).
The document presents apparatus requirements and gives guidance for standards
preparation, extract mixing, Instrument setup, and calibration. Qualitative
an»< quantitative data including Internal standard assignment, quantitation
1ons, relative retention times (RRT) values, Interlaboratory and
Intralaboratory response factors (RF) values, are presented. Furthermore,
the document provides an explicit schedule for GC/MS system calibration
Including: 1on abundance calibration, column performance testing,
sensitivity verification, system linearity, and Injection technique
reproducibility.
Since users of this document will employ different Instrumentation, and
as the state-of-the-art of sample Introduction and other areas of
Instrumentation are continuing to evolve, selected aspects of this document
may become dated 1n time. However, because a simplified 1sotop1c dilution
quantification strategy is employed, the response factor accuracy criteria as
presented represent a fair test of laboratory accuracy and precision which
will not change greatly with "time. Also, as the multilevel, multlanalyte RF
values encode the entire laboratory standardization procedure from standard
1
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preparation to data transcription, data quality monitoring 1s facilitated and
standardized. The users of this protocol or modified versions of 1t will
find RF data useful 1n diagnosing Instrument/laboratory performance. Real
time monitoring of Internal standard 1on currents provides a verification of
instrument and/or laboratory performance during data acquisition.
2
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SECTION 2
SUMMARY OF QUALITY CONTROL REQUIREMENTS
This section summarizes the quality control (QC) procedures recommended
for fused silica capillary column GC/MS analysis (FSCC GC/MS) of the
semi volatile priority pollutants. The structure of the protocol 1s such that
It will give the user guidance for Instrument setup and calibration, as well
as criteria to assess the data quality. Since such criteria were established
based on Interleboratory and Intralaboratory evaluation of this protocol, the
users of this document should be able to reproduce the data provided 1n this
document.
The QC requirements described 1n this document can be classified Into
three groups:
• QC requirements during the Initialization process
• QC requirements during sample analysis
• QC requirements during the ongoing calibration
Figure 1 outlines the various steps and decisions that have to be made prior
and during sample analysis. A brief discussion of each of the three groups
of QC requirements follows.
2.1 Initialization Process
The Initialization process begins with the analysis of the system
performance standard (Section 4.5), followed by the analysis of three
calibration standards (concentrations of 20, 100, 200 pg/mL; Section 4,4),
3
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Analyze system
performance standard!
(4.5)
•J
I
«>
*
Recalibrate,
system trouble-
shooting
Perform multilevel
calibration (6.2.3)
r 1 te r 1 aV^
1n Section 6.2.s\
No
Mioor system
troubleshooting,
remake standards,
change coli#nn
x mer i /
3
z
«z
Uw
Cl
I
Figure 1. Flowchart eunrnarlilng QC requirement! for FSCC QC/MS analysis.
4
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The purpose of the system performance standard 1s to demonstrate 1on
abundance calibration via OFTPP at 60 ng (Section 6.2.1.1), and to provide
Information regarding system linearity and ability to chromatograph acidic
and basic compounds (Section 6.2.1). Analysis of the three calibration
standards 1s performed only after the QC requirements for the system
performance standard are met (Section 6.2.1). The purpose of the multilevel
calibration 1s to demonstrate system linearity for each corpound 1n the
calibration standard. Response factors for phenol, naphthalene, anthracene,
chrysene, and benzo(a)pyrene are utilized as the principal indicators of.data
acceptability. Acceptable values of these response factors are specified 1n
Section 6.2.3. Also given 1n Section 6.2.3 are the QC criteria that must be
verified during and after the multilevel calibration.
2.2 Sample Analysis
Analysis of samples begins after the Initialization criteria are met
(Section 6.2.3). During sample analysis the absolute areas of the
quantitation 1ons of the five Internal standards (phenol-d3, naphthalene-dg,
anthracene-djo. chrysene-di2» benzo(a)pyrene-di2) are monitored. Acceptance
criteria are specified 1n Section 7.3.
2.3 Ongoing Calibration
After 8 hours of data acquisition for samples, the system performance
standard (Section 4.5) and a single level calibration standard (Section 4.4)
are analyzed to verify the system performance. After 1 week from the
Initialization procedure and after any maintenance 1s performed on the
system, the Initialization procedure must be repeated.
5
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SECTION 3
APPARATUS
3.1 Gas Chromatograph
A gas chromatograph capable of linear temperature programming and
equipped with either spHtless or on-column Injectors is required. Varfous
hardware features available on most commercially available system are
implied.
3.1.1 Column
A 25- to 30m fused silica capillary, 0.25-nm ID (narrow bo.'e) or
0.32-mm ID (wide bore)» coated wtth S percent phenyl, 95 percent raethyl
silicone (SE-54 or DBS) end coupled directly to the ion source {end of the
capillary column 1s Inserted as far as possible Into the Ion source without
Intercepting the electron beam) is required. Both physically coated and
chemically bonded liquid phases are acceptable. The column shall meet the
specifications described in Section 6.2.2.2.
3.1.2 Carrier Gas Supply
Hydrogen or helium can be used as carrier gases. If hydrogen 1s
employed, safe handling practices muse be used. Hydrogen and helium must be
o*ygen-free. UTtrapure helium (99,999 percent) can be used directly from the
cylinder; however, it is desirable to Install a de-oxo cartridge to remove
residual oxygen in case of leaks.
6
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3.2 Mass Spectrometer
A mass spectrometer with electron Impact 1on source (70 eV) 1s required.
Both quadrupole and magnetic sector Instruments can be used 1f the required,
mass range (41 to 75 amu) can be scanned repetitively 1n 1 second or less. The
mass spectrometer shall produce a unit resolution (valley between m/z 441-443
less than 10 percent of the height of the 1on at m/z 441) background corrected
spectrum from 50 ng decafluorotrlphenylphosphlne (DFTPP) Introduced through the
GC Inlet. The spectrum shall meet the 1on abundance criteria specified 1n
Section 6.2.1.1. The mass spectrometer shall be Interfaced to the GC such that
the end of the capillary *.olumn reaches the 1on source, without Intercepting
the 1on beam.
3.3 Data System
A data system that collects and records all GC/MS data, processes GC/MS
data, generates quantitation reports, library searches, records response
factors, and generates multilevel calibration curves 1s required.
3.3.1 Data Acqulsltlcn
Mass spectra shall be collected continuously throughout the analysis and
stored on a mass storage device.
3.3.2 Mass Spectral Libraries
User-created libraries cortalnlng mass spectra obtained from analysis of
authentic standards shall be used to search the GC/MS data for the compounds of
Interest.
3.3.3 Data Processing
The data system shall be capable of searching, locating, identifying,
And quantltatlng the compounds of interest. Software processing routines
shall be employed to determine the retention times, relative retention times,
7
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artd Integrate peak areas for specific ions. Display of extracted ion current
profiles, mass spectra and Kbrei-jr searches are required tt> verify t*c
results*
3.3.1 Response Tact ore ar.fi WiiUUerei Calibrations
The data system shall also be capable of recortifng and ftalntalnlng lists
of response factors and multUevef cat (bat ion curves. Computations of
relative st#t*d4nl devfaticns are also useful for verifying t>je linMT-itj of
the calibration curve. If this capability 1s not a*&1lable, ti*esa data e*jst
be produced manually for every compound Df Interest to permit an updating of
response factors to wee*; the specifications of the protocol.
8
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SECTION 4
STANDARDS AND SAMPLE EXTRACTS
4.1 Stock Solutions
Stock solutions of compounds listed 1n Table 1 can be purchased as
Individual solutions or as mixtures with certification to their purity,
concentration, solvent, or can be prepared 1n-house from materials of known
purity. When not 1n use, all stock solutions should be stored 1n the dark at
-10°C 1n sealed vials with screw caps or cr1mp-top caps. All caps should
have Teflon-Hned Hds. A mark should be made on the vial at the level of
solvent such that any solvent loss can be detected. Stock solutions should
be brought to room temperature at least 1 hour prior to use. Son1cation
shall be considered whenever the compounds do not dissolve completely at room
temperature and/or precipitate due to lower temperatures during storage. The
stability of the stock solutions has not been determined. Therefore, all
stock solut' ..»s should be made fresh once a year, or sooner, 1f comparison
with the quality control check samples indicates a significant variation.
4.2 Preparation of Stock Solutions
Stock solutions of compounds listed 1n Table 1 are prepared 1n methylene
chloride, benzene, methanol, or a mixture of these solvents following the
safety precautions given 1n Section 4.9. Compound source, purity,
concentration of stock solution, solvent, date prepared, volume prepared, and
chemist name arc required. If purity of the stock solution 1s greater than
9
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TABLE 1. TRACEABILITY OF STOCK SOLUTIONS FOR SEMIVOLATILE PRIORITY
POLLUTANTS
Compound
Source
Purity
Concentration
of stock
solution
Date
prepared
Solvent
Chemist
N-NUrosod1methy ^ ami ne
B1s(2-chloroethylJether
2-Chlorophenol
Phenol
1.3-D1chlorobenzene
1.4-D1chloroben/ene
1.2-D1ch1orobenzene
B1s(2-chloro1sopropyl)ether
Hexachloroethane
N-Nltroso-dl-n-propylamlne
Nitrobenzene
Isophorone
2-N1tropheno1
2,4-Dlmethylphenol
B1s(2-chloroethoxy)methane
2.3-Dlchlorophenol
l,?,i-Tr1chlorobenzene
Naphthalene
Hexachlorobutadlene
4-Ch1oro-3-cresol
Hexachlorocyclopentadl ene
2,4,6-Trlchlorophenol
2-Chloronaphthalene
Acenaphthlene
Dimethyl phthalate
2,6-D1n1trotoluene
Acenaphthene
2.4-Dlnltrophenol
2,4-D1n1trotoluene
4-N1trophenol
Fluorens
4-Chlorophenyl phenylether
Diethyl phthalate
4,6*D1n1tro-2-cresol
D1phenyl amine
Azobenzene
4-Bronophenyl phenyl ether
Hexachlorobenzene
Pentachlorophenol
Phenanthrene
Anthracene
Dl-n-butyl phthalate
Fluoranthene
Pyrene
Benzidine
Butyl benzyl phthalate
Benz(a)anthracene
Chryiene
J, J^-Dlchlorobenxldlne
B1s(2-ethylhexy1(phthalate
Dl-n-octyl phthalate
Benio(J*k)f1uoranthenes
Benzo(a)pyrene
Indeno(1,2,3-cd)pyrene
D1benzo(a.h)anthracone
B«nio(gh1)p«rylene
10
(continued)
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TABLE 1. CONCLUDED
Compound
Source
Purity
Concentration
of Stock
solution
Date
prepared
Solvent
Chemist
Phenol-d3
Naphthalene-dg
Anthrac«ne-dio
Chrysene-d^
B«nzo(«)pyrene-di2
TABLE la. TRACEABILITY OF STOCK SOLUTIONS FOR PESTICIDES
Compound
Source
Purity
Concentration
of stock
solution
Date
prepared
Solvent
Chemist
Alpha-BHC
Garma-BHC
Beta-BHC
Delta-BHC
Heptachlor
Aldrin
4,4'.DDE
Dleldrln
4,4'-DDD
4,4'-DDT
Beta endosulfan
Endosulfan sulfate
Endrln
Alpha endosulfan
11
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98 percent, no correction of concentration is reedecf. Because of the Urge
number of cxnpomds used for calibration, it Is desirable that the stock
solutions are prepared so that dilutions of mixtures will permit
calibration with all cor^ounds fri a single corposlte stock solution.
4.3 Internal Standards
The following Internal standards are recommended: phenol-a^
naphthalene-da» anthracene-diQ, thrysene-dif, and ben2o(a)pyrene-diz* Other
Internal standards may also be employed (Table 2). For example, phenol-dt;
may be used Instead of phenol-d3» phenanthrene-djQ Instead of anthracene-diQ,
and benzo^M Jperylene-d^ Instead of bemo(a)pyre?ie-di2* selecting the
Internal standards, considerations given in Reference 1 must be addressed.
Stock solutions of the five Internal standards are prepared In methylene
chloride-benzene (3:1) et a concentration of 5 to 10 mg/tnL each. A spiking
solution of the f1*e Internal standards Is then made In methylene chloride at
X to 2 mg/mL.
4.4 Preparation of Calibration Standards
Calibration standards are prepared from a composite stock solution
(Section 4.2} or from several composite stock solutions containing the
TABLE 2. INTERNAL STANDARDS RECOMMENDED
FOR GC/MS AKALYS1S
Phfiflol-tij or phenol ~d$
Naphthalene-dg
Anthracerie-djg or Phenanthrene-djQ
Cho
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compounds of Interest. The concentrations of the calibration standards are
20, 100, and 200 pg/mL. The 200-pg/mL calibration standard should be
employed to prepare the 20- and 100-pg/mL calibration standards. The
calibration standards should contain the Internal standards at 20 pg/mL for
phenol-d3, naphthalene-dg and anthracene-djo. and 40 pg/mL for chrysene-dj2
and benzo(a)pyrene-di2* Every calibration standard vial should be dated
accordingly (date, solvent, concentration, chemist name). All details
regarding the preparation of the calibration standards should be kept 1n a
logbook for standards traceablHty.
Calibration standards should be prepared fresh, at least nnce a month,
or more often 1f there are signs of deterioration. Standards will remain
acceptable 1f the response factors of the compounds of Interest remain v.'thln
±20 percent of the average response factors published 1n this document. When
not In use, standards should be stored at -10°C Immediately.
4.5 System Performance Standard
The system performance standard contains anthracene-djo, decafluorotrl-
phenylphosphlne (DFTPP), benzidine, and pentachlorophenol at concentrations
of 20, SO, 100, and 50 pg/mL, respectively. Additional compounds may be
added to this mixture as an Initial check on system linearity. The following
compounds are recommended: anthracene, N-n1troso-d1-n-propylam1ne, and
2-chloronaphthalene at concentrations of 200, 20, 20 pg/mL, respectively. An
instrument with adequate sensitivity and dynamic range should give response
factors of appro/1mately 1.00, 0.05, and 0.60, respectively.
4.6 Mixing of Ext racts
Composite 0.5 mL of the acid and base/neutral extracts in a precleaned
2- to 5-mL screw cap vial with Teflon-lined septum. Although th« combined
13
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acid and base/neutral standards were found to be stable for 1 north based on
response factor measurements (Reference 6), the con*pos1ting uf sample
extracts should be done i;nwed1ately prior to Injection* Note that this
compositing step Introduces a dilution factor of two which rust be considered
during data reduction,
4.7 Safety Considerations
The toxicity and carcinogenicity of each compound or reagent used in
this method have been predstly determined; however, each compound should
be treated as d potential health hazard. Exposure to these compounds should
be reduced to the lowest possible "level. Every laboratory should be
responsible for maintaining a current awareness file of OSrtA regulations
regarding the safe handling of the chemicals specified In this method. A
reference file of data handling sheets should also be made available to all
personnel involved in these analyses. Additional Information on laboratory
safety can be found In Reference 9.
The following compounds covered by this method have been classified as
known or suspected human or mammalian carcinogens; benzo(a)anthracene,
benzidine, 3,3'-d1cMorobenz1d1ne, bertzo(a}pyrene, dibenioU,h)anthracene,
and N-n1trosod1methylamiiie. Primary standards of these toxic confounds shall
be prepared 1n a hood, and a NIOSH/MESA approved toxic gas respirator should
be worn when high concentrations are handled.
14
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SECTION 5
QUALITATIVE AND QUANTITATIVE ASPECTS OF GC/MS ANALYSIS
5.1 Determination of Relative Retention Times
Relative retention times (RRT) are determined using the five Internal
standards (phenol-d3, naphthalene-dg, anthracene-dm, chrysene-di2» and
benzo(a)pyrene-di2), as shown 1n Tables 3# 3a, 4, and 4a. The values given
were obtained with new columns under the conditions noted. As columns
shorten, the RRT's change. Variations 1n columns, oven temperatures, and
temperature programs can also have effects on RRT's. In properly operated
systems this variation 1s small (relative standard deviation of consecutive
injections of standards should average approximately 0.2 percent). The
identification criteria requlras that the dally RRT of the priority pollutant
should be within ±0.01 from the average RRT determined using the multilevel
calibration data.
5.2 Determination of Response Factors
Response factors are determined using the appropriate Internal standard
as Indicated 1n Tables 3, 3a, 4, and 4a. Tables 3 and 3a give the average
response factors determined 1n an interlaboratory study. Tables 4 and 4a
give the average response factors determined by Acurex using both the
splltless and the on-column Injection techniques.
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TABLE 3. INTERLABORATORY RESPONSE FACTORS OF SEMIVOLATILE PRIORITY
POLLUTANTS {Splltless Injection)
Compound
I.S.«
Quantitation
Iond
RRTb.C
RFe.f
RSD
Pheno1-2.4.6-d3 (dO
I.S.
97
1.00
1.00
N-hltrosod1methyl amine
d3
74
0.52
0.42
46.0
B1s(2-chloroethyl}ether
d3
93
1.01
1.01
9.4
2«Chlorophenol
d3
128
1.01
0.79
11.5
Phenol
d3
94
1.01
1.10
5.5
1>3-D1chlorobenzene
d3
146
1.01
0.72
17.2
1.4-Dichlorobenzene
d3
146
1.03
0.90
17.0
1.2-D1chlorobenzene
d3
146
1.67
0.75
18.0
Ms(2-chloro1sopropyl! etktr
d3
77
1.10
0.22
20.8
Hexachloroethane
d3
117
1.12
0.35
14.1
Naphthalene-d8 (dp)
I.S.
136
1.00
1.00
N-N1troso-d1-n-propyl amine
d8
130
0.89
0.053
4.1
Nitrobenzene
dS
123
0.56
0.19
17.1
ksophorone
d8
62
0.94
0.84
19.8
2-N1tropheool
dQ
139
0.95
0.22
12.3
2.4-D1methyl phenol
da
122
0.95
0.32
11.2
B1s(2-chloroethoxy)methane
d8
93
0.98
0.51
15.6
2,4-D1chloropheno1
d8
162
0.99
0.30
13.1
1,2,4-Trlchlorobenzene
d8
180
1.00
0.32
16.0
Naphthalene
d8
128
1.00
1.08
9.7
Hexachlorobutadlene
d8
225
1.03
0.13
24.7
4-Chloro-3-cresol
08
142
1.10
0.26
15.7
Hexachlorocyc1opentadlene
d8
237
1.14
0.15
24.8
2.4.6-Trlchlorophenol
dB
146
1.16
0.19
7.9
2-Chloronaphthalene
d8
162
1.18
0.63
2.7
Acenaphthytene
d3
152
1.25
0.72
24.9
Dimethyl phthalate
d8
1.26
0.62
14. B
Z.6-Dlnltrotoluene
dS
165
1.16
0.15
16.5
Anthracene-dlO (dm)
I.S.
188
1.00
1.00
• m
Acenaphthene
dlO
154
0.84
0.81
22.4
2.4-Dinltrophenol
did
184
0.86
0.07
35.2
2.4-D1n1trotoluene
dlO
165
0.87
0.23
2.0
4-N1trophenol
dlO
139
0.89
0.10
42.0
Fluorene
dlO
166
0.90
0.96
17.9
4-Chlorophenyl phenylether
dlO
204
0.90
0.47
22.4
Diethyl phthalate
dlO
149
0.90
0.91
10.1
4,6-dInltro-2-cresol
dlO
146
0.91
0.10
35.7
D1phenyl amine
dlO
169
0.90
0.58
" 11.8
Azobenzene
dlO
77
0.92
1.05
--
4-Bromophenyl phenyl ether
dia
248
0.95
0.24
23.6
Hexachlorobenzene
dlO
2B4
0.96
0.24
20.6
(continued)
16
-------
TABLE 3. (concluded)
Compound
I.S.®
Quantitation
Ion«
RRTb»c
RFe»*
RSD
Pentachlorophenol
dlO
266
0.98
0.13
11.6
Phenanthrene
dlO
178
1.00
1.16
11.2
Anthracene
dlO
178
1.00
1.15
8.7
D1-n-butyl phthalate
dlO
149
1.07
1.28
10.6
Fluorantnene
dlO
202
1.12
1.07
7.7
Pyrene
dlO
202
1.15
1.08
9.1
Cnrysene-dl2 (di?)
I.S.
240
1.00
1.00
--
Benzidine
dl2
184
0.86
0.24
47.0
Butyl benzyl phthalate
dl2
149
0.96
0.84
63.2
Benz(a)antnracene
dl2
228
1.00
1.11
5.2
Chrvsene
dl2
228
1.00
1.02
7.8
3.3 -Dlchlorobenzldlne
dl2
252
1.00
0.28
8.3
Bls( 2-ethylhexylJphthalate
dl2
149
1.02
0.88
45.7
D1-n-octyi phtnalate
dl2
14$
1.68
1.34
44.6
Benzo,
s)pyrene-dl2
I.S.
264
1.00
1.00
mm
Benzo
|+lc)fluoranthenes
dlZB
252
0.98
1.10
17.2
Benzo
s pyrene
dl2B .
252
1.00
1.00
11.1
Indeno
1.2.3-cd)pyrene
dlZB
in
1.09
0.45
14.3
Dlbenz
a.hjanthracene
dlZB
278
1.10
0.58
15.1
Benzo gh1)perylene
dl2B
Hi
1.12
0.64
15.8
See footnotes at end of Table 3a.
17
-------
TABLE 3a. 1NTERLAB0RAT0RY RESPONSE FACTORS OF PESTICIDES
(Splitless Injection)
Compound
I.S.«
RRTb.c
Quantitation
Iond
RRF«»f
RSD
Alpha-BHC
dlQ
0.95
181
0.12
11
Gamma-BHC
dlO
0.99
181
0.11
1 i
Beta-BHC
dlO
1.02
181
0.12
ii
Delta-BHC
dlO
1.03
181
0.09
Heptachlor
did
1.07
272
0.05
26
Aldrln
dlO
1.10
263
0.14
17
Heptachloroepoxlde
<110
1.15
355
0.06
4
4.4'-DDE
dl2
0.91
245
0.23
23
Dieldrin
dt2
0.91
79
0.27
22
4.4'-DDD
dl2
0.94
235
0.40
4
4.4'-bbt
dl 2
0.96
235
0.21
16
Beta endosulfan
dlZ
0.94
195
0.04
13
Endosulfan sulfate
dl2
0.96
272
0.06
3
Endrln
dl2
0.92
81
0.07
1?
Alpha endosulfan
dl2
0.88
195
6.03
5
aI.S. is the reference Internal standard for each compound.
&RRT 1s the retention time relative to the reference Internal standard.
cRetent1on times of the I.S. are as follows: phenol-d3 5:24 min;
naphthalene-rig 8:45 itfn; anthracene-djQ 16:46 mini chrysene-dj2 23:14 min;
and benzo(a)pyrene-d^2 26:53 mln.
^Quantitation 1ons are chosen primarily for reliability of Identification
and lack of Interferences.
*RF Is the response factor relative to the reference internal standard. These
values are averages of those determined In four laboratories (Reference 2).
*Data taken from Reference 2.
18
-------
TABLE 4. INTRALABORATORY RESPONSE FACTORS OF SEMIYOLATILE PRIORITY
POLLUTANTS
Compound
SpHtless
Injection
On-Column
Injection
RF®
RSD
"KF®
RSD
RRT®
RSD
N*N1trosod1methylamine
0.72
24.5
mm m
mmm
m~m
B1s(2-chloroethyl ether
0.96
3.0
1.078
11.4
0.988
0.2
2-Chlorophenol
6. ft
4.0
1.001
9.2
0.994
6.2
Phenol
1.16
2.8
1.335
5.3
1.002
0.3
1.3-D1chlorobenzene
0.66
0.9
0.886
9.5
1.065
0.7
1,4-D1chlorobenzene
0.85
3.8
1.090
12.3
1.106
0.6
1.2-D1chlorobenzene
0.72
3.5
0.942
12.8
1.235
1.0
Bis(2-chlorolsopropyllether
0.26
7.9
0.232
6.9
1.389
1.3
Hexachloroethane
0.35
6.2
0.428
9.0
1.455
1.5
N-n1troso-d1-n-propylam1ne
0.035
3.8
0.092
7.7
0.725
0.8
Nitrobenzene
0.18
0
0.251
4.4
0.750
0.4
Isophorone
0.89
1.7
0.810
14.7
0.652
0.6
2-N1trophenol
0.23
6.7
0.326
1.4
0.872
0.3
2.4-D1methylphenol
0.30
3.3
0.367
5.6
0.940
0.3
BI s C2-chloroethoxy'methane
0.53
1.9
0.541
3.8
0.964
0.2
2,4-D1chlorophenol
0.29
0
0.388
4.4
0.980
0.1
1.2,4-Trlchlorobenzene
0.30
1.9
0.418
5.8
0.992
0.1
Naphthalene
1.01
6.5
1.089
19.2
1.008
0.1
Hexachlorobutadlene
0.12
0
0.188
5.4
1.093
0.3
4-Chloro-3-cresol
0.29
2.0
0.383
7.2
1.297
0.3
Hexachlorocyclopentadlene
0.15
7.5
0.259
6.9
1.377
0.3
2.4.6-Trlchlorophenol
0.21
5.4
0.313
7.7
1.425
0.3
2-Cnloronaphthalene
0.64
2.4
0.814
6.1
1.466
0.3
Acenaphthylene
o.ee
4.3
0.994
6.5
1.626
0.3
Dimetnyl phthalate
0.73
3.1
1.018
4.1
1.656
0.2
fc,6-D1n1trotoluene
0.18
6.3
0.275
7.4
1.671
0.2
Acenaphthene
0.61
5.0
0.757
20.0
0.745
0.0
2,4-D1n1trophenol
0.083
38.3
0.166
16.8
0.768
0.2
2,4-Dlnltrotoluene
0.23
11.1
0.325
2.5
0.795
0.2
4-N1trophenol
0.18
6.3
0.207
17.6
0.806
0.1
Fluorene
0.77
7.8
0.866
15.6
0.833
0.1
4-Chlorophenyl phenyl ether
0.38
4.1
0.524
10.7
0.843
0.1
Diethyl phthalate
0.82
4.6
1.018
14.9
0.849
0.1
4.6-D1n1tro-2-cresol
0.14
16.9
0.219
11.0
0.863
0.3
Diphenyl amine
0.52
4.0
0.587
B.O
0.870
0.1
Azobenzene
mmm
mmm
0.069
19.1
0.870
0.1
4-Bromophenyl phenyl ether
0.21
4.8
0.308
^.0
0.926
0.1
Hexachlorobenzene
0.24
4.2
0.339
5.7
0.941
0.1
Pentachlorophenol
0.14
7.1
0.205
10.4
0.931
0.1
(continued)
19
-------
TABLE 4. (concluded)
Compound
SpHtless
Injection
On-Column
Injection
RF«
RSD
RSD
RRT«
RSD
Phenanthrene
1.06
1.4
1.221
7.2
0.996
0.1
Anthracene
1.07
8.0
1.045
25.0
1.003
6.1
U1-n-butyl phthalate
1.44
9*6
1.428
22.3
1.136
0.1
Fluorantnene
1.10
6.6
1.201
12.4
1.202
0.1
Pyrene
1.13
2.8
1.160
6.4
1.237
0.1
Benzidine
0.15
24.7
b
b
b
b
Butyl-benzylphthalate
0.67
10.8
0.698
19.2
0.957
0.1
Benz1, a) anthracene
1.04
2.4
1.043
5.0
0.998
6.1
Chrysene
1.02
«.l
0.878
21.1
1.002
0.1
3.3 -D1chlorobenz1d1ne
0.31
0
b
b
b
b
Bis(2-ethylhexyl phthalate
1.07
3.9
1.012
21.7
1.637
0.1
Dl-n-octyl phtnalate
1.70
5.4
1.347
20.4
1.108
6.1
Benzo
j+k; fluoranthenest)
...
...
...
—
Benzo
a)pyrene
0.88
2.3
0.961
21.3
1.002
0.1
Indenoi1,2.3-cd)pyrene&
—
• ••
—
—
—
—
D1benzo(a. h)anthracene
0.60
2.5
0.784
17.3
1.153
0.2
Benzo(qh1 perylene
0.65
8.6
0.815
15.3
1.185
6.3
See footnotes at end of Table 4a.
20
-------
TABLE 4a.
INTRALABORATORY RESPONSE FACTORS OF PESTICIDES
Compound
Splltless
Injection
On-Column
Injection
RF«
RSD
RSD
RRT«
RSD
A1pha-BHC
0.143
9.1
0.169
7.0
0.931
0.1
Gflirma-BHC
0.154
13.7
0.269
9.2
0.982
0.1
Beta-BHC
0.188
28.4
0.247
24.3
1.024
0.1
Delta-BHC
0.103
6.2
0.127
5.0
1.024
0.1
Heptachlor
0.089
10.9
0.143
9.5
1.087
0.2
Aldrlnt)
---
• • •
m mm
...
...
...
4,4'-DDE
0.243
14.8
0.246
8.7
0.883
0.1
Dleldrln
0.242
23.3
0.204
5.0
0.880
0.1
4.41-DDD
0.458
12.1
0.476
6.0
0.923
0.1
4.4'-DDT
0.346
8.6
0.433
5.8
0.957
0.1
Beta endosulfan
0.03S
7.4
0.041
7.1
0.910
0.1
Endosulfan sulfate
b
b
0.089
12.0
b
b
Endrln
0.052
28.6
0.076
10.8
0.880
0.1
Alpha endosulfan
0.034
5.6
0.036
5.2
0.855
0.1
Heptachlor epoxide
0.059
6.2
0.077
8.5
1.196
0.2
•Number of determinations Is 3 for splltless Injection and 7 for on-
column Injection.
^Compound not present 1n the standard.
21
-------
The response factor (RF) 1s defined as follows:
o, A* • "I-S.
¦ AI.S.
where:
Ax ¦ area counts of the quantitation 1on of compound X
Ais. " area counts of the quantitation Ion of Internal standard
Wx ¦ amount of compound Injected
WI.S. " amount of Internal standard Injected
For the purposes of Interlaboratory reproducibility, all compounds must
be quantltated using the specific quantitation 1ons listed 1n Tables 3 and
3a. These 1ons have been chosen to minimize Interferences as well as to
minimize mass differences from the Internal standard. If any Interference
occurs at the primary quantitation 1on, a secondary 1on may be used as
described 1n Method 625 (Reference 7).
5.3 Mass Spectral Criteria
All fragment 1ons with Intensities greater than 10 percent of the base
peak 1n the mass spectrum of the standard must be present 1n the mass
•poetrum of the compound 1n the sample. Relative abundances must agree
within ±20 percent. Extraneous 1ons 1n the mass spectrum of the compound 1n
the sample must be accounted for 1n the Interpretation of the match.
22
-------
SECTION 6
INITIALIZATION
6.1 System Setup
6.1.1 Column Installation
Install capillary column following the recommendations described below:
• Slide fitting nuts and ferrules onto each end of the column;
graphltlzed Vespel ferrules are recommended. Cut 2 to 3 cm from
each end of the column by scoring the circumference with a diamond
cutting tool, and break off the column ends with the fingers.
• For spHtless Injectors, Insert column Into the Injection port to a
position 1.0 to 2.0 cm from the tip of a fully Inserted syringe
needle. Tighten the nut finger tight, then turn 1t one full turn
with a wrench.
• For on-column Injectors, Insert column Into the Injector as far as 1t
will go, so the column end sits 1n the taper of the alignment device.
• Insert the outlet end of the column up to the 1on source and
position 1t. Tighten the nut finger tight, then turn 1t one full
turn w1t^ a wrench.
• Leak test the column Inlet and outlet fittings with argon using the
mass spectrometer as a leak detector.
• Position the column Inside the oven such that any contact between
the oven wall and the column 1s avoided.
23
-------
6.1.2 Flow Adjustment
With narrow bore columns (0.25-mm ID) adjust the column head pressure
until the linear gas velocity 1s 25 to 30 cm/s at 30°C 1f helium 1s used as
carrier gas. If hydrogen 1s used, the desired flow 1s 40 to 60 cm/s. For
wider bore columns (0.32-mm ID) the gas flow 1s determined by the vacuum
system. Typical linear gas velocities for wider bore columns are 30 to
40 cm/s for helium and 60 to 80 cm/s for hydrogen. Whereas hydrogen gives
the ultimate 1n chromatographic performance 1t might create problems with
mass spectrometer performance (e.g., Increase the noise level).
6.1.3 GC/MS Operating Conditions
Table 5 summarizes the recommended gas chromatographic and mass
spectrometrlc operating conditions. In certain situations, 1t may be
difficult to match exactly the operating conditions suggested 1n this
document. In such cases, utilize the above parameters as guidance, but
employ the criteria published 1n this document to determine 1f a slight
variance 1s permissible.
6.1.3.1 SpHtless Injection
"Hot" needle injections are recommended for spHtless Injections.
Insert syringe containing the sample drawn Into the syringe barrel. Walt
0.1 mln for the needle to reach the Injector temperature; at 0.1 m1n a 1-pL
sample 1s Injected quickly (0.1 m1n). The needle Is removed Immediately
after 0.1 m1n. Thorough rinsing of the syringe foil owing sample injection
must be performed.
The performance of the splUless Injection 1s the most critical step 1n
the procedure. Poor Injections lead to the discrimination of either the low
or the high boiling compounds and consequently to poor analytical precision.
24
-------
TABLE 5. GAS CHROMATOGRAPHIC/MASS SPECTROMETRIC OPERATING CONDITIONS
INJECTION: SPLITLESS OR ON-COLUMN
SpHtless Injector
Dimensions:
Material:
Temperature:
Flowrates:
Sweep flow:
Split flow:
SpHtless time:
Column Temperature Program
Initial temperature:
Initial hold:
Rate:
Final temperature:
Final hold:
2 to 3 mm ID
Quartz
270°C
10 mL/mln
35 rt./m1n
30 to 6U sec
30°C for splltless; 35°C for on-column
4 m1n for splltless; 2 m1n for on-column
l0°C/m1n
270°C
Until benzo(gh1)perylene elutes
Mass Spectrometer
GC/MS transfer line temperature: 275°C
Mass range: 41-475 amu
Cycle time: Is or less
Electron energy: 70 eV
Source temperature: 280° to 300°C
Start data acquisition with start of GC temperature program.
25
-------
A correct Injection technique will yield precise absolute area counts for the
Internal standards (RSD <10 percent).
6.1.3.2 On-Column Injection
A J&W on-column Injector (J&W Scientific, Ranch Cordova, California)
was used to generate the data reported In this document. Other on-column
Injectors are available commercially. The Injector consists of a preclslon-
bore glass Insert that holds the fused silica column and the fused s111 ca
needle 1n precise alignment, facilitating a smooth Injection. Cooling of the
Injector during sample Injection 1s allowed. Injection of the sample onto the
column 1s performed via a Teflon stopcock using a fused silica needle. A Model
1701 RN syringe (Hamilton Company, Reno, Nevada) fitted with a 197 mm length x
0.15 mm ID x 0.21 mm OD fused s111ca needle was employed for the on-column data
reported from this study.
The steps that must be followed carefully when using on-column Injection
techniques are: syringe cleaning, syringe loading and sample Injection. The
syringe, fitted with the fused silica needle, 1s somewhat more difficult to
clean than a conventional syringe. The needle should be rinsed carefully. A
flight vacuum may be applied to remove residual solvent left 1n the syringe.
Prior to loading the sample, fill the syringe with solvent. Depress the
plunger all the way expelling excess solvent (unless a solvent flush technique
1s used). After loading the sample Into the fused silica needle, rinse the
outside surface of the needle with solvent to prevent possible contamination of
the Injector.
Set up the GC conditions as specified 1n Section 6.1.3, turn on the
cooling gas, and Insert the needle into the Injector. Depress the plunger at
such rate (typically 5 seconds for 1- pi Injections) to avoid pressurlzatlon of
-------
the column with excessive solvent vapor. The Initial temperature of the oven
should be set at least at the boiling point of the solvent (slightly higher
temperatures than the boiling point of solvent are preferred). The cooling
gas 1s turned off when oven temperature 1s at 40°C.
The performance of the Injection technique 1s the most critical step 1n
the procedure. Poor Injections lead to poor analytical precision. A correct
injection technique will yield precise Internal standard areas (RSD
<10 percent).
6.2 Calibration
6.2.1 System Performance Test
Prior to analysis of any calibration standards, check the GC/MS system
to see that acceptable performance criteria are achieved for DFTPP, benzidine
and pentachlorophenol. The composition of the system performance test was
given 1n Section 4.5. Figure 2 provides an example of a system performance
standard.
6.2.1.1 DFTPP Performance Test
Obtain a background corrected mass spectrum of DFTPP and check that the
1on abundance criteria specified 1n Table 6 are achieved. If all the
criteria are not achieved, the mass spectrometer needs to be retuned.
6.2.1.2 Chromatographic Performance
Response factors for pentachlorophenol and benzidine must be greater
than 0.05. The benzidine tailing factor must be less than 2. The
pentachlorophenol tailing factor must be less than 2. Calculation of the
tailing factor 1s illustrated 1n Section 6.2.2.4. Clip off few Inches from
the column or rinse column with solvent 1f the tailing factor criteria cannot
be achieved.
27
-------
12.6*
184-
100.0-
188-
22.4-
198-
277
7712
10066
,—
277
61440.
90470.
T"
313
13744.
17906.
403
3952.
12698.
I
Benzidine
Antti»cer»-d,a
DFTPP
7712.
1$4.66B
~0.600
61440.
188.066
- 0.600
13744.
198.069
£0.600
19.4-
m
266-
r
203
11904.
21664.
I I I
Pentaohlerophenol
11904.
266.000
10.800
—r~
200
3:20
I
300
6:00
I I »
400 600
6:40 8:20
—1
600
10:00
Scan
Time
Figure 2. Chromatography/sens1t1v1ty check.
28
-------
TABLE 6. DFTPP ION ABUNDANCE CRITERIA9
m/z
Ion Abundance Criteria
51
30 to 60 percent of mass 198
68
less than 2 percent of mass 69
70
less than 2 percent of mass 69
127
40 to 60 percent of mass 198
197
less than 1 percent of mass 198
198
base peak, 100 percent relative
abundance
199
5 to 9 percent of mass 198
275
10 to 30 percent of mass 198
365
greater than 1 percent of mass !
198
441
less than mass 443
442
greater than 40 percent of mass
198
443
17 to 23 percent of mass 442
aData taken
from Reference 8
29
-------
6.2.1.3 System Linearity
Demonstrate that the 20 ng anthracene-dio produces an area at m/z 188
approximately one-tenth of that required to exceed the linear range of the
system. This value must be determined by experience for each system.
6.2.2 Column Performance Testing (Optional)
Evaluate the performance of each new column Immediately following
installation Into the GC/MS system. Install the capillary column following
the procedures given 1n Section 6.1.1. Adjust the carrier flow at 40 to
60 cm/s for hydrogen or 25 to 30 cm/s for helium. Set the spHtless or the
on-column Injector as specified 1n Section 6.1.3. Inject 1 wL of a column
performance test mixture containing 20 ng each of 2,6-dlmethylphenol,
2,6-dimethylaniline, 1-octanol, nonanal, Cjq and alkanes, three fatty
acid methyl esters, etc. Such column performance test mixtures are available
commercially. Table 7 shows the contents of three commercial column test
mixtures. Figure 3 shows a chromatogram of the column performance test
mixture and how the various parameters describing column performance (Neff,
column pH, tailing factor, separation number) are determined.
Number of effective theoretical plates (Neff) 1s determined from the
Cn-fatty acid methyl ester peak using the equation:
where t'r 1s the adjusted retention time of Cn-methyl ester and Wo.5 1s the
peak width at half height. The number of effective plates for acceptable
columns must be at least 2,500 plates per meter of column.
Column pH is determined by the ratio of peak heights of 2,6-d1methyl-
aniline to that of 2,6-d1methyl phenol. A value of 0.5 to 1.5 1s acceptable.
Neff ¦ 5.454
30
-------
TABLE 7. COMPOSITION OF CAPILLARY COLUMN PERFORMANCE TEST MIXTURES
Test Mlxturs
Composition
Varlan
2-Octanone
0.2
yg/yL
P/N 82-005049-01
1-Octanol
0.2
pg/yL
(nonpolar)
Naphthalene
0.2
yg/yL
2,6-D1methyl phenol
0.2
yg/yL
2,4-D1methylan1l1ne
0.2
yg/yL
C^-Alkane
0.2
yg/yL
Ci3-Alkane
0.2
yg/yL
A1ltech
Ci3-Alkane
0.1
pg/yL
TP-5 (polar)
C^-Alkane
0.1
yg/yL
Cj5~Alkane
0.1
yg/yL
Cjfi-Alkane
0.1
yg/yL
1-Octanol
0.5
yg/yL
5-Nonanone
0.3
yg/yL
2,6-D1 methylan111ne
0.4
yg/yL
2,6-01methylphenol
0.4
yg/yL
Naphthalene
0.5
yg/yL
Analabs
Ci2-Ac1d methyl ester
41
ng/yL
Test probe LPK-013F
C^-Acld methyl ester
41
ng/yL
(general purpose)
Cio-Ac1d methyl ester
42
ng/yL
ClO-Alkane
28
ng/yL
Cn-Alkane
29
ng/yL
1-Octanol
36
ng/yL
Nonanal
40
ng/yL
2,3-Butanedlol
53
ng/yL
2,6-01 methyl aniline
32
ng/yL
2,6-01methy1phenol
32
ng/yL
01cyclohexylamine
31
ng/yL
2-Ethylhexano1c acid
38
ng/yL
31
-------
lOO.On
Column pH
Peak asymmetry
(AS) of 1-Octanol and
activity of column
RIC -
N
¦o
e
c
m
3
m
«
N
Separation
number (SN)
200
3:21
1—
400
6:42
1
600
10:03
800
13.24
Scan
Time
Figure 3. Reconstructed Ion chromatogram of column performance test mixture.
-------
Column activity toward polar compounds 1s determined as the asymmetry of
the l-:ctanol peak and as the ratio of the 1-octanol peak to that of
Cn-alkane. The tailing factor or peak asymmetry 1s evaluated by drawing a
perpendicular from the apex of the peak to the baseline and measuring the
width from the front of the peak to the perpendicular line (Wp) and from the
back of the peak to the perpendicular line (Wg); Wp and Wg are measured at
10 percent height from the baseline.
AS « JJB
w
The ratio of peak height of 1-octanol to Cji-alkane should be higher than
0.3. Tailing factors between 0.75 and 2 are acceptable.
Separation number (SN) 1s defined by equation:
SN = . L_ - 1
Wj + Wg
where D is the distance between C^q and Cn-alkanes and Wj and W2 are their
widths at half-height, respectively. A separation number of 10 or greater 1s
acceptable.
6.2.3 Multilevel Calibration
Inject 1 uL of each composite standard (concentration 20, 100, and
200 pg/mL) containing the base/neutral, acid and pesticide priority pollutants
and acquire a complete GC/MS run. Utilize the response factors for phenol,
naphthalene, anthracene, chrysene, and benzo(a)pyrene to determine the data
quality (acceptable values of these response factors must be within
1.00 ±0.2). These values should be calculated as acquired so that 1f the above
criteria are not met, corrective action can be Initiated.
33
-------
If response factors for phenol, naphthalene, anthracene, chrysene, and
benzo(a)pyrene are within 1 ±0.2, then process the data for the remaining
compounds.
The following QC criteria must be met:
o All response factors should be within 20 percent of the values given
in Tables 3, 3a, 4, and 4a. All compounds must be detected at the
20 ng level with the possible exception of benzofluoranthenes which
may overlap.
® Relative retention times of each compound in each run must be within
±0.01 from the average RRT, except for N-n1trosod1methylamine which
has to be resolved from the solvent peak.
o The relative standard deviations of the response factors of phenol,
naphthalene, anthracene, and chrysene, must be less than 10 percent
except for benzo(a)pyrene where 15 percent or less 1s acceptable.
© The average response factors of N-n1troso-d1-^-propylamine and
2-chloronaphthalene should be 0.050 and 0.60, respectively.
Reconstructed 1on chromatogrami of e standard containing all compound*
listed 1n Tables 3 and 3a, are given 1n Figures 4 and 5 for spHtless
Injection and on-column Injection, respectively. Examples of the
chromatographic resolution observed for the labeled/unlabeled compound pairs
are given in Appendix A.
34
-------
8:45 17:30 26:15 35:00 Time
Figure 4. Reconstructed 1on chromatogram of composite priority pollutant
standard (splitless Injection).
-------
100.0-1
RIC-
U
o\
1000
15:55
1500
23:52
Scan
Time
Figure 5. Reconstructed 1on chronatogram of composite priority pollutant
standard (on-colu«n Injection).
-------
SECTION 7
ONGOING QC ACTIVITIES
7.1 Dally Calibration Check
At the beginning of each day and at every 8 hours during which analyses
are performed, the system performance and calibration must be verified.
Analysis of the system performance standard (Section 4.5) shall be used to
verify the performance criteria 1n Sections 6.2.1.1 and 6.2.1.2. Analysis of
the 100-pg/mL calibration standard (Section 4.4) shall be used to verify the
following:
o All compounds Including benzidine, 2,4-d1n1trophenol, and
N-n1trosod1methyl amine must be detected,
o The RF's of phenol, naphthalene, anthracene, chrysene, and
benzo(a)pyrene must not vary by more than 16 percent from the
average RF determined 1n the multilevel calibration.
« The RF's of the other compounds 1n Tables 3 and 3a must not vary by
more than 20 percent from the average RF determined 1n the
multilevel calibration.
If these criteria are met, then the RF values for all compounds are added to
the 11st of those obtained previously. The RF values used 1n quantitation
are the average values of those RF's obtained since the most recent
multilevel calibration. If the calibration check criteria are not met,
37
-------
system recaHbration 1s required. Alternatively, preparation of fresh
standards and/or column replacement should be considered.
7.2 System RecalIbration
System recallbratlon 1s required whenever the QC criteria Indicated 1n
Section 7.1 have not been met and/or when one week has elapsed since the most
recent three level calibration was performed. Furthermore, any major system
maintenance such as source cleaning, etc., requires recallbratlon.
7.3 Internal Standards Verification
During or immediately after each data acquisition, the presence of all
five internal standards (phenol-d3, naphthalene-ds, anthracene-dig. chrysene-
dj2> benzo(a)pyrene-di2) must be verified. Absolute area counts from the
Integrated 1on currents of each Internal standard should be recorded for each
standard and sample analyzed. Graphs recommended for this purpose are
presented 1n Figure 6. The percent deviation of the absolute areas for each
Internal standard should be less than 20 percent for on-column Injections and
less than 40 percent for splltlesi. Injections. If the deviations are larger
than 20 percent for on-column Injection and 40 percent for splltless injection,
repeat analysis or perform minor system maintenance (replace quartz sleeve,
dip off column, etc.).
Clipping off 1 ft of the column and cleaning the Injector sleeve will
Improve high end sensitivity for the late elutlng compounds; repositioning
the front end of the column will Improve the chromatography of the early
elutlng compounds. Poor Injection techniques can also lead to variable
Internal standard areas.
38
-------
Figure 6. Area count* for the Internal standards versus time.
-------
7.4 Saturation Effects
Each analytical run must be checked for saturation. The level at which
a certain compound will saturate the detection system 1s a function of the
system sensitivity and the mass spectral characteristics of that compound.
The Initial method calibration requires that the system should not be
saturated for high response compounds at 200 yg/ml. If any compound 1n a
particular sample exceeds the analytical range, the sample must be diluted,
the internal standard concentration readjusted to 20 or 40 pg/mL, and the
sample reinjected. Alternatively, another fragment 1on may be used 1n
quantitation 1f 1t can be shown that for the 1on 1n question Its calibration
curve 1s linear 1n the analytical range of Interest.
40
-------
SECTION 8
DOCUMENTATION
The following QC documentation 1s required as support for the analytical
data obtained using this protocol:
• Relative Ion abundance calibration
-- all DFTPP spectra in the list and 1n bar format; complete
Table 8
• Multilevel calibration
-- all RRT, RF data obtained from multilevel calibration must be
reported. Tables 9 and 9a are supplied as a form which may be
used for this purpose.
• Single level calibration checks
-- for calibration check analysis, the RF values must be reported on
a form such as Tables 10 and 10a.
• Chromatography checks
« for the 100 ng standard of the Initial analysis and each succeeding
50 ng standard, hardcopies must be produced for benzidine (m/z
184), pentachlorophenol (m/z 266) and anthracene-djo (m/z 188).
These should Include the extracted Ion current profiles shown In
Figure 2.
41
-------
TABLE 8. OFTPP ION ABUNDANCE VERIFICATION
TUNE CHECK:
w/e
Ion Abundance Criteria
1 Relative Abundance
SI
30 > 601 of mass 198
68
less than 21 of mass 69
( H
69
mass 69 relative abundance
70
less than 21 of mass 69
I H
127
40 - 601 of mass 198
197
less than 11 of mass 198
198
buse peak, 1001 relative abundance
199
5 - 91 of mass 198
275
10 - 301 of mass 198
365
greater than 11 of mass 198
441
less'than mass 443
442
greater than 401 of mass 198
443
17 - 231 of mass 442
( 12
lvalue In parenthesis Is 1 mass 69.
-------
TABLE 9. FSCC GC/MS INITIAL QC CALIBRATION DATA ~ SEMIVOLATILE PRIORITY
POLLUTANTS
Compound
RRTa
"F20"
RF100b
RF200b
W
SB8B88SI
RSD
(*)
N-IMtrosodlmethyl ami ne
2-Chlorophenol
Phenol
<10
1.3-D1chlorobenzene
1.4-D1chlorobenzene
l.Z-Dlchlorobenzene
B1s(Z-chloro1sopropylJether
Hexachloroethane
N-H1troso-dl-n-propylamine
Nitrobenzene
Isophorone
Z-Nltrophenol
2.4-Dlmethylphenol
B1s(2-chloroethoxy)methane
Z.4-01chlorophenol
1.2.4-Trlchlorobenzene
Naphthalene
<10
Hexachlorobutadlene
4-Chloro-3-cresol
Hexachlorocyclopentadlene
2.4.6-Trlchlorophenol
5-cniorpnaphtnaiene
Acenaphthvlene
Dimethyl ohthalate
2.6-01nltrotoluene
Acenaphthene
2.4-D1n1trophenol
2.4-D1n1trololuene
4-N1trophenol
F1uorene
4-Chlorophenyl phenyl ether
Diethyl phthalate
4.6-Dlnliro-Z-cresol
Diphenylamine
Azobenzene
4-Bromophenyl phenyl ether
Hexachlorobenzene
(continued)
43
-------
TABLE 9. (concluded)
Compound
RRT®
RF20b
RFioob
RF200b
RSD
(%)
Pentachlorophenol
Phenanthrene
Anthracene
<10
Dl-n-butvl phthalate
Fluorantnene
Pyrene
Benzidine
Butyl benzyl phthalate
Benz(a)antnracene
Chrysene
<10
3.3 -Dlchlorobenzldine
B1s(2-ethylhexyl)phthalate
Dl-n-octyl phtnalate
Benzo
j+Mfluoranthenes
Benzo
a)pyrene
<15
Indeno(1.2.3-cd pyrene
D1benzol a.h)antnracene
Benzo(gh1)perylene
aRRT 1s the average relative retention time
^RF it the response factor at the level Indicated (20, 100, 200 nanograms)
1s the average response factor
44
-------
TABLE 9a. FSCC GC/MS INITIAL QC CALIBRATION DATA - PESTICIDES
Compound
RRTa
«F20b
RFioob
A1 oha-BHC
Gamma-BHC
Beta-BHC
Delta-BHC
Heptachlor
Aldrln
4.41-DDE
Dleldrln
4.4'-DDD
4"41-DDT
Beta endosulfan
Endosulfan sul'ate
Endrln
Alpha endosulfan
Notes to Tables 9 and 9a:
aRRT Is the average relative retention time.
&RF 1s the response factor at the level Indicated (20, 100, 200 nanograms).
CffF is the average response factor.
45
-------
TABLE 10. FSCC GC/MS ONGOING QC DATA ~ SEMIVOLATILE PRIORITY POLLUTANTS
sssaoastss
Compound
T?T«
RF
%D
N-N1trosod1methylamine
B1s(2-chloroethyl)ether
2-Chlorophenol
Phenol
-------
TABLE 10. (concluded)
888
Compound
TTT«
RF
ID
Pentachlorophenol
Phenanthrene
Anthracene
<15
D1-n-butyl phthalate
Fluorantnene
Pyrene
Benzidine
Butyl benzyl phthalate
Benz 'a)antnracene
Chrysene
<15
3.3 -D1chlorobenz1d1ne
Bls(2-ethy1hexyl )phthalate
D1-n-octyl phtnalate
Benzo
j+k' fluoranthenes
Benzo
a)pyrene
<15
Indeno
il,Z.3-cd)pyrene
Dlbenz
a.hjanthracenne
Benzo(qh1)perylene
8aiaai3ss33sujiiiiaasssasaaisiiattttussaaaaiassssiataaataias4laaasasassBssa
47
-------
TABLE 10a. FSCC GC/MS ONGOING QC DATA - PESTICIDES
Compound
RF
ID
A1pha-BHC
Gamma-BHC
Beta-BHC
Delta-BHC
Heptachlor
Aldrln
4.4'-DDE
Dleldrln
4,41-DDD
4.4'-DDT
Beta endosulfan
Endosulfan sulfate
Endrln
Alpha endosulfan
Notes to Tables 10 and 10a
1s average response factor from the most recent Initial calibration.
^Report the date the RF was obtained, and the data file name. Use letters
(A, B, C, etc.) for more than one data point on a single day.
48
-------
All area counts for the Internal standards must be reported for
each standard and sample analyzed. Table 11 1s supplied as a
form which may be used for this purpose.
49
-------
TABLE 11. FSCC SC/MS ONGOING QC DATA - ABSOLUTE AREA COUNTS FOR THE
INTERNAL STANDARDS
Run
Identification
Date
I.S. Area Counts
Phenol-d3
Naphthalene-dg
Anthrecene-djo
Chrysene-dj2
Benzo(a)pyrene-dj2
83aaa33ii53:::::a393sss3s:3j3ss::3s:::ss:sses88:8s8it888t338888s33::: isssssssssssssssssssssessssMssssss
-------
SECTION 9
REFERENCES
J. A. D, Sauter, L. D, Betowski, T. R. SnUh, V. A» Stricfcler, A. G. Etelmer,
B. N. Colby, and J. E. Wilkinson, "Fused Silica Capillary Column GC/MS
ftnalysis of Priority Pollutants," Journal of High Resolution
Chromatography and Chromatography Cotnmunications, A, ^65 (1981).
Z. A. Q. Sauter, et at., "Interlaboratory EC/MS Response Factor Precision
and Accuracy for Organic Cgmpoimds," sutnltted for publication 1n the
Journal of High Resolution Chromatography and Chromatography
Communications, _5, 27 (1982).
3. T. It. Smith, N. K. Mosesman, mti A. 0. Sauter, "Compositing Acid and
Base/Neutral Fractions for the FSCC GC/MS Analysis of Priority
Pollutants," paper presented at the Pittsburgh Conference and Exposition
on Analytical Chemistry and Applied Spectroscopy, March 1982.
4. A. D. Sauter, I. D, Betowskl, and 0. M. Ballard, "Comparison of Priority
Pollutants Response Factors for Triple and Single Quadruple Mass
Spectrometers," paper accepted for publication 1n Analytical Chemistry,
January 19B3 issue.
5. Method 1625, Semi volatile Organic Compounds by Isotope Dilution GC/MS,
U.S. EPA, WH55Z, Washington, Q.C. 20460
6. "Method Development for Fused Silica Capillary Colurm GC/MS," Final
Report Work Assignment SCA-OZ, EPA Contract No. 68-03-3043, Acurex
Corporation.
7. "Guidelines Establishing Test Procedures for the Analysis of Priority
Pollutants; Proposed Regulations," Method 625, Federal Register,
December 3, 1979, p. 69541.
6. J. W. Elchelberger, L. E. Harris, and W. L. Butide, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry," Anal. Chem. 47, 995 (1975).
9. "QSHA Safety end Health Stardards," General Industry (29 CFR.1910),
Occupational Safety and Health Administration, 05KA KOfc (Revised JanuAry
1S?9).
51
-------
APPENDIX A
LABELED/UNLABELED ~ COMPOUND PAIRS
52
-------
17370.
94.028.
10.BOO
11792.
97.029
10.600
142080.
320 340 360
6:36 6:67 6:18
380 400 Scan
6:39 7:00 Time
Figure A-l. Mass chromatograms for Ions at m/z 94 and m/z 97 corresponding
to phenol and phenol-d3, respectively.
53
-------
7:52 8:03 8:13 8:24 8:34 8.45 Time
Figure A-2. Mass chromatograms for Ions at ra/z 128 and m/z 136 corresponding
to naphthalene and naphthalene-dg, respectively.
54
-------
925 931
•6432.
178.063
10.600
930
16:16
940
16:27
960
16:37
1F446.
188.066
10.600
16S=M6
i
960 Scan
16:37 Tim*
Figure A-3. Mass chromatograms for 1ons at m/z 178 and m/z 188 corresponding
to phenanthrene, anthracene, and anthracene-d^g, respectively.
55
-------
22:24 22:34 22:45 22:55 23:06 Time
Figure A-4. Mass chromatograms for Ions at m/z 228 and m/z 240 corresponding
to benzo(a)anthracene, chrysene, and chrysene-d]^, respectively.
56
-------
1491
17216.
262.076
i 0.600
12032.
264.079
10.600
60048.
1400
24:60
1460
26:23
1600
26:16
1660
27:07
1600 Scon
28:00 Tims
Figure A-5. Mass chromatograms for Ions at m/z 252 and m/z 264 corresponding
to benzo(a)pyrene and benzo(a)pyrene-d12, respectively.
57
-------
|