United States Office of Water and ' /, SW-646
Environmental Protection Waste Management ..QQQ
Agency Washington, DC 20460
Solid Waste
Test Methods ^ -
for Evaluating Solid Waste
Physical/Chemical Methods
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SW846
TEST METHODS FOR EVALUATING SOLID WASTE
PHYSICAL/CHEMICAL METHODS
WASTE CHARACTERIZATION BRANCH
OFFICE OF SOLID WASTE
U.S. ENVIRONMENTAL PROTECTION AGENCY / MAY 1980
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ABSTRACT
This manual provides test procedures which may be used to
evaluate those properties of a solid waste which determine
whether the waste is a hazardous waste within the definition
of Section 3001 of the Resource Conservation and Recovery
Act (PL 94-580). These methods are approved for obtaining
data to satisfy the requirement of 40 CFR Part 261, Identifica-
tion and Listing of Hazardous Waste. This manual encompasses
methods for collecting representative samples of solid wastes,
and for determining the reactivity, corrosivity, ignitability,
and composition of the waste and the mobility of toxic species
present in the waste.
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ACKNOWLEDGEMENT
The Office of Solid Waste would like to especially thank
the following individuals and groups for the help and advice
they gave us during the preparation of this manual:
Dr. Gregory Diachenko, Food & Drug Administration,
Division of Chemical Technology, Washington, B.C.
Dr. Wayne Garrison, Environmental Research Laboratory,
Athens, GA
Dr. Donald Gurka, Environmental Monitoring and Systems
Laboratory, Las Vegas, NV
Drs. Dean Neptune and Mike Carter, Effluent Guidelines
Division, Washington, DC
Quality Assurance Division, Environmental Monitoring &
Systems Laboratory, Las Vegas, NV
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oEPA
United StatM
Environmental Protection
Agency
Office of Weter end
Wette Management
Washington, DC 20460
SW-846
Revision B
July 1981
Solid Waste
Test Methods
for Evaluating Solid Waste
Physical/Chemical Methods
Technical
U pdate
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TEST METHODS FOR EVALUATING SOLID WASTE
PHYSICAL/CHEMICAL METHODS
Technical Update
This manual (SW-846B) updates the Test Methods for Evaluating
Splid Waste (SW-846), and was written by the Hazardous and
Industrial Waste Division of the Office of Solid Waste.
U.S. ENVIRONMENTAL PROTECTION AGENCY
July 1981
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This publication (SW-846B) is the second revision to Test Methods for
Evaluating Solid Waste (SW-846). Any mention of commercial products in
the manual or this revision does not constitute endorsement by the U.S.
Government. Editing and technical content were the responsibilities of
the Hazardous and Industrial Waste Division, Office of Solid Waste.
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Revision B 4/15/81
Table of Contents
Section
Introduction
1.0 Evaluation Plan Design
2.0 Chain of Custody Considerations
3.0 Sampling Methodology
3.1 Sampling Plan Design [Reserved]
3.2 Sampling Equipment
3.3 Sample Containers
3.4 Sample Handling & Preservation [Reserved]
4.0 Ignitability (40 CFR 261.21)
5.0 Corrosivity (40 CFR 261.22)
6.0 Reactivity (40 CFR 261.23)
7.0 Extraction Procedure Toxicity (40 CFR 261.24)
7.1 Regulations
7.2 Separation Procedure
7.3 Sample Size Reduction [Reserved]
7.4 Structural Integrity Procedure
7.5 Extractors
8.0 Analytical Methodology
Gas Chromatographic Methods
8.01 Volatile organics, general
8.02 Volatile aromatics, selected ketones & ethers
8.03 Acrolein, Acrylonitrile and Acetonitrile
8.04 Phenols
8.06 Semi-volatile organics, not otherwise specified
8.08 Organochlorine pesticides and PCBs
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Revision B 4/15/81
8.09 Nitroaromatics
8.10 Polynuclear Aromatic Hydrocarbons
8.11 Haloethers [Reserved]
8.12 Semi-volatile chlorinated hydrocarbons, Not Otherwise
Specified
8.13 Chlorinated Dibenzo-p-dioxins [Reserved]
8.22 Organophosphorus pesticides
8.40 Chlorophenoxy acid pesticides
Gas Chromatographic/Mass Spectroscopy Methods
8.24 Volatile organics
8.25 Semi-volatile organics
8.27 Capillary Column GC/MS Metod for the Analysis of Wastes
High Performance Liquid Chromatographic Methods
"8.30 Polynuclear Aromatic Hydrocarbons [See method 8.10]
~8.32 Carbamates [Reserved]
Atomic Absorption Spectrographic Methods
8.49 General Requirements
8.50 Antimony
8.51 Arsenic
8.52 Barium
8.53 Cadmium
8.54 Chromium
^8.55 Cyanide
8.56 Lead
8.57 Mercury
8.58 Nickel
8.59 Selenium
8.60 Silver
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Revision B 4/15/81
Other Measurement Methods
8.55 Titrimetric Method for Cyanide
8.56 Microcoulometric Method for Total Organic Halide
8.57 Titrimetric Method for Sulfides
Sample Preparation/Introduction Techniques
8.80 Direct Injection
8.82 Headspace
8.83 Purge and Trap
8.84 Shake Out
8.85 Sonication
8.86 Soxhlet Extraction
9.0 Interference Removal Procedures
9.01 Liquid - Liquid Extraction
10.0 Quality Control/Quality Assurance
11.0 Suppliers
Appendices
I "Samplers and Sampling Procedures for Hazardous Waste
Streams", EPA-600/2-80-018
II Selected sections of "Methods for Chemical Analysis
of Water and Wastes", EPA-600/4-79-020.
Ill "Methods for Benzidine/ Chlorinated Organic
Compounds, Pentachlorophenol and Pesticides in Water and
Wastewater"
IV Selected sections from the Federal Register, "Guidelines
Establishing Test Procedures for the Analysis of Pollutants;
Proposed Regulations", 44 FR 69464-69567.
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INTRODUCTION
This first edition of "Test Methods for Evaluating Solid
Waste" contains the procedures that may be used by the regulated
community or others in order to determine whether a waste is a
hazardous waste as defined by regulations promulated under
Section 3001 of the Resource Conservation and Recovery Act
(RCRA), PL 94-580 (40 CFR Part 261). The manual provides
methodology for collecting representative samples of the
waste, and for determining the ignitability , corrosivity,
reactivity, Extraction Procedure (EP) Toxicity and composition
of the waste.
This document has been developed to:
a. provide methods which will be acceptable to the
Agency when used by the regulated community to
support waste evaluations and listing and
delisting petitions, and
b. describe the methods that will be used by the
Agency in conducting investigations under
Sections 3001, 3007, and 3008.
The practice of evaluating solid wastes for environmental
and human health hazards is new. Experience has only recently
accumulated in analyzing wastes for inorganic and organic
species, and for intrinsic properties such as pH, flash point,
reactivity and leachability . This manual will serve as a
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compilation of state-of-the-art methodology for conducting
such tests. It is meant to be a dynamic document. The
methodology descriptions will be frequently updated and
expanded in order to keep pace with the developments being
achieved by EPA, the regulated community, and others.
Standardized approved methods must be available so that
the regulated community can be certain that the data it pro-
vides will be acceptable to the Agency. This manual thus
makes available to the regulated community and others, those
methods that the Agency considers suitable.
Many of the methods presented in this manual have not been
fully evaluated by the Agency using materials characteristic of
the wastes regulated under RCRA. Such evaluations are underway.
However, until such time as the methods in this manual are
superseded, the Agency will accept data obtained by the test
methods presented in this manual. Only those data that are
obtained when Quality Control and Quality Assurance procedures
are followed by the testing organization will be accepted by
the Agency.
This manual will eventually include a second part comprised
of biological methods for determining toxic properties of
RCRA wastes. Such toxic properties may include carcinogenicity,
mutagenicity, teratogenicity, aquatic toxicity, phytotoxicity,
and mammalian toxicity. The Agency anticipates that these
methods will not be available before the later part of 1981.
ii
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Methods will be provided in this present volume for the
following specific areas:
a- design of sampling and evaluation plans;
b. collection of samples from various types of environments
(e.g., pipes, drums, pits, ponds, piles, tanks);
c. transportation and storage of samples;
d. chain-of-custody considerations to insure defensibility
of data;
e. determination of the pH, corrosivity to steel, flash point,
and explosivity;
f. conduct of the Extraction Procedure;
g. analysis of wastes and extracts for organic and inorganic
constituents;
h. safety in solid waste sampling and testing, and
i. quality control and quality assurance.
The analytical and sampling methods presented in this
manual have been derived from a number of published sources,
chiefly:
a. "Methods for the Evaluation of Water and Wastewater,"
EPA-600/4-79-020, U. S. EPA, Environmental Monitoring
and Support Laboratory, Cincinnati, OH 45268,
b. "Methods for Benzidine, Chlorinated Organic Compounds,
Pentachlorophenol and Pesticides in Water and Wastewater,"
U.S. EPA, Environmental Monitoring and Support
Laboratory, Cincinnati, OH 45268, September 1978,
iii
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c. Guidelines Establishing Test Procedures for the
Analysis of Pollutants; Proposed Regulations;
44 FR 69464-69575, and
d. "Samplers and Sampling Procedures for Hazardous Waste
Streams," EPA-600/2-80-018, U.S. EPA, Municipal Environ-
mental Research Laboratory, Cincinnati, OH 45268.
In addition, work conducted by and the assistance of
scientists of the Environmental Monitoring Systems Laboratory
at Las Vegas, NV, the Environmental Research Laboratory at
Athens, GA, and the National Enforcement Investigations Center
at Denver, CO, is gratefully acknowledgd and appreciated.
For the convenience of those using this manual, the above
publications have been appended to this manual. Users are
encouraged to review these additional sources of information
prior to initiating laboratory work.
Although a sincere effort has been made to select methods
that are applicable to the widest range of expected wastes,
significant interferences, or other problems, may be encountered
with certain samples. In these situations, the analyst is
advised to contact the Manager, Waste Analysis Program (WH-565),
Waste Characterization Branch, Office of Solid Waste, Washington,
D.C. 20460 (202-755-9187) for assistance. The manual is intended
to serve all those with a need to evaluate solid waste. Your
comments, corrections, suggestion, and questions concerning any
material contained in, or omitted from, this manual will be
gratefully appreciated. Please direct your comments to the
above address .
iv
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1.0-1
1.0 Evaluation Plan Design
Purpose
40 CFR Part 262.11 requires generators to evaluate
their waste in order to determine whether it meets the defini-
tion of a hazardous waste. On the basis of engineering
evaluations of processes and their raw materials, many genera-
tors will elect to declare their waste hazardous without
testing it. Other generators, who believe that their waste
is not hazardous, will elect to test their waste in order to
so demonstrate.
This section of the manual has two purposes. The first
is to discuss relevant factors which generators should
consider in developing a sampling and testing plan that
provides maximum information at minimum cost. The second
purpose is to outline for the regulated community standards
of proof the Agency considers sufficient to demonstrate whether
a waste does or does not possess a given property. Such
demonstrations are necessary when generators petition the
Agency under 40 CFR 260.20 and 260.22 to delist a waste.
Such demonstrations are also crucial when the Agency determines
that a waste meets the definition of a hazardous waste and
the generator elects to challenge that determination. In such
cases, conclusions regarding a waste's nature that are obtained
using the procedures and standards of proof described in this
manual will be accepted by the Agency as evidence that the
determinations were made in good faith.
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1.0-2
Why is there a need for such guidance?
The Agency has defined hazardous waste in terms of a
waste's chemical and physical properties. If all wastes were
perfectly homogeneous and if properties could be measured
with 100% accuracy, there would be no need for this manual.
In the real world, however, waste testing is far from simple
or ideal. Errors can occur:
a. during the process of collecting a representative
sample of the waste, unanticipated non-uniformity in
the process generating the waste (i.e., due to
changes in the waste's composition over time) or to
the fact that the contaminants of concern in the
waste are not uniformly distributed throughout the
particular sample of waste (i.e., non-homogeneity),
and
h. in analyzing the sample for the property of concern.
The discussion in this section of the manual pertains to how:
a. to design a sampling plan which yields a
statistically representative sample of a waste,
b. to maximize the accuracy of an evaluation while
minimizing the costs, and
c. to determine when a given waste has been adequately
characterized.
Other sections of this manual will address the mechanics of
collecting samples of a given batch or lot of waste, and of
performing the laboratory procedures required to determine
its physical or chemical properties.
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1.0-3
The decision criteria presented in this section can be
used by generators to determine if the evaluation of the
waste is adequate to demonstrate that the true value for a
specific property falls below the particular regulatory
threshold value for that property.
Statistical Calculations
A statistic is a number that describes a certain aspect
of a sample. The arithmetic mean (average) and standard
deviation (s) are all statistics computed from a part of a
populaton or universe (i.e., a sample).
The most useful statistics, and the ones to be used in
determining the adequacy of the characterization, are the arith-
metic mean of the sample (x) and the standard deviation (s),
which is a measure of the variability of the data obtained.
These terms are defined as:
r»
n
n - i
Based on these statistical measures, a confidence interval
can be developed within which the true value for the parameter
being determined can be said to lie with a high degree of
confidence. Thus if the upper bound of this interval (Upper
Confidence Level (UCL)) is below the applicable threshold
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then it can confidently be concluded to be non-hazardous. If, on
the other hand, the UCL is above the threshold, then even though
the values obtained for the samples tested fall below the
threshold, one cannot confidently conclude the waste is not a
a hazardous waste.
As will be discussed later, in developing a sampling
plan and in collecting samples of a waste, it is important to
insure that all portions of the waste have an equal chance
of being represented. If representative samples are collected,
the data obtained from testing will follow a normal distribution.
In such cases, for purposes of waste characterization, the
UCL can be calculated using the formula:
UCL = "x + (k' )(s) where
'n
k' is given in Table 1-1 and is a function of the number of
samples tested .
Example:
A waste has a true concentration of cadmium of 10 mg/kg.
The total waste was divided into 5 portions and each portion
was analyzed to yield the following concentration values:
Xli; 5mg/kg
X2 = 15mg/kg
X3 . 15 mg/kg
X4 = 8mg/kg
X5 = 7mg/kg
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1.0-5
If the first and second portions had been selected as
the random samples to test, the arithmetic mean would be
identical to the population mean:
IF = (xj_+ x2)/2 = (5 +15)/2 = 10
If however, portions 4 and 5 had been selected, the
arithmetic mean would be significantly different from the
population mean:
H - (X4 + X5/2 = (8 + 7)/2 = 7.5
Had portions 4 and 5 been selected, the UCL would still
be reasonably close to the true value (e.g., within 10%).
x = 7.5
n - 2
s - Aj[(8.0 - 7.5)* + (7.0 - 7.5)^]/l
- 0.7
UCL = 7.5 + k'(0.7)/ -
= 7.5 + (3.078)(0.7)/1.4
= 9.04
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1.0-6
Table 1-1
n-1
1
2
3
4 . . .
5 . . .
6
7
8 . . .
9
10 . . .
11
12 . . .
13
14
15
16 . . .
17
18 . . .
19
20
21
22 . . .
23 . . .
24
25 . . .
26
27 . . .
28 . . .
29
00 . . .
k'
. . . 3.078
. . . 1 .886
. . . 1.638
. . . 1.533
. . . 1 .476
. . . 1 .440
. . . 1 .415
. . . 1.397
. . . 1 .383
. . . 1.372
. . . 1 .363
. . . 1.356
. . . 1.350
. . . 1.345
. . . 1 .337
. . . 1.341
. . . 1 .333
. . . 1.330
. . . 1 .328
. . . 1.325
. . . 1 .323
. . . 1.321
. . . 1.319
. . . 1.318
. . . 1.316
. . . 1.315
. . . 1 .314
. . . 1.313
. . . 1 .311
. . . 1.282
n = Number of samples tested
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1.0-7
Sampling Cons1deration s
The physical characteristics of a waste determine the
method of collecting samples as well as the number of samples
that should be collected and tested before calculating an
initial UCL. In evaluating wastes, it is impossible to have
complete population data since only a very small fraction of
the waste will be subjected to laboratory study. The objective
of sampling, therefore, is to select a portion of the waste
which is representative of the whole. In order to use the
acceptance criteria described in this section, each component
of the waste must have an equal chance of being sampled and
tested. Therefore, before a sampling plan can be designed,
the type of contamination distribution should be determined
by an engineering analysis of the process generating the
waste. The purpose of this analysis is to determine the
appropriate sampling plan for obtaining a representative
sample of the waste in question.
Wastes can be classified into six types depending on the
uniformity of the process generating the waste and the homo-
geneity of the contaminant distribution within the waste.
These categories determine the approach that should be used
in conducting the sampling and the number of samples that
should .be tested initially. The six types of waste are:
I. Uniformly homogeneous
II. Non-uniformly homogeneous
III. Uniformly, randomly heterogeneous
IV. Non-uniformly, randomly heterogeneous
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1.0-8
V. Uniformly, non-randomly heterogeneous
VI. Non-uniformly, non-randomly heterogeneous
In these classifications uniformity is defined as constancy
of waste composition over a period of time. Homogeneity
refers to the degree to which the components of the waste are
uniformly distributed throughout the mass of waste (i.e., in a
homogeneous sample all possible subsamples have an Identical
composition).
A Type I waste (uniformly homogeneous) is the one whose
overall composition does not vary with time and for which
the constituent of concern is evenly and randomly distributed
throughout the waste (i.e., any single subsample of the
whole would be expected to contain the same proportion of a
given constituent as would any other subsample). Examples of
wastes are liquid waste products disposed of on a regular
basis (e.g., waste pesticide formulations remaining in
holding tanks at the conclusion of a production run) and
wastes generated from a well controlled manufacturing process
(e.g., spent semiconductor etching solutions).
In Type II wastes, while any given unit quantity of the
waste would be homogeneous, the nature of the process generating
the waste is such that the overall composition changes with
time. Such a situation might occur at a facility that uses
batch processes to make a variety of similar products and
which generates a well mixed, single phase liquid waste.
While any portion of the waste generated from a given production
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1.0-9
run would be equivalent, wastes from different runs would have
different properties.
Types III through VI wastes are much more complex.
Types III and V, deal with the cases where the composition
of the waste as a whole remains the same, but the components
are not uniformly distributed throughout the waste. In a
randomly heterogeneous waste (Types III and IV) the likelihood
is the same that any given sampling point will contain a
specific type of contaminant. An example of a Type III
waste is that generated by a batch plating operation that
involves a number of plating processes, where all wastes
feed into a single treatment facility, and where the sludge
is stored in a tank that holds several weeks' worth of sludge.
While the concentration of a given metal would not be uniform
thoughout the tank, no one region within the tank is more
likely to contain high concentrations of any one metal more
than any other.
With Type V waste, placement of the waste is not random
because of the nature of the storage or disposal process.
For example, when a waste is discharged into a pond or lagoon,
the heavier particles settle out first; thus, stratification
occurs on a continuous basis. Samples of waste selected
near the entrance point in the holding tank would always
the more dense material. This is the reverse of the Type IV
waste, in which no one type of waste is more likely than any
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1.0-10
other to be found at a given point.
With Types I and III wastes, a simple random sample and a
randomly prepared composite sample, respectively, will suffice
as representative samples. With Type V wastes which may be
stratified, the waste would have to be divided into a number
of strata; each stratum would then have the properties of
either a Type I or Type III waste. A random sample of each
stratum would then be used to form a composite sample. The
number of random subsamples collected from each stratum would
then be a function of the relative proportion of that particular
stratum in the waste as a whole. If, for example, dense
particles comprised 50% of the waste and 10 subsamples were
to be used in forming a composite, then 5 of those subsamples
should be randomly collected from the dense particle substratum.
At this point, it should be mentioned that when the
waste is non-uniform (i.e., Types II, IV, and VI), the generator
would have to sample the waste over time. The techniques
described herein for uniform wastes should be used in order
to determine the significance of the changes with respect to
the property of concern.
A major consideration in solid waste evaluation is the
number of samples that should be tested. The sample size is
determined by:
a. the degree of accuracy desired,
b. the cost of collecting and testing the samples,
and
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1.0-11
c. the variability of the population from which
the samples are being taken (e.g., Type I wastes
are exceedingly uniform and thus fewer samples
would have to be tested whereas Type IV wastes are
very variable and more samples would have to be
tested) .
As the number of samples tested increases, the confidence
interval gets smaller (UCL approaches the mean) and the
accuracy of the determination increases. At least 2 samples
(the recommended minimum is 3 samples) must be collected and
tested prior to calculating an initial average and upper
confidence limit.
Testing Considerations
An additional aspect of waste testing that must be
considered relates to the precision of the test methods
used to measure the parameter of concern. The standard
deviation of the values obtained when testing wastes is a
function of both the sampling variability and the measurement
variability, i.e.,:
So " flss + f2sa
where
So = overall standard deviation
Ss = standard deviation attributable to
sampling error
Sa = standard deviation attributable to
analytical error
f = complex function
In evaluating solid wastes, a number of testing procedures
are used to determine if the waste is a hazardous waste.
The precision of these methods varies not only as a function
of thet method used, but also as a function of the waste
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1.0-12
matrix and the level of the property of interest (e.g.,
concentration of toxic species).
Where prior information is available that indicates the
portion of the total testing error attributable to the measurement
phase, the generator can use this information to determine
the best (i.e., most cost effective) means of reducing the
UCL. When Sa is high, then replicated measurements on the
sample would tend to be more efficient than collecting and
testing a larger number of samples. On the other hand,
when Sa is low and Ss is high, larger numbers of samples
should be evaluated. When Sa is unknown or cannot be
estimated in advance, generators may find it cost effective
to determine Sa during the initial experiments that are con-
ducted to calculate UCL.
Once initial testing has been concluded and the UCL
calculated, the decision on whether further testing is warranted
depends upon several factors. Among these are:
a. relation of x and UCL to the threshold value,
b. cost of testing, and
c. relative cost of disposal as hazardous waste.
For example, if x is greater than the threshold value and
(UCL x) is small, there is a statistically small probability
that the waste is not a hazardous waste, and thus there is
little to be gained by further testing. If the UCL is less
than the threshold value, the Agency will accept the conclusion
drawn from such data and again there is little to be gained
from futher testing. However, this acceptance by the Agency
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1.0-13
is conditional on the generator's having used a sample
collection plan that yields a sample that Is representative
of the whole, (as was discussed previously in this section).
When the testing costs are low and the disposal costs are
high, generators may want to do additional testing in order
to increase the accuracy of the evaluation. One should keep
in mind that the standard error is inversely proportional to
the square root of the number of samples tested (e.g., increas-
ing the number of samples tested from 4 to 16 reduces the
standard error by 50%) .
The Agency believes that, after taking these factors into
account, each generator must make his own decisions as to the
size of the testing program required to adequately evaluate
a particular waste. The Agency will continue to expand
material in this manual as new information develops in order
to offer the regulated community further guidance and assistance.
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2.0-1
Section 2.0
CHAIN OF CUSTODY CONSIDERATIONS
Chain of custody establishes the documentation and control
necessary to identify and trace a sample from sample collection
to final analysis. Such documentation includes labeling to
prevent mix up, container seals to prevent unauthorized tampering
with contents of the sample containers, secure custody, and
the necessary records to support potential litigation.
Sample Labels
Sample labels (Figure 2.0-1) are necessary to prevent mis-
identification of samples. Gummed paper labels or tags are
adequate. The label must include at least the following
information:
Name of collector.
Date and time of collection.
Place of collection.
Collector's sample number, which uniquely identifies
the sample.
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2.0-2
Collector Collector's Sample No
Place of Collection
Date Sampled Time Sampled
Field Information
Figure 2 .0-1
EXAMPLE OF SAMPLE LABEL
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2.0-3
Sample Seals
Sample seals are used to preserve the integrity of the
sample from the time it is collected until it is opened in the
laboratory. Gummed paper seals may be used for this purpose.
The paper seal must include, at least, the following information:
Collector's name.
Date and time of sampling.
Collector's sample number. (This number must be
identical with the number on the sample label.)
The seal must be attached in such a way that it is necessary
to break it in order to open the sample container. An example
of a sample seal is shown in Figure 2.0-2.
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2.0-4
NAME AND ADDRESS OF ORGANIZATION COLLECTING SAMPLES
Person Collectors
Collecting Sample Sample No.
(Signature)
Date Collected Time Collected
Place Collected
Figure 2.0-2
EXAMPLE OF OFFICIAL SAMPLE SEAL
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2.0-5
Field Log Book
All information pertinent to a field survey and/or sampling
must be recorded in a log book. This must be a bound book,
preferably with consecutively numbered pages that are 21.6 by
27.9 cm (8 1/2 by 11 in.). Entries in the log book must include
at least, the following:
Purpose of sampling (e.g., surveillance, contract
number)
Location of sampling point.
Name and address of field contact.
Producer of waste and address if different than location.
Type of process (if known) producing waste.
Type of waste (e.g., sludge, wastewater)
Suspected waste composition including concentrations.
Number and volume of sample taken.
Description of sampling point and sampling methodology.
Date and time of collection.
Collector's sample identification number(s).
Sample distribution and how transported (e.g.,
name of laboratory, UPS, Fedeal Express)
References such as maps or photographs of the
s ampling site.
Field observations.
Any field measurements made (e.g., pH, flammability,
explositivity).
Sampling situations vary widely. No general rule can be
given as to the extent of information that must be entered in
the log book. A good rule, however, is to record sufficient
information so that someone can reconstruct the sampling
without reliance on the collector's memory.
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2.0-6
The log book must be protected and kept in a safe place.
Chain of Custody Record
To establish the documentation necessary to trace sample
possession from the time of collection, a chain of custody record
must be filled out and accompany every sample. This record
becomes especially important when the sample is to be intro-
duced as evidence in a court litigation. An example of a
chain of custody record is illustrated in Figure 2.0-3.
The record must contain the following minimum information:
Collector's sample number.
Signature of collector.
Date and time of collection.
Place and address of collection.
Waste type.
Signatures of persons involved in the chain of
pos session.
Inclusive dates of possession.
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2.0-7
Collector's Sample No.
CHAIN OF CUSTODY RECORD
Location of Sampling: Producer Hauler Disposal Site
Other:
Sample
Shipper Name:
Address:
number street city state zip
Collector's Name Telephone: ( )
signature
Date Sampled Time Sampled hours
Type of Process Producing Waste
Field Information
Sample Receiver:
1.
name and address of organization receiving sample
2.
3.
Chain of Possession:
.1.
signature title inclusive dates
2.
signature title inclusive dates
3.
signature title inclusive dates
Figure 2.0-3
EXAMPLE OF CHAIN OF CUSTODY RECORD
-------
2.0-8
EXAMPLE OF CHAIN OF CUSTODY RECORD
Sample Analysis Request Sheet
The sample analysis request sheet (Figure 2.0-4) is intended
to accompany the sample on delivery to the laboratory. The field
portion of this form must be completed by the person collecting
the sample and should include most of the pertinent information
noted in the log book. The laboratory portion of this form is
intended to be completed by laboratory personnel and to include at
a minimum:
Name of person receiving the sample.
Laboratory sample number.
Date of sample receipt.
Sample allocation.
Analyses to be performed.
-------
PART I: FIELD SECTION
SAMPLE ANALYSIS REQUEST
2.0-9
Collector
Affiliation of Sampler
Address
Date Sampled
number
Telephone ( )
LABORATORY
SAMPLE
NUMBER
COLLECTOR'S
SAMPLE NO.
street city
Company Contact
TYPE OF
SAMPLE*
Time
hours
state
zip
FIELD INFORMATION**
Analysis Requested
Special Handling and/or Storage
PART II: LABORATORY SECTION**
Received by
Analysis Required
Title
Date
* Indicate whether sample is soil, sludge, etc.
** Use back of page for additional information relative to sample location
Figure 2.0-4
EXAMPLE OF HAZARDOUS WASTE SAMPLE ANALYSIS REQUEST SHEET
-------
2.0-10
Sample delivery to the laboratory
Preferably, the sample must be delivered in person to the
laboratory for analysis as soon as practicable—usually within
1 or 2 days after sampling. The sample must be accompanied by
the chain of custody record (Figure 2.0-3) and by a sample anal-
ysis request sheet (Figure 2.0-4). The sample must be delivered
to the person in the laboratory authorized to receive samples
(often referred to as the sample custodian).
Shipping of Samples
A material identified in the DOT Hazardous Material Table
(49 CFR 172.101) must be transported as prescribed
in the table. All other hazardous waste samples must be trans-
ported as follows:
1. Collect sample in an appropriately sized glass or poly-
ethylene container with non-metallic teflon lined screw
cap. Allow sufficient ullage (approximately 10% by
volume) so container is not liquid full at 54°C Celsius
(130°). If sampling for volatile organic analysis, fill
container to septum but use closed cap with space to
provide an air space within the container. Large quantities,
up to 3.785 liters (1 gallon), may be collected if the
sample's flash point is >^ 23°C (73°F). In this case,
the flash point must be marked on the outside container
(e.g., carton, cooler).
-------
2.0-11
2. Seal sample and place in a 4 ml thick polyethylene bag,
one sample per bag.
Liquids
3. Place sealed bag inside a metal can with noncombustible,
absorbent, cushioning material (e.g., vermiculite or
earth) to prevent breakage, one bag per can. Pressure-
close the can and use clips, tape or other positive
means to hold the lid securely.
A. Mark the can with:
Name
Address
"Flammable Liquid N.O.S."
(or "Flammable Solid N.O.S.")
Note; Using "Flammable" does not convey the certain knowledge
that a sample is flammable but is intended to prescribe the
class of packing needed to comply with DOT regulations.
5. Place one or more metal cans in a strong outside container
such as a picnic cooler or fiberboard box. Ice or dry
ice may be used between the metal cans and the outside
container and must be contained to avoid water leakage
during transport. No other preservatives are allowed.
6. Complete carrier's certification form as shown in Figure 2.0-5
7. Samples may be transported by rented or common carrier
truck, bus, railroad, and entities such as Federal Express*
but not by normal common carrier air transport even on a
"cargo only" aircraft.
*These procedures are designed to enable shipment by entities
like Federal Express; however, they should not be construed
as an endorsement by EPA of a particular commercial carrier.
-------
2.0-12
Solids
3. Place sealed bag inside cushioned overpack. If sample
is expected to undergo change during shipment cool using
dry or wet ice. Overpack must be designed to prevent
water leakage during transport. No other preservatives
are allowed.
4. Complete carrier's certification form as shown in Figure
2.0-5.
5. Samples may be transported by rented or common carrier
truck, bus, railroad, and entities such as Federal Express* but
not by normal common carrier air transport even on a "cargo
6
only" aircraft.
Receipt and Logging of Sample
In the laboratory, a sample custodian should be assigned
to receive the samples. Upon receipt of a sample, the custodian
should inspect the condition of the sample and the sample seal,
reconcile the information on the sample label and seal against
that on the chain of custody record, assign a laboratory number,
log in the sample in the laboratory log book, and store the
sample in a secured sample storage room or cabinet until
assigned to an analyst for analysis.
The sample custodian should inspect the sample for any
leakage from the container. A leaky container containing
multiphase sample should not be accepted for analysis. This
sample will no longer be a representative sample. If the
sample is contained in a plastic bottle and the walls show
*These procedures are designed to enable shipment by entities
like Federal Express; however, they should not be construed
as,an endorsement by EPA of a particular commercial carrier.
-------
2.0-13
the sample is under pressure or releasing gases, respectively,
it should be treated with caution. The sample can be explosive
or release extremely poisonous gses. The custodian should
examine whether the sample seal is intact or broken, since a
broken seal may mean sample tampering and would make analysis
results inadmissible in court as evidence. Discrepancies
between the information on the sample label and seal and
that on the chain of custody record and the sample analysis
request sheet should be resolved before the sample is assigned
for analysis. This effort might require communication with
the sample collector. Results of the inspection should be
noted on the sample analysis request sheet and on the laboratory
sample log book.
Incoming samples usually carry the inspector's or collec-
tor's identification numbers. To further identify these
samples, the laboratory should assign its own identification
numbers, which normally are given consecutively. Each sample
should be marked with the assigned laboratory number. This
number is correspondingly recorded on a laboratory sample log
book along with the information describing the sample. The
sample information is copied from the sample analysis request
sheet and cross-checked against that on the sample label.
Assignment of Sample for Analysis
In most cases, the laboratory supervisor assigns the
sample for analysis. The supervisor should review the infor-
mation on the sample analysis request sheet, which now includes
-------
2.0-14
inspection notes recorded by the laboratory sample custodian.
The supervisor should then decide what analyses are to be
performed. The sample may have to be split with other labora-
tories to obtain the necessary information about the sample.
The supervisor should decide on the sample location and
delineate the types of analyses to be performed on each allo-
cation. In his own laboratory, the supervisor should assign
the sample analysis to at least one analyst, who is to be
responsible for the care and custody of the sample once it is
received. He should be prepared to testify that the sample
was in his possession or secured in the laboratory at all
times from the moment it was received from the custodian
until the analyses were performed.
The receiving analyst should record in the laboratory
notebook the identifying information about the sample, the
date of receipt, and other pertinent information. This record
should also include the subsequent testing data and calcu-
lations .
-------
3.0-1
Section 3.0
SAMPLING METHODOLOGY
Introduction
This section describes the equipment and procedures which
the Agency has determined are suitable for use in obtaining
a representative sample of a solid waste.
Because the types of solid waste regulated under RCRA
cover a very broad range of physical and chemical types, the
information in this section will of necessity be general in
nature. It is incumbent upon those persons conducting sampling
programs to exercise caution when using the methods described
herein.
-------
3.2-1
Sub-Section 3.2
SAMPLING EQUIPMENT
Introduction
Sampling the diverse types of RCRA regulated wastes requires
a variety of different types of samplers. A number of such
sampling devices are described in this section. While some of
these samplers are commercially available, others will have to be
fabricated by the user. Table 3.2-1 is a general guide to the types
of waste that can be sampled by each of the samplers described.
-------
3.2-2
Table 3.2-1
SAMPLING EQUIPMENT FOR PARTICULAR WASTE TYPES
Sampling
I Point
| Waste
llype
Free
(Flowing
I Liquid s
land
Slurries
Sludges
Moist
Powders
or
Granules
Dry
Powders
or
Granules
Sand or
Packed
Powders
and
Granules
Large
Grained
Solids
Drum
Coli-
wasa
Trier
Trier
Thief
Auger
arge
Trier
Sacks
and
Bags
N/A
N/A
Trier
Thief
Auger
Large
Trier
Open
Bed
Truck
N/A
Trier
Trier
Thief
Auger
Large
Trier
Closed
Bed
Truck
Coli-
wasa
Trier
Trier
Thief
Auger
Large
Trier
Storage
Tanks
or Bins
Weighted
Bottle
Trier
Trier
Thief
Large
Trier
Waste
Piles
N/A
N/A
Trier
Thief
Large
Trier
Ponds,
Lagoons
& Pits
Dipper
Trier
Thief
Large
Trier
Con-
veyor
Belt
N/A
1
N/A
Shovel
Shovel
N/A
'Large
'Trier
Pipe
Dip-
per
N/A
-------
3.2-3
3.2.1 Composite Liquid Waste Sampler (Coliwasa)
Scope and Purpose^
The Coliwasa is a device employed to sample free flowing
liquids and slurries contained in drums, shallow open top
tanks, pits and similar containers. It is especially useful
in sampling wastes which consist of a number of immiscible
liquid phases.
The Coliwasa consists of a glass, plastic, or metal tube
equipped with an end closure which can be opened and closed
while the tube is submerged in the material to be sampled.
The Coliwasa was developed by the California Department
of Health under a grant from the U.S. EPA and their report,
"Samplers and Sampling Procedures for Hazardous Waste Streams"
[Appendix I of this manual] should be consulted.
General Comments and Precautions
1. Do not use a plastic Coliswasa to sample wastes containing
organic materials.
2. Do not use a glass Coliwasa to sample liquids that contain
hydrofluoric acid.
3. If significant amounts of solid material are present
within 2 inches of the bottom of the container to be sampled,
special procedures will be necessary to obtain a representative
sample of this solid phase.
-------
Apparatus
Coliwasas are not available commercially and must be
fabricated to conform to the specifications detailed in
Figure 3.2-1. Table 3.2-2 lists the parts required to fabricate
a plastic or glass Coliwasa.
Table 3.2-2
PARTS FOR CONSTRUCTING A COLIWASA
Quantity
1
1
1
1
1
1
1
1
Item
Sample tube, translucent
PVC plastic, 4.13 cm
I.D. x 1.52 m long
x 0.4 cm wall thickness
Sample tube, borosilicate
glass, 4.13 cm I.D. x
1.52 m long
Stopper, neoprene rubber #9
Stopper rod, PVC, 0.95 cm
O.D. x 1.67 m long
Stopper rod, teflon,
0.95 cm O.D. x 1.67 m long
Locking block, PVC, 3.8 cm
O.D. x 10.2 cm long with
0.56 cm hole in center
Locking block sleeve, PVC
4.13 cm I.D. x 6.35 cm
long
T-handle, aluminum, 18 cm
long x 2.86 cm wide with
1.27 cm wide channnel
Comments
Plastic
Coliwasa
only
Glass
Coliwasa
only
Plastic
Coliwasa
only
Glass or
Plastic
Coliwasa
Fabricate by
drilling
0.56 cm hole
through
center
Fabricate
from stock
4.13 cm PVC
pipe
Fabricate
from aluminum
bar stock
Supplier
Plastic
supply
houses
Corning
Glass Works
#72-1602
Laboratory
supply house
Plastic
supply
houses
Plastic
supply
houses
Plastic
supply
houses
Plastic
supply
houses
Hardware
stores
-------
3.2-5
Quantity
1
1
1
1
1
1
1
1
Item
Swivel, aluminum bar
1.27 cm square x 5.08 cm
long with 3/8" NC inside
thread to attach stopper
rod
Nut, PVC, 3/8" NC
Washer, PVC, 3/8" NC
Nut, stainless steel,
3/8" NC
Washer, stainless steel,
3/8"
Bolt, 3.12 cm long x
3/16" NC
Nut, 3/16" NC
Washer, lock 3/16"
Comments
Fabricate
from aluminum
bar stock
Supplier
hardware
stores
Plastic
supplier
Plastic
supplier
Hardware
stores
Hardware
stores
Hardware
stores
Hardware
stores
Hardware
stores
-------
3.2-6
Assembly
Assemble sampler as follows:
1. Attach swivel to the T-handle with the 3.12cm long bolt
and secure with the 3/16" NC washer and lock nut.
2. Shape stopper into a cone by boring a 0.95 cm hole through
the center of the stopper. Insert a short piece of 0.95
cm O.D. handle through the hole until the end of the handle
is flush against the bottom (smaller diameter) surface
of the stopper. Carefully and uniformly turn the stopper
into a cone against a grinding wheel. This is done by turning
the stopper with the handle and grinding it down conically
from about 0.5 cm of the top (larger diameter) surface to
the edge of the 0.95 cm hole on the bottom surface. Attach
neoprene stopper to one end of the stopper rod and secure
with the 3/8" NC washer and lock nut.
3. Install the stopper and stopper rod assembly in the
sampling tube.
4. Secure locking block sleeve on the block with glue
or screws.
5. Position the locking block on top of the sampling tube
so that the sleeveless portion of the block fits inside
the tube, the sleeve sits against the top end of the tube,
and the upper end of the stopper rod slips through the center
hole of the block.
6. Attach the upper end of the stopper to the swivel of
the T-handle.
-------
3.2-7
7. Place the sampler In the closed position and adjust the
tension on the stopper by screwing the T-handle in or out.
8. Test the tension by filling the Coliwasa with water to
insure it is leak free.
Procedure
1. Clean Coliwasa.
2. Adjust sampler's locking mechanism to insure that the
stopper provides a tight closure. Open sampler by placing
stopper rod handle in the T-position and pushing the rod down
until the handle sits against the sampler's locking block.
3. Slowly lower the sampler into the waste at a rate which
permits the level of liquid inside and outside the sampler
to remain the same. If the level of waste in the sampler
tube is lower inside than outside, the sampling rate is
too fast and will produce a non-representative sample.
4. When the sampler hits the bottom of the waste container,
push sampler tube down to close and lock the stopper by turning
the T-handle until it is upright and one end rests on the
locking block.
5. Withdraw Coliwasa from waste and wipe the outside
with a disposable cloth or rag.
6. Place sample tube at mouth of a container and discharge
sample by slowly opening the sampler.
-------
2.86c«U-l/a»)
j
^
1
1
1
1
1
Capered
\~ ^
} 6
T
1
1
I
- Locking r
.35c«(2-l/2") block > j
1
i
i
1.52ui(5'-0")
1
1
1
1
1
1
1
1
1 1
stopper *7[lV «-
T^
I7.0cm(7")
\4"
I1-1-
1
1
€
1
1
--
I
i
1
J <
P*
Stopper rod, PVC
0.95cm(3/8")O.D.
Pipe. PVC, 4.13cM(l-5/8")l.D.
4.26cm(l~7/a")O.D.
Stopper, neoprene, #9 with
3/8" S.S. or PVG nut and washer
SAMPLING POSITION
CLOSE POSITION
Figure 3.2-1
COMPOSITE LIQUID WASTE SAMPLER (COLIWASA)
-------
3.2-9
3.2.2 Weighted Bottle
Scope and Application
The sampler consists of a glass or plastic bottle, sinker,
stopper and a line which is used to lower and raise the bottle
and to open the bottle. A weighted bottle samples liquids
and free flowing slurries.
General Comments and Precautions
1. Do not use a plastic bottle to sample wastes containing
organic materials.
2. Do not use a glass bottle to sample wastes that contain
hydrofluoric acid.
3. Before sampling insure that the waste will not corrode
the sinker, bottle holder or line.
Apparatus
A weighted bottle with line is built to the specifications in
ASTM Methods D 270 and E 300 (Figure 3,2).
procedure
1. Clean bottle.
2* Assemble weighted bottle sampler.
3. Lower the sampler to directed depth and pull out the
bottle stopper by jerking the line.
4. Allow bottle to fill completely as evidenced by cessation
of air bubbles.
5. Raise sampler, cap and wipe off with a disposable cloth.
The bottle can serve as a sample container.
-------
3.2-10
Washer
Pin
Nut
Figure 3.2
WEIGHTED BOTTLE SAMPLER
-------
3.2-11
3.2.3 Dipper
Scope and Application
The dipper consists of a glass or plastic beaker clamped
to the end of a 2 or 3 piece telescoping aluminum or fiberglass
pole which serves as the handle. A dipper samples liquids
and free flowing slurries.
General Comments and Precautions
1. Do not use a plastic beaker to sample wastes containing
organic materials.
2. Do not USQ a glass beaker to sample wastes of high pH or
which contain hydrofluoric acid.
3. Paint aluminum pole and clamp with a 2 part epoxy or
other chemical resistant paint when sampling either alkaline
or acidic wastes.
Apparatus
Dippers are not available commercially and must be fabri-
cated to confrom to the specifications detailed in Figure 3.2-3«
Table 3.2-3 lists the parts required to fabricate a dipper.
Table 3.2-3
PARTS FOR CONSTRUCTING A DIPPER
1
Quantity I Item
1
I Adjustable clamp, 6.4 to 8.9 cm
1 | (2 1/2 to 3 1/2") for 250 to
I 600 ml beakers. Heavy duty
I aluminum.
1
I Tube 2.5 to 4.5 meters long with
1 | joint cam locking mechanism.
1 Diameter 2.54 cm ID and 3.18
) cm ID.
Supplier
Laboratory
supply
houses
Swimming
pool supply
houses
-------
3.2-12
Quantity
1
4
4
Item
Polypropylene or glass beaker,
250 ml to 600 ml.
Bolts 2 1/4" x 1/4", NC
Nuts, 1/4", NC
Supplier
Laboratory
supply
houses
Hardware
stores
Hardware
stores
Procedure
1. Clean beaker, clamp, and handle.
2. Assemble dipper by bolting adjustable clamp to the pole.
Place beaker in clamp and fasten shut.
3. Turn dipper so the mouth of the beaker faces down and
insert into waste material. Turn beaker right side up when
dipper is at desired depth. Allow beaker to fill completely
as shown by the cessation of air bubbles.
4. Raise dipper and transfer sample to container.
-------
3.2-13
Varigrip clamp
Beaker
250 to 600 ml
Telescoping aluminum pole
2.5 to 4.5 meters (8 to f5 ft)
Figure 3.2-3
DIPPER
-------
3.2-14
3.2.4 Thief
Scope and Appication
A thief consists of two slotted concentric tubes usually made
of stainless steel or brass. The outer tube has a conical
pointed tip which permits the sampler to penetrate the material
being sampled. The inner tube is rotated to open and close
the sampler. A thief is used to sample dry granules or
powdered wastes whose particle diameter is less than 1/3 the
width of the slots.
Apparatus
A thief is available at laboratory supply stores. (Figure 3.2-4)
Procedure
1. Clean sampler.
2. Insert closed thief into waste material. Rotate inner
tube to open thief. Wiggle the unit to encourage material
to flow into thief. Close thief and withdraw. Place sampler
thief in a horizontal position with the slots facing upward.
Remove inner tube from thief and transfer sample to a container.
-------
3.2-15
60-100 cm —>
JL V
-HK-
1.27-2.54 cm
Figure 3.2-4
THIEF SAMPLER
-------
3.2-16
3.2.5 Trier
Scope and Application
A trier consists of a tube cut in half lengthwise with a
sharpened tip that allows the sampler to cut into sticky
solids and loosen soil. A trier samples moist or sticky solids
with a particle diameter less than 1/2 the diameter of the trier.
Apparatus
1. Triers 61 to 100 cm long and 1.27 to 2.54 cm in
diameter are available at laboratory supply stores.
2. A large trier can be fabricated to confrom to the specifications
in Figure 3-5. A metal or polyvinyl chloride pipe 1.52
m (51) long x 3.2 cm (1.4") I.D. with a 0.32 cm (1 1/8")
wall thickness is needed. The pipe should be sawed length-
wise, about 60-40 split, to form a trough stretching from one
end to 10 cm away from the other end. The edges of the slot
and the tip of the pipe are sharpened to permit the sampler
to cut into the waste material being sampled. The unsplit
length of the pipe serves as the handle.
Procedure
1. Clean trier.
2. Insert trier into waste material 0 to 45° from horizontal.
Rotate trier to cut a core of the waste. Remove trier
with concave side up and transfer sample to container.
-------
3.2-1?
C
122-183 cm
(48-72")
1
5.08-7.62 cm
\
60-100 cm
/
\
c/
k-
1.27-2.54 cm
Figure 3.2-5
SAMPUNG TKEERS
-------
3.2-18
3.2.6 Auger
Scope and Application
An auger consists of sharpened spiral blades attached to
a hard metal central shaft. An auger samples hard or packed
solid wastes or soil.
Apparatus
Augers are available at hardware and laboratory supply stores
Procedures
1. Clean sampler.
2. Bore a hole through the middle of an aluminum pie pan
large enough to allow the blade of the auger to pass
through. The pan will be used to catch the sample brought to
the surface by the auger.
3. Place pan against the sampling point. Auger through the
hole in the pan until the desired sampling depth is
reached. Back off the auger and transfer the sample in the
pan and adhering to the auger to a container. Spoon out the
rest of the loosened sample with a sample trier.
-------
3.2-19
3.2.7 Scoop and Shovel
Scope and Application
Scoops and shovels are used to sample granular or powdered
material In bins, shallow containers and conveyor belts.
Apparatus
Scoops are available at laboratory supply houses. Flat
nosed shovels are available at hardware stores.
procedure
1. Clean sampler.
2. Obtain a full cross section of the waste material with
the scoop or shovel large enough to contain the waste
collected in one cross section sweep.
-------
3.3-1
Sub-Section 3.3
SAMPLE CONTAINERS
Containers
The most important factors to consider when choosing
containers for hazardous waste samples are compatibility
with the waste, cost, resistance to breakage, and volume.
Containers must not distort, rupture, or leak as a result of
chemical reactions with constituents of waste samples. Thus,
it is important to have some idea of the properties and
composition of the waste. The containers must have adequate
wall thickness to withstand handling during sample collection
and transport to the laboratory. Containers with wide mouths
are desirable to facilitate transfer of samples from samplers
to containers. Also, the containers must be large enough to
contain the required volume of sample or the entire volume
of a sample contained in samplers.
Plastic and glass containers are generally used for
collecting and storing of hazardous waste samples. Commonly
available plastic containers are made of high-density or
linear polyethylene (LPE), conventional polyethylene, poly-
propylene, polycarbonate, teflon FEP (flourinated ethylene
propylene), polyvinyl chloride (PVC), or polymethylpentene.
Teflon FEP is almost universally usuable due to its chemical
inertness and resistance to breakage. However, its high cost
severely limits its use. LPE, on the other hand, offers the
best combination of chemical resistance and low cost when
inorganic wastes are involved.
-------
3.3-2
Glass containers are relatively inert to most chemicals
and can be used to collect and store almost all hazardous waste
samples except those that contain strong alkali and hydro-
flouric acid. Soda glass bottles are suggested due to their
low cost and ready availability. Borosilicate glass containers,
such as Pyrex and Corex, have advantages relative to Inertness
and resistance to breakage respectively but are expensive and
not always readily available. Glass containers are generally
more fragile and much heavier than plastic containers. Glass
or FEP containers must be used for waste samples that will be
analyzed for organic compounds.
The containers must have tight, screw-type lids. Plastic
bottles are usually provided with screw caps made of the same
material as the bottles. Buttress threads are recommended.
Cap liners are not usually required for plastic containers.
Teflon cap liners should be used with glass containers supplied
with rigid plastic screw caps. These caps are usually provided
with waxed paper liners. Other liners that may be suitable
are polyethylene, polypropylene, neoprene, and teflon FEP
plastics. Teflon liners may be purchased from plastic specialty
supply houses (e.g., Scientific Specialties Service, Inc.,
P.O. Box 352, Randallstown, Maryland 21133).
-------
4.0-1
Section 4
IGNITABILITY
Introduction
The objective of the Ignitability characteristic is to
identify wastes which present fire hazards due to being
ignitable under routine storage, disposal, and transportation
and wastes capable of severely exacerbating a fire once
started.
-------
Sub-Section 4.1
CHARACTERISTIC OF IGNITABILITY REGULATION
A solid waste exhibits the characteristic of
ignitability if a representative sample of the waste has any
of the following properties:
1. It is a liquid, other than an aqueous solution con-
taining less than 24 percent alcohol by volume, and has
a flash point less than 60°C (140°F), as determined
by a Pensky-Martens Closed Cup Tester, using the test
method specified in ASTM Standard D-93-79, or a Setaflash
Closed Cup Tester, using the test method specified in ASTM
standard D-3278-78, or as determined by an equivalent test
method approved by the Administrator under the procedures
set forth in §§260.20 and 260.21.*
2. It is not a liquid and is capable, under standard
temperature and pressure, of causing fire through fric-
tion, absorption of moisture or spontaneous chemical
changes and, when ignited, burns so vigorously and per-
sistently that it creates a hazard.
3. It is an ignitable compressed gas as defined in 49 CFR
173.300 an-* as determined by the test methods described
in that regulation or equivalent test methods approved
by the Administrator under §§260.20 and 260.21.
4. It is an oxidizer as defined in 49 CFR 173.151.
* ASTM Standards are available from ASTM, 1916 Race Street,
Philadelphia, PA 19103.
-------
4.1-2
A solid waste that exhibits the characteristic of
ignitabillty, but is not listed as a hazardous waste in
Subpart D, has the EPA Hazardous Waste Number of D001.
-------
Sub-Section 4.4
IGNITABLE COMPRESSED GAS
For the purpose of this regulation the following terminol-
ogy is defined:
1. Compressed gas. The term "compressed gas" shall
designate any material or mixture having in the
container an absolute pressure exceeding 40 p.s.i. at
70°F or, regardless of the pressure at 70°F, having an
absolute pressure exceeding 104 p.s.l. at 130°F, or
any liquid flammable material having a vapor pressure
exceeding 40 p.s.i. absolute at 100°F as determined by
ASTM Test D-323.
2. Ignitable compressed gas. Any compressed gas as
defined in paragraph (a) of this section shall be
classed as an "ignitable compressed gas" if any one
of the following occurs:
a. Either a mixture of 13 percent or less
(by volume) with air forms a flammable mixture
or the flammable range with air is wider than
12 percent regardless of the lower limit. These
limits shall be determined at atmospheric
temperature and pressure. The method of sampling
and test procedure shall be acceptable to the
Bureau of Explosives.
b. Using the Bureau of Explosives' Flame
Projection Apparatus (see Note 1), the flame
projects more than 18 inches beyond the ignition
-------
4.4-2
source with valve opened fully, or, the flame
flashes back and burns at the valve with any
degree of valve opening.
c. Using the Bureau of Explosives' Open Drum
Apparatus (see Note 1), there is any significant
propagation of flame away from the ignition source.
d. Using the Bureau of Explosives' Closed Drum
Apparatus (see Note 1), there is any explosion
of the vapor-air mixture in the drum.
NOTE 1: A description of the Bureau of
Explosives' Flame Projection Apparatus,
Open Drum Apparatus, Closed Drum Appa-
ratus, and method of tests may be
procured from the Bureau of Explosives.
-------
4.5-1
Sub-Section 4.5
OXIDIZER
An oxidizer for the purpose of this regulation is any
material that yields oxygen readily to stimulate the
combustion of organic matter (e.g., chlorate, permanganate,
peroxide, nitro carbo nitrate, inorganic nitrate).
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5.(XL
Section 5
CORROSIVITY
Introduction
The corrosivity characteristic, as defined in 40 CFR
261*22, is designed to identify wastes which might pose a
hazard to human health or the environment due to their ability
to:
a. Mobilize toxic metals if discharged into a landfill
environment•
b. Require handling, storage, tranportation and management
equipment to be fabricated of specially selected mater-
ials of construction, and
c. Destroy human or animal tissue in the event of inadver-
tant contact.
In order to identify such potentially hazardous materials,
the Agency has selected several properties upon which to base
the definition of a corrosive waste. These properties are
pH and corrosivity toward Type SAE 1020 steel. The following
section presents methodology for measuring the pH of aqueous
wastes and for determining whether the waste is corrosive to
steel.
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5.1-1
Sub-Section 5.1
CHARACTERISTIC OF CORROSIVITY REGULATION
1. A solid waste exhibits the characteristic of corrosivity
if a representative sample of the waste has either of
the following properties:
a. It is aqueous and has a pH less than or equal to 2
or greater than or equal to 12.5, as determined by a
pH meter using either the test method specified in the
"Test Methods for the Evaluation of Solid Waste, Physical/
Chemical Methods"* (also described in "Methods for Analysis
of Water and Wastes" EPA 600/4-79-020, March 1979), or
an equivalent test method approved by the Administrator
under the procedures set forth in §§260.20 and 260.21.
b. It is a liquid and corrodes steel (SAE 1020) at a
rate greater than 6.35 mm (0.250 inch) per year at a
test temperature of 55°C (130°F) as determined by the
test method specified in NACE (National Association of
Corrosion Engineers) Standard TM-01-69** as standardized
in "Te^t Methods for the Evaluation of Solid Waste,
Physical/Chemical Methods," or an equivalent test
method approved by the Administrator under the proce-
dures set forth in §§260.20 and 260.21.
2. A solid waste that exhibits the characteristic of corrosivity,
but is not listed as a hazardous waste in Subpart D, has
the EPA Hazardous Waste Number of D002.
*This document is available from Solid Waste Information, U.S.
Environmental Protection Agency, 26 W. St. Clair Street,
Cincinnati, Ohio 45268
**The NACE Standard is available from the National Association of
Corrosion Engineers, P.O. Box 986, Katy, Texas 77450
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5.2-1
Method 5.2
pH MEASUREMENT
Scope and Application
This method* is applicable to aqueous wastes and those
multiphasic wastes where the aqueous phase comprises at least
20% of the total volume of the waste.
Summary
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 solutions of known pH.
Interferences
1. The glass electrode, in general, is not subject to solution
interferences from color, turbidity, colloidal matter,
oxidants, reductants or high salinity.
2. Sodium error at pH levels greater than 10 can be reduced
or eliminated by using a "low sodium error" electrode.
3. Coatings of oily material or particulate matter can
impair electrode response. These coating can usually be
removed by gentle wiping or detergent washing, followed by
distilled water rinsing. An additional treatment with
hydrochloric acid (1 + 9) may be necessary to remove any
remaining film.
*This method has been adapted from Method 150.1 of "Methods
for the Analysis of Water and Wastewater", EPA-6000/4-79-020,
U.S. EPA, EMSL, Cincinnati, OH 45268
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5.2-2
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 is the change of pH inherent in the sample at
various temperatures. 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.
Apparatus
1. pH Meter-laboratory or field model. A wide variety of
instruments are commercially available with various
specifications and optional equipment.
2. Glass electrode.
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. Magnetic stirrer and Teflon-coated stirring bar.
5. Thermometer or temperature sensor for automatic compensation.
Reagents
1. Primary standard buffer salts are available from the
National Bureau of Standards and should be used in situations
-------
5.2-3
where extreme accuracy is necessary. Preparation of
reference solutions from these salts requires some special
precautions and handling(l) such as low conductivity
dilution water, drying ovens, and carbon dioxide free
purge gas* These solutions should be replaced at least
once each month.
2. Secondary standard buffers may be prepared from NBS
salts or purchased as a solution from commercial vendors.
Use of these commercially available solutions, that have
been validated by comparison to NBS standards, are recom-
mended for routine use.
Calibration
1. Because of the wide variety of pH meters and accessories,
detailed operation procedures cannot be incorporated
into this method. Each analyst must be acquainted with
the operation of each system and familiar with all instru-
ment functions. Special attention to care of the electrodes
is recommended.
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. Various instrument designs may involve use
of a "balance" or "standardize" dial and/or slope adjust-
ment as outlined in the manufacturer's instructions.
Repeat adjustments on successive portions of the two
buffer solutions as outlined in procedure 8.2 until
National Bureau of Standards Special Publication 260.
-------
readings are within 0.05 pH units of the buffer solution
value.
Procedure
1. Standardize the meter and electrode system as outlined
in Section 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
in the sample stream to an adequate depth and moved in a
manner to insure sufficient sample movement across the
electrode sensing element as indicated by drift free
«0.1 pH) readings.
• 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 tempera-
ture differences. Refer to manufacturer's instructions.
4. After rinsing and gently wiping the electrodes, if
necessary, immerse them into the sample beaker or sample
stream and stir at a constant rate to provide homogeneity
and suspension of solids. Rate of stirring should minimize
the air transfer rate at the air water interface of the
sample. Note and record sample pH and temperature.
Repeat measurement on successive volumes of sample until
values differ by less than 0.1 pH units. Two or three
volume changes are usually sufficent.
-------
5.2-5
Bibliography
1. Standard Methods for the Examination of Water and Wastevater,
14th Edition, p. 460, (1975).
2. Annual Book of ASTM Standards, Part 31, "Water", Standard
D1293-65, p. 178, (1976).
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5.3-1
Method 5.3
CORROSIVITY TOWARD STEEL
Introduction
This methodt is applicable to either aqueous or non aqueous
liquid wastes.
Summary of the Method
This test exposes coupons of SAE Type 1020 steel to the
liquid waste to be evaluated and, by measuring the degree to
which the coupon has been dissolved, determines the corrosivity
of the waste.
Precautions and Comments
1. In laboratory tests, such as this one, corrosion of
duplicate coupons is usually reproducible to within + 10%.
Occasional exceptions, in which large differences are
observed, can occur under conditions where the metal
surfaces become passivated. Therefore, at least dupli-
cate determinations of corrosion rate should be made.
2. A circular specimen of about 3.75 cm (1.5 inch) diameter
is a convenient shape for laboratory use. With a thick-
ness of approximately 0.32 cm (0.125 inch) and a 0.80 cm
(0.4 inch) diameter hold for mounting, these specimens
will readily pass through a 45/50 ground glass joint of a
distillation kettle. The total surface area of a circular
specimen is given by the following equation:
tThis method is based on NACE Standard TM-01-69(1972 Revision),
"Laboratory Corrosion Testing of Metals for the Process Industries",
National Association of Corrosion Engineers, 3400 West Loop South,
Houston, TX 77027
-------
5.3-2
A - 3.14/2(D2-d2)+(t)(3.14)(D)+(t)(3.14)(d)
where t * thickness, D - diameter of the specimen, and
d - diameter of the mounting hole. If the hole is completely
covered by the mounting support, the last term [(t) (3 .14)(d) ]
in the equation is omitted.
3. All coupons should be measured carefully to permit
accurate calculation of the exposed areas. An area cal-
culation accurate to 4^ 1% is usually adequate.
4. More uniform results may be expected if a substantial
layer of metal is removed from the coupons prior to testing
the corrosivity of the wste. This can be accomplished
either by chemical treatment (pickling), electrolytic
removal, or by grinding with a coarse abrasive. At
least 0.000254 cm (0.0001 inch ) or 2 to 3 mg/cm2 should
be removed. Final surface treatment should include
finishing with #120 abrasive paper or cloth. Final
cleaning consists of scrubbing with bleachfree scouring
powder, followed by rinsing in distilled water, then
acetone or methanol, and finally air drying. After
final cleaning the coupon should be stored in a desic-
cator until used.
5. The minimum ratio of volume of waste to area of the
metal coupon to be used in this test is 40 ml/cm2.
Equipment
1. A versatile and convenient apparatus should be used, con-
sisting of a kettle or flask of suitable size (usually 500
to 5000 millilieters), a reflex condenser with atmospheric
-------
5.3-3
seal, a thermowell and temperature regulating device, a
heating device (mantle, hot plate, or bath), and a specimen
support system. A typical resin flask set up for this type
test is shown in figure 1.
2. The supporting device and container should not he
affected by or cause contamination of the waste under test.
3. The method of supporting the coupons will vary with
the apparatus used for conducting the test but should be
designed to insulate the coupons from each other physically
and electrically and to insulate the coupons from any metallic
container or other device used in the test. Some common
support materials include: glass, fluorocarbon or coated metal.
4. The shape and form of the coupon support should assure free
contact with the waste.
Test Procedure
1. Assemble the test apparatus as described the "Equipment"
section above.
2. Fill the container with the appropriate amount of waste.
(See #5 under the "Precautions and Comments" section.)
3. Begin agitation at a rate sufficient to insure that
the liquid is kept well mixed and homogeneous.
4. Using the heating device bring the temperature of
the waste to 55° C (130° F).
5. If the anticipated corrosion rate is moderate (i.e.,
<635 mmpy), the test should be run for at leat 200 hours to
insure adequate weight loss to permit accurate results to be
obtained. If the corrosion rate is low (i.e., >100 mmpy),
-------
5.3-4
then the test duration should be on the order of 2000 hours.
in cases where the anticipated corrosion rate is completely
unknown, initial testing should be performed using a 200
hour duration.
6. In order to accurately determine the amount of material
lost to corrosion, the coupons have to be cleaned after
immersion and prior to weighing. The cleaning procedure
should remove all products of corrosion while removing a
minimum of sound metal. Cleaning methods can be divided
into three general categories: mechanical, chemical and
electrolytic.
Mechanical cleaning includes scrubbing, scraping, brushing
and ultrasonic procedures. Scrubbing with a bristle brush
and mild abrasive is the most popular of these methods; the
others are used in cases of heavy corrosion as a first step
in removing heavily encrusted corrosion products prior to
scrubbing. Care should be taken to avoid removing sound metal.
Chemical cleaning implies the removal of material from
the surface of the coupon by dissolution in an appropriate
solvent. Solvents such as acetone, dichloromethane, and
alcohol are used to remove oil, grease or resinous materials,
and are used prior to immersion to remove the products of corrosion.
Solutions suitable for removing corrosion from the steel
coupon are:
Solution Soaking Time Temperature
20% NaOH + 200g/l zinc dust 5 min Bailing
or
Cone. HC1 + 50g/l SnCl2 + 20g/l SB-^ Until clean Cold
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5.3-5
Electrolytic cleaning should be preceeded by scrubbing
to remove loosely adhering corrosion products. One method of
electrolytic cleaning that can be employed is:
Solution
Anode
Cathode
Cathode current density
Inhibitor
Temperature
Exposure Period
50 g/1 H2S04
Carbon or lead
Steel coupon
20 amp/cm2 (129 amp/in2)
2 cc organic inhibitor/liter
74°C (165°F)
3 minutes
Note; Precautions must be taken to insure good electrical
contact with the coupon, to avoid contamination of the cleaning
solution with easily reducible metal ions, and to insure
that inhibitor decomposition has not occurred. Instead of
using a proprietary inhibitor, 0.5 g/1 or either diorthotolyl
thiourea or quinolin ethiodide can be used.
Whatever treatment is employed to clean the coupons, its
effect in removing sound metal should be determined using a
blank (i.e., a coupon that has not been exposed to the waste).
The blank should be cleaned along with the test coupon and
its waste loss subtracted from that calculated for the test
coupons.
7. After corroded specimens have been cleaned and dried,
they are reweighed. The weight loss is employed as the
principal measure of corrosion. Use of weight loss as a
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5.3-6
measure of corrosion required making the assumption that
all weight loss has been due to generalized corrosion
and not localized pitting. In order to determine the
corrosion rate for purpose of this requlation, the following
formula is used:
Corrosion Rate (mmpy) =» (weight loss) (0.268)
(area) (time)
where weight loss is in milligrams, area in square
centimeters, time in hours, and corrosion rate in
millimters per year (mmpy).
-------
f!J] technical practices committsss
NACE Standard TM-01-89
(1972 Revision)
Test Method
Laboratory Corrosion Testing of Metais
for the Process Industries
Approved March, 1969
Revised August, 1972
National Association of Corrosion Engineers
2400 Wert Loop Sooth
Houston, Texas 77027
713/622-8980
-------
5.3-6
The National Association of Corrosion Engineers issues this
Standard in conformity to the best current technology
regarding the specific subject. This Standard represents
minimum requirements and should in no way be inter-
preted as a restriction on the use of better procedures or
materials. Neither is this Standard intended to apply in any
and all cases relating to the subject. Numerous external
factors may negate the usefulness of this Standard in
specific instances.
This Standard may be used in whole or in part by any party
without prejudice if recognition of the source is included.
The National Association of Corrosion Engineers assumes
no responsibility for the interpretation or use of this
Standard-
Nothing contained in this Standard of the National Asso-
ciation of Corrosion Engineers is to be construed as-
granting any right, by implication or otherwise, for manu-
facture, sale, or use in connection with any method,
apparatus, or product covered by Letters Patent, nor as
indemnifying or protecting anyone against liability for
infringement of Letters Patent.
Foreword
Unit Committee T-5A ("Corrosion in Chemical Processes")
of the National Association of Corrosion Engineers issues
this Standard with a dual purpose.
The first purpose is to standardize, as much' as possible,
simple immersion corrosion studies. In this sense, this
Standard is reasonable and effective without imposing
inflexible requirements as to apparatus, conditions, or
techniques. The actual conditions of test will be determined
by the problem at hand and limited only by the ingenuity
of the individual investigator.
The second purpose of this Standard is to present to the
user a consensus on the best current technology in this fieJd
of laboratory corrosion testing. As such, this Standard
enumerates and discusses the many factors which must be
considered, controlled, and reported in order to aid in
correlation or reproducibility of such studies.
The techniques described permit the investigator to repro-
duce to a considerable extent in the laboratory, through
judicious experimental design, the process conditions which
govern corrosion mechanisms. The tests are not to be
construed as "accelerated" tests, which are generally
unreliable. The methods described are also applicable to
materials qualification tests for quality control. However,
the latter require more rigid definition, of apparatus,
conditions, and technique.
The ultimate purpose is better correlation of results in the
future and the reduction of conflicting reports through a
more detailed recording of meaningful factors and con
ditions.
-------
TEST METHOD
Laboratory Corrosion Testing of Metals
for the Process Industries
CONTENT*
1. Genera] 2
2. Specimen Preparation 2
3. Equipment and Apparatus 3
4. Test Conditions 4
5. Methods of Cleaning Specimens
After the Test 7
6. Evaluation of Results 8
7. Calculating Corrosion Rates 9
8. Reporting the Data 10
References 10
Additional copies of this NACE Standard are available at 52 per copy from
NACE Headquarters, 2400 West Loop South, Houston, Texas 77027.
-------
1. General
1.1 This Standard describes the factors which influence
laboratory tests. These factors include specimen prepara-
tion, apparatus, test conditions (solution composition,
temperature, velocity, aeration, volume, method of
supporting specimens, duration of test), methods of
cleaning specimens, evaluation of results, and calculation of
corrosion rates. This Standard also emphasizes the impor-
tance of recording all pertinent data and provides a check
list for reporting test data.
1.2 Experience has shown that all metals and alloys do
not respond alike to the many factors that control
corrosion and that "accelerated", corrosion tests- give
indicative results only. Consequently, it is impractical to
propose an inflexible standard laboratory corrosion testing
procedure for general use except for material qualification
tests, where standardization is obviously required,
1.3 In designing any corrosion test, consideration must be
given to the various factors discussed in this test method
because these factors have been found to affect greatly the
results obtained.
,5*3
2. Specimen Preparation
2.1 In laboratory tests, corrosion rates of duplicate
specimens are usually within ± 10% under the same test
conditions. Occasional exceptions, in which a large dif-
ference is observed, can occur under conditions of border-
line passivity of metals or alloys 'that depend on a passive
film for their resistance to corrosion. Therefore, at least
duplicate specimens should be exposed in each test.
2.2 If the effects of corrosion are to be determined by
changes in mechanica] properties, untested duplicate
specimens should oe preserved in a non-corrosive environ-
n.^.ii for comparison with the corroded specimens. The
mechanical property commonly used for comparison is the
tensile strength. The*procedure for determining this value is
shown in detail in ASTM Standard E-8- latest edition.
2.3 The size and the shape of specimens will vary with the
purpose of the test, nature of the materials, and apparatus
used. A large surface-to-mass ratio and a small ratio of edge
area to total area are desirable. These ratios can be achieved
through the use of rectangular or circular specimens of
minimum thickness. Circular specimens should be cut
preferably from sheet and not bar stock to minimize the
exposed end grain.
2.3.1 A circular specimen of about 1 1/2-thch
diameter is a convenient shape for laboratory corro-
sion tests. With a thickness of approximately 1/8 inch
and a 5/16 or 7/16-inch diameter hole for mounting,
these specimens will readily pass through a 45/50
ground glass joint of a distillation kettle. The total
surface area of a circular specimen is given by the
following equation:
A= -
where t = thickness, D = diameter of the specimen,
and d = diameter of the mounting hole. If the hole is
completely covered by the mounting support, the last
term (t~d) in the equation is omitted.
2.3.2 Strip coupons fof about 4 square inches) may
be preferred as corrosion specimens, particularly if
interface or liquid line effects are to be studied by the
laboratory test, but such effects are beyond the scope
of this Standard.
2.3.3 All specimens should be measured carefuuy to
permit accurate calculation of the exposed areas.- An
area calculation accurate to plus or minus 1% is
usually adequate.
2.4 More uniform results may be expected if a substantia7
layer of metal is removed from the specimens to eliminate
variations in condition of the original metallic surface. This
can be done either by chemical treatment (pickling),
electrolytic removal, or by grinding with a coarse abrasive
paper or cloth, such as No. 50, using caie not to work
harden the surface (see Section 2.7). At least 0.0001 inch
or 10 to 15 milligrams per square inch should be removed.
If clad alloy specimens are to be used, special attention
must be given to insure that excessive metal is not removed-
After final preparation of the specimen surface-trie
specimens should be stored in a desiccator until exposure if
they are not used immediately.
2.5 Exposure of sheared edges should be avoided unless
the purpose of the test is to study effects of the shearing
operation. It may be desirable to test a surface representa-
tive of the material and metallurgical condition used in
practice.
2.6 The specimen can be stamped with an appropriate
identifying mark.
2.6.1 The stamp, besides identifying the specimen,
introduces stresses and cold work in the specimen,
that could be responsible for localized corrosion
and/or stress corrosion cracking.
2.6.2 Stress corrosion cracking at the identifying
mark is a positive indication of susceptibility to such
-------
corrosion; however, "the absence of cracking should
not be interpreted as indicating resistance. Additional
tests should be run to study specifically the effects of
stress.
2.7 Final surface treatment of the specimens should
include finishing with No. 120 abrasive paper or cloth, or
the equivalent, unless the surface is to be used in the
mill-finished condition. This resurfacing may cause some
surface work-hardening to an extent which will be deter-
mined by the vigor of the surfacing operation but is not
ordinarily significant.
2.7.1 Coupons of different alloy compositions
should never be ground on the same cloth.
2.7.2 Wet grinding should be used on alloys which
work harden quickly, such as the austenitic stainless
steels,
2.S The specimens should be finally degreased by scrub-
bing with bleach-free scouring powder, followed by
thorough rinsing in water and in a suitable solvent (such as
acetone, methanol, or a mixture of 50%methanol and ,50%
ether) and air dried. For relatively soft metals such as
aluminum, magnesium, and copper, scrubbing with abrasive
is not always needed and can mar the surface of the-
specimen. Trie use of towels for drying may introduce an
error through contamination of the specimens with grease
or lint.
2.9 The dried specimens should be weighed on an
analytical balance to an accuracy of plus or minus 0.5
milligram.
2 10 The method of specimen preparation should be
described when reporting tests results to facilitate interpre-
tation of data by other persons.
2.10.1 Reports should include trade name or
composition of specimens in the following order of
preference: (a) chemical composition determined by
analysis, (b) approximate or nominal chemical
composition, and (c) trade name or grade and
specification (if bought to MIL, ASTM, etc.)
2.10.2 Metallurgical condition of the specimens
including the degree of hot or cold working arid heat
treatment, should be described as completely as
possible.
2.11 The use of welded specimens is often desirable
because some welds may be cathodic or anodic to the base
metal and may affect the corrosion.
2.11.1 The heat-affected zone is also of importance
but should be studied separately because wtlds on
coupons do not faithfully reproduce heat input or
size effects of full-size vessels.
2.11.2 Corrosion of a welded coupon is best reported
by description 'and thickness measurements rather
than a mils-per-year rate because the attack is
normally localized and not representative of the
entire surface.
2.11.3 A complete discussion of corrosion testing of
welded coupons or the effect of heat treatment on
the corrosion resistance of a metal is not within the
scope of this Standard.
5.3-n
3. Equipment and Apparatus
3.1 A versatile and convenient apparatus should be used,
consisting of a kettle or flask of suitable size (usually 500
to 5000 mJMiters), a reflux condenser with atmospheric
seal, a sparger for controlling atmosphere or aeration, a
thermowell and temperature regulating device, a heating
device (mantle, hot plate, or bath), and a specimen support
system. If agrfation is required, the apparatus can be
modified to accept a suitable stirring mechanism such as a
magnetic stirrer. A typical resin flask set up for this type
test is shown in Figure 1.
3.2 These suggested components can be modified, simpli-
-fied or made more sophisticated to fit the needs of a
particular investigation. The suggested apparatus is basic,
and the apparatus is limited only by the judgment and
ingenuity of the investigator.
3.2.1 A glass reaction kettle can be used wnere
configuration and size of specimens will permit entry
through the narrow kettle neck.
3.2.2 In some cases, a wide mouth jar with a
suitable closure is sufficient when simple immersion
tests at ambient temperatures aie to be investigated.
3.2.3 Open beaker tests should not be used because
of evaporation and contamination.
3.2.4 In more complex tests, provisions might be
needed for continuous flow or replenishment of the
corrosive liquid while simultaneously maintaining a
controlled atmosphere.
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4. Test Conditions
4.1 Selection of the conditions for a laboratory corrosion
test will be determined by the purpose of the test.
4.1.1 If the test is to be a guide for the selection of
a material for a particular purpose, the limits of
controlling factors in service must be determined.
These factors include oxygen concentration, tempera-
ture, rate of flow, pH value, and other important
characteristics of the solution.
4.2 An effort should be made to duplicate all service
conditions in the corrosion test.
4.3 It is important that test conditions be controlled
throughout, the test in order to ensure reproducible results.
4.4 The spread in corrosion rate values for duplicate
specimens in a-given test probably should not exceed ± 105o
of the average when the attack is uniform.
4.5 Composition of soJution-
4.5.1 Test solutions should be prepared accurately
from chemicals conforming to the Standards of the
Committee on Analytical Reagents of the American
Chemical Society,1 and distilled water, except in
these cases where naturally occurring solutions or
these tzken directly from some plant,process are
used.
4.5.2 The composition of the test solution should
be controlled to the fullest extent possible and should
be described as completely and as accurately as
possible when the results are reported.
4.5.2.1 Minor constituents should not be
overlooked because they often affect corro—
sion rates-
4.5.2.2 Chemical content should be reported
zs percentage by weight of the solution.
Molarity and normality are also helpful in
defining the concentration of chemicals in the
test solution.
4.5.3 The composition of the test solution should
be checked by analysis at the end of the test to
determine the extent of change in composition, such
as might result from evaporation.
4.5.4 Evaporation losses should be controlled by a
constant level device or by frequent additions of
appropriate solution to maintain the'orieinal volume
within ± \%.
4.5.5 In some cases, composition of the test solu-
tion may change as a result of catalytic decompo-
1 - Typical resin flask that can b% useo as a versat'ri* »»d
convenient apparatus to conduct simple immersion tuts. Con-
figuration of the flask top it such th*t more sophisticated
apparatus can be added as required by the specific test b«tng
conducted. A * thermowell. B • resin flask. C* specimens hong
on supporting device, D - a*t inlet. E " heating mentlev F -
liquid interface, G - opening in flash for additional apparatus
that may be required, and H ™ reflux condenser.
sition or by reaction with the- test coupon These
changes should be determined .if possible. Where
required, the exhausted constituents should be added
or a fresh solution provided, during the course of the
test.
4.5.6 When possible, only one type of metal should
be exposed in a given test. If several different metals
are exposed in the same solution, the corrosion
products from one metal may affect the rate of attack
on another metal. For example, copper corrosion
products can reduce corrosion of stainless steel and
titanium but can accelerate corrosion of aluminum,
4.6 Temperature of solution.
4.6.1 Temperature of the corroding solution should
be controlled within ±IC (± l.S F) and must be
stated in the report of test results.
-------
4.6.2 If no specific temperature, such as boiling, is
required or if a temperature range is to be investi-
gated, the selected temperatures used in the test must
be reported.
4.6.3 For tests at ambient temperatures, the tests
should be conducted at the highest temperature
anticipated for stagnant storage in summer months.
This temperature may be as high as 40 to 45 C (104
to 113 F) in some areas. The variation in temperature
should be reported also (e.g., 40 C ± 2 C).
4.7 Aeration of solution.
4.7.1 Unless specified, the solution should not be
aerated. Most tests related to process equipment
should be run with the natural atmosphere Inherent in
the process, such as the vapors of the boiling liquid.
4.7.2 If aeration is used, the specimens should not
be located in the direct air stream from the sparger.
Extraneous effects can be encountered if the air
stream impinges on the specimens.
4.73 If complete exclusion of dissolved oxygen is
necessary, specific techniques are required such,as
prior heating of the solution and sparging with an
inert gas (usually nitrogen). A liquid atmospheric seal
is required on the test vessel to prevent further
contamination.
4.7.4 _1£ oxygen saturation of the test solution is
desired, this can best be achieved by sparging. For
other degrees of aeration, the solution should be
sparged with synthetic mixtures of air or oxygen with
an inert gas.
4.8 Solution velocity
4.8.1 The effect of velocity is not usually deter-
mined in normal laboratory tests although specific
tests have been designed for this purpose. However,
for the sake of reproducibility, some velocity control
is desirable.
4.8.2 Tests at the boiling point should be conducted
with minimum possible heat input, and boiling chips
should be used to avoid excessive turbulence and
bubble impingement.
4.8.3 In tests conducted below the boiling point,
thermal convection generally is the only source of
liquid velocity.
.4 In test solutions with high viscosities, supple-
mental controlled stirring is recommended.
4.9 Volume of test solution.
4.9.1 me volume of the test solution should b«
large enough to avoid any sppf-reiabie change in its
corrosiveness either through exhaustion of corrosive
constituents or accumulation of corrosion products
that might affect further corrosion.
4.9.2 The preferred minimum volume-to-area ratio is
250 rnillititers of solution per square inch of specimen
surface as stipulated jn ASTM Standard A-279-63 on
"Total Immersion Corrosion Test of Stainless Steels,"
4.9.3 When the test objective is to determine the
effect of a metal or a21oy on the characteristics of the
test solution (for example, to determine the effects of
rsetals on dyes), it is desirable to reproduce the ratio
of solution volume to exposed metal surface that
exists Li practice. The actual time of ^contact of the
metal with the solution also must be taken into
account. Any necessary distortion of the test con-
ditions must be considered when interpreting the
results.
4.10 Method of supporting specimens.
4.10.1 The supporting device and container should
nor be affected by or cause contamination of the test
solution.
4.10.2 The method cf supporting specimens w31 vary
with the apparatus used for conducting the test but
should be designed to insulate the specimens from
each other physically and electrically and to insulate
the specimens from any metallic container or support-
ing device used with the apparatus.
4.10.3 Shape and form of the specimen support
should assure free contact of the specimen with the
corroding solution, the liquid line, or the vapor phase
as .shown in Figure 1. If clad alloys are exposed,
special procedures will be required to insure that only
the cladding is exposed unless the purpose is to test
the ability of the cladding to protect cut edges in the
test solution.
4.10.4 Some common supports are glass or ceramic
rods, glass saddles, glass hooks, fluorocarbon plastic
strings, and various insulated or coated metallic
supports.
4.11 Duration of test.
4.11.1 Although duration of any test will be deter-
mined by the nature and purpose of the test» an
-------
excellent procedure for evaluating the effect of time
on corrosion of the metal and also on the corrosive-
ness of the environment in laboratory tests has been
presented by Wachter and Treseder.2 This technique
is calied the "Planned Interval Test," and the proce-
dure and evaluation of results are given in Table 1.
Other procedures that require the removal of solid
corrosion products between exposure periods will not
measure accurately the normal changes of corrosion
with time.
4.1J.2 Materials which experience severe corrosion
generally do not need lengthy tests to obtain accurate
corrosion rates. Although this assumption is valid in
many cases, there are cases where it is not valid. For
example, lead exposed to sulfuric acid corrodes at an
extremely high rate at first whiJe building a protective
film, then the rates decrease considerably so that
further corrosion is negligible. Tne phenomenon of
forming a protective film is observed with many
corrosion resistant materials, and therefore short tests
on such materials would indicate a high corrosion rate
and woiJd be completely misleading.
4.113 Short time tests also can give misleading
results on alloys that form passive films, such as
stainless steels. With borderline conditions, a pro-
longed test may be needed to permit breakdown of
the passive film and subsequently more rapid attack.
Consequently, tests run for long periods are con-
siderably more -realistic than those conducted for
short durations. This.statement must be qualified by
stating that corrosion should notproceed'to the point
where the original specimen size or the exposed area
is drastically reduced or where the metal is per-
forated.
4.11.4 If anticipated corrosion rates are moderate or
low the following equation2 gives a suggested test
duration:
2000
TABl_£ 1 - =M»nned IntervM
Duration of test (hr)- COITOsion rate (mpy)
Examples: Where the corrosion rate is 10 mpy, the
test should run for at least 200 hours. If the rate is 1
mpy, the duration should be at least 2000 hours.
4.11.4.1 This method of estimating test
duration is useful only as an aid in deciding,
after a test has been made, whether or not it is
desirable to repeat the test for a longer period.
Tne most common testing periods are 48 to
168 hours (2 to 7 days).
4.11.5 In some cases, it may be necessary to know
the degree of contamination caused by the products
of corrosion; this can be accomplished by analysis of
(Reprinted by permission from "Chemical Engineering Progress.*"
June, 1947.)
'5.3-4*
•u
8 A,
oo " "
- 1 I
H 0 1
A,
Time
B
t H
Identical specimens-all placed in the same corrosive Quid. Impend
conditions of the test kept constant for entire time t •+• 1. Letters,
AI, At, AJ+J, B, represent corrosion damage experienced by each
test specimen. A; is calculated by subtracting At from Al+ j.
Occurrences During Corrosion Test
Liquid corrosiveness
Metal corrodibility
Criteria
unchanged
decreased
increased
unchanged
decreased
increased
A,=B
B B
AI A2
-------
The solution after corrosion has occurred. The corro-
sion rate can be calculated from the concentration of
the matrix metal found in the solution, and it can be
compared to that determined from the weight loss of
the specimens. However, scrr.s of the corrosion
products usually adhere to ihe specimen as a. scale,
and the corrosion rate calculated from the metal
content in the solution is not always correct.
5.3-15
5. Methods of Cleaning Specimens After the Test
S.I Before specimens are cleaned, their appearance should 5.2 Cleaning specimens after the test is a vital step.in the
be observed and recorded. Location of deposits, variations corrosion test procedure and, if not done properly, can
in types of deposits, or variations in corrosion products are cause misleading results.
extremely important in evaluating localized corrosion, such
as pitting and concentration cell attack. 5.2.1 Generally, the cleaning procedure should.
TABLE 2—Method*for Chemical Cleaning of Corrosion Test Specimens After Exposjre
Material
Chemical
Time
Tempera-
ture
Remarks
Aluminum and
Aluminum Alloys
Copper and
Copper ADoys
Lead and
Lead Alloys
Iron and Steel
Magnesium and
Magaesium ADoys
Nickel and
Nickel Alloys
Stainless Steel
Tin and Tin AlJoys
Zinc
70% HSO3
2r»Cr03>5%H3P04, Sob.
15-20% KC1
5-10%H2S04.
1% acetic acid
5% ammonium acetate
80g/lNaOH,
50 g/1 majuiitol,
0.62 g/1 hydrazine sulfate
20% NaOH, 200 g/1 zinc dast
cone. HO, 50 g/1.
Snd2 + 20 g/1 SbCl3
15%CrO3, !%AgCrO4 Spin.
15-20% HC1
10%H2SO4
10%HNO3
lS#Na3PO«
10%NH4a
followed by
5%CrO3. 1% AgN03 Sob.
Saturated amnionium
acetate
lOOj/lNaCN
2-3 mia
lOrr.b
2-3 nib
2-3 min
10 min
5 min
30 rr.in, or
until clem
5 mia
LJntfl ctean
15 min
Unu! cl:an
Until clsLi
UntD ck£n
10 -in
5 min
20 we
Untfl clean
15 nib
Room
175-185 F
(79-S5 Q
Room
Room
Boiling
Hot
Boiling
Bofling
Cold
Boiling
Room
Room
140 F
(60Q
Boiling
Room
Boiling
Room
Room
FoDow by light scrub.
Used when oxide film
resists HNO3 treat- •
ment. Follow by 70S.
HNO3 treatment pre-
viously described.
Follow by light scrub.
•
Follov by light scrub.
Follow by light scrub.
Removes PbO.
FoUow by light scrub.
Removes PbO and/or
PbSO«.
Follow by light scrub.
...
—
—
...
...
Avoid contamination
with chlorides
Follow by scrubbing.
Follow by light
scrubbing.
Follow by light scrub.
• • •
-------
remove all corrosion products from specimens with a
minimum removal of sound metal.
5.2.2 Set rules cannot be applied to specimen
cleaning because procedures will vary depending on
the type of metal being cleaned and on the degree of
adherence of corrosion products.
5.3 Cleaning methods can be divided into three general
categories: mechanical, chemical, and electrolytic.
5.3.1 Mechanical cleaning includes scrubbing,
scraping, brushing, mechanical shocking, and ultra-
sonic procedures. Scrubbing with a bristle brush and
mild abrasive is the most popular of these methods;
the others are used principally as a supplement to
remove heavily encmsted corrosion products before
scrubbing. Care should be used to avoid the removal
of sound metal.
5.3.2 Chemical cleaning implies the removal of
material from the surfs ce of the specimen by dissolu-
tion in an appropriate chemical solution. Solvents
such as acetone, carbon tetrachlorice, and alcohol.-^re
used to remove oil.-grease, or resin and are usually
applied prior to other methods of cleaning. Chemicals
are chosen for application to a specific material. Some
of these treatments in general use are outlined in
Tabie 2.
5.3.3 Electrolytic cleaning should be preceded by
scrubbing to remove loosely adhering corrosion
products. One method of electrolytic cleaning that
has been found to ^>e useful for many metals and
alloys is as followr
Solution 5% (by weight) H, SO4
Anode Carbon or lead
Cathode Test specimen.
Cathode CD 20 amp/dm3
(129amp/sqin)
Inhibitor 2 cc organic inhibitor
per liter
Temperature 74 C (165 F)
Exposure period... 3 minutes
53.3.1 Precautions must be taken to insure
good electrical contact with the specimen, to
avoid contamination of the solution with
easily reducible metal ions, and to insure that
inhibitor decomposition has not occurred.
Instead of using~2 milliiiters of any proprie-
tary inhibitor, 0.5 gram per liter of inhibitors
such as dionhotolyl thiourea or quinoline
ethiodide can be used.
5.4 Whatever treatment is used to clean specimens after a
corrosion test, its effect in removing metal should be
determined, and the weight loss should be corrected
accordingly: A "blank" specimen should be weighed
before and after exposure to the cleaning procedure to
establish this weight loss.
5.4.1 Following removal of all scale, the specimen
should be treated as discussed in Section 2.8.
5.4.2 A description of the cleaning method should
be included with the data reported.
5.3-1
6. Evaluation of Results
6.1 After corroded specimens have been cleaned, they
should be reweighed with an accuracy corresponding to
that of the original weighing. The weight loss during the
test period can be used as the principal measure of
corrosion.
6.2 After the specimens have been reweighed, they should
be examined carefully for the presence of pits. If there are
pits, the average and maximum depths of pits are deter-
mined after measurement with a pit gauge or a calibrated
microscope which can be focused first on the edge and then
on the bottom of the pit. An excellent discussion of pitting
corrosion has been published.3
6.2.1 Pit depths should be reported in millimeters
or thousandths of an inch for the test period and not
interpolated or extrapolated to millimeters per year
or thousandths of an inch per year or any other
arbitrary period because rarely, if ever, is the rate of
initiation or propagation of pits uniform.
6.2.2 The size, shape, and distribution of pits
should be noted. A distinction should be made
between those occurring underneath the supporting
devices (concentration cells) and those on surfaces
that were freely exposed to the test solution..
6.3 If the material being tested is suspected of being
subject to dealloying forms of corrosion such as dezincifi-
cation, or to intergranular attack, a cross section of the
specimen should be microscopically examined to determine
the type and depth of such attack.
6.4 The specimen may be subjected to simple bending
tests to determine whether any embrittlement has occurred.
6.5 It may be desirable to make quantitative mechanical
tests to compare the exposed specimens with uncorroded
specimens reserved for the purpose, as described in Section
2.2.
-------
7. Calculating Corrosion Rates
7.1 The calculation of corrosion rates requires several
pieces of information and several assumptions.
7.1.1 The use of corrosion rates implies that all
weight loss has been due to general corrosion and not
to localized corrosion, such as pitting or sensitized
areas on welded coupons. Localized corrosion is
reported separately.
7.1.2 The use of corrosion rates also implies that
the material has not been internally attacked as by
dezincification or intereranular corrosion.
7.1.3 Internal attack can be expressed as a corrosion
rate if desired. However, the calculations must not be
based on weight loss, which is usually small, but on
rnicrosections which show depth of attack.
7.2 Assuming that localized or internal corrosion is not
present or are recorded separately in the report, the
corrosion rate expressed as mils penetration per year (rr.py)
or millimeters per year (rnmpy) can be calculated by thu
equations:
wt loss x 534
mpy = (area) (time) (metal density)
wtlossx 13.56
(area) (time) (metal density)
where weight loss is in milligrams, area is square inches of
metal surface exposed, and time is hours exposed.
Metal density of many common alloys (expressed in grams
per cubic centimeter) is listed in Table 3. The density for
new or unlisted alloys can be obtained from the producer
or from various metal handbooks.
5.3-1?
TABLE 3 — Density of Common Metals for Use in Corrosion Rate Calculations9
Alloy
Density,
&/cc
Aluminum
99.0 + A1 . , 2.71
AU-2Mn . 2,73
AU.OMg.O.«-Si;0.25& 2.70
Brass
85 Cu. 15 Zn 8.75
71 Cu, 28 Zn, I Sn . 8.53
65 Cu, 35 Zn 8.47
60Cu,39.25Zn,0.75Sn 8.41
Bronze
95 CU, 5Sn 8.86
90 Cu, 10 Sn 8.78
85Cu..SSn, 5Zn,SPb - 8.80
94.8 Cu, 3 Si , 8.53
95Cu,SAJ 8J7
85-90 Cu, 10 Al 7.58
Copper
99.90 Cu. 0.01 P
8.91
Cu pro-Nickel
90 Cu, 10 Ni 8.93
70 Cu, 30 Ni 8.94
Iron
94 Fe, 3.5 C, 2.5 Si 7.00
96 Fe, 3.0 C 7.60
99 ~ Fe, 0.025 S, 0.017 Mn, 0.012 C, 0.005 P .... 7.86
J 14.5 Si, 0.35 Mn, 0.85 C 7.00
Alloy
Dsnsitv
g/cc
Lead
99.90 + Pb 11.34
Nickel
99.4Ni + Co 8.89-
67Ni,30Cu 8.84
62 Ni, 30 Mo, 5 Fe 9.24
58 Ni, 17 Mo, 15Cr,5W, 5 Fft £.94
80 Ni, 14 Cx, 6 Fe 8.51
Steel
0.20C,Mn,P,S
7.85
Stainless Steel
ll.50-13.50Cr, 0.15 C _ . . 7.75
14.00-18.00 Cr, 0.12 C 7.70
18.00-20.00 Ci, 8.00-12.00 Ni, 0.08 C 7.93
16.00-18.00 Ci, 10.00-14.00 Ni,
2.00-3.00 Mo, 0.08 C 7.98
17.00-19.00 Cr, 9.00-12.00 Ni, 0.08 C'Ti 8.02
17.00-19.00 Cr, 9.00-12.00 Ni, 0.08 C, Co 8^
19.00-21.00 Cr, 24.00-30.00 Ni, ZOO-3.00 Mo,
3.00-4.00 Cu 8.02.
Tantalum
16.60
Tin 730
Titanium 4.54
Zirconium 6.53
-------
S. Reporting the Data
8.1 The importance of reporting all data as completely as
possible cannot be overemphasized.
S.2 Expansion of the testing program in the future or
correlating the results with tests of other investigators will
be possible only if all pertinent information is properly
recorded.
8.3 The following checklist is a recommended guide for
reporting all important information and data:
8.3.1 Corrosive media and concentration (changes
during test).
8.3.2 Volume of test solution.
8.3.3 Temperature (maximum, minimum, average).
8.3.4 Aeration (describe conditions or technique).
8.3.5 Agitation (describe conditions or technique).
8.3.6 Type of apparatus used for test.
8.3.7 Duration of each test.
8.3.8 Chemical composition or trade name of
metals tested.
8.3.9 Form and metallurgical conditions of speci-
mens.
83.10 Exact size, shape, and aita of specimens.
8.3.11 Treatment used to prepare specimens for test-
8.3.12 Number of specimens of each material tested,
and whether specimens were tested separately or
which specimens were tested in the same container.
8.3.13 Method used to clean specimens after expo-
sure and the extent of any error expected by this
treatment.
8.3.14 Actual weight losses for each specimen..
8.3.15 Evaluation of attack if other than genera],
such as crevice corrosion under support rod, pit depdi
and distribution, and results of microscopic examina-
tion or bend tests.
8.3.16 Corrosion rates for each specimen expressed
as mils per year.
8.4 Minor occurrences or deviations from the proposed
test program often can have significant effects and should
be reported if known.
8.5 Statistics can be a valuable tool for analyzing tie
results from test programs designed to generate adequate
data and should be used wherever possible. Exteilent
references for the use of statistics in corrosion studies
induce References 4 through 8.
5.3-1*
References
1. Joseph Rosin. Reagent Chemicals and Standard*, Fifth Edition.
D. Van Nostrand Co., Inc., 120 Alexander St., Princeton, N. J.,
1966.
2. A. Vv'achter and R. S. Trtseoer. Corrosion Testing Evaluation of
Metals for Process Equipment. Chemical Engineering Progress.
43, 315-326(1947) June.
3. N. D. Greene and M« G. Fontana. A i_nucaJ Analysis of Pining
Corrosion. Corrosion. IS, 25t (1959) January.
4. Harold S. MieWey, Thomas K. Sherwood, and Charles E. Reed
(editors). Applied Mathematics in Chemical Engineering, (second
edition). McGraw-Hill, New York, N. Y... 1957.
5. D. M. Douglas, Are You Using or Abusing Data? Paper presented
at the Northeast-North Central Region Conference, National
J . B. Ajots
W. G. Ashbaugh
R. L Becker
G. L. Bedford
J.M. Bialosky
J. F. Bosich
F. C. Brauuean
B. Brod\vin
W. B. Brooks
A. A, Brouwe*
T. V. Bruno
H. G. Burbidge
W. H. Burton"
J. W. Cangi
R. F. Catlett
A. S. Couper
T. F. Degnan
H. L. Dickey
C. P. Dillon
L E. Drake, Ji.
G. B. Elder
J. L. English
O. H. Fenner
A. O.'Fisher
M. G. Fontana
J. M. Ford
R. E. Gackenbach
P. J. Gegner
L. V>'. Gieekman
D. L. Graver
Membership of Unit
J. J. HaJbig
F. B. Hamel
A. C. Hamstead
P. R. Handt
F. A. Kencershot
D. R. Hise
T. L. Hoffman
P. R. Hoffmann
R. L. Horst.Jr.
E. C. Hoxie
J. R. Jeter
R. L. Kane
J. M. Keyes
R. W.Kirch ncr
G. Kobrin
Committee T-5A
A. S.Krisher
K. F.Krysiak
J. C. Kuli
E. V. Kunkel
J. Q. Lackey
P. T. Lahi
R. P. Lee
R. B. Leonard
W. A. Luce
R. McFailand
S. W. McHrath
J. M. MaDory
D. S. Neill
G. T. Paul
J. H. Peacock
A. J. Pietrzak
H. D. Rice
L. M, Rogers
J. W. Rollins
G. A. Saltzman
R, C. Scarberry
L- R. Scharfstein
J. H. Schemel
J. R. Schiey
N. B. Schmidt
S. W. Shepaid
O. W. Siebcrt
G. L. Snair
S. S. Spate:
J. M. Stammeft
Association of Corrosion Engineers, October 4-6, 1965, Pitts-
burgh, Pa.
6. W. J. You den. Experimentation and Measurement. National
Science Teachers Association, Washington, D. C, 1962.
7. F. F. Booth and G. E. G. Tucker. Statistical Distribution of
Endurance in Electrochemical Stress-Corrosion Tests. Corrosion,
21,173 (1965) May.
8. Compilation of Examples of Applying Statistics to Corrosion
Problems. ASTM Standard G-16, ASTM, 1916 Race St., Phila-
delphia, Pa.
9. Metals Handbook, American Society for Metals, Metals Park,
Novelry, Ohio, Sth Edition, Volume 1,1961.
L. S. Surteej
H. C. Templeton
A. Tesmen
R. L. Thompwn
A. P. C. Thomson
E. A. Tice
J. W. Tracht
R. S. Treseder
J. M. A. Van dei Horst
A. Wachter
W. W. Wheeler
H. Wijsman
C. P. Williams
C. A. Zimmerman
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6.0-1
Section 6.0
REACTIVITY
Introduction.
The regulation (40 CFR 261.23) defines reactive wastes
to include wastes which (1) readily undergo violent chemical
change; (2) react violently or form potentially explosive
mixtures with water; (3) generate toxic fumes when mixed
with water or, in the case of cyanide or sulfide bearing
wastes, when exposed to mild acidic or basic conditions; (4)
explode when subjected to a strong initiating force; (5)
explode at normal temperatures and pressures; or (6) fit
within the Department of Transportation's forbidden explosives,
Class A explosives, or Class B explosives classifications.
This definition is intended to identify wastes which,
because of their extreme instability and tendency to react
violently or explode, pose a problem at all stages of the
waste management process. The definition is to a large extent
a paraphrase of the narrative definition employed by the
National Fire Protection Association. The Agency chose to
rely on a descriptive, prose definition of reactivity because
the available tests for measuring the variegated class of
effects embraced by the reactivity definition suffer from a
number of deficiencies.
-------
6.1-1
Sub-Section 6.1
CHARACTERISTIC OF REACTIVITY REGULATION
A solid waste exhibits the characteristic of reactivity
If a representative sample of the waste has any of the following
properties:
1. It is normally unstable and readily undergoes
violent change without detonating.
2. It reacts violently with water.
3. It forms potentially explosive mixtures with water.
4. When mixed with water, it generates toxic gases,
vapors or fumes in a quantity sufficient to present a
danger to human health or the environment.
5. It is a cyanide or sulfide bearing waste which,
when exposed to pH conditions between 2 and 12.5,
can generate toxic gases, vapors or fumes in a
quantity sufficient to present a. danger to human
health or the environment.
6. It is capable of detonation or explosive reaction
if it is subjected to a strong initiating source or
if heated under confinement.
7. It is readily capable of detonation or explosive
decomposition or reaction at standard temperature
and pressure.
8. It is a forbidden explosive as defined in 49 CFR
173.51, or a Class A explosive as defined in 49 CFR
173.53 or a Class B explosive as defined in 49 CFR 173.88.
-------
6.1-2
A solid waste that exhibits the characteristic of
reactivity, but is not listed as a hazardous waste in Subpart D,
has the EPA Hazardous Waste Number of D003.
-------
6.2-1
Sub-Section 6.2
DEFINITION OF EXPLOSIVE MATERIALS
For purposes of this regulation a waste which is a
reactive waste by reason of explosivity is one which meets
one or more of the following descriptions:
1. Is explosive and ignites spontaneously or undergoes
marked decomposition when subjected for 48 consecutive
hours to a temperature of 75°C (167°F).
2. Firecrackers, flash crackers, salutes, or similar
commercial devices which produce or are intended to
produce an audible effect, the explosive content of
which exceeds 12 grains each in weight, and pest control
bombs, the explosive content of which exceeds 18 grains
each in weight; and any such devices, without respect
to explosive content, which on functioning are liable
to project or disperse metal, glass or brittle plastic
fragments.
3. Fireworks that combine an explosive and a detonator
or blasting cap.
4. Fireworks containing an ammonium salt and a chlorate.
5. Fireworks containing yellow or white phosphorus.
6. Fireworks or fireworks compositions that ignite
spontaneously or undergo marked decomposition when
subjected for 48 consecutive hours to a temperature of
75°C (167°F).
7. Toy torpedoes, the maximum outside dimension of
which exceeds 7/8 inch, or toy torpedoes containing a
mixture of potassium chlorate, black antimony and sulfur
with an average weight of explosive composition in each
torpedo exceeding four grains.
8. Toy torpedoes containing a cap composed of a mixture
of red phosphorus and potassium chlorate exceeding an
average of one-half (0.5) grain per cap.
9. Fireworks containing copper sulfate and a chlorate.
10. Solid materials which can be caused to deflagrate
by contact with sparks or flame such as produced by safety
fuse or an electric squib, but cannot be detonated (see Note
1) by means of a No. 8 test blasting cap (see Note 2).
Example: Black powder and low explosives.
-------
6.2-2
11. Solid materials which contain a liquid ingredient,
and which, when unconfined (see Note 3), can be detonated
by means of a No. 8 test blasting cap (see Note 2); or
which can be exploded in at least 50 percent of the
trials in the Bureau of Explosives' Impact Apparatus
(see Note 4) under a drop of 4 inches or more, but cannot
be exploded in more than 50 percent of the trials under
a drop of less than 4 inches Example: High explosives,
commercial dynamite containing a liquid explosive ingredient
12. Solid materials which contain no liquid ingredient
and which can be detonated, when unconfined (see Note 3),
by means of No. 8 test blasting cap (see Note 2); or
which can be exploded in at least 50 percent of the
trials in the Bureau of Explosives' Impact Apparatus
(see Note 4) under a drop of 4 inches or more, but cannot
be exploded in more than 50 percent of the trials under
a drop of less than 4 inches. Example: High explosives,
commercial dynamite containing no liquid explosive ingre-
dient, trinitrotoluene, amatol, tetryl, picric acid,
ureanitrate, pentolite, commercial boosters.
13. Solid materials which can be caused to detonate
when unconfined (see Note 3), by contact with sparks or
flame such as produced by safety fuse or an electric
squib; or which can be exploded in the Bureau of Explosives'
Impact Apparatus (see Note 4), in more than 50 percent
of the trials under a drop of less than 4 inches. Example:
Initiating and priming explosives, lead azide, fulminate
of mercury, high explosives.
14. Liquids which may be detonated separately or when
absorbed in sterile absorbent cotton, by a No. 8 test
blasting cap (see Note 2); but which cannot be exploded
in the Bureau of Explosives' Impact Apparatus (see Note
4), by a drop of less than 10 inches. The liquid must
not be significantly more volatile than nitroglycerine
and must not freeze at temperatures above minus 10°F.
Example: High explosives, desensitized nitroglycerine.
15. Liquids that can be exploded in the Bureau of
Explosives' Impact Apparatus (see Note 4) under a drop
of less than 10 inches. Example: Nitroglycerine.
16. Blasting caps. These are small tubes, usually
made of an alloy of either copper or aluminum, or of
molded plastic closed at one end and loaded with a charge
of initiating or priming explosives. Blasting caps (see
Note 5) which have been provided with a means for firing
by an electric current, and sealed, are known as electric
blasting caps.
-------
6.2-3
17. Detonating primers which contain a detonator and an
additional charge of explosives, all assembled in a suit-
able envelope.
18. Detonating fuses, which are used in the military
service to detonate the high explosive bursting charges
of projectiles, mines, bombs, torpedoes, and grenades.
In addition to a powerful detonator, they may contain
several ounces of a high explosive, such as tetryl or
dry nitrocellulose, all assembled in a heavy steel enve-
lope. They may also contain a small amount of radioactive
component. Those that will not cause functioning of
other fuses, explosives, or explosive devices in the
same or adjacent containers are classed as class C explo-
sives and are not reactive waste.
19. A shaped charge, consisting of a plastic, paper, or
other suitable container comprising a charge of not to
exceed 8 ounces of a high explosive containing no liquid
explosive ingredient and with a hollowed-out portion
(cavity) lined with a rigid material.
20. Ammunition or explosive projectiles, either fixed,
semi-fixed or separate components which are made for use
in cannon, mortar, howitzer, recoilless rifle, rocket,
or other launching device with a caliber of 20 mm or
larger.
21. Grenades. Grenades, hand or rifle, are small metal
or other containers designed to be thrown by hand or pro-
jected from a rifle. They are filled with an explosive
or a liquid, gas, or solid material such as a tear gas
or an incendiary or smoke producing material and a bursting
charge.
22. Explosive bombs. Explosive bombs are metal or
other containers filled with explosives. They are used in
warfare and include airplane bombs and depth bombs.
23. Explosive mines. Explosive mines are metal or compo-
sition containers filled with a high explosive.
24. Explosive torpedoes. Explosive torpedoes, such as
those used in warfare, are metal devices containing a means
of propulsion and a quantity of high explosives.
25. Rocket ammunition. Rocket ammunition (including
guided missiles) is ammunition designed for launching from
a tube, launcher, rails, trough, or other launching device,
in which the propellant material is a solid propellent
explosive. It consists of an igniter, rocket motor, and
projectile (warhead) either fused or unfused, containing
high explosives or chemicals.
-------
6.2-4
26. Chemical ammunition. Chemical ammunition used in
warfare is all kinds of explosive chemical projectiles,
shells, bombs, grenades, etc., loaded with tear, or
other gas, smoke or incendiary agent, also such miscel-
laneous apparatus as cloud-gas cylinders, smoke generators,
etc., that may by utilized to project chemicals.
27. Boosters, bursters, and supplementary charges.
Boosters and supplementary charges consist of a casing
containing a high explosive and are used to increase the
intensity of explosion of the detonator of a detonating fuse
Bursters consist of a casing containing a high explosive and
are used to rupture a projectile or bomb to permit release
of its contents.
28. Jet thrust units or other rocket motors containing a
mixture of chemicals capable of burning rapidly and produc-
ing considerable pressure.
29. Propellant mixtures (i.e, and chemical mixtures
which are designed to function by rapid combustion with
little or no smoke).
Note 1; The detonation test is performed by placing the
sample in an open-end fiber tube which is set on the end
of a lead block approximately 1 1/2 inches in diameter
and 4 Inches high which, in turn, is placed on a solid
base. A steel plate may be placed between the fiber
tube and the lead block.
Note 2; A No. 8 test blasting cap is one containing two
grams of a mixture of 80 percent mercury fulminate and 20
percent potassium chlorate, or a cap of equivalent strength.
Note 3; "Unconfined" as used in this section does not
exclude the use of a paper or soft fiber tube wrapping to
facilitate tests.
jjo t e 4; The Bureau of Explosives' Impact Apparatus is a
testing device designed so that a guided 8-pound weight
may be dropped from predetermined heights so as to impact
specific quantities of liquid or solid materials under
fixed conditions. Detailed prints may be obtained from
the Bureau of Explosives, 2 Pennsylvania Plaza, New York,
New York, 10001.
Note 5; Blasting caps, blasting caps with safety fuse,
or electric blasting caps in quantities of 1,000 or
less are classified as class 0 explosives and not subject
to regulation as a reactive waste.
-------
7.0-1
Section 7.0
EXTRACTION PROCEDURE TOXICITY
Introduction^
The Extraction Procedure (EP) is designed to simulate
the leaching a waste will undergo if disposed of in an
improperly designed sanitary landfill. It is a laboratory
teat in which a representative sample of a waste is extracted
with distilled water maintained at pH » 5 using acetic acid.
The extract obtained from the EP (the "EP Extract") is then
analyzed to determine if any of the thresholds established
for the 8 elements (i.e., arsenic, barium, cadmium, chromium,
lead, mercury, selenium, silver), four pesticides (i.e.,
Endrin, Lindane, Methoxychlor, Toxaphene), and two herbicides
(i.e., 2,4,5-Trichlorophenoxypropionic acid, 2,4-Dichloro-
phenoxyacetic acid) have been exceeded. If the EP Extract
contains any one of the above substances in an amount equal
to or exceeding the levels specified in 40 CFR 261.24, the
waste possesses the characteristic of Extraction Procedure
Toxicity and is a hazardous waste.
The Extraction Procedure consists of 5 steps:
1. Separation Procedure
A waste containing unbound liquid is filtered and if
the solid phase is less than 0.5% of the waste, the solid
phase is discarded and the filtrate analyzed for trace
elements, pesticides, and herbicides (step 5). If the waste
contains more than 0.5% solids, the solid phase is extracted
and the liquid phase stored for later use.
-------
7.0-2
2. Structural Integrity Procedure/Particle Size Reduction
Prior to extraction, the solid material must
either pass through a 9.5 mm (0.375 in) standard sieve,
have a surface area per gram of waste of 3.1 cm^ , or if it
consists of a single piece, be subjected to the Structural
Integrity Procedure. The Structural Integrity Procedure Is
used to demonstrate the ability of the waste to remain intact
after disposal. If the waste does not meet one of these
conditions it must be ground to pass the 9.5 mm sieve.
3. Extraction of Solid Material
The solid material from step 2 is extracted for
24 hours in an aqueous medium whose pH is maintained at or
below 5, using 0.5 N acetic acid. The pH is maintained
either automaticlly or manually. Acidification to ph 5
is subject to a specification as to total amount of acid
to be added to the system.
4. Final Separation of the Extraction from the Remaining
Solid
After extraction, the liquid: solid ratio is adjusted
to 20:1 and the mixture of solid and extraction liquid are
separated by filtration, the solid discarded and the liquid
combined with the filtrate obtained in step I. This is the
EP Extract that is subjected to the evaluation requirements
in 40 CFR 261.24.
-------
7.0-3
5• Testing (Analysis) of EP Extract
Inorganic and organic species are identified and
quantified using the appropriate methods in Section 8 of
this manual.
-------
Figure 7.G
EXTRACTION PROCEDURE FLOWCHART
7.0-4
Wet Waste Sample
contains < 0.5% ^
non-filterable
solids
Liquid Solid
Separation
V
Representative
Waste Sample
> 100 grams
Dry Wasi
— ^Solld
Discard
/ >
:e Sample
0.5%
^ non-filterable
solids
1
Liquid Solid
Separation
\
Particle size
Liquid
> 9.5 mm
< 9.5 mm monolithic
.quid
Sample Size
Reduction
Structural
Integrity
Procedure
Extraction of Solid Waste
Solid {
Store at
4° C and/or
at pH = 2
Liquid Solid Separation
Discard
Liquid
EP Extract
1
Analysis Methods
-------
7.1-1
Sub-Section 7.1
CHARACTERISTIC OF EP TOXICITY REGULATION
A solid waste exhibits the characteristic of EP
toxicity if, using the test methods described in Appendix II
of 40 CFR Part 261 or equivalent methods approved by the
Administrator under the procedures set forth in 40 CFR 260.20 and
260.21, the extract from a representative sample of the waste
contains any of the contaminants listed in Table 7.1-1 at a concen-
tration equal to or greater than the respective value given in
that Table. Where the waste contains less than 0.5 percent filter-
able solids, the waste itself, after filtering, is considered
to be the extract for the purposes of this section.
A solid waste that exhibits the characteristic of
EP toxicity, but is not listed as a hazardous waste in
Subpart D, has the EPA Hazardous Waste Number specified in
Table 7.1-1 which corresponds to the toxic contaminant
causing it to be hazardous.
-------
7.1-2
Table 7.1-1
MAXIMUM CONCENTRATION OF CONTAMINANTS
FOR CHARACTERISTIC OF EP TOXICITY
EPA
Hazardous Waste
Number
D004
D005
D006
D007
D008
D009
D010
D011
D012
D013
D014
D015
D016
D017
Contaminant
Maximum
Concentration
(milligrams per liter)
Arsenic 5.0
Barium 100.0
Cadmium 1.0
Chromium 5.0
Lead 5.0
Mercury 0.2
Selenium...... 1.0
Silver 5.0
Endrin (1,2,3,4,10,10-Hexachloro-l
7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-1
4-endo, endo-5,8-dimethanonaph-
thalene 0.02
Lindane (1,2,3,4,5,6-
Hexachlorocyclohexane, gamma
isomer 0-4
Methoxychlor (1,1, l-Trichloro-2,2-bis
[p-methoxyphenyl]ethane). 10.0
Toxaphene (C^QHiQCl3, Technical
chlorinated camphene, 67-69
percent chlorine).... 0.5
2,4-D (2,4-Dichlorophenoxyacetic
acid) 10.0
2,4,5-TP [Silvex] (2,4,5-
Trichlorophenoxypropionic acid) 1.0
-------
7.1-3
APPENDIX II
«
EP TOXICITY TEST
Procedure
1. A representative sample of the waste to be tested
(minimum size 100 grams) should be obtained using the
methods specified in Appendix I of 40 CFR 261 or any
other method capable of yielding a representative sample
within the meaning of 40 CFR 260.
2. The sample should be separated into its component
liquid and solid phases using the method described in
"Separation Procedure" below. If the dry weight of
the solid residue* obtained using this method totals
less than 0.5% of the original wet weight of the waste,
the residue can be discarded and the operator should
treat the liquid phase as the extract and proceed immedi-
ately to Step 8.
3. The solid material obtained from the Separation
Procedure should be evaluated for its particle size. If
the solid material has a surface area per gram of material
equal to, or greater than, 3.1 cm^ or passes through a
9.5 mm (0.375 inch) standard sieve, the operator should
proceed to Step 4. If the surface area is smaller or the
*The percent solids is determined by drying the filter pad at
80°C until it reaches constant weight and then calculating
the percent solids using the following equation:
(weight of pad + solid) - (tare weight of pad) X 100 - % solids
initial wet weight of sample
-------
7-1-4
particle size larger than specified above, the solid
material would be prepared for extraction by crushing,
cutting or grinding the material so that it passes through
a 9.5 mm (0.375 inch) sieve or, if the material is in a
single piece, by subjecting the material to the "Structural
Integrity Procedure" described below.
4. The solid material obtained in Step 3 should be weighed
immediately and placed in an extractor with 16 times its
weight of deionized water. Do not allow the material
to dry prior to weighing. For purposes of this test, an
acceptable extractor is one which will impart sufficient
agitati'on to the mixture to not only prevent stratification
of the sample and extraction fluid but also insure that
•
all sample surfaces are continuously brought into contact
with well mixed extraction fluid.
5. After the solid material and deionized water are placed
in the extractor, the operator should begin agitation
and measure the pH of the solution in the extractor.
If the pH is greater than 5.0, the pH of the solution
should be decreased to 5.0 + 0.2 by adding 0.5 N acetic
acid. If the pH is equal to or less than 5.0, no acetic
acid should be added. The pH of the solution should be
monitored, as described below, during the course of the
extraction and if the pH rises above 5.2, 0.5N acetic
acid should be added to bring the pH down to 5.0 +0.2.
However, in no event shall the aggregate amount of acid
-------
7.1-5
added to the solution exceed 4 ml of acid per gram of
solid. The mixture should be agitated for 24 hours and
maintained at 20°-40°C (68° 104°F) during this time. It
is recommended that the operator monitor and adjust the
pH during the course of the extraction with a device
such as the Type 45-A pH Controller manufactured by
Chemtrix, Inc., Hillsboro, Oregon 97123 or its equivalent,
in conjunction with a metering pump and reservoir of 0.5N
acetic acid. If such a system is not available, the
following manual procedure shall be employed:
a. A pH meter should be calibrated in accordance
with the manufacturer's specifications.
»
b. The pH of the solution should be checked and,
if necessary, 0.5N acetic acid should be
manually added to the extractor until the
pH reaches 5.0 ± 0.2. The pH of the solution
should be adjusted at 15, 30, and 60 minute
intervals, moving to the next longer interval
if the pH does not have to be adjusted more
than 0.5 pH units.
c. The adjustment procedure should be continued
for at least 6 hours.
d. If at the end of the 24-hour extraction period,
the pH of the solution is not below 5.2 and
the maximum amount of acid (4 ml per gram of
solids) has not been added, the pH should be
adjusted to 5.0 +_ 0.2 and the extraction
continued for an additional four hours, during
which the pH should be adjusted at one hour
intervals.
At the end of the 24 hour extraction period, deionized
water should be added to the extractor in an amount
determined by the following equation:
-------
7.1-6
V- (20)(W) - 16(W) - A
V- ml delonlzed water to be added
W» weight in grams of solid charged to extractor
A= ml of 0.5N acetic acid added during extraction
7. The material in the extractor should be separated
into its component liquid and solid phases as described
under "Separation Procedure."
8. The liquids resulting from Steps 2 and 7 should be combined.
This combined liquid (or the waste Itself if it has less
than 0.5% solids, as noted in step 2) is the extract and
should be analyzed for the presence of any of the contami-
nants specified in Table I of 40 CFR 261.24 using the
Analytical Procedures designated below.
Separation Procedure
Apparatus
A filter holder, designed for filtration media having a
nominal pore size of 0.45 micrometer and capable of applying
a 5.3 kg/cm^ (75 psig) hydrostatic pressure to the solution
being filtered shall be used. For mixtures containing non-
absorptive solids, where separation can be effected without
imposing a 5.3 kg/cm^ pressure differential, vacuum filters
employing a 0.45 micrometer filter media can be used.
-------
7.1-7
Procedure*
1. Following manufacturer's directions, the filter unit
should be assembled with a filter bed consisting of
a 0.45 micrometer filter membrane. For difficult
or slow to filter mixtures a prefilter bed consisting
of the following prefilters in increasing pore size
(0.65 micrometer membrane, fine glass fiber prefilter,
and coarse glass fiber prefilter) can be used.
2. The waste should be poured into the filtration unit.
3. The reservoir should be slowly pressurized until liquid
begins to flow from the filtrate outlet at which point
the pressure in the filter should be immediately lowered
to 10-15 psig. Filtration should be continued until
liquid flow ceases.
4. The pressure should be increased stepwise in 10 psig
increments to 75 psig and filtration continued until
flow ceases or the pressurizing gas begins to exit from
the filtrate outlet.
*This procedure is intended to result in separation of the "free"
liquid portion of the waste from any solid matter having a
particle size >0.45um. If the sample will not filter, various
other separation techniques can be used to aid in the filtration.
As described above, pressure filtration is employed to speed up
the filtration process. This does not alter the nature of the
separation. If liquid does not separate during filtration, the
waste can be centrifuged. If separation occurs during centrifuga-
tion, the liquid portion (centrifugate) is filtered through the
0.45um filter prior to becoming mixed with the liquid portion of
the waste obtained from the initial filtration. Any material
that will not pass through the filter after centrifugation is
considered a solid and is extracted.
-------
7.1-8
5. The filter unit should be depressurized, the solid material
removed and weighed and then transferred to the extraction
apparatus, or, in the case of final filtration prior to
analysis, discarded. If the solid is to be extracted do
not allow the material retained on the filter pad to dry
prior to weighing.
6. The liquid phase should be stored at 4°C for subsequent
use in Step 8.
Structural Integrity Procedure
Apparatus
A Structural Integrity Tester having a 3.18 cm (1.25
in.) diameter hammer weighing 0.33 kg (0.73 Ibs.) and having
a free fall of 15.24 cm (6 in.) shall be used. This device
is available from Associated Design and Manufacturing
Company, Alexandria, VA., 22314, as Part No. 125, or it may
be fabricated to meet the specifications shown in Figure 7-2.
Procedure
1. The sample holder should be filled with the material
to be tested. If the sample of waste is a large mono-
lithic block, a portion should be cut from the block
having the dimensions of a 3.3 cm (1.3 in.) diameter x
7.1 cm (2.8 in.) long cylinder. For a fixated waste,
samples may be cast in the form of a 3.3 cm (1.3 in.)
diameter x 7.1 cm (2.8 in.) cylinder for purposes of
conducting this test. In such cases, the waste may be
allowed to cure for 30 days prior to further testing.
-------
7.1-9
2. The sample should be placed into the Structural Integrity
Tester, then the hammer should be raised to its maximum
height and dropped. This should be repeated fifteen
times.
3. The material should be removed from the sample
holder, weighed, and transferred to the extraction
apparatus for extraction.
Procedures for Analyzing Extract
The test methods for analyzing the extract are as
follows:
1. For arsenic, barium, cadmium, chromium, lead, mercury,
selenium or silver: "Methods for Analysis of Water and
Wastes," Environmental Monitoring and Support Laboratory,
Office of Research and Development, U.S. Environmental
Protection Agency, Cincinnati, Ohio 45268 (EPA-600/4-79-020,
March 1979).
2. For Endrin; Lindane; Methoxychlor; Toxaphene; 2,4-D;
2,4,5-TP (Silvex): in "Methods for Benzidine, Chlorinated
Organic Compounds, Pentachlorophenol and Pesticides in
Water and Wastewater," September 1978, U.S. Environmental
Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268.
as standardized in "Test Methods for the Evaluation of
Solid Waste, Physical/Chemical Methods."
For all analyses, the method of standard addition shall
be used for the quantification of species concentration.
-------
7.1-10
This method is described in "Test Methods for the Evaluation
of Solid Waste." (It is also described in "Methods for Analysis
of Water and Wastes.")
-------
o
7.1-11
t
15.25cm
(6")
V -. /:
3.3cm
(1.3")
9.4cm
(3.7")
COMBINED
WEIGHT
33Kg
(.73lb|
(3.15cm)
(1.25")
SAMPLE
ELASTOMERIC*
SAMPLE HOLDER
7.1cm
(2.8")
I
^ELASTOMERIC SAMPLE HOLDER FABRICATED OF
MATERIAL FIRM ENOUGH TO SUPPORT THE SAMPLE
Figure 7-2.
COMPACTION TESTER
-------
7.2-1
Method 7.2
SEPARATION PROCEDURE
Scope and Application
This procedure is used to separate a waste into its liquid and
solid phases both prior to and after extraction.
Summary of Method
The Separation Procedure involves vacuum or pressure filtration
of a waste or extraction mixture. To minimize filtration time,
pressure, settling, centrifugation and prefilters may be employed as
an adjunct to filtration. Pressure filtration is required when vacuum
filtration is inadequate for complete separation.
Apparatus
1. Filter holder: A filter holder capable of supporting a 0.45
urn filter membrane and able to withstand the pressure needed
to accomplish separation. Suitable filter holders range from
simple vacuum units to relatively complex systems that can
exert up to 5.3 kg/cm^ (75 psi) of pressure. The type of
filter holder used depends upon the properties of the mixture
to be filtered. Filter holders known to the Agency and deemed
suitable for use are listed in Table 7.2-1
2. Filter membrane: Filter membrane suitable for conducting the
required filtration shall be fabricated from a material which:
a. is not physically changed by the waste material to
be filtered.
b. does not absorb or leach the chemical species for
which a waste's EP Extract will be analyzed.
-------
7.2-2
Table 7.2-2 lists filter media known to the Agency and generally
found to be suitable for solid waste testing.
3. In cases of doubt contact the filter manufacturer to determine
if either membrane or prefilter are adversely affected by the
particular waste. If no information is available, submerge
the filter in the waste's liquid phase. After 48 hours a
filter that undergoes visible physical change (i.e. curls,
dissolves, shrinks, or swells) is unsuitable for use.
Use the following procedure to establish if a filter
membrane will leach or adsorb chemical species.
a. Prepare a standard solution of the chemical species of
interest.
b. Analyze the standard for its concentration of the chemical
species.
c. Filter the standard and re-analyze. If the concentration
of the filtrate differs from the original standard, the
filter membrane leaches or absorbs one or more of the
chemical species.
General Procedure
1. Weigh filter membrane and prefilter to + 0.01 gram. Handle
membrane and prefilters with blunt curved tip forceps or
vacuum tweezers, or by applying suction with a pipette.
2. Assemble filter holder, membranes, and prefilters following
the manufacturer's instructions. Place the 0.45 urn membrane
on the support screen and add prefilters in ascending order of
pore size. Do not pre-wet filter membrane.
-------
7.2-3
3. Allow slurries to stand to permit the solid phase to settle.
Slow to settle wastes may be centrifuged prior to filtra-
tion.
4. Wet the filter with a small portion of the waste's or extraction
mixture's liquid phase. Transfer the remaining material to the
filter holder and apply vacuum or gentle pressure (10-15 psi)
until all liquid passes through the filter. Stop filtration when
air or pressurizing gas moves through the membrane. If this point
is not reached under vacuum or gentle pressure slowly increase the
pressure in 10 psi increments to 75 psi. Halt filtration when
liquid flow stops.
5. Remove solid phase and filter media and weigh to +_ 0.01 gram.
Discard solid if it comprises less than 0.5% of the mixture
(see below). If the sample contains >0.5% solids use the wet
weight of the solid phase obtained in this separation for
purposes of calculating amount of liquid and acid to employ
for extraction using the following equation:
W = Wf - Wt
W = wet weight in grams of solid to be charged to extractor
Wf - wet weight in grams of filtered solids and filter media
Wt = weight in grams of tared filters.
Procedure for Determining Percent Solids of a Waste
1. Determine percent solids of a waste sample by:
a. separately weighing the waste sample and filters.
b. filtering the waste material.
c. drying the solid and filters at 80°C until two
successive weighings yield the same value. Cal-
-------
7-2-4
culate the percent solids using the following
equation:
weight of filtered solid and filters - tared weight of filters x 100 = % solids
initial weight of waste material
NOTE; This procedure is only used to determine if the solid
must be extracted or if it can be discarded unextracted. It
is not used in calculating the amount of water or acid to use
in the extraction step. Do not extract solid material that
has been dried at 80°C. A new sample will have to be used
for extraction if a % solids determination is performed.
-------
7.2-5
Table 7.2-1
APPROVED FILTER HOLDERS
Vacuum Filters
Manufacturer
Size
Model No
Comments
Nalgene
500 ml
Systems
Millipore
142 mm
45-0045
p
Nuclepore
Millipore
ressure Filters
Nuclepore
Micro Filtration
47 mm
4 7 mm
142 mm
142 mm
410400
XX10 047 00
420800
302300
YT30 142 HW
Disposable plastic unit,
includes prefilter and
filter pads, and reservoir
Should only be used when
solution is to be analyzed
for inorganic constituents
-------
7.2-6
Table 7.2-2
APPROVED FILTRATION MEDIA
Filter
Type
Supplier
Filter To Be Used
For Aqueous Systems*
Filter To Be Used
For Organic Systems*
Coarse
Pref liter
Medium
Pref liters
Fine
Pref liters
Fine
Filters
(0.45um)
Gelman
Nuclepor e
Millipore
Nuclepore
Millipore
Nuclepore
Millipore
Gelman
Pall
Nuclepore
Millipore
Selas
61653
61669
210907
211707
AP25 042 00
AP25 127 50
21095
211705
AP20 042 00
AP20 124 50
210903
211703
AP25 042 00
AP25 127 50
60173
60177
047NX50
142NX25
111107
112007
HAWP 047 00
HAWP 142 50
83485-02
83486-02
61652
61669
210907
211707
AP25 042 00
AP25 127 00
21095
211705
AP20 042 00
AP20 124 50
210903
211703
AP25 042 00
AP25 127 50
60540
60544
181107
182007
FHLP 047 00
FHLP 142 00
83485-02
83486-02
-------
7.4-1
Method 7.4
STRUCTURAL INTEGRITY PROCEDURE
Application
The Structural Integrity Procedure (SIP) is employed
to approximate the physical degradation a monolithic waste
undergoes in a landfill or when compacted by earthmoving
equipment.
Equipment
1. Structural Integrity Tester meeting the specifications
detailed in Figure 7.4-1.
2 . Sample holders of elastomeric material firm enough
to support a cylindrical waste sample 3.3 cm (1.32 in.) in
diameter and 7.1 cm (2.84 in.) long.
Procedure^
1. Cut a 3.3 cm in diameter by 7.1 cm long cylinder
from the waste material. For wastes which have been treated
using a fixation process the waste may be cast in the form
of a cylinder and allowed to cure for 30 days prior to testing.
2. Place waste into sample holder and assemble the
tester. Raise the hammer to its maximum height and drop.
Repeat 14 times.
3. Remove solid material from tester and scrape off
any particles adhering to sample holder. Weigh the waste to
the nearest 0.01 gram and transfer its to the Extractor.
-------
7.5-1
Sub-Section 7.5
EXTRACTORS
Introduction
An acceptable extractor is one which will prevent
stratification of a waste sample and extraction fluid and
will insure that all sample surfaces continuously contact
well mixed extraction fluid. There are two types of acceptable
extractors: 1) stirrers and 2) tumblers. Stirrers consist of a
container in which the waste/extraction fluid mixture is
agitated by spinning blades. Rotators agitate by turning a
sample container end over end through a 360° revolution.
Stirrer
Scope and Application
One such stirrer approved for use in evaluating solid
waste is illustrated in Figure 7.5-1. It is a container in
which a waste/extraction fluid mixture is agitated by 2
blades spinning at >_ 40 rpm. This extractor can be used
with either automatic or manual pH adjustment.
Precautions
1. Large particles (>_ 0.25 in. in diameter) may be
ground by the spinning blades or abrade the container.
If metal containers are employed this may result in
contamination of the EP Extract.
2. Monolithic wastes should not be extracted in the stirrer
as they may bend or break the stirring blades.
-------
7.5-2
Summary of Operation
Place waste In extractor, add extraction fluid and
stir for the required period of time. Adjust pH while stirrer
is in operation by addition of acid through port in cover.
pH may be continuously monitored using port in cover designed
to accept a pH electorde.
Manufactures
Extractors of this design may be fabricated by the user
or are known to be available commercially from Associated
Design and Manufacturing Co. and Millipore Corporation.
Rotary Extractor
Scope and Application
The rotary extractor consists of a rack or box type
device holding a number of plastic or glass bottles which
rotate at approximately 29 rpm. Rotary extractors are used
with manual pH adjustment.
Precautions
1. Use glass or fluorocarbon bottles for wastes whose
EP Extract will be analyzed for organic compounds.
For extracts to be analyzed only for metals, poly-
ethylene bottles may be substituted.
2. Be careful not to tighten the screws too far and
shatter the bottle when using the design in Figure
7.5-2.
3. Do not use glass bottles for extracting large blocks
of waste as these may cause the bottles to shatter.
-------
7.5-3
4. It is recommended that the bottles be alternated In
an opposing manner in the apparatus to minimize torque
(e.g., when one bottle faces up, the next bottle faces
down.) When extracting an odd number of samples,
balance the extractor by adding a bottle containing
an amount of water approximately equal to the volume
in the other bottles.
Equipment
1. Rotary extractors approved for use in evaulating the
EP toxicity of solid wastes are illustrated in Figure
7.5-2 and 7.5-3.
2. Plastic or glass bottles sized to fit the particular
extractor.
3. The equipment illustrated in Figure 7.5-2 may be
fabricated by the user or is available commercially
from Associated Design and Manufacturing Co.
4. The equipment illustrated in Figure 7.5-3 is available
from the Acurex Corporation.
Summary of Operation
Fill plastic or glass bottles with the solid material.
Add distilled deionized water to each bottle and start extractor.
Stop extractor after 1 minute and adjust pH. Restart extractor
and continue pH adjustment for the first six hours of agitation
as described in the "Manual pH Adjustment Procedure" (Section
7.1). After 24 hours of agitation stop extractor, check pH as
described and, if within range specified, adjust volume
of fluid and remove for liquid solid separation.
-------
7.5-4
NON CLOGGING SUPPORT BUSHING
1 inch BLADE AT 30° TO HORIZONTAL
Figure 7.5-1
EXTRACTOR
-------
7.5-5
-------
7.5-6
-------
8.01-1
Method 8.01
METHOD FOR VOLATILE ORGANICS
Scope and Application
The following volatile organic compounds may be determined
by this method:
Volatile Compound Detector
Acrylamide FID
Benzyl Chloride HSD
Bis(2-chloroethoxymethane) HSD
Bis(2-Chloroisopropyl) ether HSD
Carbon Disulfide FID
Carbon Tetrachloride HSD
Chloroacetaldehyde HSD
Chlorobenzene HSD, FID
Chloroform HSD
Chloromethane HSD '
Dichlorobenzene HSD
Dibromomethane HSD
Ethyl ether FID
Formaldehyde FID
Methanol FID
Methyl Ethyl Ketone FID
Methyl Isobutyl Ketone FID
Paraldehyde FID
Tetrachloroethane HSD
Tetrachloroethene HSD
Trichloroethene HSD
Trichlorofluoromethane HSD
Trichloropropane HSD
Vinyl Chloride HSD
Vinylidene Chloride HSD
-------
8.01-2
Summary of Method
1. The volatile compounds are introduced to the gas chromato-
graph by direct injection (Method 8.80), the Headspace
Method (Method 8.82), or the Purge and Trap method (Method
8.83). A temperature program is used in the gas chromato-
graph to separate the organic compounds. Detection is
achieved by either a halide specific detector (HSD) or a
flame ionization dectector (FID). The detector used
depends on the compound(s) of interest.
2. If interferences are encountered, the method provides an
optional gas chromatographic column that may be helpful
in resolving the compounds of interest from the inter-
ferences .
Interferences
Samples can be contaminated by diffusion of volatile
organics (particularly chlorofluorocarbons and methylene
chloride) through the sample container septum during shipment
and storage. A sample blank carried through sampling and
subsequent storage and handling can serve as a check on such
contamination.
Apparatus
It Vial with cap - 40 ml capacity screw cap (Pierce #13075 or
equivalent). Detergent wash and dry at 105°C before use.
2. Septum - teflon - forced silicone (Pierce #12722 or
equivalent). Detergent wash, rinse with tap and distilled
deionized water, and dry at 105°C for one hour before use.
-------
8.01-3
3. Sample introduction apparatus (Methods 8.80, 8.82 and 8.83)
4. Gas chromatograph - Analytical system complete with
programmable gas chromatograph suitable for on-column
injection and all required accessories, including HSD or,
FID, column supplies, recorder and gasses. A data system
for measuring peak area is recommended.
5. Carbopack B 60/80 mesh coated with 1% SP-1000 packed
in an 8 ft. x 0.1 in. ID stainless steel or glass column
(Column 1) or ?orisil-C 100/200 mesh coated with n-octane
packed in a 6 ft. x 0.1 in. ID stainless steel or glass
column (Column 2).
6. Syringes - 5 ml glass hypodermic with Luerlok tip (2 each).
7. Micro syringe - 10, 25, 100 ul.
8. 2-way syringe valve with Luer ends (3 each).
9. Syringe - 5 ml - gas tight with shut-off valve.
10. Bottle - 15 ml screw-cap, with teflon cap liner.
Reagents
1. Activated carbon - Filtrosorb 200 (Calspan Corp.) or
equivalent.
2. Organic-free water generated by passing tap water through
a carbon filter bed containing about 1 Ib. of activated
carbon. A water purification system (Millipore Super -
Q or equivalent) 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 one hour.
-------
8.0M.
3. Stock standards - prepare stock standard solutions in
methyl alcohol using assayed liquids or gas cylinders
as appropriate. Because of the toxicity of many of
the compounds being analyzed, primary dilutions of these
materials should be prepared in a hood . A NIOSH/MESA
approved toxic gas respirator should be used when
the analyst handles high concentrations of such
materials.
a. Place about 9.8 ml of methyl alcohol into a 10 ml
ground glass stoppered volumetric flask. Allow
about 10 minutes or until all alcohol wetted surfaces
have dried. Weigh the flask to the nearest 0.1 mg.
b. Add the assayed reference material:
i. Liquids
Using a 100 ul syringe, immediately add 2 drops
of assayed reference material to the flask, then
reweigh. Be sure that the 2 drops fall directly
into the alcohol without contacting the neck of
the flask.
ii . Gases
To prepare standards from any of the organic
compounds that boil below 30°C, fill a 5 ml valved
gas-tight syringe with the reference standard to the
5 ml mark. Lower the needle to 5 mm above the methyl
alcohol meniscus. Slowly inject the reference
standard above the surface of the liquid (the heavy
gas will rapidly dissolve into the methyl alcohol).
-------
8.01-5
c. Reweigh, dilute to volume, stopper, then mix by
by inverting the flask several times. Transfer the
standard solution to a 15 ml screw cap bottle with a
teflon cap liner.
d. Calculate the concentration in mg/1 from the net
gain in weight.
e. Store stock standards at 40° C. Prepare fresh standards
weekly for the 4 compounds whose BP <^ 30° C. All other
standards must be replaced with fresh standards each
month.
Calibration
1. Using stock standards, prepare secondary dilution standards
in methyl alcohol that contains the compounds of interest,
either singly or mixed together.
2. Assemble necessary gas chromatographic apparatus and establish
operating parameters equivalent to those indicated in the
Procedure section. By injecting secondary standards, adjust
the sensitivity of the analytical system for each compound
being analyzed so as detect _< 1 ug.
Quality Controls
1. Before processing any samples, the analyst should daily
demonstrate through the analysis of an organic-free water
or solvent blank that the entire analytical system is
interference free.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to
validate the precision of the sampling technique.
-------
8.01-6
Laboratory replicates should be analyzed to validate the
the precision of the analysis. Fortified samples should
be analyzed to validate the accuracy of the analyses.
Where doubt exists over the identification of a peak
on the gas chromatogam, confirmatory techniques such as
mass spectroscopy should be used.
3. The analyst should maintain constant surveillance of
both the performance of the analytical system and the
effectiveness of the method in dealing with each sample
matrix by spiking each sample with known amounts of the
compounds the waste is being analyzed for. Using these
spiked samples, readjust the sensitivity of the instrument
such that 1 ug/gm of sample can be readily detected
(see Quality Control).
Procedure
1. Assemble gas chromatograph with either Column 1 or 2.
(See Apparatus section.)
Column 1:
a) Set helium gas flow at 40 ml/min flow rate.
b) Set column temperature at 45° C for 3 minutes
then program a 8° C/minute temperature rise to
220° C and hold for 15 minutes.
Column 2:
a) Set helium carrier gas flow at 40 ml/minute flow
rate.
b) Set column temperature at 50° C for 3 minutes, then
-------
8.01-7
program a 6° C/min temperature rise to 170° C and
hold for 4 minutes.
2. Introduce volatile compounds to the gas chromatograph
using direct injection, headspace, or purge and trap.
3. Table 8.01-1 summarizes the estimated retention times for
a number of organic compounds analyzable using this
method. An example of the separation achieved by Column 1
is shown in Figure 8.01-1.
4. Calibrate the system immediately prior to conducting any
analysis and recheck as in Quality Control for each type
of waste. Calibration should be done no less frequently
than at the beginning and end of each analyses session.
C aleu1a t i o n s
1. If a response for the contaminant being analyzed for is greater
than 2x background, then the waste does not meet
the criteria for delisting of being fundamentally different
than the listed waste. If a response is not noted, then
prior to concluding that the sample does not contain the
specific contaminant, the analyst must demonstrate, using
the spiked samples, that the instrument sensitivity is
< 1 ug/gm of sample.
-------
Table 8.01-1
ORGANOHALIDES TESTED USING PURGE AND TRAP METHOD
8.01-S
Compound
Acrylamlde
Bis(2-chloroethoxymethane)
Bis(2-chlorolsopropyl) ether
Carbon disulfide
Carbon tetrachloride
Chloroacetaldehyde
Chlorobenzene
Chloroform
Chloromethane
1,3-Dichlorobenzene
1,2-Dlchlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
Dichloromethane
Ethyl ether
Formaldehyde
Methanol
Methyl ether ketone
Methyl isobutyl ketone
Paraldehyde (trimer of
acetaldehyde)
Retention Time (Min)
Col. 1 Col. 2
13.0
14.4
24.2
10.7
1.50
34.00
34.9
35.4
9.30
11.4
18
12
5
22
12
15
12
15
.8
.1
.28
.4
.6
.4
.6
.4
-------
Compound
1,1,2,2-Tetrachloroethane
1,2,2,2-Tetrachloroethane
1,1,1,2-Tetrachloroethane
Tetrachloroethene
1,1,1-Tetrachloroethane
1.1,2-Trlchloroethane
Trichloroethene
Trichlorofluoromethane
Trichloropropane
Vinyl Chloride
Vinylidene Chloride
8.01-9
Retention Time (Mln)
Col. 1 Col. 2
21.6
21.7
12.6
16.5
15.8
7.18
15.0
13.1
18.7
13.1
2.67
5.28
-------
8.01-10
NS
oo
1S
O
Ol
U>
BROMOMETHANE
CHLORO ETHANE
CHLOROMETHANE
1.1-DJCHLOROETHENL
cis-1.2- DICHLORETHENE
1.1,1- TRICHLOROETHANE
1, 2-OICHLOROPROPANE
i^
trans -1, 3-D1CHLOROPROPENE
cisi 1, 3-DICHLOROPROPENE
1. 2-DIBROMOETHANE
1.1.1.2.TETRACHLOROETH ANt
1, 2. 3-TR1CHLOROPROPANE
MMHK
1.1, 2. 2-TETRACHLOROETHANE
CHLOROBENZENE
1-CHLOROHEXANE
BROMOBENZENE
O2O
ggo
— o
O
2.CHLOROTOLUENE
1, 4-DI CHLORO BENZENE
is
§1
-------
8.02-1
Method 8.02
METHOD FOR VOLATILE AROMATICS, KETONES AND ETHERS
Scope and Application
This method covers the determination of certain volatile
aromatics, some methyl ketones, and ethyl ether. The following
compounds may be determined by this method:
Benzene
Chlorobenzene
Dichlorobenzene
Ethyl ether
Methyl ethyl ketone
Methyl isobutyl ketone
Toluene
Xylene
Summary of Method
The volatile compounds are introduced to the gas chroma-
tograph by direct injection (Method 8.80), the headspace
method (Method 8.82), or the purge and trap technique (Method
8.83). A temperature program is used in the GC system to
separate the compounds prior to detection with a photoionization
detector (PID) or flame ionization detector (FID).
Interferences
Samples can be contaminated by diffusion of volatile
organics (particularly chlorofluorcarbons and methylene
chloride) through the sample container septum during shipment
and storage. A sample blank carried through sampling and
subsequent storage and handling can serve as a check for such
contamination.
-------
a.02^2
Apparatus
1. Gas chromatograph - Analytical system complete with
programmable gas chromatograph suitable for on-column injection
and all required accessories including Model PI-51-02 photo-
ionization detector (P1D), column supplies, recorder and gases.
A data system for measuring peak areas is recommended.
2. Supelcoport 100/200 mesh coated with 5% SP-2100 and
1.75% Bentone 34 packed in a 6 ft. x 0.085 inch ID stainless
steel column.
3. Syringes - 5 ml glass hypodermic with Luerlok tip
(2 each).
4. Micro syringes - 10, 25, 100 ul .
5. 2-way syringe valve with Luer ends (3 each),
6. Syringe - 5 ml gas-tight with shut-off valve.
7. Bottle - 15 ml screw-cap, with teflon cap liner.
Reagents
1. Activated carbon - Filtrosorb 200 (Calgon Corp.)
or equivalent.
2. Organic-free water generated by passing distilled or deionized
water through a filter bed containing of activated carbon.
A water purification system (Millipore Super-Q or equivalent)
may be used to generated 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 one hour.
-------
8.02-3
3. Stock standards - Prepare stock standard solution in
methyl alcohol using assayed liquids. Because of the
toxicity of many of the compounds being analyzed primary
diluting of these materials should be prepared in a
hood. A NIOSH/MESA approved toxic gas respirator should
be used when the analyst handles high concentrations of
such materials .
a. Place about 9.8 ml of methyl alcohol into a
10 ml 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 0.1 mg.
b. Using a 100 ul syringe, immediately add 2 drops
of assayed reference material to the flask, then
reweigh. Be sure that the 2 drops fall directly
into the alcohol without contacting the neck of the
flask.
c. Dilute to volume, stopper, then mix by inverting
the flask several times. Transfer the standard solution
to a 15 ml screw-cap bottle with a teflon cap liner.
d. Calculate the concentration in ug/ml from the net
gain in weight .
e. Store stock standards at 4°C• All standards
must be replaced with fresh standard each month.
-------
8.02-4
Calibration
1. Using stock standards, prepare secondary dilution
standards in methyl alcohol that contains the compounds
of interest, either singly or mixed together.
2. Assemble necessary gas chromatographic apparatus and
establish operating parameters equivalent to those
indicated in the Procedure section. By injecting secondary
standards adjust the sensitivity limit and the linear
range of the analytical system for each compound being
analyzed for to a sensitivity of <_ 1 ug/1 (2 x background).
Quality Control
1. Before processing any samples the analyst should
daily demonstrate through the analysis of an organic-
free water or solvent blank that the entire analytical
system is interference free.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to
validate the precision of the sampling technique. Labora-
tory replicates should be analyzed to validate the precision
of the analysis. Fortified samples should be analyzed
to validate the accuracy and sensitivity of the analysis.
Where doubt exists over the identification of a peak on
the gas chromatogram, confirmatory techniques such as
mass spectroscopy should be used.
3. The analysis should maintain constant surveillance of
both the performance of the analytical system and the
effectiveness of the method in dealing with each sample
-------
8.02-5
matrix by spiking each sample with known amounts of the
compounds the waste is being analyzed for. Using these
spiked samples readjust the sensitivity of the instrument
such that 1 ug/gm of sample can be readily detected.
Procedure
1. Assemble gas chromatograph with column specified
in Apparatus section and detector indicated in
Table 8.02-1.
2. Set helium carrier gas at 36 ml/min flow rate.
3. Hold column temperature at 50°C for 2 minutes then
program a 6°C/minute rise to 90°/C for a final hold of
40 minutes.
4. Table 8.02-1 summarizes the estimated retention times for
a number of organic compounds analyzable using this method.
An example of the separation achieved by this column is
shown in Figure 8.02-1.
5. Calibrate the system immediately prior to conducting
any analyses and recheck as in Quality Control for each
type of waste. Calibration should be done no less frequently
than at the beginning and end of each analysis session.
Calculations
1. If a response for the contaminant being analyzed for
is greater than 2 x background, then the waste
does not meet the criteria for delisting of being funda-
mentally different than the listed waste. If a response
is not noted, then prior to concluding that the sample
-------
8.02-6
IU
Ul
(SI
LU
UJ 2
O
i
o
- |2
S Id
LU
LU
f«J u3
LU Z
S S
O 03
u z
5H
«r 9
en
O
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28
RETBYTIORI TIME-MINUTES
Figure 8.02-1
GAS CHROMATOGRAM OF VOLATILE AROMATICS
-------
8.02-7
does not contain the specific contaminant, the analyst
must demonstrate, using spiked samples, that the method
sensitivity is _<_ 1 ug/gm of sample.
2. When duplicate and
spiked samples are analyzed, all data obtained 'should be
reported.
3. If one desires to determine the actual concentration of
the compound in the waste, the method of standard addition
should be used.
Table 8.02-1
CHROMATOGRAPHY
Retention Time
Compound (min)*
Benzene 3.33
Chlorobenzene 9.17
1,4 Dichlorobenzene 16.8
1,3 Dichlorobenzene 18.2
1,2 Dichlorobenzene 25.9
Ethyl ether
Methyl ethyl ketone
Methyl isobutyl ketone
Toluene 5.75
Xylene
Detector
Methods
PID
PID
PID
PID
PID
FID
FID
FID
PID
PID
* 6' x 0.085 in. column packed with 1.75% Bentone 34 and 5%
SP-2100 on Supelcoport 100/200.
-------
8.03-1
Method 8.03
METHOD FOR ACRYLONITRILE, ACETONITRILE
AND ACROLEIN
Scope and Application
This method is applicable to the determination of
acrylonitrile, acetonitrile and acrolein in waste samples.
Summary of Method
The compounds are introduced into the gas chromatograph
by either direct injection (Method 8.80), the headspace
technique (Method 8.82) or the purge and trap technique
(Method 8.83).
A temperature program is used in the gas chromatograph
to separate the organic compounds for detection by a flame
ionization detector (FID).
Interference
Samples can be contaminated by diffusion of volatile
organics (particularly chlorofluorocarbons and methylene
chloride) through the sample container septum into the sample
during shipment and storage. A sample blank carried through
sampling and subsequent storage and handling protocols can
serve as a check on such contamination.
Apparatus
1. Vial, 40 ml capacity screw cap (Pierce #13075
or equivalent). Detergent wash and dry at 105°C before
use.
2. Septum, teflon faced silicone (Pierce #12722 or
equivalent). Detergent wash, rinse with tap and
-------
8.03-2
distilled deionized water, and dry at 105°C for one hour
before use.
3. Sample introduction apparatus (see Methods 8.80, 8.82 or
8.83).
4. Gas chromatograph - Analytical system complete with
programmable gas chromatograph suitable for on-column
injection and all required accessories, column supplies,
recorder and gasses.
A data system for measuring peak areas is recommended.
5. Chromosorb 101 80/100 mesh packed in a 6' x 1/8" O.D.
stainless steel or glass column.
6. Syringes, 5 ml glass hypodermic with Luerlock tip (2
each) .
7. Micro syringe, 10, 25, 100 ul.
8. 2-way syringe valve with Luer ends (3 each).
9. Syringe - 5 ml gas tight - with shut-off valve.
10. Vial, 15 ml screwcap, with teflon cap liner.
Reagents
1. Activated carbon, Filtrosorb - 200 (Calspan Corp.) or
eqivalent.
2. Organic-free water generated by passing tap water through
a carbon filter bed containing about 1 Ib of activated
carbon. A water purification system (Millipore Super -
Q or equivalent) may be used to generate organic-free
deionized water. Organic free water may also be prepared
by boiling distilled deionized water for about 15 minutes.
Subsequently, while maintaining the temperature at 90°C,
-------
8.03-3
bubble a contaminant-free inert gas through the water
for one hour.
3. Stock standards - prepare stock standard solutions in
propanol using assayed liquids or gas cylinder as appro-
priate. Because of the toxicity of many of the compounds
being analyzed, primary dilutions of these materials
should be prepared in a hood. A NIOSH/MESA approved
toxic gas respirator should be used when the analyst
handles high concentrations of such materials.
a. Place about 9.8 ml of propanol into a 10 ml ground
glass stoppered volumetric flask. Allow about 10
minutes or until all alcohol wetted surfaces have
dried. Weigh the flask to the nearest 0.1 mg.
b. Using a 100 ul syringe, immediately add all 2 drops
of assayed reference material to the flask, then
reweigh. Be sure that the 2 drops fall directly
into the propanol without contacting the neck of the
flask.
c. Dilute to volume, stopper, then mix by inverting the
flask several times. Transfer the standard solution
to a 15 ml screw-cap bottle with a teflon cap liner.
d. Calculate the concentration in micrograms/liter
from the net gain in weight.
e. Store stock standards at A°C. All standards must be
replaced with fresh standard each month.
-------
8.03^
Calibration
1. Using stock standards, prepare secondary dilution standards
in propanol that contain the compounds of interest,
either singly or mixed together.
2. Assemble necessary gas chromatographic apparatus and
establish operating parameters equivalent to those indicated
in the Procedure section. By injecting secondary standards,
adjust the sensitivity limit and the linear range of the
analytical system for each compound being analyzed for a
sensitivity of <_ 1 ug ( 2 x background).
Quality Control
1. Before processing any samples, the analyst should demon-
strate through the analysis of an organic-free water or
solvent blank that the entire analytical system is inter-
ference free.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to
validate the precision of the sampling technique. Labora-
tory replicates should be analyzed to validate the accuracy
of the analysis. Where doubt exists over the identification
of a peak on the gas chromatogram, confirmatory techniques
should be used.
3. The analyst should maintain constant surveillance of both
the performance of the analytical system and the effective-
ness of this method in dealing with each sample matrix by
-------
8.03-5
spiking each sample with known amounts of the compounds
the waste is being analyzed for. Using these spiked
samples, readjust the sensitivity of the instrument such
that 1 ug/gm of sample can be readily detected.
Procedure
1. Assemble gas chromatograph with column specified in
Apparatus section.
2. Set helium carrier gas at 45 ml/minute flow rate.
3. Hold column temperature at 80°C for 5 minutes, then
program an 8°C/minute rise to 150°C and hold until all
species have eluted.
4. Calibrate the system immediately prior to conducting any
analyses and recheck as in Quality Control for each type
of waste. Calibration should be done no less frequently
than at the beginning and end of each analysis session.
See Figure 8.03-1 for an example of the chromatogram
to be expected using these conditions.
-------
8.03-6
COLUMN: CHROMOSORB 101
PROGRAM: 80'C -5 MINUTES.
8CC/MINUTE TO 150"C
DETECTOR: FLAME IONIZAT10N
LU
2468
RETENTION TIME-MINUTES
10
Figure 8.03-1
GAS CHROMATOGRAM OF ACROLECN AND ACRYLONITRILE
-------
8.04-1
Method 8.04
PHENOLS
Scope and Application
The following compounds may be determined by this
method:
2-Chlorophenol
Cresol(s)
Cresylic acid(s)
2,4,-Dimethylphenol
4,6-Dinitro-o-cresol
4-Nitrophenol
Pentachlorophenol
Tetrachlorophenol
Trichlorophenol(s)
Summary of Method
The sample or sample extract is injected onto the gas
chromatograph column following appropriate sample preparatioi
procedures: (Shake out (Method 8.84), Soxhlet extraction
(Method 8.86), or Sonication (Method 8.85)). A general
clean-up procedure employing liquid-liquid extraction is
described in Method 9.1. An optional clean-up procedure
specific for this group of compounds is described at the end
of this method, for samples or extracts which may require
further clean-up. A temperature program is used in the GC
system to separate the compounds before detection with a
Flame lonization Detector (FID) or Halogen Specific Detector
(HSD).
-------
8.04-2
The method also provides for the preparation of penta-
fluorobenzylbromide (PFB) derivatives for electron capture gas
chromatography with additional clean-up procedures to aid the
analyst in the elimination of interferences.
Interferences
1. Solvents, reagents, glassware, and other sample processing
hardware may yield discrete artifacts and/or elevated base-
lines causing misinterpretation of gas chromatograms. 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 and
purification of solvents by distillation in all-glass
systems may be required.
2. Interferences coextracted from s-amples will vary considerably
from source to source depending upon the waste being sampled.
While general cleanup techniques are provided in Section 9
and as part of this method, unique samples may require additional
cleanup.
Apparatus
1. Vial with cap - 40 ml capacity screw cap (Pierce #13075
or equivalent). Detergent wash and dry at 105°C before use.
2. Septum, teflon faced silicone (Pierce #12722 or equivalent).
Detergent wash, rinse with tap and distilled deionized
water, and dry at 105°C for one hour before.
3. Sample introduction apparatus (Methods 8.80 or 8.82 or 8.83).
-------
8.04-3
4. Gas chromatograph - Analytical system complete with
programmable gas chromatograph suitable for on-column
injection and all required accessories, including FID or
HSD, column supplies, recorder and gases. A data system
for measuring peak area is recommended.
5. Supelcoport 80/100 mesh coated with 1% SP-1240 DA in
1.8 meter long 2 mm ID glass column (Column 1) or Chromosorb
W-AWDMCS 80/100 mesh coated with 5% OV-17 packed in a 1.8
meter long X mm ID glass column (Column 2).
6. Syringes - 5 ml glass hypodermic with L-uerlok tip (2each).
7. Micro syringe - 10, 25, 100 ul.
8. 2-way syringe valve with Luer ends (3 each).
9. Syringe - 5 ml gas tight with shut-off valve.
10. Bottle - 15 ml screw-cap, with teflon cap liner.
11. Kuderna-Danish apparatus (K-D) [Kontes K-570000 or equivalent]
with 3 ball Snyder column.
12. Water bath - heated with concentric ring cover capable
of temperature control (± 2°C). The bath should be used
in a hood.
13. Chromatographic column - 10mm ID by 100mm length with
teflon stopcock.
14. Reaction vial - 20 ml with teflon - lined cap.
Reagents
1. 2 - propanol - pesticide quality or equivalent
2. Stock standards - prepare stock standard solutions at
a concentration of 1.0 ug/ul by dissolving 0.100 grams
-------
8.04-4
of assayed reference material in pesticide quality 2-propanol
and diluting to volume in a 100 ml ground glass stopped
volumetric flask. The stock solution is transferred to
ground glass stoppered reagent bottles, stored in a refrigera-
tor, and checked frequently for signs of degradation or
evaporation, especially just prior to preparing working stan-
dards from them.
3. PFB derivative reagents:
a. Hexane and toluene - pesticide quality or equivalent
b. Sodium gulfate - (ACS) granular, anhydrous (purified
by heating at 400°C for 4 hours in a shallow tray).
c. Potassium carbonate - (ACS) powdered.
d. Silica gel - (ACS) 100/200 mesh, grade 923; activated
at 130°C and stored in a dessicator.
e. Pentafluorobenzyl bromide - 97% minimum purity.
f. 1,4,7,10,13,16-Hexaoxacylooctadecane (18
crown 6) - 98% minimum purity.
Calibration
1. Using stock standards, prepare secondary dilution standards
in 2-propanol that contain the compounds of interest, either
singly or mixed together.
2. Assemble necessary gas chromatographic apparatus and
establish operating parameters equivalent to those in the
"procedure section." By injecting secondary standards
adjust the sensitivity limit and the linear range of the
analytical system for each compound being analyzed for to
a sensitivity of < 1 ug (2X background).
-------
8.04-5
Quality Control
1. Before processing any samples the analyst should demon-
strate through the analysis of an organic - free water
or solvent blank that the entire analytical system is
interference free.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to vali-
date the precision of the sampling technique. Laboratory
replicates should be analyzed to validate the accuracy of
the analysis. Where doubt exists over the identification
of a peak on the gas chromatogram confirmatory techniques
such as mass spectroscopy should be used.
3. The analyst should maintain constant surveillance of both
the performance of the analytical system and the effective-
ness of this method in dealing with each sample matrix by
spiking each sample with known amounts of the compounds
the waste is being analyzed for and using these spiked
samples readjust the sensitivity of the instrument such
that 1 ug/gm of sample can be readily detected (see Quality
Control).
Flame lonization Gas Chromatography Procedure
1. Assemble gas chromatograph with column 1 and Flame lonization
Detector (apparatus section).
2. Set nitrogen carrier gas at 30 ml/min flow rate.
3. Set column temperature at 80°C at injection and program
to immediately rise at 8°C/mln to 150°C.
-------
S.04-6
Derivatization and Electron Capture Gas Chromatography Procedure
1. Pipet a 1.0 ml aliquot of the 2-propanol solution of standard
or sample extract into a glass reaction vial. Add 1.0 ml
derivatization reagent. This is a sufficient amount of
reagent to derivatize a solution whose total phenolic
content does not exceed 0.3 mg/ml.
2. Add about 3 mg of potassium carbonate to the solution and
shake gently.
3. Cap the mixture and heat for 4 hours at 80°C in a hot
water bath.
4. Remove the solution from the hot water bath and allow it
to cool.
5. Add 10 ml hexane to the reaction vial and shake vigorously
for one minute. Add 3.0 ml of distilled deionized water
to the reaction vial and shake for two minutes.
6. Decant organic layer into a concentrator tube and cap with
a glass stopper.
7. Pack a 10mm ID chromatographic column with 4.0 grams of
activated silica gel. After settling the silica gel by
tapping the column, add about two grams of anhydrous sodium
sulfate to the top.
8. Pre-elute the column with 6 ml hexane. Discard the eluate
and just prior to exposure of the sulfate layer to air
pipet onto the column 2.0 ml of the hexane solution that
contains the derivatized sample or standard. Elute the
column with 10.0 ml of hexane (Fraction 1) and discard
-------
8.04-7
this fraction. Elute the column, in order with 10.0 ml
15% toluene in hexane (Fraction 2), 10.0 ml 40% toluene
in hexane (Fraction 3), 10.0 ml 75% toluene in hexane
(Fraction 4), and 10.0 ml 15% 2-propanol in toluene (Fraction
5). Elution patterns for the phenolic derivatives are
shown in Table 8.04-2 Fractions may be combined as desired
depending upon the specific phenols of interest or level
of interferences. Collect the fractions in appropriate
sized K-D apparatus and concentrate each fraction to 10 ml.
9. Assemble gas chromatograph with column 2 and HSD (apparatus
section).
10. Using 5% methane/95% argon as the carrier gas adjust flow to
30 ml/min.
11. Set column temperature at 200°C.
12. Inject 2-5ml of the appropriate fraction using the solvent -
flush technique. Smaller (1.0 ml) volumes can be injected
if automatic devices are employed. Record volume injected
to the nearest 0.05 ml and the resulting peak size in area
units. If the peak area exceeds the linear range of the
system dilute the extract and reanalyze.
Calibrate the system immediately prior to conducting any
analyses and recheck as in Quality Control for each type of
waste. Calibration should be done no less frequently than at
the beginning and end of each session.
-------
Calculations
1. If a response for the contaminant being analyzed for is
greater than 2X background is noted, then the waste does
not meet the criteria for delisting of being fundamentally
different than the listed waste. If a response is not noted,
then prior to concluding that the sample does not contain
the specific contaminant, the analyst must demonstrate, using
spiked samples, that the instrument sensitivity is _<_ 1 ug/gm.
2. When duplicate and spiked samples are analyzed, all data
obtained should be reported.
3. If one desires to determine the actual concentration of
the compound in the waste, the method of standard addition
should be used.
-------
8.04-9
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PHENOL
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2. 4. 6-TRICHLOROPHENOL
4-CHLORO-3-METHYLPHENOL § § p
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-------
8.04-10
Table 8.04-1
FLAME IONIZATION GAS CHROMATOGRAPHY OF PHENOLS
Retention Time*
Compounds (minutes)
2-Chlorophenol 1.70
Cresol(s)
Cresylic acid
2,4-Dimethylphenol 4.03
4 ,6-Ditvitro-O-cresol
4-Nitrophenol 24.25
Pentachlorophenol 12.42
Phenol 3.01
Tetrachlorophenol
2,4,6-Trichlorophenol 6.05
*Column 1
-------
8.04-11
Table 8.04-2
ELECTRON CAPTURE GAS CHROMATOGRAPHY OF PFB DERIVATIVES
Retention Recovery percent by fraction
Parent compound time
(minutes)*
2 , 4 , 6— T ri chlor opheno 1 ...
I
3.3
1.8
2.9
7.0
28.8
. 14.0
2345
90 >1
90 10
95 7
50 50
75 20
>1 90
Retention times included for qualitative information only.
The lack of accuracy and precision of the derivatization
reaction precludes the use of this approach for quantitative
purposes.
*Column 2
-------
8.06-1
Method 8.06
GAS CHROMATOGRAPHY GENERAL METHOD
FOR EXTRACTABLE ORGANICS
Scope and Application
This is a general gas chromatographic method suitable for
the determination of the presence of the following compounds in
RCRA materials:
Formic acid
Heptachlor
Hexachlorethane
Hexachloracyclopentadiene
Maleic anhydride
Naphthoquinone
Phosphorodithioic acid esters
Phthalic anhydride
2-Picoline
Pyridine
Toluene Diisocyanate(s)
Prior to using this method waste samples should be
prepared for chromatography (if necessary) using the appropriate
sample preparation method (e.g., shake out, soxhlet extraction,
sonication).
Summary of Method
Chromatographic conditions are described which allow
for the measurement of the presence of the compounds in the
extract at levels sufficient to determine the presence of the
compounds in the original waste at a concentration of 1 ug/gram.
-------
8.06-2
If interferences are encountered during chromatography
the method provides a general purpose cleanup procedure to aid
the analyst.
Precaution
Solvents, reagents, glassware, and other sample process-
ing hardware may yield discrete artifacts and/or elevated baseline
causing misinterpretation of gas chromatograms. All of these
materials should be demonstrated to be free from interferences
under the conditions of the analysis by running method blanks in
order to prevent difficulties during the analysis. Specific
selection of reagents and purification of solvents by distillation
in all-glass systems may be required to eliminate interferences.
Interfering substances coextracted from the sample will
vary considerably from source to source, depending upon the
diversity of the waste being sampled. While general cleanup
techniques are provided as part of this method, unique samples
may require additional cleanup approaches to achieve the
sensitivities required.
Phthalate esters contaminate many types of products
commonly found in the laboratory. The analyst should demonstrate
that no phthalate residues contaminate the sample or solvent
extract under the conditions of the analysis before deciding that
the compound being analyzed for is actually present. Of particular
importance is the avoidance of plactics such as polyvinyl chloride
because phthalates are commonly used as plasticizers and are
easily extracted.
-------
8.06-3
Apparatus
1. Vial with cap - 40 ml capacity screw cap (Pierce #13075 or
equivalent). Detergent wash and dry at 105°C before use.
2. Septum - teflon - forced silicone (Pierce #12722 or equiva-
lent). Detergent wash, rinse with tap and distil'ed dionized
water, and dry at 105°C for one hour before.
Gas chromatograph - Analytical system complete with program-
mable gas chromatograph suitable for on-column injection and
all required accessories, including Halogen Specific or
Flame lonization Detector, column supplies, recorder and
gases. A data system for measuring peak area is recommended.
3. Supelcoport 100/120 mesh, coated with 1.5% SP-2250 + 1.95%
SP-2401 in a 180 cm long x 4 mm ID glass column (column 1) or
Supelcoport, 100/120 mesh, coated with 3% OV-1 in a 180 cm
long x 4 mm ID glass column (column 2).
4. Syringes - 5 ml glass hypodermic with luerlok tip (2 each).
5. Micro syringe - 10, 25, 100 ul.
6. 2-way syringe valve with Luer ends (3 each).
7. Syringe - 5 ml gas-tight with shut-off valve.
8. Bottle - 15 ml screw-cap, with teflon cap liner.
9. Drying column - 20 mm ID pyrex chromatograph column with
•
coarse frit.
10. Kuderna-Danish (K-D) Apparatus equipped with the appropriate
Synder columns [Kontes K-570000 or equivalent].
11. Boiling chips - solvent extracted, approximately 10/40 mesh.
-------
12. Water Bath - heated, with concentrating cover, capable of
temperature control (2°C). The bath should be used in a
hood.
13. Chromatography column-300 mm long x 10 mm ID with coarse
fritted disc at bottom and Teflon stopcock. (Kontes K-420540-
0213 or equivalent).
Reagents
1. Sodium sulfate - (ACS) Granular, anhydrous (purified by
heating at 400°C for 4 hours in a shallow tray).
2. Stock standards - Prepare stock standard solutions at a
concentration of 1.00 ug/ul by dissolving 0.100 grams of
assayed reference material in pesticide quality iso-octane
or other appropriate solvent and diluting to volume in a 100
ml ground glass stoppered volumetric flask. The stock
solution is stored in a refrigerator, and checked frequently
for signs of degradation or evaporation, especially just
prior to preparing working standards from them.
3. Diethyl Ether - Nanograde, redistilled in glass if necessary.
Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM Laboratories,
Inc., 500 Executive Boulevard, Elmsford, N.Y. 10523.)
Procedures recommended for removal of peroxides are provided
with the test strips. After cleanup, 20 ml ethyl alcohol
preservative must be added to each liter of ether.
-------
8.06-5
4. Florisil - PR grade (60/200 mesh); purchase activated at
1250°F and store in dark in gl^ss container with ground
glass stoppers or foil-lined screw caps.
5. Hexane - Pesticide Quality
Calibration
1. Prepare calibration standards that contain the compounds of
interest either singly or mixed together.
2. Assemble the necessary gas chromatographic apparatus and
establish operating parameters equivalent to those Indicated
in the Gas Chromatography section. By injecting calibration
standards, adjust the sensitivity of the detector and the
analytical system for each compound being analyzed for so as
to detect <^ 1 ug of the compound.
3. Before using any cleanup procedure, the analyst must process
a series of calibration standards through the procedure to
determine elution patterns and the absence of interferences
from the reagents.
Quality Control
1. Before processing any samples, the analyst should demonstrate
through the analysis of a distilled water method blank, that
all glassware and reagents are interference free. Each time
a set of samples is extracted or there is a change in
reagents, a method blank should be processed as a safeguard
against chronic laboratory contamination. The blank samples
should be carried through all stages of the sample preparation
and measurement steps.
-------
8.06-6
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to validate
the precision of the sampling technque. Laboratory replicates
should be analyzed to validate the precision of the analysis.
Fortified samples should be analyzed to validate the sensi-
tivity and accuracy of the analysis. If the fortified samples
do not indicate sufficient sensitivity to detect jC 1 ug of
the compound per gm of sample then the sensitivity of the instru-
ment should be increased or the sample should be subjected to
additional clean up. The fortified samples should be carried
through all stages of the sample preparation and measurement
steps. Where doubt exists over the identification of a peak
on the chromatogram, confirmatory techniques such as mass
spectroscopy should be used.
Cleanup
1. If the entire extract is to be cleaned up to remove inter-
ferences it must first be concentrated to about 2 ml:
a. To the K-D apparatus, add a clean boiling chip and attach a
two-ball micro-Snyder column. Prewet the column by adding
about 0.5 ml hexane through the top. Place the K-D apparatus
on a hot 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 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 volume of
-------
8.06-7
liquid reaches about 0.5 ml, remove the K-D apparatus
and allow It to drain for at least 10 minutes while, cooling.
Remove the micro-Synyder column and rinse its lower joint
into the concentration tube with 0.2 ml of hexane.
2. Florisil Column Clean-up
In order to employ the Florisil column cleanup procedure the
analyst will have to determine using standards the elution pattern
of each compound to be analyzed for. Once this has been done the
following procedure can be followed to effect cleanup of the
sample.
a. Place 100 g of Florisil into a 500 ml beaker and heat
for approximately 16 hours at 400°C. After heating
transfer to a 500 ml reagent bottle. Tightly seal and
cool to room temperature. When cool add 3 ml of
distilled water which is free of phthalates and inter-
ferences. Mix thoroughly by shaking or rolling for 10
minutes and let it stand for at least 2 hours. Keep
the bottle sealed tightly.
b. Place lOg of this Florisil preparat-ion into a 10 mm ID
chromatography column and tap the column to settle the
Florisil. Add 1 cm of anhydrous sodium sulfate to the
top of the Florisil.
c. Preelute the column with 40 ml of hexane. Discard this
eluate and, just prior to exposure of the sodium sulfate
layer to the air, transfer the sample onto the column
using an additional 2 ml of hexane to complete the transfer.
-------
8.06-8
d. Just prior to exposure of the sodium sulfate layer to
the air add 40 ml hexane and continue the elution at
the rate of 2 ml/minute. This eluate is Fraction 1.
Concentrate the fraction by standard K-D technque. No
solvent exchange is necessary. After concentration and
cooling, transfer the 10 ml volumetric flask, dilute to
10 ml and analyze by gas chromatography.
e. Next elute the Florisil with 100 ml of 5 percent ethyl
ether/95% hexane (v/v) and concentrate as in step d.
[Fraction 2] .
f. Next, elute with 100 ml of 15% ethyl ether/85% hexane
(v/v) and concentrate Fraction 3 as in step d.
g. Elute with 100 ml of 50% ethyl ether/50% hexane (v/v),
and concentrate, Fraction A as in step d.
h. Finally, elute with 100 ml of ethyl ether, and concen-
trate, Fraction 5 as in step d.
Gas Chromatography
1. Assemble gas chromatograph with either Column 1 or 2 (see
Apparatus).
Column 1 (Supelcoport 100/120 with 1.5% SP 2250 + 1.95% SP 2401)
a. Set carrier gas at 60 ml/minute flow rate.
b. Column temperatures will vary from 180°C to 220°C depending
on the compound.
Column 2 (Supelcoport 100/120 with 3% OV-1)
a. Set carrier gas at 60 ml/min flow rate.
b. Column temperature will vary from 200°C to 220°C depending
on the compound.
-------
8.06-9
2. Calibrate the system at the beginning and end of an analytical
session by spiking allquots of the extract with calibration
standards.
3. Inject 2-5 ul of the sample extract or appropriate Florisil
eluate using the solvent-flush technique. Smaller (1.0 ul)
volumes can be injected if automatic devices are employed.
Record the volume injected to the nearest 0.05 ul, and the
resulting peak size, in area units.
4. If a response for the contaminant being analyzed for is greater
than 2x background, then the waste does not meet the criteria
for delisting of being fundamentally different than the
listed waste. If a response is not noted, then prior to
concluding that the sample does not contain the specific
contaminant, the analyst must demonstrate, using the spiked
samples, that the method sensitivity ±e <_ I \ig of compound
per gm of sample.
5. If the peak area measurement is prevented by the presence of
interferences, further cleanup is required.
-------
8.06-10
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-------
8.08-1
Method 8.08
GC METHOD FOR ORGANOCHLORINE PESTICIDES AND PCB's
Scope and Application
This method covers the determination of certain organochlorine
pesticides and polychlorinated biphenyls (PCB's). The following
compounds may be determined by this method:
Chlordane
Chlorinated biphenyls
Endrin
Heptachlor
Lindane
Methoxychlor
Toxaphene
Summary of Method
Prior to using this method, the waste samples should be
prepared for chromatography (if necessary) using the appropriate
sample preparation method (i.e. shake out, sonication, or soxhlet
extraction). This method provides chromatographic conditions
for the detection of _>. 1 n»8/l of the compounds in the extract.
If interferences are encountered, the method provides a
selected general purpose cleanup procedure to aid the analyst
in their elimination.
Interferences
Solvents, reagents, glassware, and other sample processing
hardware may yield discrete artifacts and/or elevated baselines
causing misinterpretation of gas chromatograms. All of these
-------
8.08-2
materials must be demonstrated to be free from Interferences
under the conditions of the analysis by running method blanks.
Specific selection of reagents and purification of solvents by
distillation in all-glass systems may be required.
Interferences coextracted from the samples will vary con-
siderably from source to source, depending upon the diversity of
the waste being sampled. While a general cleanup technique is
provided as part of this method, unique samples may require
additional cleanup.
Glassware must be scrupulously clean. Clean all glassware
as soon as possible after use by rinsing with the last solvent
used. This should be followed by detergent washing in hot water.
Rinse with tap water, distilled water, acetone and finally
pesticide quality hexane. Heavily contaminated glassware may
o
require treatment in a muffle furnace at 400 C for 15 to 30
minutes. Some high boiling materials, such as PCBs, may not be
eliminated by this treatment. Volumetric ware should not be
heated in a muffle furnace. Glassware should be sealed/stored
in a clean environment immediately after drying or cooling to
prevent any accumulation of dust or other contaminants. Store
inverted or capped with aluminum foil.
Interferences by phthalate esters can pose a major problem
in pesticide analysis. These materials elute in the 15% and 50%
fractions of the Florisil cleanup. They usually can be minimized
by avoiding contact with any plastic materials. The contamination
from phthalate esters can be completely eliminated with the use
of a microcoulometric or electrolytic conductivity detector.
-------
8.08-3
Apparatus
1. Kuderna-Danish (K-D) Apparatus, equipped with a 3 ball
Snyder column (Kontes K-570000 or equivalent).
2. Boiling chips—extracted with extraction solvent,
approximately 10/40 mesh.
3. Water .bath, heated, with concentric ring cover, capable
of temperature control (~*"2 C). The bath should be used in
a hood.
4. Gas chromatograph—Analytical system complete with gas
chromatograph suitable for on-column injection and all
required acessories including electron capture or halogen
specific detector, column supplies, recorder, gases, syringes.
A data system for measuring peak areas is recommended.
5. Chromatographic column-Pyrex, 400 mm X 25 mm OD, with coarse
fritted plate and Teflon stopcock (Kontes K-42054-213 or
equivalent).
R.eagents
1. Methylene chloride-Pesticide quality or equivalent.
2. Sodium Sulfate (ACS) Granular anhydrous (purified by heating
o
at 400 C for 4 hrs. in a shallow tray).
3. Stock standards--Prepare stock standard solutions at a
concentration of 1.0 ug/ul by dissolving 0.100 grams of
reference material in pesticide quality isooctane or other
appropriate solvent and diluting to volume in a 100 ml ground
glass stoppered volumetric flask. Store the solution in a
refrigerator. Check stock solutions frequently for signs
-------
of degradation or evaporation, especially just prior to
preparing working standards from them.
4. Mercury, triple distilled.
5. Hexane, pesticide residue analysis grade.
6. Isooctane (2,2,4-trimethyl pentane), pesticide residue
analysis grade.
7. Acetone, pesticide residue analysis grade.
8. Diethyl ether, Nanograde, redistilled in glass if necessary.
Must be free of peroxides as indcated by EM Quant Test Strips
(Test strips are available from EM Laboratories, Inc.,
500 Executive Blvd, Elmsford, N.Y., 10523).
9. Florisil, PR grade (60/100 mesh); purchase activated at
o
1250 F and store in glass containers with glass stoppers or
foil lined screw caps. Before use activate each batch at
least 16 hours at 130°C in foil covered glass container.
Calibration
1. Prepare calibration standards that contain the compounds
of interest, either singly or mixed together.
2. Assemble the necessary gas chromatographic apparatus and
establish operating parameters equivalent to those indicated
in Table 8.08-1. By injecting calibration standards, adjust
the sensitivity of the detector and of the analytical system
for each compound being analyzed for so as to detect <^ 1 ug.
3. The cleanup procedure utilizes Florisil chromatography.
Florisil from different batches or sources may vary in absorp-
tion capacity. To standardize the amount of Florisil which is
used, the use of lauric acid value (Mills, 1968) is suggested.
-------
8.08-5
The referenced procedure determines the adsorption from
hexane solution of lauric acid (mg) per gram Florisil.
The amount of Florisil to be used for each column is calculated
by dividing this factor into 110 and multiplying by 20 grams.
4. Before using any cleanup procedure, the analyst must process
a series of calibration standards through the procedure to
validate elution patterns and the absence of interferences
from the reagents.
Quality Control
1. Before processing any samples, the analyst should demonstrate
through the analysis of a distilled water method blank, that
all glassware and reagents are interference-free. Each time
a set of samples is extracted or there is a change in reagents
a method blank should be processed as a safeguard against
chronic laboratory contamination. The blank samples should be
carried through all stages of the sample preparation and
measurement steps.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to validate
the precision of the sampling technique. Laboratory replicates
should be analyzed to validate the precision of the analysis.
Fortified samples should be analyzed to validate the sensitivity
and accuracy of the analysis. If the fortified samples do not
indicate sufficient sensitivity to detect _< 1 ug/gm of sample
then the sensitivity of the instrument should be increased or
the extract subjected to additional cleanup. The fortified
samples should be carried through all stages of the sample
-------
8.08-6
preparation and measurement steps. Where doubt exists over
the identification of a peak on the chromatogram, confirmatory
techniques such as mass spectroscopy should be used.
Cleanup and Separation
Cleanup procedures are used to extend the sensitivity of a
method by minimizing or eliminating interferences that mask or
otherwise disfigure the gas chromatographic response to the
pesticides and PCB's. The Florisil column allows for a select
fractionation of the compounds and will eliminate polar materials.
Elemental sulfur interferes with the electron capture gas chro-
matography of certain pesticides but can be removed by the
techniques described below.
Florisil Column Cleanup:
1. Add a weight of Florisil, (nominally 21g,) predetermined by
calibration to a chromatographic column. Settle the Florisil
by tapping the column. Add sodium sulfate to the top of the
Florisil to form a layer 1-2 cm deep. Add 60 ml of hexane to
wet and rinse the sodium sulfate and Florisil. Just prior to
exposure of the sodium sulfate to air, stop the elution of the
hexane by closing the stopcock on the chromatography column.
Discard the eluate.
2. Adjust the sample extract volume to 10 ml and transfer it from
the K-D concentrator tube to the Florisil column. Rinse the
tube twice with 1-2 ml hexane, adding each rinse to the column.
3. Place a 500 ml K-D flask and clean concentrator tube under the
chromatography column. Drain the column into the flask until
the sodium sulfate layer is nearly exposed. Elute the column
-------
8.08-7
with 200 ml of 6% ethyl ether in hexane (Fraction 1) using a
drip rate of about 5 ml/min. Remove the K-D flask and set
aside for later concentration.
4. Elute the column again, using 200 ml of 15% ethyl ether in
hexane (Fraction 2), into a second K-D flask. Perform the
third elution using 200 ml of 50% ethyl in hexane (Fraction
3). The elution patterns for the pesticides and PCB's are
shown in Table 8.08-2.
5- Concentrate the eluates by standard K-D techniques, substituting
hexane for the glassware rinses and using the water bath at
about 85°C. Adjust final volume to 10 ml with hexane. Analyze
by gas chromatography.
6. Elemental sulfur will usually elute entirely in Fraction 1.
To remove sulfur interference from this fraction or the
original extract, pipet 1.00 ml of the concentrated extract
into a clean concentrator tube or Teflon-sealed vial. Add 1-3
drops of mercury and seal. Agitate the contents of the vial
for 15-30 seconds. Place the vial in an upright position on
a reciprocal laboratory shaker for 2 hours. Analyze by gas
chromatography.
Gas Chromatography
Table 8.08.1 summarizes some recommended gas chromatographic
column materials, operating conditions for the instrument, and
stifle estimated retention times. Examples of the separations
achieved by these columns are shown in Figures 8.08-1 through
8.08-10. Calibrate the system at the beginning and end of an
analytical session by spiking aliquots of the extract with the
-------
8.08-8
compound of Interest in order to insure a sensitivity of
_<_ 1 ug/gm of original waste tested.
1. Inject 2-5 ul of the sample extract using the solvent flush
technique. Smaller (1.0 ul) volumes can be injected if auto-
matic devices are employed. Record the volume injected to the
nearest 0.05 ul, and the resulting peak size, in area units.
2. If the peak area exceeds the linear range of the system, dilute
the extract and reanalyze. If the peak area measurement is
prevented by the presence of interferences, further cleanup is
required.
3. If detectable amounts of the compounds of interest are detected
the waste does not meet the criteria for dellsting of being
fundamentally different from that of the listed waste.
Bibliography
1. Mills, P.A., "Variation of Florisil Activity: Simple
Method for Measuring Absorbent Capacity and Its Use in
Standardizing Florisil Columns," Journal of the Association
of Official Analytical Chemists, 51, 29(1968).
-------
8,08-9
Table 8.08.1
GAS CHROMATOGRAPHY OF PESTICIDES AND PCB's
Compound
Chlordane
Endrin
Lindane
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
Retention
(min)
Col. I1
(3)
6.55
(3)
(3)
(3)
(3)
(3)
(3)
(3)
(3)
zone
Col. 22
(3)
8.10
(3)
(3)
(3)
(3)
(3)
(3)
(3)
(3)
1. Supelcoport 100/120 mesh, coated with 1.5% SP-2250/1.95%
SP-2401 packed In a 180 cm long x 4 mm ID glass column with 5%
Methane/95% Argon carrier gas at 60 ml/minute flow rate. Column
temperature is 200°C.
2. Supelcoport 100/120 mesh, coated with 3% OV-1 in a 180 cm
long x 4 mm ID glass column with 5% Methane/95% Argon carrier gas
at 60 ml/minute flow rate. Column temperature is 200°C.
3. Multiple peak response. See Figures 8.08-2 through 8.08-10,
-------
8.08-10
Table 8.08.2
DISTRIBUTION AND RECOVERY OF CHLORINATED PESTICIDES
AND PCBs USING FLORISIL COLUMN CHROMATOGRAPHY
Recovery (percent) by
fraction
Compound
ethyl ether
15% 50%
ethyl ether ethyl ether
Chlordane
Endrin
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
100
4 96
96
97
97
95 4
97
103
90
95
-------
8.08-11
COLUMN: 1.5% SP-2250*
1.95% SP-2401 ON SUPH.COPORT
TEMPERATURE: 200'C.
DETECTOR: ELECTRON CAPTURE
LU
c
4 8 12
RETENTION TIME-MINUTES
16
Figure 8.08-1
GAS CHROMATOGRAM OF PESTICIDES
-------
8.08-12
COLUMN: 1.5K SP-2250*
1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 200'C.
DETECTOR: ELECTRON CAPTURE
4 8 12
RETENTION TIME-MINUTES
16
Figure 8.08-2
GAS CHROMATOGRAM OF CHLORDANE
-------
8.08-13
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01
79
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2
m
2
c
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00
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O>
3
m
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2
C
m
ts>
oo
8.08-14
O
CO
-------
8.08-15
COLUMN: 1.5% SP-2250 +1.95% SP-2401 ON SUPB.COPORT
TEMPERATURE: 160'C.
DETECTOR: ELECTRON CAPTURE
10 14 18
RETENTION TIME-MINUTES
22
Figure 8.08-7
GAS CHROMATOGRAM OF PCB-1242
-------
8.08-16
COLUMN: 1.5% SP-2250+ 1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
& 10 14 18
RETENTION TIME-MINUTES
22
26
Figure 8.08-8
GAS CHROMATOGRAM OF PCB-1248
-------
8.08-17
K)
O>
i s
s
m
oo
K>
CD
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m
Is
m
C/I
i
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1
-------
8.09-1
Method 8.09
GC METHOD FOR SEMIVOLATILE AROMATICS AND
PHOSPHORODITHIOIC ACID ESTERS
Scope and Application
This method covers the determination of the following
compounds:
Compounds Detector
Dinitrobenzene FID
2,4-Dinitrotoluene ECD
Naphthoquinone FID
Nitrobenzene FID
Phosphorodithioic acid esters FID
Phthalic anhydride FID
2-Picoline FID
Pyridine FID
Prior to using this method, the waste samples should be prepared
for chromatography,if necessary, using the appropriate sample
preparation method (i.e. shake out, sonication, or soxhlet
extraction). This method provides gas chromatographic
techniques for measurement of the compound(s) of interest.
Flame ionization or electron capture detection may be used,
depending on the compound(s) of interest.
Summary of Method
If interferences are encountered the method provides a
general purpose cleanup procedure to aid the analyst in
their elimination.
-------
8.09-2
Interferences
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 running method blanks.
Specific selection of reagents and purification of solvents by
distillation in all-glass systems may be required.
Interferences coextracted from the samples will va~y
considerably from source to source, depending upon the diversity
of the waste being sampled. While general cleanup techniques are
provided as part of this method, unique samples may require
additional cleanup.
Apparatus
1. Kuderna-Danish (K-D) Apparatus including:
a. Concentrator tube-lOml. graduated (Kontes K-570050-1025 or
equivalent). Calibration must be checked. Ground glass
stopper (size 19/22 joint) is used to prevent evaporation
of extracts.
b. Evaporative flask-500 ml (Kontes K-57001-0500 or
equivalent). Attach to concentrator tube with springs.
(Kontes K503000-0121 or equivalent).
c. Snyder column-three-ball macro (Kontes K503000-0121 or
equivalent).
d. Snyder column-two ball micro (Kontes K-569001-0219
or equivalent).
-------
2. Boiling chips - solvent extracted, approximately
10/40 mesh.
3. Water bath - heated, with concentric ring cover, capable
of temperature control (+2°C). The bath should be used
in a hood.
4. Gas chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injection and all
required accessories including both electron capture and
flame ionization detectors, column supplies, recorder,
gases, syringes. A data system for measuring peak
areas is recommended.
5. Chromatography column - 400 mm long x 10 mm ID, with coarse.
fritted plate on bottom and Teflon stopcock.
Reagents
1. Methylene chloride - Pesticide quality or equivalent.
2. Sodium sulfate - (ACS) Granular anhydrous (purified by
heating at 400°C for 4 hrs. in a shallow tray).
3. Stock standards - prepare stock standard solutions at a
concentration of 1.0 ug/ul by dissolving 0.100 grams of
reference material in pesticide quality isooctane
or other appropriate solvent and diluting to volume in a
100 ml ground glass stoppered volumetric flask. The
stock solution is transferred to ground glass stoppered
reagent bottles, stored in a refrigerator, and checked
frequently for signs of degradation or evaporation,
especially just prior to preparing working standards
from them.
-------
8.09-^
4. Acetone, hexane, methanol, toluene - pesticide quality or
equivalent.
5. Florisil-PR grade (60/100 mesh); purchase activated at
1250°F and store in glass containers with glass stoppers
or foil lined screw caps. Before use, activate each
batch overnight at 200°C in glass containers loosely
covered with foil.
Calibration
Prepare calibration standards that contain the compounds
of interest, either singly or mixed together.
Assemble the necessary gas chromatographic apparatus
and establish operating parameters equivalent to those indicated
in Table 8.09-1. By injecting calibration standards adjust
sensitivity of the detector and the analytical system for each
compound being analyzed for so as to detect <_ 1 ug.
Before using any cleanup procedure, the analyst must
process a series of calibration standards through the procedure
to validate elution patterns and the absence of interferences
from the reagents.
Quality Control
Before processing any samples, the analyst should
demonstrate through the analysis of a distilled water method
blank that all glassware and reagents are interference-free.
Each time a set of samples is extracted or there is a change in
reagents, a method blank should be processed as a safeguard
against chronic laboratory contamination. The blank samples
-------
8.09-5
should be carried through all stages of the sample preparation
and measurement steps.
Standard quality assurance practices should be used with
this method. Field replicates should be collected to validate
the precision of the sampling technique. Laboratory replicates
should be analyzed to validate the precision of the analysis.
Fortified samples should be analyzed to validate the precision
and accuracy of the analysis. If the fortified samples do
not indicate sufficient sensitivity to detect _< 1 ug/gram of
sample, the sensitivity of the instrument should be increased
or the sample subjected to additional clean up. The fortified
samples should be carried through all stages of the sample
preparation and measurement steps. Where doubt exists over
the identification of a peak on the chromatogram, confirmatory
techniques such as mass spectroscopy should be used.
Cleanup and Separation
1. Prepare a slurry of lOg of activated Florisil in 10%
methylene chloride in hexane (V/V). Use it to pack a 10 mm
chromatography column gently tapping the column to settle
the Florisil. Add 1 cm anhydrous sodium sulfate to the top
of the Florisil.
2. Just prior to exposure of the sodium sulfate layer to
the air, transfer the 1 ml sample extract onto the column
using an additional 2 ml of toluene to complete the
transfer.
-------
8.09-6
3. Just prior to exposure of the sodium sulfate layer
to the air, add 30 ml 10% methylene chloride in hexane and
continue the elution. Elution should be at a rate of about
2 ml per minute. Discard the eluate from this fraction.
4. Next elute the column with 30 ml of 10% acetone/90%
methylene chloride (V/V) into a 500 ml K-D flask equipped
with a 10 ml concentrator tube. Concentrate the collected
fraction by the K-D technique described in Method 9.01
including the solvent exchange into 1 ml toluene. This
fraction should contain the compounds of interest.
5. Analyze by gas chromatography.
Gas Chromatography
1. Dinitrotoluene is analyzed by a separate injection
•
into an electron capture gas chromatograph. The other
compounds covered by this method are analyzed by injection of
a portion of the extract into a gas chromatograph with a flame
ionization detector. Table 8.09-1 summarizes some recommended
chromatographic column materials and operating conditions
for the instruments. Included in this table are estimated
retention times. Examples of the separations achieved by
the,primary column are shown in Figures 8.09-1 and 8.09-2.
Calibrate the system at the beginning and end of an analytical
session by spiking aliquots of the extract with each of the
compounds of interest.
2. Inject 2-5 ul of the sample extract using the solvent
flush technique. Smaller (1.0 ul) volumes can be injected
if automatic devices are employed.
-------
8.09-7
3. If a response for the contaminant being analyzed is
greater than 2X background, then the waste does not meet
the criteria for delisting of being fundamentally different
than the-listed waste. If a response is not noted, then
prior to concluding that the sample does not contain the
specific contaminant, the analyst must demonstrate,
using spiked samples, that the instrument sensitivity
is <1 ug/gm of sample.
-------
8.09-8
Table 8.09-1
GAS CHROMATOGRAPHY RETENTION TIMES
Retention time
(min)
Compound
Column Column
A B
Nitrobenzene
2,4
2,6
-Dinitrotoluene
-Dinitrotoluene
3
5
3
.31
.35
.52
4
6
4
.31
.54
.75
A. Gas-Chrom Q, 80/100 mesh, coated with 1.95% OF-1/1.5%
OV-17 packed in a 4' x 1/4" OD. glass column. FID analysis
requires nitrogen gas at 44 ml/minute and 85°C column temper-
ature. EDC analysis requires 10% Methane/90% argon carrier
gas at 44 ml/minute flow rate and 145°C column temperature.
B. Gas-Chrom Q, 80/100 mesh, coated with 3% OV-101 packed in
a 10' x 1/4" OD glass column. FID analysis requires nitrogen
carrier gas at 44 ml/minute flow rate and 100°C column temper-
ature. BCD analysis requires 10% methane/90% argon carrier
gas at 44 ml/minute flow rate and 150°C column temperature.
-------
COLUMN: 1.5X OV-17+ 1.95% QF-1 ON GAS CHROM Q
TEMPERATURE: 85*C.
DETECTOR: FLAME IONIZAT10N
COLUMN: 1.5% OV-17* 1.95% QF-1 ON GAS CHROM 0
TEMPERATURE: 145*C.
DETECTOR: ELECTRON CAPTURE
Ul
s
i
2468
RETENTION TIME-MINUTES
Figure 8.09-1
GAS CHRQMATOGRAM OF NITROBENZENE AND ISOPROPANE
2 46 8 10
RETENTION TIME-MfNUTES
Figure 8.09-2
GAS CHROMATOGRAM OF DINITROTOLUENES
-------
8.10-1
Method 8.10
GC AND HPLC METHODS FOR POLYNUCLEAR
AROMATIC HYDROCARBONS
Scope and Application
This method covers the determination of certain polynuclear
aromatic hydrocarbons (PAH). The following compounds may be
determined by this method:
Benz(a)anthracene
Benz(b)fluoranthene
Chrysene
Creosote (Phenanthrene and Carbazole)
Naphthalene
This method contains both liquid and gas chromatographic
approaches, depending upon the needs of the analyst. However,
the gas chromatographic procedure cannot adequately resolve the
following three pairs of compounds: Anthracene and phenanthrene;
chrysene and benzo(a)anthracene; benzo(b)fluoranthene and benzo(k)-
fluoranthene. If one wishes to resolve these pairs, the liquid
chromatographic approach must be used for these compounds.
Summary of Method
Prior to using this method, the waste samples should be
prepared for chromatography (if necessary) by using the appropriate
sample preparation method (i.e., Shake Out, Sonication, or Soxhlet
Extraction). This method provides chromatograpahic conditions
which allow for the detection of the compounds in the extract by
-------
8.10-2.
either High Performance Liquid Chromatograph (HPLC) or gas
chromatography.
If interferences are encountered, the method provides a
selected general purpose cleanup procedure to aid the analyst in
their elimination.
Interferences
Solvents, reagents, glassware, and other sample processing
hardware may yield discrete artifacts and/or elevated baselines
causing misinterpretation of the chromatograms. All of these
materials should be checked to insure freedom from interferences
under the conditions of the analysis by running method blanks.
Specific selection of reagents and purification of solvents by
distillation in all-glass systems may be required.
Interferences coextracted from the samples will vary consi-
derably from source to source, depending upon the diversity of
the waste sample. While a general clean-up technique is provided
as part of this method, unique samples may require additional
clean-up. The extent of interferences that may be encountered
using liquid chromatographic techniques has not been fully assessed.
Apparatus
1. Kuderna-Danish (K-D) Apparatus equipped with the appropriate
•Snyder columns [Kontes K-570000 or equivalent]
2. Boiling chips - solvent extracted, approximately 10/40 mesh.
3. Water bath - Heated, with concentric ring cover, capable
of temperature control (+2° G). The bath should be used
in a hood.
4. HPLC Apparatus:
-------
8.10-3
a. Gradient pumping system, constant flow.
b. Reverse phase column, 5 micron HC-ODS Sil-X,
250 mm x 26 mm ID (Perkin Elmer No. 809-0716 or
equivalent).
c. Fluorescence detector, for excitation at 280 nm
and emission at 389 nm.
d. UV detector, 254 nm, coupled to fluorescence detector.
e. Strip chart recorder compatible with detectors.
(A data system for measuring peak areas is recommended).
5. Gas chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injection and all
required accessories including dual flame ionization detectors,
column supplies, recorder, gases, syringes. A data system
for measuring peak areas is recommended.
6. Chromatographic column - 250 mm long x 10 mm ID with
coarse fritted disc at bottom and Teflon stopcock.
Reagents
1. Methylene chloride, pentane, cyclohexane, high purity water -
HPLC quality, distilled in glass.
2. Sodium sulfate - (ACS) Granular, anhydrous (purified by
heating at 400° C for 4 hrs, in a shallow tray).
3. Stock standards - Prepare stock standard solutions at a
concentration of 1.0 mg/ml by dissolving 0.100 grams of
assayed reference material in pesticide quality isooctane
or other appropriate solvent and diluting to volume in a
100 ml ground glass stoppered volumetric flask. The stock
solution should be stored in a refrigerator, and checked
-------
8.10-4
frequently for signs of degradation or evaporation, especially
just prior to preparing working standards from them.
4. Acetonitrile - Spectral quality.
5. Silica gel - 100/200 mesh desiccant (Davison Chemical
grade 923 or equivalent). Before use, activate for at
least 16 hours at 130° C in a foil covered glass container.
Calibration
Prepar'e calibration standards that Contain the compounds of
interest, either singly or mixed together.
Assemble the necessary HPLC or gas chromatographic apparatus
and establish operating parameters equivalent to those indicated
in Table 8.10-1 or 8.10-2. By injecting calibration standards
adjust the sensitivity of the detectors and the analytical systems
for each compound being analyzed for so as to detect ^1 ug.
Before using any cleanup procedure, the analyst must process
a series of calibration standards through the procedure to validate
elution patterns and the absence of interferences from the
reagents.
Quality Control
Before processing any samples, the analyst should demonstrate
through the analysis of a distilled water method blank, that all
glassware and reagents are interference-free. Each time a set
of samples is extracted or there is a change in reagents, a
method blank should be processed as a safeguard against laboratory
contamination. The blank samples should be carried through all
stages of the sample preparation and measurement steps.
Standard quality assurance practices should be used with
-------
8.10-5
this method. Field replicates should be collected to validate
the precision of the sampling technique. Laboratory replicates
should be analyzed to validate the precision of the analysis.
Fortified samples should be analyzed to validate the sensitivity
and accuracy of the analysis. If the fortified samples do not
indicate sufficient sensitivity to detect _<_ 1 ug of contaminant/
gm of sample then the sensitivity of the instrument should be
increased or the sample subjected to additional clean up. The
fortified samples should be carried through all stages of the
sample preparation and measurement steps.
Cleanup and Separation
(If the sample does not require cleanup, this section may
be omitted.) Before the silica gel cleanup technique can be
utilized, the extract solvent must be exchanged to cyclohexane.
1. Add a 1-10 ml aliquot' of sample extract (in methylene
chloride) and a boiling chip to a clean K-D concentrator
tube.
2. Add 4 ml cyclohexane and attach a micro-Snyder column.
Prewet the micro Snyder column by adding 0.5-ml raethylene
chloride to the top.
3. Place the micro-K-D apparatus on a boiling (100° C) 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.
-------
8.10-6
4. When the apparent volume of the- liquid reaches 0.5 ml, remove
K-D apparatus and allow it to drain for at least 10 minutes
while cooling.
5. Remove the micro-Snyder column and rinse its lower joint
into the concentrator tube with a minimum of cyclohexane.
6. Adjust the extract volume to about 2 ml.
Silica Gel Column Cleanup for PAHs
1. Prepare a slurry of lOg activated silica gel in methy-
lene chloride and place this in a 10 mm ID chromatography
column. Gently tap the column to settle the silica gel and
elute the methylene chloride. Add 1-2 cm of anhydrous
sodium sulfate to the top of the silica gel.
2. Preelute the column with 40-ml pentane. 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 of cyclo-
hexane to complete the transfer.
3. Just prior to exposure of the sodium sulfate layer to
the air, add 25 ml pentane and continue elution of the
column. Discard the pentane eluate.
4. Elute the column with 25 ml of 40% methylene chloride/60%
pentane (v/v) and collect the eluate in a 500 ml K-D flask equipped
with a 10 ml concentrator tube. Elute the column at a rate
of about 2 ml/minute.
5. Concentrate the collected fraction to less than 10 ml
by K-D techniques, using pentane to rinse the walls of the
glassware. Proceed with HPLC or gas chromatographic analysis.
-------
8.10-7
High Performance Liquid Chromatography (HPLC)
1. To the extract in the concentrator tube, add 4 ml aceto-
nitrile and a new boiling chip, then attach a micro-Snyder
column. Increase the temperature of the hot water bath to
95-100° C. Concentrate the solvent to less than 0.5 ml.
After cooling, remove the micro-Snyder column and rinse its
lower joint into the concentrator tube with about 0.2 ml
acetonitrile. Adjust the extract volume to 1.0 ml.
2. Calibrate the system at the beginning and end of an
analytical session by spiking aliquots of the extract with
the compound of interest in order to insure a sensitivity
of ^ 1 ug. Table 8.10-1 summarizes the recommended HPLC
column materials and instrument operating conditions. Included
in this table are estimated retention times. An example of
the separation achieved by this column is shown in Figure
8.10-1.
3. Inject 2-4 ml of the sample extract with a high pressure
syringe or sample injection loop. Record the volume injected
to the nearest 0.05 ml, and the resulting peak size, in
area units.
4. If the peak area exceeds the linear range of the system,
dilute the extract and reanalyze.
5. If measurement of the peak area measurement is prevented
by the presence of interfering species, further cleanup is
required.
6. The UV detector is recommended for the determination of
napthalene and the fluorescence detector is recommended for
-------
8.10-8
the remaining PAHs.
Gas Chromatography
The gas chromatographic procedure will not resolve certain
isomeric pairs as was previously indicated. The liquid chroma-
tographic procedure must be used for these materials.
1. To achieve maximum sensitivity with this method, the
extract must be concentrated to 1.0 ml. Add a clean boiling
chip to the methylene chloride extract in the concentrator
tube. Attach a two-ball micro-Snyder column. Prewet the
micro-Snyder column by adding about 0.5 ml of methylene
chloride to the top. Place this micro-K-D apparatus on a
hot water bath (60-65° C) so that the concentrator tube is
partially Immersed in the hot water. Adjust the vertical
position of the apparatus and water temperature as required
to complete the concentration in 5 to 10 minutes. At the
proper rate of distillation the balls will actively chatter
but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 ml, remove the K-D apparatus and allow
it to drain for at least 10 minutes while cooling. Remove
the micro-Snyder column and rinse its lower Joint into the
concentrator tube with a small volume of methylene chloride.
Adjust the final volume to 1.0 ml and stopper the concentrator
tube.
2. Table 8.10-2 describes the recommended gas chromatographic
column material and operating conditions for the instrument.
Included in this table are some estimated retention times
-------
8.10-9
that should be achieved by this method. Table 8ilO-3 indicates
the appropriate chromatographic detectors. Calibrate the
gas chromatographic system at the beginning and end of an
analytical session by spiking aliquots of the extract with
the compound of interest in order to insure a sensitivity of
< 1 ug.
3. Inject 2-5 ml of the sample extract using the solvent-
flush technique. Smaller (1.0 ml) volumes can be injected
if automatic devices are employed. Record the volume injected
to the nearest 0.05 ml, and the resulting peak size, in area
units.
4. If the peak area measurement is prevented by the presence
of interferences, further cleanup is required. If a response
for the contaminant being analyzed for is greater than 2x
background is noted; then the waste does not meet the
criteria for delisting of being fundamentally different
than the listed waste. If a response is not noted, then
prior to concluding that the sample does not contain the
specific contaminant, the analyst must demonstrate, using
the spiked samples, that the method sensitivity is
< 1 ug of contaminant/gm of sample.
-------
8.10-10
Table 8.10-1
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF PAH'S
Compound Retention time
(min)
Naphthalene 16.17
Phenanthrene 22.32
Benzo(a)anthracene 29.26
Benzo(b)fluoranthene 32.44
Chrysene 30 .14
Chromatographlc conditions:
Reverse phase HC-ODS Sil-X 2.6 x 250 mm Perkin-Elmer column;
isocratic elution for 5 minute using 40% acetonitrile/60% water,
then linear gradient elution to 100% acetonitrile over 25 minutes;
flow rate is 0.5 ml/minute.
-------
8.10-11
Table 8.10-2
GAS CHROMATOGRAPHY OF PAH'S
Compound Retention time
(min)
Nephthalene 4.5
Phenanthrene 15.9
Benzo(a)anthracene 20.6
Benzo(b)fluoranthene 28.0
Chrysene . . 24.7
Chromatographic conditions:
Chromosorb W-AW-DCMs 100/120 mesh coated with 3% OV-17 ,
packed in a 6' x 2 mm ID glass column, with nitrogen carrier gas
at 40 ml/minute flow rate. Column temperature was held at 100°
C for 4 minutes, then programmed at 8°/minute to a final hold at
280° C.
-------
8.10-12
Table 8.10-3
APPROPRIATE GAS CHROMATOGRAPHIC DETECTORS
Compound Detector
Benz(a)anthracene FID
Benz(b)fluoranthene FID
Carbazole ECD
Chrysene FID
Phenanthrene ECD
Naphthalene FID
-------
COLUMN: HC-ODS SIL-X
MOBILE PHASE: 40* TO 100% ACETONITRILE IN WATER
DETECTO& FLUORESCENCE
1!
Ol S"
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!.._.!_
8.10-13
i L
8 12 16 20 24
28
36
40
RETENTION TIME-MINUTES
Figure 8.10-1
LIQUID CHROMATOGRAPHI OF POLYNUCLEAR AROMATICS
-------
8.12-1
Method 8.12
GC METHOD FOR CHLORINATED HYDROCARBONS
Scope and Application
This method covers the determination of certain chlorinated
hydrocarbons. The following compounds may be determined by
this method.
Benzotrichloride
Benzyl Chloride
Dichlorobenzene
Dichloropropanol
Hexachlorobutadiene
Tetrachlorobenzene
Summary of Method
Prior to using this method, the waste samples are prepared
for chromatography (if necessary) using the appropriate
sample preparation method (i.e. shake out, sonication, or
soxhlet extraction). This method describes chromatographic
conditions which allow the accurate measurement of the compounds
using electron capture detection.
If interferences are encountered or expected, the method
provides a selected general purpose cleanup procedure to aid
the analyst in their elimination.
Interferences
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
-------
3.12-2
under the conditions of the analysis by running method blanks.
Specific selection of reagents and purification of solvents
by distillation in all glass systems may be required.
Interferences coextracted from the samples will vary
considerably from source to source, depending upon the diversity
of the waste being sampled. While general cleanup techniques
are provided, unique samples may require additional cleanup.
Apparatus
1. Kuderna-Danish (K-D) Apparatus (Kontes K-570000 or
equivalent).
a. Snyder column-three-ball macro (Kontes K503000-0121
or eqivalent).
b. Snyder column-two-ball micro (Kontes K503000-0219
or eqivalent).
2. Boiling chips-solvent extracted, approximately 10/40 mesh.
3. Water bath -heated, with concentric ring cover, capable
of temperature control (+2°C). The bath should be used in
a hood.
4. Gas chromatograph-Analytical system complete with gas
chromatograph suitable for on-column injection and all
%
required accessories including electron capture detector
column supplies, recorder, gases, syringes. A data
system for measuring peak areas is recommended.
5. Chromatography column-300 mm long x 10 mm ID with coarse
fritted disc at bot'tom and Teflon stopcock.
-------
8.12-3
Reagents
1. Methylene chloride, hexane isooctane and petroleum
ether (boiling range 30-60°C)-pesticide quality or
equivalent.
2. Sodium sulfate (ACS) granular, anhydrous (purified by
heating at 400°C for A hours in a shallow tray).
3. Stock standards - Prepare stock standard solutions at
a concentration of 10 ug/ul by dissolving 0.100 grams of
assayed reference material in pesticide quality isooctane
or other appropriate solvent and diluting to volume in
a 100 ml ground glass stoppered volumetric flask. The
stock solution is stored in a refrigerator, and checked
frequently for signs of degradation or evaporation,
especially just prior to preparing working standards from
them.
4. Florisil-PR grade (60/100 mesh); purchase activated at
1250°F and store in the dark in glass containers with
glass stoppers or foil lined screw caps. Before use,
activate each batch at 130°C in covered glass containers
loosely covered with foil.
Calibration
1. Prepare calibration standards that contain the compounds
of interest, either singly or mixed together.
2. Assemble the necessary gas chromatographic apparatus
and establish operating parameters equivalent to those
indicated in Table 8.12-1. By injecting calibration
-------
8.12-4
standards adjust the sensitivity of the detector and
the analytical system for each compound being analyzed
for to detect £ 1 ug.
3. The cleanup procedure utilizes "Florisil chromatography.
Florisil from different batches or sources may vary in
adsorption capacity. To standardize the amount of
Elorisil which is used, the use of lauric acid value
(Mills, 1968) is suggested. The referenced procedure
determines the adsorption from hexane solution of lauric
acid (mg) per gram Florisil. The amount of Florisil to
be used for each column is calculated by dividing this
ratio by 110 and multiplying by 20 grams.
4. Before using any cleanup procedure, the analyst must
process a series of calibration standards through the
procedure to validate elution patterns and the absence of
interferences from the reagents.
Quality Control
1. Before processing any samples, the analyst should demonstrate
through the analysis of a distilled water method blank, that
all glassware and reagents are interference-free. Each
time a set of samples is extracted or there is a change
in reagents, a method blank should be processed as a
safeguard against chronic laboratory contamination.
The blank samples should be carried through all stages
of the sample preparation and measurement steps.
-------
8.12-5
2. Standard quality assurance practices should be used
with this method. Field replicates should be collected
to validate the precision of the sampling technique.
Laboratory replicates should be analyzed to validate the
precision of the analysis. Fortified samples should be
analyzed to validate the sensitivity and accuracy of
the analysis. If the fortified samples do not indicate
sufficient sensitivity to detect _< 1 ug/gm of the sample
then the sensitivity of the instrument should be increased
or the sample subjected to additional cleanup. The
fortified samples should be carried through all stages
of the sample preparation and measurement steps. Where
doubt exists over the identification of a peak on the
chromatogram, confirmatory techniques such as mass
spectroscopy should be used.
Cleanup and Separation
Unless the sample is known to require cleanup omit this
section and proceed to analysis by gas chromatography.
1. Adjust the sample extract to 10 ml.
2. Place a 12 gram charge of activated Florisil in a 10 mm
ID chromatography column. After settling the Florisil
by tapping the column add a l-2cm layer of anhydrous
granular sodium sulfate to the top.
3. Pre-elute the column, after cooling, with 100 ml of
petroleum ether. Discard the eluate and just prior to
-------
8.12-6
exposure of the sulfate layer to air, quantitatively
transfer the sample extract into the column by decantatlon
and subsequent petroleum ether washings. Just prior to
exposure of the sodium sulfate layer to the air, begin
eluting the column with 200 ml petroleum ether and
collect the eluate in a 500 ml K-D flask equipped with
a 10 ml concentrator tube. This fraction should contain
all of the chlorinated hydrocarbons.
4. Add 1-2 clean boiling chips to the flask and attach a
three-ball Snyder column. Prewet the Snyder column
by adding about 1 ml hexane to the top. Place the K-D
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 l-2ml, remove the K-D apparatus
and allow it to drain for at least 10 minutes while cooling.
Note; The dichlorobenzenes have a sufficiently high
volatility that significant losses may occur in concen-
tration steps if care is not exercised. It is important
to maintain a constant gentle evaporation rate and not
to allow the liquid volume to fall below l-2ml before
-------
8.12-7
removing the K-D from the hot water bath. When the
apparatus is cool, remove the Snyder column and rinse
the flask and its lower joint into the concentrator
tube with 1-2 ml hexane.
5. Transfer to a 10 ml volumetric flask and dilute to
volume.
Gas Chromatography
1. Table 8.12-1 lists the recommended gas chromatographic
column materials and operating conditions for the instrument.
Included in this table are some estimated retention times.
Examples of the separations achieved by this column are
shown in Figures 1 and 2. Calibrate the system at the
beginning and end of an analytical session by spiking
aliquots of the sample being analyzed with the compound
of interest in order to insure a sensitivity of (1
ug/gm) of original waste or extract tested.
2. Inject 2-5 ul of the sample extract using the solvent flush
technique. Smaller (1.0 ul) volumes can be injected if
automatic devices are employed.
3. If the peak area exceeds the linear range of the system
the extract can be diluted and reanalyzed.
4. If peak detection is prevented by the presence of inter-
ferences, further cleanup is required. If detectable
amounts of the compounds of interest are detected the
waste does not meet the criteria for delisting of being
fundamentally different from that of the listed waste.
-------
8.12-8
Calculations
1. If a response for the contaminant being analyzed for
greater than 2x background is noted, then the waste does
not meet the criteria for delisting of being fundamentally
different than the listed waste. If a response is not
noted, then prior to concluding that the sample does not
contain the specific contaminant, the analyst must
demonstrate, using the spiked samples, that the instrument
sensitivity is <_ 1 ug/gm of sample.
Bibliography
1. Mills, P.A., "Variation of Florisil Activity: Simple
Method for Measuring Absorbent Capacity and Its Use in
Standardizing Florisil Columns," Journal of the Association
of Official Analytical Chemists, 51,29 (1968).
-------
8.12-9
Table 8.12-1
RETENTION TIMES FOR SOME CHLORINATED HYDROCARBONS
Compound
1, 3 dichlorobenzene
1, 4 dichlorobenzene
1, 2 dichlorobenzene
Hexachlorobutadiene
Benzotrichloride
Benzyl chloride
Dichloropropanol
Tetrachlorobenzene(s)
Retention time (min)*
4.0
4.3
5.3
11.6
(1) Gas chrom Q, 80/100 mesh, coated with 1.5% OV-1/1.5%
OV-225 packed in a 1.8 m long x 2 mm ID glass column
with 5% methane/95% Argon carrier gas at 30 ml/min
flow rate. Column temperature is 75°C. Under these
conditions Retention Time of Aldrin is 18.8 min at
160°C.
-------
8.12-10
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-------
8.22-1
Method 8.22
PHORATE
Scope and Application
This method covers the determination of the presence of
phorate in RCRA materials.
Prior to using this method, the waste samples should be
prepared for chromatography (if necessary) using the appropriate
sample preparation method (i.e., shake out, sonication, or
soxhlet extration).
Summary of Method
This method provides chromatographic conditions which
allow for the detection of the compounds in the extract.
The detector of choice is the flame photometric detector
operated in the phosphorus mode. A thermionic detector
operated in the phosphorus-nitrogen mode may also be used.
If interferences are encountered, it may be necessary to
apply cleanup procedures.
Interferences
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 running method blanks.
Specific selection of reagents and purification of solvents
by distillation in all-glass systems may be required.
Interferences coextracted from the samples will vary
considerably from source to source, depending upon the
-------
8.22-2
diversity of the waste sample. Unique samples may require
clean-up to achieve the required sensitivity in which case
the analyst must devise appropriate cleanup procedures.
Elemental sulfur may interfere with the determination of
organophosphorus pesticides by flame photometric gas
chromatography.
Apparatus
1. Gas chromatograph-Analytical system complete with gas
chromatograph suitable for on-column injection and programmed
temperature operation. Required accessories include: A
flame photometric or phosphorous-nitrogen detector, column
supplies, recorder, gases, and syringes. A data system for
measuring peak areas Is recommended. A 6 foot long x 4mm
ID glass column packed with 5% SP-2401 on 100/120 mesh
Supelcoport shall be used.
Reagents
1. Hexane, isooctane, methylene chloride - pesticide quality
or equivalent.
2. Stock standards - Prepare stock standard solutions at a
concentration of 1.00 ug/ul by dissolving 0.0100 grams of
assayed reference material in pesticide quality Isooctane
or other appropriate solvent and diluting to volume In a
10.0 ml ground glass stoppered volumetric flask. Transfer
the stock solution to a small glass vial and seal with a
Teflon lined screw cap and store in a refrigerator.
Check frequently for signs of degradation or evaporation,
especially just prior to preparing working standards from
them.
-------
8.22-3
Calibration
1. Prepare calibration standards that contain the compounds
of interest, either singly or mixed together. The
secondary standards should be prepared at concentrations
that will bracket the working range of the chromatographic
sys tern.
2. Establish operating parameters equivalent to those
indicated on Figure 8.22-1. By injecting calibration
standards, adjust the limit of the detection so as to
detect < 1 ug of phorate.
Quality Control
1. Before processing any samples, the analyst should
demonstrate through the analysis of a distilled water
method blank, that all glassware and reagents are
interference free. Each time a set of samples is extracted
or there is a change in reagents, a method blank should
be processed as a safeguard against chronic laboratory
contamination.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected to vali-
date the precision of the sampling technique. Laboratory
replicates should be analyzed to validate the precision
of the analysis. Fortified samples of waste should be
analyzed to validate the accuracy of the analysis.
Where doubt exists over the identification of a peak on
the chromatogram, confirmatory techniques such as mass
spectrometry should be used.
-------
8.22-4
Procedure
1. The recommended gas chromatographic column materials and
operating conditions for the instrument are described under
"Apparatus" and on Figure 8.22-1. The retention time for
phorate using this column is 1.43 min. An example chroma-
togram for phorate and other organophosphorus pesticides
is shown in Figure 8.22-1. Calibrate the system at the
beginning and end of an analytical session by spiking aliquots
of the extract with phorate in order to insure an analytical
sensitivity of ^ 1 ug/gm of original waste tested.
2. Inject 2 to 5 ul of the sample extract using the solvent-
flush technique. Smaller (1.0 ul) volumes can be
injected if automatic devices are employed. Record the
volume injected to the nearest 0.05 ul, and the resulting
peak size, in area units.
3. If the peak area exceeds the linear range of the system,
dilute the extract and reanalyze.
4. If peak detection is prevented by the presence of
interferences, further cleanup is required.
Results
If a response for the contaminant being analyzed for is
greater than 2x background is noted; then the waste does not
meet the criteria for delisting of being fundamentally differ-
ent than the listed waste. If a response is not noted, then
prior to concluding that the sample does not contain the
specific contaminant, the analyst must demonstrate, using
the spiked samples, that the method sensitivity is _< 1 ug of
phorate/gm of sample.
-------
N9
8.22-5
DISULFOTON
PHORATE
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-------
8.40-1
Method 8.40
METHOD FOR CHLOROPHENOXY ACID HERBICIDES
Scope and Application
This method can be used for the determination of various
chlorinated phenoxy acid herbicides in EP extracts and other
RCRA materials.
The following pesticides may be determined individually
by this method:
Compound Detector
Dichlorophenoxy acetic acid HSD
2,4,5-TP (Silvex) [2,4,5-Trichlorophenoxy HSD
proponic acid]
Summary of Method
Prior to using this method, the waste samples should be
prepared for chromatography, if necessary, using the appropriate
sample preparation method (i.e., Sonication or Soxhlet extraction)
The esters are hydrolyzed to acids and extraneous organic
material is removed by a solvent wash. The acids are converted
to methyl esters which are extracted from the aqueous phase.
The extract is cleaned by passing it through a micro-adsorption
column. Identification of the esters is made by selective
gas chromatographic separations and may be corroborated
through the use of two or more unlike columns. Detection and
measurement is accomplished by electron capture, microulometric
or electrolytic conductivity gas chromatography. Results
are reported in micrograms per liter.
-------
8.40-2
Interferences
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 should be checked to insure freedom from interferences
under the conditions of the analysis. Specific selection of
reagents and purification of solvents by distillation in all-
glass systems may be required.
The interferences in extracts of solid waste are often
high and varied and may pose great difficulty in obtaining
accurate and precise measurement of chlorinated phenoxy acid
herbicides. Sample cleanup procudures are generally required
and may result In loss of certain of these herbicides. It
is not possible to describe procedures for overcoming all of
the interferences that may be encountered in solid waste
samples.
Organic acids, especially chlorinated acids, cause the
most direct interferences with the determination. Phenols
including chlorophenols will also interfere with this procedure.
The herbicides, being strong organic acids, react readily
with alkaline substances and may be lost during analysis.
Glassware and glass wool should be acid-rinsed and sodium
sulfate should be acidified with sulfuric acid to avoid this
possiblity.
Apparatus
1. Gas Chromatograph - Equipped with glass lined injection port.
2. Detector Options:
-------
8.40-3
a. Electron Capture - Radioactive (Tritium or Nickel-63)
b. Microcoulometric Titration
c. Electrolytic Conductivity
3. Recorder - Potentiometric strip chart (10 in.) compatible
with the detector.
4. Gas Chromatographic Column Materials
a. Tubing - pyrex (180 cm long x 4 mm ID)
b. Glass Wool - Silanized
c. Solid Support - Gas-Chrom-Q (100-120 mesh)
d. Liquid Phase - Expressed as weight percent coated on
solid support.
i. OV-210,5%
ii. OV-17, 1.5% plus QF-1 or OV-210, 1.95%
5. Kuderna-Danish (K-D) Glassware
a. Snyder Column - three-ball (macro) and two-ball
(micro)
b. Evaporative Flasks - 250 ml
c. Receiver Ampules - 10 ml, graduated
d. Ampule Stoppers
6. Blender - High speed, glass or stainless steel cup.
7. Graduated cylinders - 100 ml and 250 ml.
8. Erlenmeyer flasks - 125 ml, 250 ml ground glass 24/40
9. Microsyringes - 10, 25, 50 and 100 1.
10. Pipets - Pasteur, glass disposable (140 mm long x 5 mm ID).
11. Separatory Funnels - 60 ml and 2000 ml with Teflon stopcock.
12. Glass wool - Filtering grade, acid washed.
-------
8.40-4
13. Diazald Kit - Recommended for the generation of diazomethane
(available from Aldrich Chemical Co., Cat. #210,025-2)
Reagents
1. Boron Trifluoride-methanol-esterification-reagent, 14 percent
boron trifluoride by weight.
2. N-Methyl-N-nitroso-p-toluenesulfonamide (Diazald) - High
purity, melting point range 60-62°C. Precursor for the
generation of diazomethane (see Appendix IV).
3. Potassium Hydroxide Solution - A 37 percent aqueous solution
prepared from reagent grade potassium hydroxide pellets and
reagent water.
4. Sodium Chloride - (ACS) Saturated solution (pre-rinse NaCl
with hexane) in distilled water.
5. Sodium Hydroxide - (ACS) 10 N in distilled water.
6. Sodium Sulfate, Acidified - (ACS) granular sodium sulfate,
treated as follows: Add 0.1 ml of concentrated sulfuric
acid to lOOg of sodium sulfate slurried with enough ethyl
ether to just cover the solids. Remove the ether with the
vacuum. Mix 1 g of the resulting solid with 5 ml of
reagent water and ensure the mixture has a pH below 4.
Store at 130°C.
7. Sulfuric Acid - (ACS) concentrated, Sp. Gr. 184
8. Florisil - PR grade (600-100 mesh) purchased activated
at 1250°F and stored at 130°C.
9. Carbitol (diethylene glycol monoethyl ether).
10. Diethyl Ether - Nanograde, redistilled in glass, if necessary.
a. Must be free of peroxides as indicated by EM Quant
-------
8.40-5
test strips. (Test strips are available from EM
Laboratories, Inc. 500 Executive Blvd., Elmsford,
N.Y. 10523.) Procedures recommended for removal
of peroxides are provided with the test strips.
11. Benzene, hexane - Nanograde, redistilled in glass, if
necessary.
12. Pesticide Standards - Acids and methyl esters, reference
grade.
a. Stock standard solutions - Dissolve 100 mg of each
herbicide in 60 ml ethyl ether; then make to
100 ml with redistilled hexane. Solution contains
1 mg/ml.
b. Working standard - Pipet 1.0 ml of each stock
solution into a single 100 ml volumetric flask.
Make to volume with a mixture of ethyl ether and
hexane (1:1). Solution contains 10 ug/ml of each
standard.
c. Standards for Chromatography (Diazomethane Procedure) -
Pipet 1.0 ml of the working standard into a glass stoppered
test tube and evaporate the solvent using a steam bath.
Add 2 ml diazomethane to the residue. Let stand 10
minutes with occasional shaking, then allow the
solvent to evaporate spontaneously. Dissolve the
residue in 200 ul of hexane for gas Chromatography.
d. Standard for Chromatography (Boron Trifluoride
Procedure) - Pipet 1.0 ml of the working standard
into a glass stoppered test tube. Add 0.5 ml
-------
8.40-6
of benzene and evaporate to 0.4 ml using a two-ball
Snyder microcolumn and a steam bath. Proceed as in
the Esterificatlon section (3)(a). Esters are then
ready for gas chromatogrphy.
Calibration
1. By injecting secondary standards, adjust the sensitivity
limit and the linear range of the analytical system for
each compound being analyzed for to a sensitivity of
jC 1 ug (2 x background).
2. Standards, prepared from methyl esters of phenoxy acid
herbicides calculated as the acid equivalent are injected
frequently as a check on the stability of operating
conditions. Gas chromatograms of several chlorophenoxys
are shown in Figure 8.40-1.
3. The elution order and retention ratios of methyl esters
of chlorinated phenoxy acid herbicides are provided in
Table 8.40-1, as a guide.
Quality Control
1. Duplicate and spiked sample analyses are recommended as
quality control checks. Quality control check samples
and performance evaluation samples should be analyzed on
a regular basis.
2. Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
Hydrolysis
1. Add 15 ml of distilled water and a small boiling stone to
-------
8.40-7
the flask containing the ether extract (prepared by
approriate sample preparation method) and fit the flask
with a three-ball Snyder column. Evaporate the ether in a
steam bath and continue heating for a total of 60 minutes.
2. Transfer the concentrate to a 60-ml separatory funnel.
3. Acidify the contents of the separatory funnel by adding
2 ml of cold (4°C) 25 percent sulfuric acid. Extract
the herbicides once with 20 ml of ether and twice with 10 ml
of ether. Collect the extracts in a 125 ml Erlenmeyer
flask containing about 0.5 g of acidified anydrous sodium
sulfate. Allow the extract to remain in contact with
the sodium sulfate for approximately two hours.
Easter if ication
1. Transfer the ether extract through a funnel plugged with
glass wool into a Kuderna-Danish flask equipped with a
10 ml graduated ampule. Use liberal washings of ether.
Using a glass rod crush any caked sodium sulfate during
the transfer.
a. If esterification is to be done with diazomethane,
evaporate to approximately 4 ml on a steam bath (do
not immerse the ampule in water) and proceed as directed
under "Diazomethane Esterification". Prepare diazo-
methane as directed in manufacturer's instructions.
b. If esterification is to be done with boron trifluoride,
add 0.5 ml benzene and evaporate to about 5 ml on a
steam bath. Remove the ampule from the flask and
further concentrate the extract to 0.4 ml using a
-------
8.40-8
two-ball Snyder microcolumn and proceed as directed
under "Boron Trifluoride Esterification.
2. Diazomethane Esterification:
a. Disconnect the ampule from the K-D flask and place
•
in a hood away from steam bath. Adjust volume to
4 ml with ether, add 2 ml diazomethane, and let stand
10 minutes with occasional swirling.
b. Rinse inside wall of ampule with several hundred
microliters of ethyl ether. Take sample to
approximately 2 ml to remove excess diazomethane
by allowing solvent to evaporate spontaneously
(room temperature).
c. Dissolve residue in 5 ml of hexane. Analyze by
gas chromatography.
d. If further cleanup of the sample is required, proceed
as in 3(c), substituting hexane for benzene.
3. Boron Trifluoride Ester ification:
a. After the benzene solution in the ampule has cooled,
add 0.5 ml of borontrifluoride-methanol reagent.
Use the two-ball Snyder microcolumn as an air-cooled
condenser and hold the contents of the ampule at 50°C
for 30 minutes on the steam bath.
b. Cool and add about 4.5 ml of a neutral 5 percent
aqueous sodium sulfate solution so that the benzene-
water interface is in the neck of the Kuderna-Danish
ampule. Seal the flask with a ground glass stopper
and shake vigorously for about one minute. Allow
-------
8.40-9
to stand for three minutes for phase separation.
c. Pipet the solvent layer from the ampule to the top
of a small column prepared by plugging a disposable
Pasteur pfpet with glass wool and packing with 2.0
cm of sodium sulfate over 1.5 cm of Florisil adsorbent.
Collect the elute in in a graduated ampule. Complete
the transfer by repeatedly rinsing the ampul with
small quantities of benzene and passing the rinses
through the column until a final volume of 5.0 ml
of eluate is obtained. Analyze by gas chromatography.
Calculation and Results
In analyzing EP extracts to determine if a waste is a
hazardous waste by reason of EP toxicity, calculate results
in milligrams per liter as the acid equivalent after correcting
for recovery data. When duplicate and spiked samples are
analyzed, all data obtained should be reported.
In determining if a waste should be delisted, if a
response for the contaminant being analyzed for greater than
2 x background is noted, then the waste does not meet the
criteria for delisting of being fundamentally different than
the listed waste. If a response is not noted, then prior to
concluding that the sample does not contain the specific
contaminant, the analyst must demonstrate, using that spiked
samples, that the instrument sensitivity is < lug of contaminant
/gm of sample.
-------
8.24-1
Method 8.24
GC/MS METHOD FOR VOLATILE ORGANICS
Scope and Application
This method is designed to determine volatile organic
compounds.
Summary of Method
The volatile compounds are introduced to the gas chromato-
graph by direct injection (Method 8.80), the Headspace Method
(Method 8.82) or the Purge and Trap Method (Method 8.83). 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 spectrometer operating
parameters are given.
Interferences
Interferences coextracted from the samples will vary con-
siderably from source to source, depending upon the particular
waste or extract being tested. The analytical system, however,
should be checked to insure freedom from interferences under
the conditions of the analysis by running method blanks.
Method blanks are run by analyzing organic-free water in the
normal manner. The use of non-TFE plastic tubing, non-TFE
thread sealants, or flow controllers with rubber components
in the purging device should be avoided.
Samples can be contaminated by diffusion of volatile
-------
8.24-2
organics (particularly methylene chloride) through the septum
seal into the sample during shipment and storage. A field
blank prepared from organic-free water and carried through
the sampling and handling protocol can serve as a check on
such contamination.
Cross contamination can occur whenever high level and
low level samples are sequentially analyzed. To reduce cross
contamination, it is recommended that the purging device and
sample syringe be rinsed out twice, between samples, with
organic-free water. Whenever an unusually concentrated sample
is encountered, it should be followed by an analysis of
organic-free water to check for cross-contamination. For
samples containing large amounts of water soluble materials,
suspended solids, high boiling compounds, or high organohalide
levels, it may be necessary to wash out the purging device
with a soap solution, rinse with distilled water, and then
dry in a 105°C oven between analyses.
Apparatus and Materials
1. Gas chromatograph—Analytical system complete with a
temperature programmable gas chromatograph suitable for
on-column injection and all required accessories including
an analytical column.
a. Column l--An 8 ft. stainless steel column (.125 in.
OD x 2 mm ID) packed with 1% SP-1000 coated on 60/80
mesh Carbopack B proceeded by a 1 ft. stainless steel
column (.125 in. OD x 2mm ID) packed with 1% SP-1000
coated on 60/80 mesh Chromosorb W. A glass column .25 i
x 2mm ID may be substituted for the stainless steel. The
-------
8.24-3
glass precolumn is necessary only during conditioning.
b. Column 2—An 8 ft. stainless steel column (.125 in.
OD x 2 mm ID) packed with 0.2% Carbowax 1500 coated
on 60/80 mesh Carbopack C preceded by a 1 ft. stainless
steel column (.125 in. OD x 2 mm ID) packed with 3%
Carbowax 1500 coated on 60/80 mesh Chromosorb W. A
glass column (1/4 in. OD x 2mm ID) may be substituted
for the stainless steel. The precolumn is necessary
only during conditioning.
2. Syringes—glass, 5 ml hypodermic with Luer-Lok tip
(3 each) .
3. Micro syringes--10, 25, 100 ul .
4. 2-way syringe valve with Luer ends (3 each, Teflon or
Kel-F) .
5. Syringe--5 ml gas-tight with shut-off valve.
6. 8-inch, 20 gauge syringe needle—one needle for each
5-ml syringe.
7. Mass Spectrometer--capable of scanning from 20-260 a.m.u.
in six seconds or less at 70 volts (nominal), and producing
a recognizable mass spectrum at -unit resolution from 50 ng
of DFTPP when injected through the GC inlet. The mass
spectrometer must be interfaced with a gas chromatograph
«
equipped with an all-glass, on-column injector system
designed for packed column analysis. All sections of
the transfer lines must be glass or glass-lined and
deactivated. Use Sylon-CT, Supelco, (or equivalent)
to deactivate. The GC/MS interface can utilize any
-------
8.24-4
separator that gives recognizable mass spectra (back-
ground corrected) and acceptable calibration points
at the limit of detection specified for each compound
in Table 8.24-1 .
8. A computer system should be interfaced to the mass
spectrometer to allow acquisition of continuous mass
scans for the duration of the chromatographic program.
The computer system should also be equipped with mass
storage devices for saving all data from GC-MS runs.
There must be computer software available to allow
searching any GC/MS run for specific ions and plotting
the intensity of the ions with respect to time or
scan number. The ability to integrate the area under
a specific ion plot peak is essential for quantification.
Reagents
1. Activated carbon--Filtrasorb-200 (Calgon Corp.) or
equivalent.
2. Organic-free water.
a. Organic-free water is defined as water free of
interference when employed in the purge and trap
procedure described herein. It is generated by
passing distilled deionized water through a carbon
filter bed containing activated carbon.
b. A water system (Millipore Super-Q or equivalent) may
be used to generate organic-free deionized water.
c. Organic-free water may also be prepared by boiling
deionized distilled water 15 minutes. Subsequently,
-------
8.24-5
while maintaining the temperature at 90°C, bubble a
contaminant-free inert gas through the water for
one hour. While still hot, transfer the water to a
narrow mouth screw cap bottle equipped with a Teflon
seal .
3. Stock standards (2 mg/ml)--Prepare stock standard solutions
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 toxic gas respirator
should be worn when the analyst handles high concentrations
of such materials.
a. Place about 9.8 ml of methanol into a 10 ml ground
glass stopped volumetric flask. Allow the flask to
stand, unstoppered, for about 10 minutes or until
all alcohol wetted surfaces have dried. Tare the
flask to the nearest 0.1 mg .
b. Add the assayed reference material:
Liquids—using a 100 ul syririge, immediately add
2 to 3 drops of assayed reference material to the
flask, then reweigh. Be sure that the drops fall
directly into the alcohol without contacting the
neck of the flask.
Gases—To prepare standards of bromomethane, chloro-
ethane, chloromethane, and 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 methyl alcohol menicus . Slowly inject
-------
8.24-6
the reference standard into the neck of the flask
(the heavy gas will rapidly dissolve into the methyl
alcohol).
c. Reweigh the flask, dilute to volume, stopper, then
mix by inverting the flask several times. Transfer
the standard solution to a 15 ml screw-cap bottle
equipped with a Teflon cap liner.
d. Calculate the concentration in mg per ml (equivalent
to ug per ul) from the net gain in weight.
e. Store stock standards at 4°C. Prepare fresh standards
every second day for the four gases and 2-chloroethyl-
vinyl ether. All other standards must be replaced
with fresh standards each week.
Surrogate Standard Dosing Solution—From stock standard
solutions prepared as above, add a volume to give 1000 ug
each of bromochloromethane, 2-bromo-l-chloropropane, and
1,4-dichlorobutane to 40 ml of organic-free water contained
in a 50 ml volumetric flask, mix and dilute to volume.
Prepare a fresh surrogate standard dosing solution weekly.
Dose the surrogate standard mixture into every 5 ml sample
and reference standard analyzed.
-------
8.24-7
Calibration
1. Using the stock standards, prepare secondary dilution
standards of the compounds of interest, either singly
or mixed together in methanol. The aqueous standards
must be prepared fresh daily. Standards should be at
concentrations such that the aqueous standards to be
prepared will bracket the working range of the
chromatographic system. If the limit of detection
listed in Table 8.24-1 is 10 ug/1, for example, prepare
secondary methanolic standards at 100 ug/1, and 500 ug/1,
so that aqueous standards prepared from the secondary
calibration standards, and the primary standards will
define the linearity of the detector in the working
range.
2. Assemble the necessary gas chromatographic and mass
spectrometer apparatus and establish operating parameters
equivalent to those indicated in Table 8.24-1. By inject-
ing secondary dilution standards, adjust the sensitivity
of the analytical system for each compound to <1 ug.
Quality Control
1. Before processing any samples, the analyst should daily
demonstrate through the analysis of an organic-free
water method blank, that the entire analytical system
is interference-free. The blank samples should be carried
through all stages of the sample preparation and measurement
steps .
-------
2. Standard quality assurance as indicated in Section 10 should
be followed. Field replicates should be collected to validate
the precision of the sampling technique. Laboratory
replicates and fortified samples should be analyzed to
validate the sensitivity and accuracy of the analysis.
If the fortified samples do not indicate sufficient
sensitivity to detect £ 1 ug/gm of sample then the sensitivity
of the instrument should be increased or the sample
subjected to additional clean up. The fortified samples
should be carried through all stages of the sample
preparation and measurement steps.
3. The analyst should maintain constant surveillance of
both the performance of the analytical system and the
effectivenesss of the method in dealing with each sample
matrix •
Gas Chromatography-Mass Spectrometry
1. Table 8.24-1 summarizes the recommended gas chromatographic
column materials and operating conditions for the instru-
ment. Included in this table are some estimated retention
times. An example of the separation achieved by Column 1
is shown in Figure 8.24-4.
2. GC-MS Determination--Suggested analytical conditions are
given below. Operating conditions vary from one system
to another, therefore, each analyst must optimize the
conditions for his particular GC/MS system.
-------
8.24-9
3. Mass Spectrometer Parameters:
Electron energy—70 volts (nominal).
Mass, rang.e--20-27, 33-260 amu.
Scan time—6 seconds or less.
4. Calibration of the gas chromatography-mass spectrometry
GC-MS system—Evaluate the system performance each day
that it is to be used for the analysis of samples or
blanks by examining the mass specrum of DFTPP or BFB .
a. To use DFTPP, remove the analytical column and
substitute a column more appropriate to the boiling
point of the reference compound (e.g. 3% SP-2250 on
Supelcoport). Inject a solution containing 50 ng
DFTPP and check to insure that the performance
criteria listed in Table 8.24-2 are met.
b. To use BFB, inject a solution containing 20 ng BFB
and check to insure that the performance criteria
listed in Table 8.24-3 are met.
c. If the system performance criteria are not met for
either test, the analyst must retune the spectrometer
and repeat the performance check. The performance
criteria must be met before any samples or standards
may be analyzed.
5. Analyze an internal or external calibration standard to
develop response factors for each compound.
Quantitative Determination
1. To qualitatively identify a compound, obtain an Extracted
Ion Current Profile (EICP) for the primary ion and at
least two other ions (if available) listed in Table
-------
8.24-10
8.24-4. The criteria below must be met for a quantitative
identification.
a. The characteristic ions for the compound must be found
to maximize in the same or within one spectrum of each
other.
b. The retention time at the experimental mass spectrum
must be within 60 seconds of the retention time of the
authentic compound.
c. The ratios of the three EICP peak heights must agree
with + 20% with the ratios of the relative intensities
for these ions in a reference mass spectrum. The
reference mass spectrum can be obtained from either a
standard analyzed through the GC-MS system or from a
reference library.
d. Structural isomers that have very similar mass spectra
can be explicitly identified only if the resolution
between the isomers in a standard mix is acceptable.
Acceptable resolution is achieved if the valley height
between isomers is less than 25% of the sum of the two
peak heights. Otherwise structural isomers are
identified as isomeric pairs.
2. The primary ion listed in Table 8.24-4 is to be used to
quantify each compound. If the sample produces an inter-
ference for the primary ion, use a secondary ion to
quantify.
3. For low concentrations, or direct aqueous injection of
acrylonitrlie and acrolein, the characteristic masses
-------
8.24-11
listed for the compounds in Table 8*24-4 may be used for
selected ion monitoring (SIM). SIM is the use of a mas.s
spectrometer as a substance selective detector by
measuring the mass spectrometric response at one or several
characteristic masses in real time.
Results
If a response for the contaminant being analyzed for
greater than 2 X background is noted; then the waste does not
meet the criteria for delisting of being fundamentally different
than the listed waste. If a response is not noted, then prior
to concluding that the sample does not contain the specific
contaminant, the analyst must demonstrate, using the spiked
samples, that the instrument sensitivity is < 1 ug/gm of
sample.
-------
g.24-12
References
1. "The Analysis of Halogenated Chemical Indicators of
Industrial Contamination in Water by the Purge and
Trap Method," U.S. EPA, Environmental Monitoring and
Support Laboratory, Cincinnati, OH, 45268, Dec. 1978.
2. Determining Volatile Organics at Microgram-per-Liter
Levels by Gas Chromatography," T.A. Bellar and J.J.
Lichtenberg, Jour. AWWA, 66, 739-744, Dec. 1974.
3. ASTM Annual Standards--Water, Part 31, Method D2908
"Standard Recommended Practice for Measuring Water by
Aqueous-Injection Gas Chromatography."
4. "Direct Analysis of Water Samples for Organic Pollutants
with Gas Chromatography-Mass Spectrometry," Harris, L.E.,
Budde, W.L., and Eichelberger, J.W. Anal. Chem., 46, 1912
(1974).
5. "Sampling and Analysis Procedures for Screening of
Industrial Effluents for Priority Pollutants," March 1977
(revised April 1977). USEPA, Effluent Guidelines Division,
Washington, D.C. 20460.
6. "Proceedings: Seminar on Analytical Methods for Priority
Pollutants," Volume 1 - Denver, Colorado, November 1977;
Volume 2 - Savannah, Georgia, May 1978; Volume 3 - Norfolk,
Virginia, March 1979; USEPA, Effluent Guidelines Division,
Washington, D.C. 20460.
-------
8.24-13
Table 8.24-1
GAS CHROMATOGRAPHY OF ORGANICS
Compound
Retention Time (min)
Column 1 Column 2
chlorome thane
vinyl chloride
trichlorofluorom ethane
1,1-dichloroethane
chloroform
1,2-dichloroethane
1 ,1,1-trichloroethane
carbon tetrachlor ide
trichloroethene
1,1,2-trichloroethane
benzene
1,1,2,2-tetrachloroethane
tetrachloroethene
toluene
chlorobenzene
acrolein
acrylonitr lie
1 .50
2 .67
7 .18
9.30
10.68
11 .40
12.60
13 .02
15.80
16.52
21,62
21 .67
24.18
2.10
2.57
5.14
6.48
7.70
8.29
9.28
9.45
11 .98
12.86
12.95
17.70
17.44
18.53
20.57
*
Column 1 Eight ft. stainless steel column (.125 in OD x 0.1 in.
ID) packed with 1% SP-1000 coated on 60/80 mesh
Carbopack B preceded by a 1 ft. stainless steel column
(.125 in. OD x 0.1 in. ID) packed with 1% SP-1000 coated
on 60/80 mesh Chromosorb W. (A glass column (.25 in.
OD x 2 mm ID) may be substituted). Carrier gas helium
at 40 ml/min. Temperature program: 3 min isothermal
at 45°C, then 8°/min to 220°, hold at 220° for 15 minutes
Column 2 Eight ft. stainless steel column (.125 in
ID) packed with 0.2% Carbowax 1500 coated
Carbopack C preceded by a 1 ft. stainless
(.125 in. OD x 0.1 in. ID) packed with 3%
OD x 0.1 in
on 60/80 mesh
steel column
Carbowax 1500
coated on 60/80 mesh
( .25 in. OD x mm ID)
helium at 40 ml/min.
isothermal at
all compounds
60°C then
elute.
Chromosorb W. (A glass column
may be substituted. Carrier
Temperature program: 3 min
8°/min to 160°, hold at 160 until
-------
8.24-14
Table 8.24-2
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA
Mass Ion Abundance Criteria
51 30-60% of mass 198
68 less than 2% of mass 69
70 less than 2% of mass 69
127 40-60% of mass 198
197 less than 1% of mass 198
198 base peak, 100% relative abundance
199 5-9% of mass 198
275 10-30% of mass 198
365 greater than 1% of mass 198
441 present but less than mass 443
442 greater than 40% of mass 198
443 17-23 of mass 442
-------
8.24-15
Table 8.24-3
BFB KEY IONS AND ION ABUNDANCE CRITERIA
Mass Ion Abundance Criteria
50 20-40% of mass 95
75 50-70% mass 95
95 base peak, 100% relative abundance
96 5-9% of mass 95
173 less than 1% of mass 95
174 70-90% of mass 95
175 5-9% of mass 95
176 70-90% of mass 95
177 5-9% of mass 95
-------
Table 8.24-4
8.24-16
CHARACTERISTIC
Compound
chlorome thane
vinyl chloride
trichlorof lu or ome thane
1 , 1 -d i ch lor o ethane
chloroform
1,2-dichloroethane
1,1,1-trichloroethane
carbon tetrachloride
trichloroethene
1,1,2-trichloroethane
benzene
tetrachloroethene
1,1,2,2-tetrachloroethane
toluene
chlorobenzene
acrolein
acrylonitrile
IONS OF
E
50
62
101
63
85
83
62
97
117
95
83
99
78
252
129
83
166
91
112
26
26
VOLATILE ORGANICS
I Ions
52
64
103
65 83
98 100
85
64 98 100
99 177 199
119 121
97 130 132
85 97
132 134
254 256
131 164 166
85 131 133
168
92
114
27 55 56
51 52 53
Primary Ion
50
62
101
63
83
98
117
130
97
78
164
168
92
112
56
53
-------
8.24-17
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1.1.1- TRICHLOROETHANE
1. 2-DICHLOROPROPANE
trans -1. 3-DICHLOROPROPENE
cis-1. 3-D1CHLOROPROPENE
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-------
8.25-1
Method 8.25
GC/MS METHOD, GENERAL
Scope and Application
The following compounds may be determined by this method!
Benzo(a)anthracene
Benzo(a)pyrene
Benzotrichloride
Benzyl chloride
Benzo(b)fluoranthene
Chlordane
Chlorinated dibenzodioxin
Chlorinated biphenyls
2-Chlorophenol
Chrysene
Creosote
Cresol(s)
Cresylic acid(s)
Dichlorobenzene(s)
Dichlorophenoxyacetic acid
Dichloropropanol
2,4-Dimethylphenol
Dinltrobenzene
4,6-Dinitro-o-cresol
2,4-Dinitrotoluene
Endrin
Formic acid
Heptachlor
-------
8.25-2
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Hexachloroeyelopentadiene
Lindane
Maleic anhydride
Methyl ethyl ketone
Methyl isobutyl ketone
Naphthalene
Napthoquinone
Nitrobenzene
Pentachlorophenol
Phenol
Phthalic anhydride
2-Picoline
Pyridine
Tetrachlorobenzene(s)
Toluenediamine
Toluene diisocyanate(s)
Toxaphene
Trichlorophenol(s)
2,4,5-TP(Silvex)
Summary of Method
prior to using this method, the waste samples should be
prepared for chromatography (if necessary) using the appropriate
•
sample preparation method (i.e., shake out, sonication, or
soxhlet extraction). If emulsions are a problem, continuous
-------
8.25-3
extraction techniques should be used. This method describes
chromatographlc conditions which allow for the separation of
the compounds In the extract.
Interferences
Solvents, reagents, glassware, and other sample processing
hardware may yield discrete artifacts and/or elevated baselines
causing misinterpretation of chromatograms. 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 and purification of solvents
by distillation in all-glass systems may be required.
Interferences coextracted from the samples will vary
considerably from source to source, depending upon the
diversity of the industrial complex or waste being
sampled.
The recommended analytical procedure may not have
sufficient resolution to differentiate between certain isomeric
pairs. These are anthracene and phenanthrene, chrysene,
and benzo(a)anthracene, and benzo(b)fluoranthene and
benzo(k)fluoranthene. The GC retention time and mass spectral
data are not sufficiently unique to make an unambiguous
distinction between these compounds. If identification of
these specific compounds is required Method 8.10 should be
used.
Apparatus
1. Separatory funnel - 20 mm, with Teflon stopcock (Ace
Glass 7228-T072 or equivalent).
-------
3.25-4
2. Drying column - 20 mm ID pyrex chromatographic column
equipped with coarse glass frit or glass wool plug.
3. Kuderna-Danish (K-D) Apparatus equipped with 3-ball Snyder
column (Kontes K-570000 or equivalent).
4. Water bath - Heated with concentric ring cover, capable
of temperature control (+_ 2°C). The bath should be used
in a hood.
5. Gas chromatograph - Analytical system complete with gas
chromatograph capable of on-column injection and all
required accessories including column supplies, gases, etc.
6. Column 1 - For Base/Neutral and Pesticides a 6-foot glass
column (1/4 in OD x 2 mm ID) packed with 3% SP-2250 coated
on 100/120 Supelcoport (or equivalent).
7. Column 2 - For Acids, a 6-foot glass column (1/4 in OD x
2mm ID) packed with 1% SP-1240 DA coated on 100/120 mesh
Supelcoport (or equivalent).
8. Mass Spectrometer - Capable of scanning from 35 to 450
a.m.u. every 7 seconds or less at 70 volts (nominal) and
producing a recognizable mass spectrum at unit resolution
from 50 ng of DFTPP when the sample is introduced through
the GC inlet (References 2). The mass spectrometer must
be interfaced with a gas chromatograph equipped with an
injector system designed for splitless injection and
glass capillary columns or an injector system designed
for on-column injection with all-glass packed columns.
All sections of the transfer lines must be glass or
-------
8.25-5
glass-lined and must be deactivated. (Use Sylon-CT,
Supelco, Inc., or equivalent to deactivate.)
*
Note; - Systems utilizing a jet separator for the GC effluent
are recommended since membrane separators may lose sensitivity
for light molecules and glass frit separators may inhibit the
elution of polynuclear aromatics. Any of these separators
may be used provided that it gives recognizable mass spectra
and acceptable calibration points at the required limit of
detection.
9. A computer system must be interfaced to the mass spectro-
meter to allow acquisition of continuous mass scans for
the duration of the chromatographic program. The computer
system should also be equipped with mass storage devices
for saving all data from GC-MS runs. There must be
computer software available to allow searching any GC-MS
run for specific ions and plotting the intensity of the
ions with respect to time or scan number. The ability
to integrate the area under any specific ion plot peak
is essential if quantification is to be attempted.
10. Continuous liquid-liquid extractors-Teflon or glass
connecting joints and stopcocks, no lubrication. (Hershberg-
Wolf Extractor-Ace Glass Co., Vineland, N.J. P/N 6841-10 or
eqivalent).
Reagents
1. Sodium hydroxide - (ACS) 6N in distilled water.
2. Sulfuric acid - (ACS) 6N in distilled water.
-------
8.25-6
3. Sodium sulfate - (ACS) granular anhydrous (rinsed with
methylene chloride (20 ml/g) and conditioned at 400°C
for 4 hrs . ) .
4. Methylene chloride - Pesticide quality or equivalent.
5. Stock standards - Prepare stock standard solutions at a
concentration of 1.00 ug/ul. For example, dissolve 0.100
grams of assayed reference material in pesticide quality
isooctane or other appropriate solvent and dilute to
volume in a 100 ml ground glass stoppered volumetric
flask. The stock solution is transferred to 15 ml Teflon
lined screw cap vials, stored in a refrigerator, and
checked frequently for signs of degradation of evaporation,
especially just prior to preparing working standards
from them. Protect standards from light.
Procedure
1. Calibrate instrument using the conditions indicated in
Table 8.25-1, so that a response greater than 2 times
background is obtained for 1 ug of each species being
analyzed for.
2. Inject the extract derived from 1 gm of the sample being
analyzed. The extract should be diluted and range finding
studies conducted prior to injection of concentrated extract
in order to prevent instrument overload.
Results
If a response for the contaminant being analyzed for is
greater than 2x background is noted; then the waste does not
-------
8.25-7
meet the criteria for delisting of being fundamentally
different than the listed waste. If a response is not noted,
then prior to concluding that the sample does not contain
the specific contaminant, the analyst must demonstrate, using
the spiked samples, that the instrument sensitivity is
_£ 1 ug/gm of sample.
guallty Control
1. Before processing any samples, demonstrate through the
analysis of a method blank, that all glassware and
reagents are interference-free. Each time a set of
samples Is extracted or there is a change in reagents,
a method blank should be processed as a safeguard against
chronic laboratory contamination.
2. Standard quality assurance practices should be used with
this method. Field replicates should be collected and
analyzed to determine the precision of the sampling
technique. Laboratory replicates should be analyzed to
determine the precision of the analysis. Fortified
samples should be analyzed to determine the accuracy
of the analysis. Field blanks should be analyzed to
check for contamination introduced during sampling
and transportation.
-------
8.25-8
Table 8.25-1
CHROMATOGRAPHIC CONDITIONS
Benz(a)anthracene BN
Benzo(a)pyrene BN
Benzotrichloride BN
Benzyl chloride BN
Benz(b)fluoanthene BN
Chlordane BN
Chlorinated dibenzodioxin BN
Chlorinated biphenyls BN
2-Chlorophenol BN
Chrysene BN
Creosote BN
Cresol(s) A
Cresylic acid(s) A
Dichlorobenzene(s) BN
Dichlorophenoxyacetic acid A
Dichloropropanol BN
2,4-Dimethylphenol A
Dinitrobenzene BN
4 ,6-Dinitro-o-cresol A
2,4-Dinitrotoluene BN
Endrin P
Formic acid BN
Heptachlor P
Hexachlorobenzene BN
Hexachlorobutadiene BN
Hexachloroethane BN
Hexachlorocyclopentadiene BN
Lindane P
Maleic anhydride BN
Methyl ethyl ketone A
Methyl isobutyl ketone A
Naphthalene BN
Napthoquinone BN
Nitrobenzene B
Pentachlorophenol A
Phenol BN
Phthalic anhydride BN
2-Picoline BN
Pyridine BN
-------
8.25-9
Table 8.25-1 (Cont.)
Tetrachlorobenzene(s) BN
Toluenediamine BN
Toluene dlisocyanate(s) BN
Toxaphene P
Trichlorophenol(s) A
2,4,5-TP(Silvex) A
(A) °8 foot glass column (1/4 in. OD x 2 mm ID) packed with
1% SP-1240 DA coated on 100/120 mesh Supelcoport. Carrier
gas helium at 30 ml per min. Temperature program: 2 min
isothermal at 70°, then 8° per min to 200°C. If desired,
capillary or SCOT columns may be used.
(BN) #Six foot glass column (1/4 in. OD x 2 mm ID) packed with
3% SP-2250 coated on 100/120 mesh Supelcoport. Carrier gas
helium at 30 ml per min. Temperature program: isothermal
for 4 minutes at 50°C, then 8° per min to 270°C. Hold at
270°C for 30 minutes. If desired, capillary or SCOT columns
may be used.
(P) °6 foot glass column (1/4 in. OD x 2 mm ID) packed with 3%
SP-2250 coated on 100/120 mesh Superlcoport. Carrier gas
helium at 30 ml per min. Temperature program: isothermal
for 4 minutes at 50°C, then 8° per minute to 270°. Hold at
270°C for 30 minutes. If desired, capillary or SCOT columns
may be used
-------
8.25-10
,1,3-DICHLOROBENZENE
1,4-DICHLOROBENZENE
1.2-DICHLOROBENZENE + HEXACHLOROETHANE
BIS(METHYL-2CHLOROETHYL)ETHER + BISI2-CHLOBOETHYDETHER
.HEXACHLOROBUTADIENE
.1.2.4-TRICHLOROBENZENE
ISOPHORONE
NAPTHALENE
HEXACHLOROCYCLOPENTADIENE
C1
TO
za
rn
2-CHLORONAPTHALENE
ACENAPHTHYLENE
ACENAPHTHENE
DIMETHYL PHTHALATE
2,6-DINITROTOLUENE
4-CHLOROPHENYL PHENYL ETHER
IYLPHTHALATE
DIPHENYLHYDRAZINE
( DIETHYLPHTHALATE
VIEl •»- 1.2-
HEXACHLOROBENZENE
"4-BROMO PHENYL PHENYL ETHER
PHENANTHRENE ANTHRACENE
ro
«n
m
CO
v>
d-10 ANTHRACENE
DIBUTYL PHTHALATE
FLUORANTHENE
PYRENE
BENZYL BUTYL PHTHALATE
B1S|2-ETHYLHEXYL)PHTHALATE
4,4'-DICHLOROBENZIDtNE
U)
W
BENZO(b)FLUORANTHENE
BENZOdOFLUOfiANTHENE
D1OCTYL PHTHALATE
O 2 O
3o£
BENZO(a)PYRENE
C
S
•2.
_ 01
53
t» O
>^ fO
co ro
TJ 3 01
m ~ o
i
JNOENO{1.2.3-cd)PYRENE
/DIBENZO(ah) ANTHRACENE
2 oo
S
BENZO(ghi)PERYLENE
3S
o
-------
8.25-11
o
m
o
m
^j
O
IE
m
2 _»
C ro
-H
m
co
ro
o
ro
ro
oSi
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§8"
co O %'
to ro 1.
c/> S Ifa
m =; o
O . P
§5
2-CHLOROPHENOL m 1
m
3D
2-NITROPHENOL ro
o
o
0°
PHENOL -°
2,4-DIMETHYLPHENOL
O
z
co
C
-o
P
O
O
-o
O
so
2,4-DICHLOROPHENOL
2,4,6-TRICHLOROPHENOL
4-CHLORO-3-METHYLPHENOL
2,4-DIWITROPHEWOL
2-METHYL-4.6-DINITROPHEWOL
PEiMTACHLOROPHEMOL
4-NITROPHEIMOL
-------
8.25-12
m
m
Z
c
to
o
/?-BHC & HEPTACHLOR
ro
en
ALDRIN
HEPTACHLOR EPOXIDE
ENDOSULFAN I
=====— DIELDRIN & 4.4' DDE
-4,4'-DDT
ENDOSULFAN SULFATE
ENDOSULFAN E & 4,4'-DDD
-------
8.25-13
COLUMN: 3% SP-2250 ON SUPacOPORT
PROGRAM: 50'C, 4 WIN, 8°PER IVIIN TO 270'C
DETECTOR: MASS SPECTROMETER
'PEAKS GIVING THE THREE
CHARACTERISTIC IONS
RETENTION TIME-MINUTES
Figure 8.25-4
GAS CHROMATOGRAM OF CHLORDANE
-------
8.25-14
COLUMN: 3% SP-2250 DIM SUPELCOPORT
PROGRAM: 50°C. 4 MIN. 8°PER MIN TO 270°C
DETECTOR: MASS SPECTROMETER
25 30 35
RETENTION TIME-MINUTES
Figure 8.25-5
GAS CHRQMATOGRAM OF TOXAPHENE
-------
8.25-15
COLUMN: 3% SP-2250 ON SUPELCOPORT
PROGRAM: 50°C. 4 MIIM, 8°PER MIN TO 270°C
DETECTOR: MASS SPECTROMETER
A m/e 224 PRESENT
B m/e 260 PRESENT
C m/e 294 PRESENT
20
25 30*
RETENTION TIME-MINUTES
35
Figure 8.25-6
GAS CHROMATOGRAM OF AROCHLOR 1248
-------
8.25-16
COLUMN-: 3% SP-2250 ON SUPELCOPERT
PROGRAM: 50°C, 4 MIN, 8° PER MIN TO 270°C
DETECTOR: MASS SPECTROMETER
A m/e 294 PRESENT
B m/e 330 PRESENT
C m/e 362 PRESENT
20 25 30
RETENTION TIME-ftflfWUTES
Figure 8.25-7
GAS CHHOMATOGRAM OF AROCHn5R 1254
-------
8.49-1
Method 8.49
GENERAL REQUIREMENTS FOR METALS ANALYSIS
I. It is essential to prevent contamination of the sample.
Reagents must be of the highest grade and all laboratory
equipment must be kept scrupulously clean and protected.
Pipets with disposable tips are recommended. Glassware,
plastic containers, and sample tubes should be subjected
to the following series of washes in the order given:
a. Thoroughly scrub with detergent and water.
b. Rinse with a solution of one part concentrated nitric
acid to one part water.
c. Rinse with water.
d. Rinse with a solution of one part hydrochloric acid
and one part water.
e. Rinse with water.
f. Rinse with deionized distilled water.
g. Dry the plastics at 50° C, the glassware at 105° C.
2. Sample bottles may be of borosilicate glass, polyethylene,
polypropylene or teflon. It is recommended that they be
used one time and discarded. If this is not feasible,
they must be subjected to the series of washes listed in
1.
3. Deionized distilled water is made by passing distilled
water through a mixed bed ion exchange resin column.
All reagents, standards and dilutions shall be prepared
using this water.
-------
Revision B 4/15/81 /«.*27-l
Method 8.27
Capillary Column GC/MS method for the analysis of Wastes
Scope and Application
This method may be used to determine the presence and
concentration of the volatile and extractable organic compounds
which are listed in Appendix VIII, 40 CFR 261.33 in wastes. The
method employs capillary column gas chromatography-mass
spectrometry. Quantitation can be performed in two ways depending
on the level of information required.
Summary of Method
The waste is categorized by its physical makeup into one
of the following three classes.
0 Liquid (either single or multi-phase systems)
0 Solid
0 Combination of Liquid and Solid
Liquids are analyzed in their "as received" form except that
if more than one phase is present the organic and aqueous phases
are separated and the two phases analyzed separately. The
organic phases are analyzed by direct injection onto the
capillary column using either the split or splitless technique.
Aqueous phases are determined by a combination of the purge
and trap technique for volitiles and a series of extractions
for the base/neutrals and acids. The extracted fractions
are then combined and analyzed as a single solution using
the splitless technique.
Solids are analyzed using purge and trap technique for
volatiles and a soxhlet extraction for the extractables.
Samples containing both liquid and solid phases are first
separated into their component liquid and solid phases
using centrifugation. The separated phases are then analyzed
as either liquids or solids as described above.
The components of the sample are quantitated in either
of two ways, depending on the degree of quantitation necessary.
The first way estimates concentration and assigns these
estimated concentrations into ranges. Since this is not a
rigorous quantitative procedure it may only be used for
order of magnitude type estimates of concentration. These
ranges are:
0 Greater than 50%
0 -Between 10 and 50%
0 Between 1 and 10%
0 Between 100 ppm and 1%
0 Between 1 and 100 ppm
-------
Revision B 4/15/81 8.27-2
For determining the precise concentration of a component in a
sample the method of standard additions is employed.
Procedure
The analyst must first determine which category the
sample belongs to.
0 If the sample is a liquid go to the section labled
"Liquids" (I) of this procedure.
0 If the sample is a solid go to the section labled
"Solids" (II) of this procedure.
0 If the sample contains both liquid and solid go to
the section labled "Mixtures of Liquids and Solids"
(III) of this procedure.
I. Liquids
The analyst should determine if more than one liquid
phase is present in the sample. If more than one phase is
present the sample should be separated into its organic and
aqueous phases respectively. This separation can be achieved
by using either gravity or cenrifugation. The separated
phases should be weighed. 10.0 gm of well mixed sample
should be used.
A. Organic Liquids
1. Summary
Organic Liquids are injected directly onto the
capillary column using either the split or splitless
mode. The liquid may be diluted if necessary to
facilitate sample handling or to accomodate the
linear range of the mass spectrometer.
2. Apparatus and Materials
a. Sample Vials - 10 dram vials with teflon lined caps
b. Gas Chromatograph. - Analytical system capable of
split and splitless injections and all required
accessories including column supplies, gases, etc.
c. Column - 30m SE-30, SE-52, SE-54, or equivalent,
.2 to .25mm internal diameter with a film thickness
between .15 to .40u.
d. Mass Spectrometer - Capable of scanning from 35 to
450 daltons every 1 second or less. The mass-
spectrometer must be able to operate at 70
volts for electron ionization and must produce
a recognizable mass spectrum for 50 ng or less
of DFTPP when the sample is introduced through
the GC column. The GC column should be directly
interfaced (i.e. no separator) to the mass
spectrometer through either an all glass or all
glass lined system. If a fused silica capillary
column is used, the analyst is required to
complete the interface by placing the end of
-------
Revision B 4/15/81 8.27-3
the column in the ion source.
e. Computer System - The computer system interfaced
to the mass spectrometer should be capable of
continuously acquiring mass spectra for the duration
of the gas chromatographic program, (about 1 hr.)
All data must be stored either within the data
system or on line mass storage devices such as
disk or tape. The system must have software
available capable of searching GC/MS runs for
the following:
1) selected ion chromatograms
2) total ion chromatograms
3) reverse and forward search for any compound
from the EPA/NIH Mass Spectral Data Base.
3. Reagents
a. Methylene chloride - Pesticide quality
b. Ethyl Ether - "
c. Ethylacetate - " "
d. Methanol - "
e. Standards - Standards can be made up as necessary
if appropriate reagents are available. Naphthalene-dg
or phenanthrene-dio may be used as internal standards.
4. Calibration
a. The mass spectrometer is calibrated with either
PFK or FC-43 ovver the scan range. The mass spec-
trometer should be scaned from 35 to 450 daltons
in 2 sec or less. 50ng or less of DFTPP should
be injected in the splitless mode using the
conditions given in Table 8.27-1.
b. The DFTPP spectrum obtained from the top of the
chromatographic peak (backgroud subtracted)
should meet the criteria listed in Table 8.24-2.
5. Sample Preparation
a. If the liquid can be convieniently drawn into a
10 ul syringe, then no sample preparation is
necessary. Weigh 1 gm of the liquid into a
a pre-tared 10 dram vial. Add the internal
standard at a level that would give 50 ng on
column when injected. (The amount added will
vary with split ratio)
b. If it is necessary to dilute the sample, a weighed
portion of the organic phase should be transfered
to an appropriate volumetric flask and diluted
to volume with one of the solvents listed in
the reagent section. The internal standard is
added at a level that would give 50 ng on column
when injected prior to dilution. Record the
dilution volume.
-------
Revision B 4/15/81 8.27-4
6. Gas Chromatography/Mass spectrometry
a. Establish the chromatographic conditions given
in table 8.27-1.
b. Set the Gas Chromatograph for either split
or splitless injection depending on estimated
concentration. For example, an organic liquid
that can be conveniently drawn up in a syringe
can be analyzed using the split mode. An oily
sample that needs to be diluted to 1:100 might
best be handled using the splitless mode. If
using the split mode, record the split ratio.
Record both linear and volume column flow.
c. Inject sample, start the chromatographic program,
and acquire data. Record amount of sample
injected. (1 to 5 ul when using split mode and
1 to 2 ul when the splitless mode is employed).
d. Inject appropriate standards and acquire data using
sample conditions as employed in c.
7. Qualitative and Quantitative Determination
a. A compound will be judged to have been identified
if either three or more characteristic ions of
the compound maximize within one scan of the
apex of the peak and the integrated ion areas
agree with a library or standard mass spectrum
within + 20%; or, a reverse search yields a numerical
value equivalent to the criteria stated above.
b. Samples can be quantitated in two ways. The first
is by the method of standard additions. This
method is always acceptable and must be used
when the actual concentration is needed. The
second method is used when order of
magnitude estimates of concentration are
needed. This is done by comparing the Total
Ion Chromatogram of the compound in the sample
with a standard. For example, if 100 ng of
benzene gives a total of 10,000 integrated area
counts then a peak corresponding to toluene
with 25,000 counts would be expected to correspond
to about 250 ng. When using this method the
analyst should try to use standards which resemble
the compounds in question as closely as possible.
The internal standard is used as a method check.
For example, if 50 ng of the internal standard
normally gives 5000 integrated area counts,
this condition should be met in the sample
+20%
c. Example Calculation
5 ul of a 5 mg/ml solution of benzene was injected
with a split ratio of 100:1 to produce the
-------
Revision B 4/1S/81 C.27-5
chromatogram in figure 8.27-1. The integrated
area of the benzene peak from the total ion
chromatogram was 7840 counts. 10 gm of an
organic liquid sample was disolved in methylene
chloride in a volumetric flask to a final volume
of 100 ml. 5 ul of this solution was injected
with the same split ratio to produce the chromatogram
in figure 8.27-2. The integrated area for
benzene in this sample was 4235 counts. The
peak for toluene gave an area of 4827 counts.
The estimated concentration of benzene and
toluene in this sample are:
Benzene Standard
5 mg/ml = 5 ug/ul
5 ug/ul x 5 ul = 25 ug injected
7840 counts/25 ug = 313.6 counts/ug
Benzene in Sample
4235 counts/5 ul injected x 1 ug/ 313.6 counts =
13.5 ug/5 ul
13.5 ug/5 ul x 1000 ul/ml -
2700 ug/ml
2700 ug/ml x 100 ml/10 gm dilution = 27000 ug/gm
27000 ug/gm = 27 mg/gm
27 mg/gm x 1 gm/1000 mg = .027 = 2.7%
The sample is 2.7% benzene
Toluene in sample
By an analogus method the sample is calculated to
be 3.1% toluene.
8. Report
a. Report the results of each analysis giving
the method used to quantify each comound.
Report the scan number of each compound.
b. Example:
-------
Revision B 4/15/81 8.27-6
Compound Quantitation Scan Amount Range
Method #
Benzene Estimate/Benzene 500 2% 1-10%
Toluene Estimate/Benzene 622 3% 1-10%
B. Aqueous Liquid
1. Summary
Aqueous liquids are analyzed by purge and trap and
extraction methods given in Methods 8.83 and 8.84.
After the aqueous sample is purged and traped and
extracted by Methods 8.83 and 8.84 the traped material
and the extracts (which have been combined) are
analyzed by capillary column gas chromatography-mass
spectrometry.
2. Apparatus and Materials
See the appropriate sections in Methods 8.83, 8.84, and
the apparatus and materials section for organic liquids
in this method
3. Reagents
See the appropriate sections in Methods 8.83, 8.84, and
the reagents section for organic liquids in this method.
4. Calibration
a. The mass spectrometer is calibrated with PFK or
FC-43 over the scan range of interest. For the
volatiles scan over the range 20 to 260 daltons,
and scan over the range 35 to 450 daltons for
the base/neutrals and acid extractables. The
scan rates should be 2 sec. or less. 50 ng or
less of bromoflurobenzene or DFTPP should be injected
for the volitiles and extractables respectively.
Chromatographic conditions are given in Tables
8.27-2 and 8.27-3.
b. The specta obtained from the top of the Chromatographic
peak (background subtracted) should meet the criteia
listed in Tables 8.24-2 and 8.24-3.
5. Sample Preparation
a. Follow the purge and trap and extraction methods
given in Methods 8.83 and 8.84 of this manual
for preparation of the sample.
b. Base/neutral and acid extractable fractions
may be combined and analyzed in a single GC/MS
analysis.
6. Gas Chromatography/Mass Spectrometry
a. Establish the chromatographic conditions described
in Tables 8.27-2 or 8.27-3, whichever is appropriate.
b. Set gas chromatograph in either the split or
-------
Revision B 4/15/81 8.27-7
splitless mode. If using the split mode record
the split ratio. Record both the linear and
volume column flow.
c. When analyzing volatiles it may be necessary to
adjust desorption time or cool the first few
cm of the column with a flurocarbon spray
in order to maintain chromatographic resolution.
d. Inject sample and acquire data/ recording the amount
injected. Follow the same procedure for any standards.
7. Quantitative and Qualitative Determination
a. A compound can be qualitativly identified in either
of two ways. At least three characteristic ions of the
compound must maximize within one scan of the apex
of the peak and the integrated ion areas agree with
a library or standard mass spectrum within jH 20%;
or, a reverse search yeilds a numerical value
equivalent to the criteria stated above.
b. Samples can be quantitated in two ways. The first
is by the method of standard additions. This
method is always acceptable and should be used
when the exact concentration is needed. The
second method is to be used only for order of
magnitude estimates of concentartion. This is
done by comparing the Total Ion Chromatogram of
the compound in the sample with a standard. For
example, if 100 ng of benzene gives a total of
10,000 integrated area counts then a peak corresponding
to toluene with 25,000 counts would be expected
to correspond to about 250 ng. When using this
method the analyst should try to use standards
which resemble the compounds in question as
closely as possible. The internal standard is used
as a method check. For example, if 50 ng of
the internal standard normally gives 5000 integrated
area counts, this condition should be met in
the sample +20%
c. Example Calculation
A 10 gm sample contained 3.5 gm of organic liquid
with a volume of 3.9 ml. 5 ul of the organic
liquid was injected with a split ratio of 100
to 1. The integrated area of benzene gave 3582
counts. Benzene in the organic phase is calculated:
3582 counts/5 ul x 1 ug/313.6 counts =11.4 ug/5 ul
11.4 ug/5 ul x 1000 ul/ml = 2280 ug/ml
the density is 3.5gm/3.9ml « .9 gm/ml
2280 ug/ml = 2.28mg/ml
2.28mg/ml x Igm/lOOOmg x 1 ml/.9gm « .0025 = .25%
-------
Revision B 4/15/81 8.27-8
The purge and trap analysis of the aqueous phase
was performed on 6.5 gm of liquid. Benzene gave
12562 counts. The benzene in the aqueous phase
is:
12562 counts/6.5 gm x 1 ug/313.6 counts =
40.0 ug/6.5 gm = 6.2 ppm
This is insignificant compared to .25%
The total amount of benzene in the sample is
calculated:
.25% x .35 of total = .0875% or 875 ppm
8. Report
a. Report the results of each analysis giving
each compound identified, the scan number, the
quantity of the compound, and the method used
to calculate that quantity.
b. Example
Compound Quantitation Scan Amount Range
Method #
Benzene Estimate/Benzene 687 875ppm 100ppm-l%
-------
Revision B 4/15/81 8.27-9
II. Solids
Two samples of well mixed solid should be used in this
analysis. One sample is used for the purge and trap analysis
of volatiles and one for soxhlet extraction analysis.
A. Purge and Trap Determination of Volatiles in Solids
1. Summary
An appropriate weight of sample (1-10 gin) is diluted
with 10 ml of organic-free water. The diluted
sample is purged for 12 min. with inert gas at
room temperature. The gaseous phase is passed
through a sorbent trap where the organic compounds
are concentrated. The contents of the trap are
desorbed into the GC/MS by heating and backflushing
the trap.
2. Apparatus and Materials
a. See the apparatus section of Method 8.83 of this
manual.
b. Gas Chromatograph - Analytical system capable of
split and splitless injections and all required
accessories including column supplies, gases, etc.
c. Column - 30m SE-30, SE-52, SE-54, or equivalent,
.2 to ,25mm internal diameter with a film thickness
between .15 to .40u.
d. Mass Spectrometer - Capable of scanning from 20 to
260 daltons every 1 second or less. The MS must
be able to operate at 70 volts for electron
ionization and must produce a recognizable mass
spectrum for 50 ng or less of BFB when the sample
is introduced through the GC column. The GC column
should be directly interfaced (i.e. no separator)
to the mass spectrometer through an all glass
or all glass lined system. If a fused silica
capillary column is used, the analyst is required
to complete the interface by directly connecting
the end of the column to the ion source.
e. Computer System - The computer system interfaced
to the mass spectrometer should be capable of
continuously acquiring mass spectra for the duration
of the gas chromatographic program, (about 1 hr.)
All data must be s,tored either within the data
system or on line mass storage devices such as
disk or tape. The system must have software
available capable of searching GC/MS runs for
the following:
1) selected ion chromatograms
2) total ion chromatograms
-------
Revision B 4/15/81 8.27-10
3) reverse and forward search for any compound
from the EPA/NIH Mass Spectral Data Base
3. Reagents - Standards as necessary (See Methods 8.24,
8.83, and I-A-3e of this Method)
4. Calibration
a. The mass spectrometer is calibrated with either
PFK or FC-43 over the scan range. 50ng or less of
BFB should be injected in the splitless mode
using the conditions given in table 8.27-3.
b. The spectrum obtained from the top of the chro-
matographic peak (backgroud subtracted) should
meet the criteria listed in table 8.24-2.
5. Sample Preparation
a. Weigh an appropriate sample into a pretared 10 to
15 ml Teflon lined, screw-capped vial.
b. Dilute the sample with 10 ml distilled water.
Disperse the sample into the water. Transfer
the total sample to the purging device using a
syringe with an 1/8 in. gauge Teflon needle.
Seal the sample in the purging device. Add
the internal standard and purge with 40
ml/min (He or N2) for 12 min. at room temperature.
6. Gas Chromatography/Mass spectrometry
a. Establish the chromatographic conditions given
in table 8.27-1.
b. Set up the Gas Chromatograph for either split
or splitless injection. If using the split
mode, record the split ratio, linear and volume
column flow.
c. The first few inches of the column should be
cooled using flurocarbon spray. Heat the trap
to 200°C. Backflush it for 4 min in the
desorb mode into the gas chromatograph.
7. Qualitative and Quantitative Determination
a. A compound can be qualitativly identified in either
of two ways. At least three characteristic ions of the
compound must maximize within one scan of the apex
of the peak and the integrated ion areas agree with
a library or standard mass spectrum within + 20%;
or, a reverse search yeilds a value equivalent to
the criteria stated above.
b. Samples can be quantitated in two ways. The first
is by the method of. standard additions. This
-------
Revision B 4/15/81 8.27-11
method is always acceptable and should be used
when the exact concentration is needed.
The second method is to be used only for order
of magnitude estimates of concentration as
given on page 1 of this method. This is done
by comparing the Total Ion Chromatogram of the
compound in the sample with a standard. For
example, if 100 ng of benzene gives a total of
10,000 integrated area counts then a peak corresponding
to toluene with 25,000 counts would be expected
to correspond to about 250 ng. When using this
method the analyst should try to use standards
which resemble the compounds in question as
closely as possible. The internal standard is
used as a method check. For example, if 50 ng
of the internal standard normally gives 5000
integrated area counts this condition should
be met in the sample jf20%.
Example Calculation
5.0 gm of a solid sample was mixed with 10 ml
water and purged and traped by the procedure
specified. A splitless injection gave 29,043
integrated area counts for toluene. 1 ul of
a standard solution of Toluene 100 ug/ml gave
16,290 integrated counts.
16290 counts/ 1 ul x 1000 ul/100 ug = 162900 counts/ug
29043 counts/5 gm x 1 ug/162900 counts = .18 ug/5 gm
.18 ug/5 gm = .036 ug/gm = .036 ppm
-------
Revision B 4/15/81 8.27-12
8. Report
a. Report the results of each analysis giving
each compound identified, the scan number/ the
quantity of the compound, and the method used
to calculate that quantity.
b. Example
The level of toluene in the sample is very low
and for the purpose of this analysis is reported
at less than 1 ppm
-------
Revision B 4/15/81 8.27-13
B. Soxhlet Extraction for Solids
1. S umma ry
The sample is mixed with anhydrous sodium sulfate,
placed in an extraction thimble or between two
plugs of glass wool and extracted using methylene
chloride. The extract is reserved. The remaining
contents of the thimble are mixed with distilled
water and the pH is adjusted to 2 or less. This
aqueous mixture is extracted with ethyl ether.
The two extracts are dried, combined, and anaylzed
in one GC/MS analysis.
2. Apparatus and Materials
a. Soxhlet extractor - 40 mm id, with 500 ml round-
bottom flask.
b. Kuderna-Danish Apparatus [Kontes K-570000 or equivalent]
with 3-ball snyder column
c. Gas Chromatograph - Analytical system capable of
split and splitless injections and all required
accessories including column supplies, gases, etc.
d. Column - 30m SE-30, SE-52, SE-54, or equivalent,
.2 to .25mm internal diameter with a film thickness
between .15 to .40u.
e. Mass Spectrometer - Capable of scanning from 35 to
450 daltons every 1 second or less. The MS must
be able to operate at 70 volts for electron
ionization and must produce a recognizable mass
spectrum for 50 ng or less of DFTPP when the sample
is introduced through the GC column. The GC column
should be directly interfaced (i.e. no separator)
to the mass spectrometer through an all glass
or all glass lined system. If a fused silica
capillary column is used, the analyst is required
to complete the interface by directly connecting
the end of the column to the ion source.
f. Computer System - The computer system interfaced
to the mass spectrometer should be capable of
continuously acquiring mass spectra for the duration
of the gas chromatographic program, (about 1 hr.)
All data must be stored within the data system.
Mass storage devices such as disk or tape are
accepable. The system must have software available
to allow searching GC/MS runs for the following:
1) selected ion chromatograms
2) total ion chromatograms
3) reverse and forward search for any compound
from the EPA/NIH Mass Spectral Data Base
3. Reagents
a. Methylene chloride - Pesticide grade
b. Ethyl Ether - Pesticide grade
-------
Revision B 4/15/81 8.27-14
c. Anhydrous Sodium Sulfate, ACS grade, purified
by heating at 400°C for 4 hr. in a shallow tray.
4. Calibration
a. The mass spectrometer is calibrated with either
PFK or FC-43 over the scan range. 50ng or less of
DFTPP should be injected in the splitless mode
using the conditions given in table 8.27-1.
b. The DFTPP spectrum obtained from the top of the
chromatographic peak (backgroud subtracted)
should meet the criteria listed in table 8.24-2.
5. Sample Preparation
a. Blend 10.0 gm of the solid sample with 10.0 gm
of anhydrous sodium sulfate. Weigh this mixture
to the nearest 0.1 gm. Place in either a paper
(pre-washed with methylene chloride and dried)
or glass extraction thimble.
b. Place the thimble in the extractor. (If any
problems arise when using the thimble, i.e. if
the sample clogs the thimble, an alternative
would be to place a plug of glass wool in the
extraction chamber, transfer the sample into
the chamber, then cover the sample with another
plug of glass wool.)
c. Place 250 ml of methylene chloride into the 500 ml
roundbottom flask, add a boiling chip and attach
the flask to the extractor. Extract the sample
for 16 hours.
d. After the extraction is complete, cool the extract;
rinse extractor flask and thimble with fresh
solvent. Combine the extract and rinse.
Dry the extract by passing it through a 4 inch
column of sodium sulfate that has been washed
with solvent. Collect the dried extract
in a 500 ml Kuderna-Danish (KD) flask fitted with
a 10 ml graduated concentartor tube.
Empty the contents of the thimble into a pre-weighed
250 ml Erlinmeyer flask. Add 100 ml distilled
water to the flask.
e. Adjust the pH to 2 or less with sulfuric acid
solution. Extract three times with fresh 60 ml
portions of ethyl ether. Combine the three
extracts and dry by passing through a 4 inch
column of sodium sulfate. Rinse column with
fresh solvent. The dried extract is added to
the KD.
f. Evaporate the aqueous solution in the erlinmeyer
flask to dryness; cool the flask and weigh the
residue. Determine the weight difference between
-------
Revision B 4/15/81 8.27-15
the residue in the erlinmeyer flask and the
original sample.
g. Concentrate the dried extracts in the KD. A
level that would give a final concentration of
about 1 mg/ml is generally appropriate for
GC/MS.
h. The concentrated extract should be placed in a
volumetric flask and made up to the appropriate
volume.
6. Gas Chromatography/Mass spectrometry
a. Establish the chromatographic conditions given
in table 8.27-1.
b. Set the Gas Chromatograph for either split
or splitless injection. If using the split
mode, record the split ratio. Record both
liniar and volume column flow.
c. Inject sample and acquire data. Record amount
of sample injected. (2 to 5 ul for split
and 1 to 2ul for splitless)
d. Inject appropriate standards and acquire data as
in c.
7. Qualitative and Quantitative Determination
a. A compound can be qualitativly identified in either
of two ways. At least three characteristic ions of the
compound must maximize within one scan of the apex
of the peak and the integrated ion areas agree with
a library or standard mass spectrum within + 20%;
or, a reverse search yeilds a value equivalent to
the criteria stated above.
b. Samples can be quantitated in two ways. The first
is by the method of standard additions. This
method is always acceptable and should be used
when the exact concentration is needed. The
second method is to be used only for order of
magnitude estimates of concentartion. This is
done by comparing the Total Ion Chromatogram of
the compound in the sample with a standard. For
example, if 100 ng of benzene gives a total of
10,000 counts then a peak corresponding to toluene
with 25,000 counts would be expected to correspond
to about 250 ng. When using this method the
analyst should try to use standards which resemble
the compounds in question as closely as possible.
The internal standard is used as a method check.
For example, if 50 ng of the internal standard
normally gives 5000 integrated area counts this
condition should be met in the sample +20%.
c. Example Calculation
10 gm of solid sample was extracted with methylene
chloride and ethyl ether as in the procedure. The
-------
Revision B 4/15/81 8.27-16
sample lost about 4 gm during the extraction. The
combined extracts were diluted to 500 ml with
methylene chloride. 5 ul was injected with a
split ratio of 100 to 1. Hexachlorobenzene was
found in the extract with a total of 7,121
total area counts.
Dichlorobenzene was used as a standard
5 ul x 1 mg/ml x 1 ml/1000 ul = 5 ug
Total counts for Dichlorobenzene was 3760
3760 counts/5 ug = 752 counts/ug
Hexachlorobenzene in sample
7121 counts/5 ul x 1 ug/752 counts = 9.47 ug/5 ul
9.47 ug/5 ul x 1000 ul/1 ml x 500 ml = 947000ug
947000 ug = .947 gm
.947 gm/10 gm = .0947 = 9.5% hexachlorobenzene
8. Report
a. Report the results of each analysis giving
each compound identified, the scan number, the
quantity of the compound, and the method used
to calculate that quantity.
b. Example
Compound Quantitation Scan Amount Range
Method #
Hexachloro Estimate/dichloro 693 9.5 % 1-10%
benzene benzene
-------
Revision B 4/15/81 8.27-17
III. Mixtures of Liquids and Solids
A 10 to 20 gm sample of well mixed waste is used. The sample
is divided into its component phases and the procedures oulined
in sections I and II of this Method are employed for analysis.
A. Separation Procedure for Liquids and Solids
1. Summary
A 10 to 20 gnv sample of the waste is separated into
its component phases by centrifugation. The Liquid
Phases are either decanted or pipeted for analysis
using section I and the solid residue is analyzed
using section II.
2. Apparatus and Materials
a. Centrifuge tubes - 10-20 ml pyrex glass or equivalent
with ground glass stopper.
b. Centrifuge - Capable of 2400 RPM
3. Reagents - Reserved
4. Calibration - See calibration sections in parts
I and II of this method
5. Sample Preparation
a. Alliquot a 10 to 20 gm sample of well mixed waste
into a pre weighed cenrifuge tube. Weigh.
b. Place tube into centrifuge and spin at 2400 RPM
for 15 min. or until the solids and liquid phases
are separated.
c. Pour off liquid phase and weigh. Proceed to section
I of this method.
d. Weigh remaining solids and proceed to section II of
this method. The purge and trap method for the
determination of volatiles in solids may be
omitted since the volatiles are determined in the
liquid phase of the sample.
B. Report
1. Report the results as a weighted average of the
liquid phases and solid phase.
2. Example calculation
See sections I and II
-------
Revision B 4/15/81 8.27-18
Table 8.27-1 (Liquids)
Column: SE-30, SE-52, SE-54 (30 m)
Linear Flow Rate: 50 cm/sec H2 or 30 cm/sec He
Temperature Program: Inject at 25°C then 50°C
Program 50° to 280° C at 8°/min
Hold at 280°C for 15 min.
Table 8.27-2 (Extractables)
Column: SE-30, SE-52, SE-54 (30 m)
Linear Flow Rate: 50 cm/sec H2 or 30 cm/sec He
Temperature Program: Inject at 50°C hold 2 min.
Program to 280°C at 8°C/min
Hold at 280°C for 15 min
Table 8.27-3 (Volatiles)
Column: Same as 8.27-2
Linear Flow Rate: Same as 8.27-2
Temperature Program: Inject at 25°C (cool head of column with
flurocarbon spray) then to 50°C
Program 50°C to 200°C at 4°C/mir
Hold at 200°C for 10 min
-------
Revision B 4/15/81 8.27-19
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8.49-2
4. Acids must be Spectrograde. If metals are found to be
present the acid they should be diluted 1:1 with distilled
water and redistilled in a hood.
5. An Atomic Absorption Spectrophotometer provides the
simplest and most accurate means of determining metal
concentrations in the low ranges required. In principle,
a prepared solution is vaporized in a flame. Metal
atoms in the flame are capable of absorbing light of a
particular and characteristic wavelength. When a light
beam, which emanates from the pure metal in a cathode
lamp, is passed through a flame, the absorption is propor-
tional to the concentration. Through various detecting
devices, this absorption may be read directly or be re-
corded as a peak on a strip chart recorder.
Most metals may be determined in an air plus acetylene
flame. However, some metals require more energy in
order to dissociate and a nitrous oxide - acetylene
flame must be used. Additionally, when the fuel itself
absorbs a particular wavelength, then the fuel may be
changed to a non-interfering one.
The most common type of burner is known as a "premix"
in which the spray is mixed with air or oxidant. It may
be fitted with a conventional head (4 inch single slot),
a 3 slot "Boling" head for air-acetylene fuel, or a 2
inch short slot for use with nitrous oxide-acetylene
fuel. These various heads influence the burning rate,
carbon build-up and sensitivity. The instrument manual
-------
8.49-3
will list the head that has been found to work best for
a given element.
A graphite furnace is especially useful for measuring
low levels. In this device the sample is evaporated in
a graphite tube, charred and vaporized in a confined and
inert atmosphere. However, this method is susceptible
to interference from chlorides and other dissolved solids.
If the samples smoke, light scattering can occur which
causes erroneously high results. A deuterium background
corrector should be employed to correct for non-specific
interference.
The spectrophotometer chosen should have a wavelength
range of 190 to 800 nanometers, and use an output device
with high sensitivity and fast response time. The instru-
ment may provide direct readout or use a 10 millivolt
strip chart recorder. The use of a recorder will furnish
a permanent record of the analysis and the operating
condit ions.
Single element hollow cathode lamps are preferred.
Electrodeless discharge lamps may also be used when
available.
Venting
A vent must be installed 6 to 12 inches above the burner
to remove toxic fumes and vapors. The slight vacuum insures
a constant air flow which tends to stabilize the flame. A
variable speed blower is an advantage in that the air flow
may be adjusted to prevent disturbance of the flame.
-------
7. Instrument Operation .
Procedures vary with the particular model and manufacturers'
instructions should be followed. In general, the following
should be carried out:
a. Install the selected cathode lamp.
b. Install the proper burner head.
c. Set the wavelength dial and align the lamp.
d. Set the slit width.
e. Turn on the instrument and apply correct current.
f. Allow the instrument to warm up. This time varies
but is usually around 20 minutes.
g. Turn on the air and adjust flow rate.
h. Turn on the acetylene, adjust flow rate and ignite
flame. Note that it may be necessary to reduce the
fuel flow if the sample fluid acts as as a fuel.
i. Atomize deionized distilled water acidified with 1.5
ml concentrated nitric acid per liter, and check the
aspiration rate. Adjust the rate to between 3 and 5
ml per minute. Zero the instrument.
j. Atomize a solution containing a known amount of test
element and adjust the burner up, down or sideways
to get maximum response.
k. When analyses are finished, turn off the acetylene
first and then the air.
Optimum Concentration Range
The most accurate results may be obtained in this range
-------
8.49-5
and it varies for different elements and for different instru-
ments. It may be necessary to dilute a sample of higher con-
centration in order to keep it in this range, in which case
the value obtained must be doubled, tripled, etc. accordingly.
In the case of lower concentrations it may be sufficient
to show that the sample is below the required limit and
therefore non-hazardous. When the generator wishes to know
the exact level, it may be possible to concentrate a sample
to bring it into the optimum range.
It is necessary to correct for background absorption
for precise work. A double beam instrument or dual double
beam system permits a wider range of selectivity and precision
A background corrector is frequently useful to minimize
background absorption.
Calibration
The concentration of metal in the sample is determined
by comparing its absorbance to that of solutions of known
concentration. The samples are preserved at a pH of less
than 2 with nitric acid, and calibration is performed with
distilled deionized water acidified in the same way.
Quantification
When analyzing industrial wastes or the Toxicant Ex-
traction Procedure extract, where unknown dissolved materials
may interfere with the determination, the Method of Standard
Additions must be used:
-------
8.49-6
Method of Standard Additions
In this method, equal volumes of sample are added
to a deionized distilled water blank and to three stan-
dards containing different known amounts of the test
element. The final volume of the blank and of the stan-
dards must be the same so that the interfering substance
is present in the same amount. The absorbance of each
solution is determined and then plotted on the vertical
axis of a graph with the concentrations of the standard
plotted on the horizontal axis. When the resulting line
is extrapolated back to zero absorbance, the point of
interception of the abscissa is the concentration of the
unknown. The abscissa on the left of the ordinate is
scaled the same as on right side, but in the opposite
direction from the ordinate. An example of a plot so
obtained is shown below:
Concentration
I Cone, of
Sample
Addn 0
No Addn
Addn I
Addn of 50%
of Expected
Amount
Addn 2 Addn 3
Addn of 100% Addn of 150%
of Expected of Expected
Amount Amount
Figure 8.49-1
PLOT OF METHOD OF STANDARD ADDITIONS
-------
8.49-7
Note; For this method to be valid, the plot must be linear.
The slope of this line should not differ by more than 20%
from the slope of the standard solutions. The effect of the
assumed interference must not change as the proportion of
sample to standard changes.
Solids, Sludges and Slurries
Solids, sludges and slurries may be analyzed by these
methods by weighing out suitable portions and digesting as
described for each metal. The material is then filtered
through a 0.45 micron filter while washing down the sides of
the beaker and rinsing the filter with distilled deionized
water. The filtrate is then made up to a suitable volume
and analyzed in the ususal manner. Results can be related
back to the original sample weight and reported as mg/kg.
Conclusion
The details of the following approved methods are examples
of acceptable techniques. Dilutions and concentrations may
have to be varied to suit the instrument being used. It is
important not to overwhelm the instrument with very high
concentrations above the optimum recommended range. Contami-
nation can result which is difficult to remove. At the same
time, many dilutions introduce error which can be avoided by
some knowledge of the waste beforehand. If nothing is known,
caution is advised.
For additional information the applicable sections of
"Methods for Chemical Analysis of Water and Wastes", EPA
600/4-79-020 (Appendix II of this manual) may be consulted.
-------
8.50-1
Method 8.50
ANTIMONY
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of antimony in a
waste or Extraction Procedure Extract
Summary of Method
A sample is digested using nitric acid and the concentra-
tion of antimony measured using either a flame or graphite
furnace equipped atomic absorption spectrometer. The flame
method is most accurate when employed with solutions containing
1-40 mg/liter antimony while the graphite furnace procedure
is best suited for solutions containing 20 - 300 ug/liter.
Apparatus
Atomic absorption spectrometer equipped with either a
graphite furnace or flame burner head as described in
Section 8.49.
Reagents
1. When using the flame procedure air and acetylene are
required. Air should be cleaned and dried through a
suitable filter to remove oil, water, and other foreign
substances. The source may be a compressor or a cylinder
of industrial grade compressed air. High purity acetylene
should be used. Acetone which is always present in
acetylene cylinders, can be prevented from entering and
damaging the burner head by replacing a cylinder when
its pressure has fallen to 7 kg/era-^ (100 psig) acetylene.
-------
3.50-2
2. Nitric Acid, J.T. Baker Ultrex Grade or equivalent.
3. Antimony potassuim tartrate, analytical reagent grade.
4. Hydrochloric Acid Solution, J.T. Baker Ultrex Grade or
equivalent diluted 1:1 with distilled deionized water.
Procedure
Standard Solutions
1. Prepare Antimony Standard Stock Solution by dissolving
2.7426 g antimony potassium tartrate, analytical reagent
grade, in distilled deionized water containing 1.5 ml
HN03/liter and bringing to volume in a 1 liter volumetric
flask (1 ml - 1 mg Sb).
2. Prepare working standards from stock solution. If it is
desired to work in the optimum concentration range, using
the flame technique, the following is suggested:
a. Transfer 0, 0.1, 1.0, 2.0, 3.0, and 4.0 ml of stock
solution to separate 100 ml volumetric flasks. Bring
t*o volume with distilled deionized water containing
2 ml HN03/liter. The concentrations of these working
standards are 0, 1, 10, 20, 30 and 40 mg Sb/liter.
3. When using the graphite furnace procedure working standards
should be prepared to cover the range 20 to 300 ug Sb/liter
by preparing dilutions of the above working standards.
Sample Preparation
1. Transfer 100 ml of well-mixed sample to a 250 ml beaker.
Add 3 ml cone. HN03. Place the beaker on a hot plate
and evaporate to near dryness, cautiously, so that the
sample does not boil. Cool, and add another 3 ml cone.
-------
8.50-3
HN03. Cover the beaker with a watch glass and return
to the hot plate. Heat so as to produce a gentle
refluxing. Continue adding 3 ml portions of HNOg until
the sample is light in color and no longer changes in
color. Evaporate to near dryness and cool. Add 1 ml
of 1:1 HC1 and warm the beaker in order to dissolve any
precipitates. Cool the beaker. Transfer to a 100 ml
volumetric flask and bring to volume using distilled
deionized water containing 2 ml HN03/liter. Note that
hydrochloric acid is used to aid in dissolving of antimony
residues .
2. If particulates such as silicate remain in the sample, it
must be centrifuged and the supernatant sampled.
Standard Addition
a. Take the 50, 75, and 100 ug standards and pipet 5 ml from
each into separate 10 ml volumetric flasks. Add to each
2 ml of the prepared sample. Bring to volume with
distilled deionized water.
b. Add 2 ml of prepared sample to a 10 ml volumetric flask.
Bring to volume with distilled deionized water. This is
the blank.
Note; The absorbance from the blank will be 1/5 that
produced by the prepared sample. The absorbance from the
spiked standards will be 1/2 that produced by the standard
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optimum
absorbance can be judged.
-------
8.50-4
Instrument Operation
Wavelength: 217.6 nanometers
Note: The presence of lead in the waste may
interfere since lead absorbs at this
wavelength. In this case, use the 231.1
nanometer antimony line.
Optimum Concentration Range: 1-40 ing/liter
Lower Detectable limit: 0.2 mg/liter
Fuel: Acetylene
Oxidant: Air
Type of Flame: Lean
A general outline for instrument operation is given in
Section 8.49. Follow the manufacturer's instructions for
the Spectrophotometer being used.
Graphite Furnace Method
Wavelength: 217.6 nanometers
Optimum Concentration Range: 20-300 ug/liter
Lower Detection Limit: 3 ug/liter
Purge gas: Argon or nitrogen
Drying time and temperature: 30 sec at 215° C
Ashing time and temperature: 30 sec at 800° C
Atomizing time and temperature: 10 sec at 2700° C
The conditions listed above are based on a 20 ul injection;
continuous flow purge gas and non-pyrolytic graphite on a
Perkin Elmer model HGA 2100 furnace. Other equipment will
have different requirements. Follow the manufacturer's manual.
Note: If the chloride presents a matrix problem, add an excess
of 5 mg ammonium nitrate to the furnace and ash using a ramp
-------
8.50-5
accessory; or with incremental steps, until ashing temperature
is reached.
Quantification
The absorbances of spiked samples and blank vs. the
concentrations are plotted according to the Method of Standard
Additions as described in Section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line, a non-
linear interference is present. This can sometimes be over-
come by dilution, or addition of other reagents if there is
some knowledge about the waste.
-------
8.51-1
Method 8.51
ARSENIC
Scope and Application
The following atomic absorption procedures are approved
for determining the concentration of arsenic in a waste or
Extraction Procedure Extract. Both methods are generally
acceptable for RCRA type samples. However, the graphite
furnace method is sensitive to the presence of chlorides and
dissolved solids in the sample, while the hydride method requires
greater preparation and is sensitive to high concentrations
of chromuim, copper, mercury, silver, cobalt, and molybdenum.
Summary of Methods
In the furnace method the sample is digested using nitric
acid and hydrogen peroxide. The method is most accurate for
solutions containing 5 - 100 ug Arsenic/liter.
In the gaseous hydride method the sample is digested
using nitric and sulfuric acids. The arsenic is then reduced
to the trivalent form with stannous chloride, and converted
to arsine, AsH3, using zinc metal. The hydride is swept into
an argon entrained hydrogen flame for analysis. It is most
accurate when employed for solutions containing 2 - 20 ug
As/liter. However, mercury, silver, cobalt and molybdenum
can interfere in the analysis.
Preoxidation (digestion) is required in both methods in
order to convert organic forms of arsenic to the inorganic
form, and to remove organic interferences.
-------
8.51^2
Precaution
Severe poisoning can result from ingestion of as little
as 100 mg Arsenic. Chronic effects can appear with constant
Intake of lower levels. Exercise care in the handling and
cleanup of the materials and work area.
Graphite Furnace Method
Apparatus
1. Atomic absorbtion spectrometer equipped with a non-pyrolytic
graphite furnace.
Reagents
1. Standard Stock Solution
Stock solution may be purchased, or prepared as follows:
a. Dissolve 4g NaOH in 100 ml distilled deionized water.
Dissolve 1.3203 g of analytical reagent grade arsenic
trioxide (AS203) in the solution.
b. Acidify this solution with 20 ml concentrated HN03.
c. Transfer to a 1 liter volumetric flask with careful
rinsing, and bring to volume with distilled deionized
water. The concentration of this solution is 1000
mg arsenic/liter (1 ml^l mg As).
2. Nickel Nitrate Solution, 5%
Dissolve 24.780 g of ACS Reagent Grade Ni(N03>2•6H20
in distilled deionized water and make up to 100 ml.
3. Nickel Nitrate Solution, 1%
Dilute 20 ml of the 5% solution to 100 ml with distilled
deionized water.
-------
8.51-3
4. 30% Hydrogen Peroxide, ACS Reagent Grade
5. Sodium Hydroxide, ACS Reagent Grade
6. Concentrated Nitric Acid, J.T. Baker Chemical Co. Ultrex
Grade or equivalent.
Procedure
1. Transfer 100 gm (or in the case of EP Extract analysis 100
ml) of well mixed sample to a 250 ml beaker and add 2 ml
of 30% H202 and 1 ml of concentrated HN03. Heat on
a hot plate for 1 hour at 95°C or until the volume is
less than 50 ml. It may be necessary to repeat this
operation to digest all of the material present.
2. Cool, transfer to 50 ml volumetric flask and bring to
volume with distilled deionized water.
3. If particulate matter remains, after oxidation, it
must be removed by centrifugation and only the supernatant
used.
4. Pipet 25 ml of this solution into a 50 ml volumetric
flask. Add 5 ml of 1% nickel nitrate solution and bring
to 50 ml with distilled deionized water. The sample is
ready for injection.
5. Prepare standards from stock solution. The following
provides standards covering the optimium working range:
6. Pipet 1 ml stock solution into a 1 liter volumetric
flask. Bring to volume with distilled deionized water
containing 1.5 ml cone. HN03 per liter. The concentration
of this solution is 1 mg As/liter (1 ml=l ug As).
7. Prepare 6 working standards by transferring 0, 1.0.
-------
2.5, 5, 7.5, and 10 ml from (a) into 100 ml volumetric
flasks. Add to each flask 1 ml concentrated HN03, 2
ml 30% H202 and 2 ml 5% nickel nitrate solution, bring
to volume with distilled deionized water. The concen-
trations of these working standards are 0, 25, 50, 75,
and 100 ug As/liter. They are ready for injection.
8. Take the 50, 75, and 100 ug/liter standards and pipet 5 ml
from each into separate 10 ml volumetric flasks. Add to
each 2 ml of the prepared sample. Add 0.3 ml of 1% of
nickel nitrate solution, and 0.3 ml H202 solution.
Bring to volume with distilled deionized water.
9. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Add 0.8 ml nickel nitrate solution and 0.8 ml
H202 solution. Bring to volume with distilled deionized
water. This is the blank.
Note; The absorbance from the blank, will be 1/5 that
produced by the prepared sample. The absorbance from the
spiked standards will be 1/2 that produced by the standards
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optiraium
absorbance can be judged.
Instrument Operation:
Wavelength: 193.7 nanometer
Optimum Concentration Range: 5-100 ug/liter
Lower Detection Limit: 1 ug/liter
Drying time and temperature: 30 sec at 125°C
Ashing time and temperature: 30 sec at 1100°C
Atomizing time and temperature: 10 sec at 2700°C
-------
8.51-5
Purge gas: Argon
These conditions are based on a 20 ul injection; con-
tinuous flow purge gas and non-pyrolytic graphite on a Perkln
Elmer Model HGA 2100 furnace. Other equipment will have
different operating requirements. For specific information,
the manufacturer of the instrument should be consulted.
Quantification
The absorbances of spiked samples and blank vs. the con-
centrations are plotted according to the Method of Standard
Additions as described in Section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a non-linear
interference is present. This can sometimes be overcome by
dilution, or addition of other reagents if there is some
knowledge about the waste.
Gaseous Hydride Method
Reagents
1. Potassium iodide solution: Dissolve 20 g KI in 100 ml
deionized distilled water.
2. Stannous chloride solution: Dissolve 100 g SnCl2 in
100 ml cone HC1.
3. Zinc slurry: Add 50 g zinc metal dust (200 mesh) to 100
ml deionized distilled water.
4. Diluent: Add 100 ml 18N H2S04 and 400 ml concentrated
HC1 to 400 ml deionized distilled water in a 1-liter
volumetric flask and bring to volume with deionized
distilled water.
-------
8.51-6
5. Arsenic Solutions:
a. Stock arsenic solution, (1000 mg/liter):
Dissolve 1.3203 g arsenic trioxide, AS203, in 100 ml
distilled water containing 4 g NaOH. Acidify with
20 ml concentrated HN03 and dilute to 1000 ml with
deionized distilled water (1 ml = 1 mg As).
b. Intermediate arsenic solution: Pipet 1 ml stock
arsenic solution into a 100-ml volumetric flask and
bring to volume with deionized distilled water con-
taining 1.5 ml concentrated HN03/1 ("1 ml = 10 "8 As).
c. Standard arsenic solution: Pipet 10 ml inter-
mediate arsenic solution into a 100-ml volumetric
flask and bring to volume with deionized distilled
water containing 1.5 ml concentrated HN03/1 (1 ml »
1 ug (As).
6. Concentrated nitric acid, J.T. Baker Chemical Co. Ultrex
grade or equivalent.
7. Concentrated sulfuric acid, J.T. Baker Chemical Co. Ultrex
grade or equivalent.
Apparatus
1. Atomic Absorption Spectrophotometer with Boling burner.
2. Flow meter, capable of measuring 1 liter/minute, such
as that used for auxiliary argon.1
3. Medicine dropper, capable of delivering 1.5 ml, fitted
into a size "0" rubber stopper.
4. Reaction flask, a pear-shaped vessel with side arm and
50 ml capacity, both arms having 14/20 joint.1
5. Special gas inlet-outlet tube, constructed from a micro
^Gilmont No. 12 or equivalent
-------
8.51-7
cold finger condenser^ by cutting off the portion below
the 14/20 ground glass joint.
6. Magnetic stirrer, strong enough to homogenize the zinc
slurry.
7. Drying tube, 10 cm polyethylene tube filled with glass
wool to keep particulate matter out of the burner.
Apparatus Set-Up
Argon
Flow
JM-3325
Medicine
Dropper in
Size "0"
Rubber
Stopper
(Auxiliary Air)
Argon
- (Nebulizer
Air)
• JM-5835
1. Assemble the apparatus as shown above.
2. Connect the outlet of the reaction vessel to the auxiliary
oxidant input of the burner with tygon tubing.
3. Connect the inlet of the reaction vessel to the outlet
side of the auxiliary oxidant (Argon supply) control
valve of the instrument.
Procedure
1. To a 50 gm sample (or in the case of analysis of EP extracts
50 ml) of the material to be analyzed in a 100 ml beaker
add 10 ml concentrated HN03 and 12 ml 18 N H2S04.
Evaporate the sample in the hood on an electric hot
^Scientific Glass JM-5835 or equivalent.
2Scientific Glass JM-3325 or equivalent.
-------
plate until white 803 fumes are observed (a volume of
about 20 ml). Do not let the sample char. If charring
occurs, immediately turn off the heat, cool, and add an
additional 3 ml of HNC>3 . Continue to add additional HN03
in order to maintain an excess (as evidenced by the forma-
tion of brown fumes). Do not let the solution darken
because arsenic may be reduced and lost. When the sample
remains colorless or straw yellow during evolution of
SOg fumes the digestion is complete. Cool the sample,
add about 25 ml distilled deionized water and again
evaporate until 803 fumes are produced in order to
expel oxides of nitrogen. Cool. Transfer the digested
sample to a 100 ml volumetric flask. Add 40 ml of concen-
trated HC1 and bring to a volume with distilled deionized
water.
2. Prepare working standards from the standard Arsenic
solution under Reagents (5c). Transfer 0, 0.5, 1.0, 1.5,
2.0, and 2.5 ml standard to 100 ml volumetric flasks and
bring to volume with diluent. These concentrations will be
0, 5, 10, 15, 20 and 25 ug As/liter.
3. Take the 15, 20, and 25 rag/liter standards and transfer
quantitatively 25 ml from each into separate 50 ml
volumetric flasks. Add to each 10 ml of the prepared
sample. Bring to volume with distilled deionized
water containing 1.5 ml HN03/liter.
4. Add 10 ml of prepared sample to a 50 ml volumetric
flask. Bring to volume with distilled deionized water
-------
8.51-9
containing 1.5 ml HN03 per liter. This is the blank.
Note; The absorbance from the blank, will be 1/5 that
produced by the prepared sample. The absorbance from the
spiked standards will be 1/2 that produced by the standards
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optimum
absorbance can be judged.
5. Transfer a 25-ml portion of the digested sample or
standard to the reaction vesse-1, and add 1 ml potassium
iodide solution. Add 0.5 ml SnCl2 solution. Allow at
least 10 min for the metal to be reduced to its lowest
oxidation state. Attach the reaction vessel to the
special gas inlet-outlet glassware. Fill the medicine
dropper with 1.50 ml zinc slurry that has been kept in
suspension with the magnetic stirrer. Firmly insert the
stopper containing the medicine dropper into the side
»
neck of the reaction vessel. Squeeze the bulb to introduce
the zinc slurry into the sample or standard solution. The
metal hydride will produce a peak almost immediately. After
the recorder pen begins to return to the base line, the
reaction vessel can be removed. Caution: Arsine is very
toxic. Caution must be taken to avoid inhaling arsine gas.
Instrument Operation
Fuel: Argon-hydrogen flame
Wavelength: 193.7 nanometer
Optimium Concentration Range: 2-20 ug As/liter
Lower Detection Limit: 2 ug/liter
-------
8.51-10
1. Turn on the Argon and adjust the flow rate to about
8 liters/minute, with an auxiliary Argon flow of 1 liter/minute
2. Turn on the hydrogen, adjust flow rate to about 7
liters/minute and ignite. The flame is colorless. The hand
may be passed 1 ft above the burner to detect heat, in order
to insure ignition.
Follow the instructions given with the Atomic Absorption
Spectrophotometer.
Quantification
The absorbances of spiked samples and blank vs. the con-
centrations are plotted according to the Method of Standard
Additions as described in Section 8.49. The extrapolated
value will be 1/10 the concentration of the original sample.
If the plot does not result in a straight line a non-
linear interference is present. This can sometimes be overcome
by dilution, or addition of other reagents if there is some
knowledge about the waste.
-------
8.52-1
Method 8.52
BARIUM
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of Barium in a waste
or an Extraction Procedure Extract.
Summary of Method
The sample is digested using nitric and/or hydrochloric
acids and the concentration of Barium measured using either a
flame or graphite furnace equipped atomic absorption spectro-
photometer. The flame method is most accurate when employed
with solutions containing 1-20 mg Ba/liter while the graphite
furnace procedure is best suited for solutions containing 10-
200 ug Ba/liter.
Precaution
Barium affects the heart muscle. A dose of 550 to 600
mg is considered fatal to man. Afflictions resulting from
ingestion, inhalation or absorption involve the heart, blood
vessels and nerves.
Interferences
If a nitrous oxide-acetylene flame is used with Direct
Aspiration chemical interferences are virtually eliminated.
In this method Potassium (1000 mg/1) is added to prevent
ionlzation of barium in this flame. If the air-acetylene
flame must be used then the presence of phosphate, silicon
and aluminum will lower the barium absorbance. This may be
overcome by addition of lanthanum.
-------
8.52-2
The graphite furnace method Is sensitive to the presence
of chlorides and dissolved solids.
Direct Aspiration Method
Reagents
1. Air, cleaned and dried through a suitable filter to remove
oil, water, and other foreign substances. The source may
be a compressor or a cylinder of industrial grade compressed
air.
2. Acetylene, should be of high purity. Acetone, which is
always present in acetylene cylinders, can be prevented
from entering and damaging the burner head by replacing
the cylinder when its pressure has fallen to 100 psig.
3. Deionized distilled water: Use deionized distilled water
for the preparation of all reagents and calibration
standards, and as dilution water.
4. Hydrochloric acid, HC1, concentrated, Ultrex grade.
5. Nitric acid, HN03, concentrated, Ultrex grade.
6. Nitrous oxide, commercially available cylinders.
7. Potassium Chloride solution: Dissolve 95 g KC1 in
distilled deionized water and bring to volume in a 1
liter volumetric flask.
8. Lanthanum Chloride solution if needed. Dissolve 25 g,
reagent grade, La203 slowly in 250 ml concentrated.
HC1. (Reaction is violent.) Dilute to 500 ml with distilled
deionized water.
9. Barium Standard Stock Solution
-------
8.52-3
1000 mg/liter solution may be purchased or prepared as
follows: Dissolve 1.7787 g barium chloride dihydrate,
(BaCl2 • 2H20), analytical, reagent grade, in about
200 ml distilled deionized water. Add 1.5 ml cone.
HN03« Add to 1 liter volumetric flask and bring to
volume. 1 ml » 1 mg Ba.
Apparatus
1. Atomic Absorption Spectrophotometer with nitrous oxide
burner head with 2" slot. Note: A razor blade is required
to dislodge, about every 20 minutes, the carbon crust
that forms along the slot surface.
2. T-junction valve for rapidly changing from nitrous oxide
to air, so the flame can be turned on or off with air as
oxidant to prevent flashbacks.
Procedure
1. Sample preparation
a. Transfer 100 ml of sample to a 250 ml beaker and add
3 ml concentrated HN03. Place it on a hot plate and
evaporate to near dryness, slowly, so that the sample
does not boil. Cool the beaker and add another 3 ml
HN03. Cover with a watch glass and return to the
hot plate. Continue heating gently, and add acid as
necessary until the material is light in color.
Evaporate to near dryness and cool. Add 2 ml of 1:1
HC1 and warm the beaker to dissolve any precipitate.
Wash down the beaker and watch glass with distilled
-------
8.52-^
deionized water and transfer quantitatively to a 100
ml volumetric flask, add 2 ml Potassium Chloride
solution and bring to volume with distilled deionized
water containing 1.5 ml HN03/liter.
Notet If air-acetylene is being used add 4.7 ml lanthamum
chloride solution before bringing to volume.
b. If insoluble silicates or other material is present
the sample must be filtered or centrifuged and the
supernatant sampled.
2. Prepare working standards from the Standard Stock Solution.
If it is desired to bracket the optimum concentration
range the following is suggested:
a. Transfer 0, 0.1, 0.5, 1.0, 1.5, and 2 ml of
Standard Stock Solution to 100 ml volumetric
flasks. Add 2 ml potassium chloride solution.
If lanthanum chloride was added to the sample, 4.7
ml must be added to the standards. Bring to volume
with distilled deionized water containing 1.5 ml
concentrated HN(>3/liter. The concentrations of these
standards will be 0, 1, 5, 10, 15 and 20 mg Ba/liter.
Standard Addition
1. a. Take the 5, 10, and 15 mg standards and pipet 5 ml
from each into separate 10 ml volumetric flasks. Add to
each 2 ml of the prepared sample and 0.06 ml KC1
solution. Bring to volume with distilled deionized
water containing 1.5 ml HN03/liter.
-------
8.52-5
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Add 0.16 ml KC1 solution and bring to
volume with distilled deionized water containing
1.5 ml HN03 per liter. This is the blank.
Note; The graph peak from the blank will be 1/5
that produced by the prepared sample. The peaks from the
spiked standards will be 1/2 that produced by the standards
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optimum
absorbance can be judged.
Instrument Operation
Wavelength: 553.6 nanometers
Fuel: acetylene
Oxidant: Nitrous Oxide
Type of flame: Fuel rich
Optimum Concentration Range: 1-20 mg/liter Lower
Detection Limit: 0.1 mg/liter
Follow the instructions given with the Atomic Absorption
Spectrophotometer. The following is included as a guide:
a. Install a nitrous oxide burner head.
b. Turn on the acetylene (without igniting the
flame), and adjust the flow rate to the value specified
by the manufacturer for a nitrous oxide-acetylene flame.
c. Turn off the acetylene.
d. With both air and nitrous oxide supplies turned on,
set the T junction valve to nitrous oxide and adjust
-------
3.52-6
the flow rate according to the specifications of the
manufacturer.
e. Turn the switching valve to the air position and
verify that the flow rate is the same.
f. Turn the acetylene on and ignite to a bright yellow
flame.
g. With a rapid motion, turn the switching valve to
nitrous oxide. The flame should become rose-red; if
it does not, adjust fuel flow to obtain a red cone in
flame.
h. Atomize deionized distilled water containing 1.5 ml
cone. HN03/1 and check the aspiration rate. Adjust
if necessary to a rate between 3 and 5 ml/min.
i. Atomize a 1-mg/l standard of the metal and adjust the
burner (both sideways and vertically) in the light
path until maximum response is obtained.
j. The instrument is now ready to run standards and
samples•
k. To extinguish the flame, turn the switching valve
from nitrous oxide to air, and turn off the acetylene.
This procedure eliminates the danger of flashback
that may occur on direct ignition or shutdown of
nitrous oxide and acetylene.
Quantification
The absorbance of spiked samples and blank vs. concentrations
are plotted according to the Method of Standard Additions
defined in Section 8.49. The extrapolated value will be 1/5
-------
8.52-7
the concentration of the original sample. If the plot does
not result in a straight line a non-linear interference is
present. This can sometimes be overcome by dilution, or
addition of other reagents if there is some knowledge about
the waste.
Graphite Furnace
Reagents
1. Barium Standard Stock Solution 1000 mg/liter solution may
be purchased or prepared as follows: Dissolve 1.7787 g
barium chloride dihydrate, BaCl2»2H20 analytical reagent
grade, in about 200 ml distilled deionized water. Add
1.5 ml concentrated HN03 and bring to volume in a 1 liter
volumetric flask. 1 ml = 1 mg Ba.
2. Concentrated Nitric Acid
Procedure
Sample Preparation
(Note: All chloride and hydrochloric acid is avoided.)
1. a. Transfer 100 ml of sample to a 250 ml beaker and
add 3 ml concentrated HN03. Place it on a hot plate and
evaporate to near dryness, slowly, so that the sample
does not boil. Cool the beaker and add another 3 ml
HN03. Cover with a watch glass and return to the hot
plate. Continue heating gently, and add acid as
necessary until the material is light in color.
Evaporate to near dryness and cool. Add a small quantity
of 1:1 HN03 and warm the beaker to dissolve any
precipitate.
-------
8.52-8
Wash down the beaker and watch glass with distilled
deionized water and transfer quantitatively to a 100
ml volumetric flask. Bring to volume with distilled
deionized water containing 1.5 ml HN03/liter.
b. If insoluble silicates or other material is present
the sample must be filtered or centrifuged and the
supernatant sampled.
2. a. Prepare working standards from the Standard Stock Solution.
If it is desired to bracket the optimum concentration
range the following is suggested:
b. Transfer 1 ml stock solution to a 1 liter volumetric
flask. Bring to volume with distilled deionized
water acidified with 1.5 ml cone. HN03/liter.
Concentration: 1000 ug Ba/liter.
c. Transfer 0, 5, 10, 15, and 20 ml from a) to separate
100 ml volumetric flasks and bring to volume with
distilled deionized water acidified with 1.5 ml concentrated
HN03/liter. The concentrations of these working
standards will be 0, 50, 100, 150, and 200 ug Ba/liter.
Standard Addition
1. a. Take the 100, 150, and 200 ug standards and pipet 5
ml from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample. Bring to
volume with distilled deionized water containing 1.5
ml HN03/liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric
-------
8.52-9
flask. Bring to volume with distilled deionlzed
water containing 1.5 ml HNC>3 per liter. This is
the blank.
Note; The absorbance of the blank will be 1/5 that
produced by the prepared sample. The absorbance of the
spiked standards will be 1/2 that produced by the standards
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optimum
absorbance can be judged.
Instrument Operation
Wavelength: 553.6 nanometers
Optimum Concentration range: 10 - 200 ug/liter Lower
Detection limit: 2 ug/liter
Drying time and temperature: 30 seconds at 125°C
Ashing time and temperature: 30 seconds at 1200°C
Atomizing time and temperature: 10 seconds at 2800°C
Purge gas: Argon. Do not use nitrogen.
These conditions are based on a 20 ul injection, continuous
flow purge gas and non-pyrolytic graphite on a Perkin Elmer
model HGA 2100 furnace. Other equipment will have different
requirements. Follow the manufacturer's manual.
Quantification
The absorbance of spiked samples and blank vs. the
concentrations are plotted according to the Method of Standard
Additions as defined in Section 8.49. The extrapolated
value will be the concentration of the original sample.
If the plot does not result in a straight line a non-
-------
8.52-10
linear interference is present. This can sometimes be overcome
by dilution, or addition of other reagents if there is some
knowledge about the waste.
-------
S.53-1
Method 8.53
CADMIUM
Scope and Application
The following procedures are approved methods for
determining the concentration of Cadmium in a waste or Extraction
Procedure Extract.
Precaution
Cadmium is highly toxic. Minute quantities are suspected
of causing adverse changes in human kidneys.
Cadmium is particularly susceptible to contamination of
the work area. Special care should be taken in the handling
of the material.
Comments
Two Atomic Absorption methods are described.
1. Direct Aspiration
This method is suitable for higher concentrations (0.05-2
ug/liter optimum).
2. Graphite Furnace
This method is suitable for lower levels (0.5-10 ug/liter
optimum).
Direct Aspiration
Reagents
1. Air free of oil, water, and other foreign substances. The
source may be purified air from a compressor or a cylinder
of industrial grade compressed air.
-------
S.53-2
2. Acetylene, standard commercial grade. Acetone, which is
always present in acetylene cylinders, can be prevented
from entering and damaging the burner head by replacing
a cylinder when its pressure has fallen to 7 kg/cm^
(100 psig) acetylene.
3. Nitric Acid, concentrated, ACS.
A. Cadmium Standard Stock Solution (1000 mg/liter):
Carefully weigh 3.1368 g Cadmium Sulfate Octahydrate
(Cd SO^.S H20), analytical reagent grade, and dissolve
in distilled delonized water containing 1.5 ml HN03/liter.
Transfer quantitatively to a 1 liter volumetric flask
and bring to volume with the same water. 1 ml - 1 mg Cd.
5* Hydrochloric acid solution: Dilute concentrated HC1 (ACS)
1:1 with distilled deionized water.
Procedure
Sample Preparation
1. Transfer 100 gm of well-mixed sample (use 100 ml when analyzing
an EP extract) to a 250 ml beaker. Add 3 ml concentrated
HNf>3. Place the beaker on a hot plate and evaporate
to near dryness, cautiously, so that the sample does not
boil. Cool, and add another 3 ml concentrated HN03-
Cover the beaker with a watch glass and return to the
hot plate. Heat so as to produce a gentle refluxing.
Continue adding 3 ml portions of HN03 until the sample
is light in color and no longer changes in color. Evaporate
to near dryness and cool. Add 5 ml of 1:1 HC1 and warm
the beaker just to dissolve any precipitates.
-------
8.53-3
b. Cool and bring to 100 ml with distilled deionized
water.
2. If particulates such as silicates remain in the sample
it must be centrifuged and the supernatant aspirated.
3. Prepare working standards from stock solution. If it
is desired to work in the optimum concentration range
the following is suggested:
a. Transfer 10 ml stock solution to a 100 ml volumetric
flask. Bring to volume with distilled deionized
water containing 1.5 ml concentrated HN03/ liter.
Concentration: 100 mg/liter.
b. Transfer 0, 0.1, 0.5, 1.0, 1.5, and 2.0 ml solution
from a) to separate 100 ml volumetric flasks. Bring
to volume with distilled deionized water containing
1.5 ml HN03/liter. The concentrations of these
standards will be 0, 0.1, 0.5, 1.0, 1.5, and 2.0 mg
Cd/liter.
Standard Addition
a. Take the 1.0, 1.5, and 2.0 mg/liter standards and pipet 5 ml
from each into separate 10 ml volumetric flasks. Add
to each 2 ml of the prepared sample. Bring to volume
with distilled deionized water containing 1.5 ml
HN03/liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Bring to volume with distilled deionized
water containg 1.5 ml HN03 per liter. This is the
blank.
-------
Note! The absorbance from the blank will be 1/5 that
produced by the prepared sample. The absorbance from
the spiked standards will be 1/2 that produced by the
standards plus the contribution from 1/5 of the prepared
sample. Keeping these In mind the correct dilutions to
produce optimum absorbance can be judged.
Instrument Operation
Wavelength: 228.8 nanometers
Optimum Concentration Range: 0.05 - 2 mg/liter
Lower Detection Limit: 0.005 mg/liter
Fuel: Acetylene
Oxidant: air
Type of flame: oxidizing.
A general outline for instrument operation is given in Section
8.49. Follow the manufacturer's instructions for the Spectro-
photometer being used.
Quantification
The absorbance of spiked samples and blank vs. the
concentrations are plotted according to the Method of Standard
Additions as defined in Section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a non-
linear interference is present. This can sometimes be
overcome by dilution, or addition of other reagents if
there is some knowledge about the waste.
-------
8.53-5
Graphite Furnace
Reagents
1. Cadmium Standard Stock Solution (1000 mg/liter). Carefully
weigh 3.1368 g Cadmium Sulfate Octahydrate (Cd SO^.S H2<>),
analytical reagent grade, and dissolve in distilled
deionized water containing 5 ml concentrated HN03/liter.
Transfer quantitatively to a 1 liter volumetric flask
and bring to volume with the same water. 1 ml « 1 mg Cd.
2. Concentrated Hfl(>3 (ACS)
3. Concentrated HN03 diluted 1:1 with distilled deionized
water.
4. Ammonium phosphate solution (40%) Dissolve 40 grams
Ammonium mono hydrogen phosphate, (NH4)2HP04, analytical
reagent grade, in distilled deionized water and bring
to 100 ml.
Procedure
1. Sample Preparation
a. Transfer 100 gm of well-mixed sample (or when analyzing
an EP extract 100 ml) to a 250 ml beaker. Add 3 ml
concentrated HN03. Place the beaker on a hot
plate and evaporate to near dryness, cautiously, so
that the sample does not boil. Cool, and add another
3 ml concentrated HN03. Cover the beaker with a
watch glass, and return to the hot plate. Heat so
as to produce a gentle refluxing. Continue adding
3 ml portions of concentrated HN03 until the
-------
8.53-6
sample is light in color and no longer changes in
color. Evaporate to near dryness and cool. Add 1
ml of 1:1 HN(>3 and warm the beaker just to dissolve
any precipitates. Cool the beaker.
b. Add 2 ml Ammonium phosphate solution and bring to
100 ml with distilled deionized water.
2. If particulates such as silicates remain in the sample
it must be centrifuged and the supernatant sampled.
3. Prepare working standards from the stock solution. If
it is desired to work in the optimum concentration range
the following is suggested:
a. Transfer 1 ml stock solution to a 1 liter volumetric
flask. Bring to volume with distilled deionized
water containing 5 ml concentrated HN03/liter.
Concentration: 1000 ug/liter.
b. Transfer 10 ml from a) to a 100 ml volumetric flask
and bring to volume with distilled deionized water
containing 5 ml HN03/liter. Concentration: 100
ug/liter.
c. Transfer 0, 0.5, 1.0, 2.5, 5.0, and 10 ml from b)
to 100 ml volumetric flasks. Add 2 ml Ammonium
phosphate solution. Bring to volume with distilled
deionized water containing 5 ml HN03/liter. The
concentrations of these working standards will be 0,
0.5, 1.0, 2.5, 5.0, and I'O ug Cd/liter.
-------
8.53-7
Standard Addition
1. a. Take the 2.5, 5.0 and 10 ug/liter standards and pipet 5 ml
from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample and 0.06 ml
ammonium phosphate solution. Bring to volume with
distilled deionized water.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask and add 0.16 ml ammonium phosphate solution.
Bring to volume with distilled deionized water.
This is the blank.
Note; The absorbance from the blank will be 1/5 that produced
by the prepared sample. The absorbance from the spiked
standards will be 1/2 that produced by the standards plus
the contribution from 1/5 of the prepared sample. Keeping
these in mind the correct dilutions to produce optimium
absorbance can be judged.
Instrument Operation
Wavelength: 228.8 nanometers
Optimum Concentration Range: 0.5 - 10 ug/liter
Lower Detection Limit: 0.1 ug/liter
Purge Gas: Argon
Drying time and temperature: 30 sec at 125°C
Ashlngtime and temperature: 30 sec at 500°C
Atomizing time and temperature: 10 sec at 1900°C
These conditions are based on a 20 ul injection continuous
flow purge gas and non-pyrolytic graphite on a Perkin Elmer
-------
8.53-8
model HGA 2100 furnace. Other equipment will have different
requirements. Follow the manufacturer's manual.
(Quantification
The absorbance of spiked samples and blank vs. the
concentrations are plotted according to the Method of
Standard Additions as defined in Section 4.49. The extra-
polated value will be 1/5 the concentration of the original
sample.
If the plot does not result in a straight line a non-
linear interference is present. This can sometimes be
overcome by dilution, or addition of other reagents if
there is some knowledge about the waste.
-------
8.54-1
Method 8.54
CHROMIUM
Scope and Application
The following procedures are approved methods for determining
the concentration of chromium in an Extraction Procedure Extract,
an industrial liquid waste, or landfill liquid component or leachate
Precaution
Hexavalent chromium is both acutely toxic and carcinogenic
to man. While the trivalent form is less toxic to humans,
care should be exercised in its handling.
Summary of Method
The sample is digested using nitric and/or hydrochloric
acids and the concentration of chromium measured using either a
flame or graphite furnace equipped atomic absorption spectre-
photometer. The flame method is most acurate when employed
with solutions containing 0.5-10 mg Cr/liter while the graphite
furnace procedure is best suited for solutions containing
5-100 ug Cr/liter.
Comments
1. The direct aspiration method is suitable for a concentration
range of 0.5-10 mg/1. The nitrous oxide-acetylene flame is
recommended bacause, while less sensitive, it eliminates
interferences from other common metals such as iron and nickel.
If the air-acetylene flame must be used it should be lean.
Interference from other ions has reportedly been overcome by
addition of ammonium biflouride.
-------
8.54^2
2. In the furnace method the presence of calcium interferes.
However, since at concentrations above 200 mg/1 its
effect becomes constant, it is added to insure this
minimum level. Hydrogen peroxide is used to bring all
chromium present in the sample to the +6 state.
Direct Aspiration
Reagents
1. Nitrous oxide, commercially available cylinders
2. Acetylene should be of high purity standard commercial
grade •
3. Concentrated Nitric Acid (HN03)
4. Chromium Stock Solution (1000 mg/liter):
Dissolve 1.9231 grams chromium trioxide (Cr03) analytical
reagent grade, in deionized distilled water. Add 1.5 ml
HNO^ and bring to volume in a 1 liter volumetric flask
with this same water (1 ml = 1 mg Cr).
5. 1% ammonium biflouride solution if needed for air-acetylene
flame:
Dissolve 0.2 grams sodium sulfate in distilled deionized
water. Add 1 gram ammonium biflouride (NH4HF2). Bring
to volume in a 100 ml volumetric flask with distilled
deionized water.
Apparatus
1. Atomic Absorption Spectrophotometer with nitrous oxide
burner head with 2" slot.
2. T-junction valve for rapidly changing from nitrous oxide
to air, so the flame can be turned on or off with air as
oxidant to prevent flashbacks.
-------
8.54-3
Procedure
Sample preparation
1. a. Transfer 100 ml of well-mixed sample to a 250 ml
beaker. Add 3 ml cone HN03. Place the beaker on a
hot plate and evaporate to near dryness, cautiously,
taking care not to let the sample boil. Cool, and
add another 3 ml cone HNC>3. Cover the beaker with
a watch glass and return to the hot plate. Heat so
as to produce a gentle refluxing. Continue adding
3 ml portions of HN03 until the sample is light in
color and no longer changes in color. Evaporate to
near dryness and cool. Add 1 ml HN03 and warm the
beaker until any precipitates present dissolve.
b. Cool and bring to 100 ml with distilled deionized water.
Note; If the air-acetylene flame is used add 1 ml
ammonium biflouride solution just before making up
to 100 ml volume.
2. If particulates, such as silicates, remain in the digested
sample, then centrifuge the digestate and only aspirate the
supernatant.
3. Prepare working standards from stock solution. To bracket
the optimum concentration range the following is suggested:
a. Transfer 0, 0.1, 0.2, 0.5, 0.8, and 1.0 ml stock solution
to separate 100 ml volumetric flasks. Bring to volume
with distilled deionized water containing 1.5 ml HN03/liter
These working standards will contain 0, 1, 2, 5, 8 and
10 mg Cr/liter.
-------
Note; If the air-Acetylene flame must be used add
1 ml ammonium biflouride solution just before
making up to 100 ml volume.
4 . Standard Addition
a. Take the 5, 8, and 10 mg standards and pipet 5 ml
from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample. Bring to
volume with distilled deionized water containing 1.5
ml HN03/liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Bring to volume with distilled deionized
water containing 1.5 ml HN03 per liter. This is the
blank.
Note; The absorbance from the blank (b), will be
1/5 that produced by the prepared sample. The absorb-
ance from the spiked standards will be 1/2 that
produced by the standards plus the contribution from
1/5 of the prepared sample. Keeping these in mind
the correct dilutions to produce optimum absorbance
can be judged .
Instrument Operation
Wavelength: 357.9 nanometers
Optimum concentration range: 0.5-10 mg/liter
Lower detection limit: 0.05 mg/liter
Fuel: Acetylene
Oxident: Nitrous Oxide
Type of Flame: Fuel rich
-------
8.54-5
Follow the instructions given with the Atomic Absorption
Spectrophotometer. The following is included as a guide:
a. Install a nitrous oxide burner head.
b. Turn on the acetylene (without igniting the flame),
and adjust the flow rate to the value specified by
the manufacturer for a nitrous oxide-acetylene flame.
c. Turn off the acetylene.
d. With both air and nitrous oxide supplies turned on,
set the T-junction valve to nitrous oxide and adjust
the flow rate according to the specifications of the
manufacturer.
e. Turn the switching valve to the air position and
verify that the flow rate is the same.
f. Turn the acetylene on and ignite to a bright yellow
flame.
g. With a rapid motion, turn the switching valve to
nitrous oxide. The flame should become rose-red; if
it does not, adjust fuel flow to obtain a red cone
in flame.
h. Atomize deionized distilled water containing 1.5 ml
conc«HNC>3/l and check the aspiration rate. Adjust
if necessary to a rate between 3 and 5 ml/min.
Atomize a 1 mg/1 standard of the metal and adjust
the burner (both sideways and vertically) in the
light path until maximum response is obtained.
The instrument is now ready to run standards and
samples •
-------
8.5^-6
i. To extinguish the flame, turn the switching valve
from nitrous oxide to air, and turn off the acetylene.
This procedure eliminates the danger of flashback
that may occur on direct ignition or shutdown of
nitrous oxide and acetylene.
Quantification
The absorbance of spiked samples and blank vs the concen-
tration are plotted according to the Method of Standard Additions
as defined in General Requirements #11. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a non-linear
interference is present. This can sometimes be overcome by
dilution, or addition of other reagents if there is some
knowledge about the waste.
-------
S.54-7
Graphite Furnace
Reagents
1. Chromium Standard Stock Solution (1000 mg/liter):
a. Dissolve 1.9231 grams Chromium trioxide (Cr03> analytical
reagent grade, in deionized distilled water. Add 5 ml
HN03 and bring to volume in a 1 liter volumetric flask
with this same water (1 ml « 1 mg Cr).
2. Concentrated HN03
3. Calcium nitrate solution:
Dissolve 11.8 grams Calcium nitrate tetrahydrate
(Ca(N03>2.4H20), analytical reagent grade in deionized
distilled water and dilute to 100 ml (1ml « 20mg Ca).
4. 30% hydrogen peroxide
Procedure
Sample Preparation
1. a. Transfer 100 ml of well-mixed sample to a 250 ml
beaker. Add 3 ml cone HN03. Place the beaker on a
hot plate and evaporate to near dryness, taking care
not to let the sample boil. Cool, and add another
3 ml cone HNC^. Cover the beaker with a watch glass
and return to the hot plate. Heat so as to produce
a gentle refluxing. Continue adding 3 ml portions
of HN03 until the sample is light in color and no
longer changes in color. Evaporate to near dryness
and cool. Add 1 ml of 1:1 HN03 and warm the beaker
until any precipitate present dissolves. Cool the
beaker .
-------
b. Add 1 ml of 30% hydrogen peroxide and 1 ml Calcium
nitrate solution. Transfer to a 100 ml flask and
bring to volume with distilled deionized water.
2. If any particulates such as silcates remain in the digested
sample, then centrifuge the digestate and only aspirate
the supernatant.
3. Prepare working standards from stock solution: To bracket
the optimum working range the following is suggested:
a. Transfer 1 ml stock solution to a 1 liter volumetric
flask. Bring to volume with distilled deionized
water containing 5 ml HN03/liter. Concentration
lOOOug/liter.
b. Transfer 0, 1, 2.5, 5, 7.5 and 10 ml from a) to
separate 100 ml volumetric flasks. Add 1 ml 30%
hydrogen peroxide and 1 ml calcium nitrate solution.
Bring to volume with distilled deionized water
containing 5 ml HN03/liter. The concentrations of
these working standards are 0, 10, 25, 50, 75, and
100 ug Cr/Hter.
Standard Addition
1. a. Take the 50, 75, and 100 ug standards and pipet 5 ml
from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample. Add
0.03 ml CaN03 solution and 0.03 ml 1^02 solution.
Bring to volume with distilled deionized water.
-------
8.54-9
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Add 0.08 ml CaN03 solution and 0.08 ml
H202 solution. Bring to volume with distilled
deionized water. This is the blank.
Note; The absorbance from the blank (b), will be
1/5 that produced by the prepared sample. The absorb-
ance from the spiked standards will be 1/2 that
produced by the standards plus the contribution from
1/5 of the prepared sample. Keeping these in mind
the correct dilutions to produce optimum absorbance
can be judged.
Instrument Operation
Wavelength: 357.9 nanometers
Optimum Concentration Range: 5-100 ug/liter
Lower detection limit: 1 ug/liter
Purge gas: Argon
Drying time and temp.: 30 secs-125°C
Ashing time and temp.: 30 secs-1000°C
Atomizing time and temp.: 10 secs-2700°C
The conditions listed above are based on a 20 ul injection;
continuous flow purge gas and non-pyrolytic graphite on a
Perkin Elmer model HGA 2100 furnace. Other equipment will
have different requirements. Follow the manufacturer's manual.
Quantification
The absorbances of spiked samples and blank vs concentra-
tions are plotted according to the Method of Standard Additions
as defined in General Requirements #11. The extrapolated
-------
8.54-10
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a non-linear
interference is present. This can sometimes be overcome by
dilution, or addition of other reagents if there is some know-
ledge about the waste.
-------
8.56-1
Method 8:56
LEAD
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of Lead in a waste
or an Extraction Procedure Extract.
Precaution
Lead is poisonous and accumulates in the body. Drinking
water does not usually contain more than 20 ug/liter. It can
be leached from old lead plumbing by acid water.
Summary of Method
The sample is digested using nitric and/or hydrochloric
acids and the concentration of lead measured using either a
flame or graphite furnace equipped atomic absorption spectre-
photometer. The flame method is most accurate when employed
with solutions containing 1-20 mg Pb/liter while the graphite
furnace procedure is best suited for solutions containing
5-100 ug Pb/liter.
Comment s
1. Two wavelengths are suitable and may be tested for
best operating conditions. The 217.0 nm wavelength is
more sensitive but may produce more background noise.
2. The Graphite Furnace method is susceptible to sulfate
interference. Up to 1500 ppm sulfate can be suppressed
by addition of Lanthanum.
-------
8.56-2
3. Contamination of the working environment is a special
problem. All glassware should be cleaned immediately
prior to use and then covered.
Direct Aspiration
Reagents
1. Air, cleaned and dried through a suitable filter to
remove oil, water, and other foreign substances. The
source may be a compressor or a cylinder of industrial
grade compressed air. Breathing quality air can cause
flashback.
2. Acetylene should be of high quality grade. Avoid "purified
grade" acetylene. Acetone, which is always present in
acetylene cylinders, can be prevented from entering and
damaging the burner head by replacing a cylinder when
its pressure has fallen to 7 kg/cm^ (100 psig) acetylene.
3. Nitric Acid cone.
4. Lead Standard Stock Solution 1000 mg/liter stock solution
may be purchased or prepared as follows: Dissolve 1.5985
g lead nitrate (Pb(N03)2) analytical reagent grade, in
about 200 ml distilled deionized water. Acidify with 10
m 1 cone. HN03 and bring to volume in a 1 liter volumetric
flask. (1 ml = 1 mg Pb.)
5. Hydrochloric acid solution: Dilute cone. HC1 1:1 with
distilled deionized water.
-------
8.56-3
Procedure
Sample Preparation
1. a. Transfer 100 ml of well-mixed sample to a 250 ml
beaker. Add 3 ml cone. HN03. Place the beaker on
a hot plate and evaporate to near dryness, cautiously,
in order to keep the sample from boiling. Cool,
and add another 3 ml cone. HN03. Cover the beaker
with a watch glass and return to the hot plate.
Heat so as to produce a gentle refluxing. Continue
adding 3 ml portions of HN03 until the sample is
light in color and no longer changes In color.
Evaporate to near dryness and cool. Add 5 ml of 1:1
HC1 and warm the beaker just to dissolve any precipi-
tates .
b. Cool and bring to 100 ml with distilled deionized
water .
2. If particulates such as silicates remain in the sample
it must be centrifuged and the supernatant aspirated.
3. Prepare working standards from stock solution. If it is
desired to work in the optimum concentration range
the following is suggested:
a. Transfer 0, 0.1, 0.5, 1.0, 1.5, and 2.0 ml of
stock solution to separate 100 ml volumetric
flasks. Bring to volume with distilled deionized
water containing 1.5 ml HN03/liter. The concentrations
of these working standards are 0, 1, 5, 10, 15, and
20 mg Pb/liter.
-------
8.56-4
Standard Addition
1. a. Take the 10, 15, and 20 mg standards and pipet 5
ml from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample. Bring to
volume with distilled deionized water containing 1.5
ml HN03/liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Bring to volume with distilled deionized
water containing 1.5 ml HN03 per liter. This is
the blank.
Note; The absorbance from the blank (b), will be 1/5
that produced by the prepared sample. The absorbance
from the spiked standards will be 1/2 that produced by
the standards plus the contribution from 1/5 of the
prepared sample. Keeping these in mind the correct
dilutions to produce optimum absorbance can be judged.
Instrument Operation
Wavelength: 283.3 nanometers
217.0 nanometers (more sensitive)
Optimum Concentration Range: 1-20 mg/liter
Lower Detectable limit: 0.1 mg/liter Fuel: Acetylene
Oxidant: Air
Type of flame: Oxidizing
This method is especially sensitive to turbulance in the
flame. Special care should be taken to position the
burner so that the light beam passes through the most
-------
8.56-5
stable center portion of the flame. The absorbance
should be maximized with a lead standard.
A general outline for instrument operation is given in
Section 8.49. Follow the manufacturer's instructions for
the Spectrophotometer being used.
Quantification
The absorbances of spiked samples and blank vs. the
concentrations are plotted according to the Method of
Standard Additions as defined in Section 8.49. The extrapo-
lated value will be 1/5 the concentration of the original
sample.
If the plot does not result in a straight line a
nonlinear interference is present. This can sometimes be
overcome by dilution, or addition of other reagents if there
is some knowledge about the waste.
-------
8.56-6
Graphite Furnace
Reagents
1. Lead Standard Stock Solution 1000 mg/llter stock solution
may be purchased or prepared as follows: Dissolve 1.5985 g
lead nitrate (Pb(N03>2) analytical reagent grade, in
about 200 ml distilled deionized water. Acidify with 10
ml cone HN03 and bring to volume in a 1 liter volumetric
flask. 1 ml - 1 mg Pb.
2. Concentrated HNC-3
3. Lanthanum Nitrate Solution: Dissolve 58.64 g analytical
reagent grade Lanthanum oxide (La2 03) in 100 ml cone.
HN03 and dilute to 1000 ml with distilled deionized
water. 1 ml - 50 mg La.
Procedure
Sample Preparation
1. a. Transfer 100 ml of well-mixed sample to a 250 ml
beaker. Add 3 ml cone. HNC>3. Place the beaker on
a hot plate and evaporate to near dryness, cautiously,
so that the sample does not boil. Cool, and add
another 3 ml cone. HN03- Cover the beaker with a
watch glass, and return to the hot plate. Heat so
as to produce a gentle refluxing. Continue adding 3
ml portions of HN03 until the sample is light in
color and no longer changes in color. Evaporate to
near dryness and cool. Add 1 ml of 1:1 HN03 and
warm the beaker just to dissolve any precipitates.
Cool the beaker. Add 10 ml of Lanthanum nitrate
-------
8.56-7
solution and bring to volume in a 100 ml volumetric
flask using distilled deionized water.
2. If particulates such as silicates remain in the sample
it must be centrifuged and the supernatant sampled.
3. Prepare working standards from the Stock solution. If
it is desired to work in the optimum concentration range
the following is suggested:
a. Transfer 1 ml stock solution to a 1 liter volumetric
flask. Bring to volume with distilled deionized
water containing 5 ml cone. HN03/liter. Concentra-
tion: 1000 ug/liter.
b. Transfer 0, 0.5, 2.5, 5.0, 7.5, and 10 ml from
(a) to separate 100 ml volumetric flasks. Add 10
ml Lanthanum Nitrate solution and bring to volume
using distilled deionized water. The concentra-
tions will be 0, 5, 25, 50, 75, and 100 ug/liter.
Standard Addition
1. a. Take the 50, 75, and 100 ug standards and pipet
5 ml from each into separate 10 ml volumetric
flasks. Add to each 2 ml of the prepared sample.
Add 0.8 ml lanthanum nitrate solution and bring to
volume with distilled deionized water.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Add 0.8 ml lanthanum nitrate solution and
bring to volume with distilled deionized water.
This is the blank.
-------
8.56-8
Note; The absorbance from the blank (b), will be 1/5
that produced by the prepared sample. The absorbance
from the spiked standards will be 1/2 that produced by
the standards plus the contribution from 1/5 of the
prepared sample. Keeping these in mind the correct
dilutions to produce optimium absorbance can be judged.
Instrument Operation
Wavelength: 283.3 nanometers
217.0 nanometers
Optimum concentration range: 5-100 ug/liter
Lower detection limit: 1 ug/liter.
Purge gas: Argon
Drying time and temp: 30 sec - 125°C
Ashing time and temp: 30 sec - 500°C
Atomizing time and temp: 10 sec - 2700°C
The 217.0 nanometer wavelength is more sensitive.
The electrodeless discharge lamp has been reported
helpful at this wavelength. The sample may also be
successfully atomized at a lower temperature. (2400°C)
The conditions listed above are based on a 20 ul
injection; continuous flow purge gas and non-pyrolytic
graphite on a Perkin Elmer model HGA 2100 furnace. Other
equipment will have different requirements. Follow the
manufacturer's manual.
Quantification
The absorbances of spiked samples and blank vs. the
concentrations are plotted according to the Method of Standard
-------
8.56-9
Additions as defined in Section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a nonlinear
interference is present. This can sometimes be overcome
by dilution, or addition of other reagents if there is
some knowledge about the waste.
-------
8.57-1
Method 8.57
MERCURY
Scope and Application
The flameless, cold vapor, atomic absorption procedure
is approved for use in determining the concentration of
mercury in an Extraction Procedure Extract, a RCRA waste, or
landfill leachate.
Precaution
The extreme toxiclty of mercury is well known. Care
should be taken in the handling of materials and to avoid
breathing any vapors. A trap should be constructed to receive
the exhaust gas.
Summary of Method
The sample is pretreated to break down organic mercury
compounds, and the mercury is then reduced to the zero oxi-:
dation state. In this form it is swept from the solution by
a purge gas and enters a glass or plastic absorption cell.
This cell has been aligned with the light beam from the
mercury hollow cathode lamp in the atomic absorption spectro-
photometer, and the mercury absorption is proportional to
the concentration.
Comments
1. The method is subject to interference from sulfide above
20 mg/1. Oxidation with potassium permanganate can be
used to remove this interference.
-------
8.57-2
2. Chloride containing wastes require excess permanganate
for removal. Chlorine gas is generated and must be
purged from the sample before analysis.
3. Copper may also interfere. It is expected that the
Method of Standard Addition will compensate for this.
4. In this method it is possible that some organic substance
will not be oxidized and will absorb at the given wavelength.
In this case, a second sample can be prepared and analyzed
without adding any stannous sulfate to reduce the mercury.
The true mercury value can then be found from the difference.
5. Disposal of residues:
The wastes from mercury analyses can be saved and treated
to recover the mercury. (See "Disposal".) The contaminated
mercury may be returned to the manufacturer for recycling.
Apparatus
1. An atomic absorption spectrophotometer with an open
presentation area so that the cold vapor cell may be
attached to it.
2. Absorption cell 10 cm. long and about 1" diameter; composed
of glass or plexiglass and having quartz end windows.
3. Air pump capable of delivering 2 liters of air/minute
with rotameter for monitoring air flow.
4. Tygon aeration tubing and glass tube with fritted end
(coarse porosity glass).
5. Drying tube 6" x 3/4" dia. containing 20 grams magnesium
perchlorate.
-------
8.57-3
60 W light bulb positioned to shine on the absorption
cell so that the temperature is maintained 10°C above
ambient. This prevents condensation by moisture. If
the drying tube is used this may not be necessary.
Scrubber bottle containing a glass frit immersed in
absorbing solution:
a. 1 part 0.1 molar potassium permanganate to 1 part
10% sulfuric acid. or
b. 3% potassium iodide solution containing 0.25% iodine
I
c
n t
u t
AIR PUMP
r f t
r I ^
1 DESICCANT ,, ,';
! 1
ABSORPTION
3 •* -BUBBLER CELL
\
/
T
c
•
3
T
^^N
-4
SAMPLE SOLUTION
IN BOD BOTTLE
Figure 8.57-1
PLAMELESS MERCURY A.A. SET-UP
SCRUBBER
CONTAINING
A MERCURY
ABSORBING
MEDIA
-------
8.57-4
Procedure
1. Assemble equipment as in the diagram.
2. Set the wavelength at 253.7 nanometer.
3. Install the absorption cell and align it in the light
beam using 2 cards, each having a 1" diameter hole.
These are placed over the ends of the cell for ease in
centering the beam.
4. Turn on the air and adjust flow rate to 1 liter/minute.
and allow the air to flow continuously.
5. Follow manufacturer's instructions for instrument warm
up and operation.
6. Prepare working standards from stock solution:
To bracket the optimum concentration range (0.2-10
ug/liter) the following is suggested:
a. Transfer 1 ml of stock solution to a 1 liter
volumetric flask and bring to volume with distilled
deionized water containing 1.5 ml HN03/liter. Con-
centration: 1000 ug/liter. (1 ml - 1 ug Hg)
b. Transfer 0, 1, 2, 5, 8, and 10 ml from a. to separate
100 ml volumetric flasks and bring to volume with
distilled deionized water containing 1.5 ml HN03/
liter. The concentrations of these working standards
will be 0, 10, 20, 50, 80 and 100 ug Hg/liter. (10
ml aliquots transferred for analysis will contain
0, 0.1, 0.2, 0.5, 0.8, and 1.0 ug Hg.)
-------
8.57-5
Reagents
1. Sulfurlc acid solution (0.5N):
Dilute 14 ml concentrated H2S04 to 1 liter with distilled
deionized water.
2. Nitric Acid, concentrated
3. Stannous Sulfate solution:
Add 25 grams SnSO^ to 250 ml sulfuric acid solution
(0.5N). This mixture should be stirred continuously
during analysis.
4. Sodium chloride - Hydroxylamine sulfate solution:
Dissolve 12 grams NaCl and 12 grams hydroxylamine sulfate
in distilled deionized water and dilute to 100 ml.
5. Potassium Permanganate solution: (5% w/v)
Dissolve 5g KMn(>4 in 100 ml distilled deionized water.
6. Potassium Persulfate solution: (5% w/v) Dissolve 5g
K2S2°8 in 10° ml distilled deionized water.
7. Mercury stock solution (1000 mg/liter):
Dissolve 0.1354 grams mercuric chloride (HgCl2) in
distilled deionized water containing 1.5 ml concentrated
HN03/liter. Bring to volume in a 100 ml volumetric
flask. (1 ml - 1 mg Hg.)
-------
8.57-6
7. Standard Addition
a. Take the 50, 80 and 100 ug standards and transfer
quantitatively 25 ml from each into separate 50 ml
volumetric flasks. Add to each 10 ml of the sample.
Bring to volume with distilled deionized water con-
taining 1.5 ml HN03/liter.
b. Add 10 ml of sample to a 50 ml volumetric flask.
Bring to volume with distilled deionized water con-
taining 1.5 ml HN03/liter. This is the blank.
Note; The absorbance from the blank will be 1/5 that
produced by the prepared sample. The absorbance from the
spiked standards will be 1/2 that produced by the standards
plus the contribution from 1/5 of the prepared sample.
Keeping these in mind the correct dilutions to produce optimium
absorbance can be judged.
8. Treatment of sample and standards:
a. Transfer a 10 ml aliquot to a 300 ml BOD bottle.
Add distilled deionized water to make 100 ml. Add 5
ml sulfuric acid solution and 2.5 ml concentrated
HN03. After mixing add 15 ml potassium permanganate
solution. Additional permanganate may be required
for some wastes. Shake and add enough so that the
purple color persists for at least 15 minutes. Then
add 8 ml potassium persulfate solution and heat for
2 hours in a water bath at 95°C. Cool. The sample
is now fully oxidized.
-------
8.57-7
b. During oxidation chlorides, if present, are converted
to free chlorine which interferes in the analysis*
Additionally any excess permanganate must be destroyed,
Add 25 ml hydroxylamine sulfate solution and allow to
stand for at least 30 seconds.
c. Purge the dead air space in the upper portion of the
BOD bottle, add 5 ml well-mixed stannous sulfate
solution, and immediately attach the bottle to the
bubbler apparatus as in the diagram, thus forming a
closed system. The pumps must run continuously.
The absorbance will increase and reach a maximum
within 30 seconds.
d. After about 1 minute, when measurement is complete,
open the bypass valve and allow the gas to flow
through the mercury trap. When absorbance returns
to baseline remove the sample bottle and replace
it with a BOD bottle containing distilled deionized
water. Flush the system for a few minutes. Close
the valve and run the next sample or standard.
Quantification
The absorbances of spiked samples and blank vs. the
number of micrograms mercury are plotted according to the
Method of Standard Addition as defined in Section 8.49. If
the additions were made as suggested in the method, the extra-
polated value will be the number of micrograms contained
-------
8.57-8
in 2 ml of the sample. From this the concentration in the
original sample can be calculated.
If the plot does not result in a straight line, a non-
linear interference is present. This can sometimes be overcome
by dilution.
-------
8.57-9
APPENDIX
DISPOSAL OF IONIC MERCURY IN SOLUTION
1. Bring the pH of the solution to neutral or basic by
adding sodium carbonate. Sodium hydroxide may have to
be added if neutralization cannot be achieved with sodium
carbonate.
2. Add granular zinc or magnesium as follows: For every
100 grams of either mercurous or mercuric chloride
present in the solution, add 110 grams zinc or 40 grams
magnesium. This achieves a 4 molar excess.
3. Stir the solution for 24 hours in a hood. CAUTION:
Hydrogen gas will be released during this process.
4. After 24 hours the solid material (mercury amalgam)
will have separated. Decant and discard the supernatant
*
liquid to the sewer.
5. Quantitatively transfer the solid material to a
convenient container and allow to dry.
The following companies are among those that have
recycled mercury in the past:
These companies may be able to supply a steel flask of
76 Ibs. capacity which can be used for storage and shipment
of contaminated metal.
1) Bethlehem Apparatus Co., Inc.
Front and Depot Sts.
Hellertown, PA 18055 Tel: (215) 838-7034
-------
8.57-10
2) Goldsmith Division, National Lead Co.
Ill North Wabash
Chicago, 111. 60602 Tel: (312) 726-0232
3) Wood Ridge Chemical Corp.
Park Place East
Wood Ridge, N.J. 07075 Tel: (201) 939-4600
4) Quicksilver Products, Inc.
350 Brannon Street
San Francisco, Cal. 94107 Tel: (415) 781-1988
-------
8.58-1
Method 8.58
NICKEL
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of nickel in a waste
or Extraction Procedure Extract.
Summary of Methods
A sample is digested using nitric acid and the concen-
tration of nickel is measured using either a flame or graphite
furnace equipped atomic absorption spectrophotometer . The
flame method is most accurate when employed with solutions
containing 0.3-5 mg Ni/liter, while the graphite furnace
procedure is best suited for solutions containing 5-100 mg/Ni
liter.
-------
8.58-2
Direct Aspiration
Reagents
1. Air, cleaned and dried through a suitable filter to
remove oil, water and other foreign substances. The
source may be a compressor or a cylinder of industrial
grade compressed air.
2. Acetylene should be of high purity. Acetone, which is
always present in acetylene cylinders, can be prevented
from entering and damaging the burner head by replacing
a cylinder when its pressure has fallen to 7 kg/cm^ (100
psig) acetylene.
3. Nitric acid, concentrated
4. Nickel Standard Stock solution, (1000 mg/liter):
Dissolve 4.9532 g nickel nitrate hexahydrate (Ni(NO.;j) *
6H20), analytical reagent grade, in distilled deionized
water containing 1.5 ml concentrated HN3/liter. Bring to
volume in a 1 liter volumetric flask using the same
water. (1 ml * 1 mg Ni)
Procedure
Sample Preparation
1. a. Transfer 100 gms (or in the case of EP extracts and
100 ml) of well-mixed sample to a 250 ml beaker .
Add 3 ml concentrated HN03. Place the beaker on
a hot plate and evaporate to near dryness, cautiously,
so that the sample does not boil. Cool, and add
another 3 ml concentrated HN03. Cover the beaker with a
watch glass and return to the hot plate. Heat so as
-------
8.58-3
to produce a gentle refluxing. Continue adding 3 ml
portions of HN03 until the sample is light in color
and no longer changes in color. Evaporate to near
dryness and cool. Add 5 ml of 1:1 HNC>3 and warm
the beaker in order to dissolve any precipitates.
b. Cool and bring to 100 ml with distilled deionized
water .
2. If particulates such as silicates remain in the sample,
it must be centrifuged and the supernatant aspirated.
3. Prepare working standards from stock solution. If it
is desired to work in the optimum concentration range, the
following is suggested:
a. Transfer 10 ml stock solution to a 100 ml volu-
metric flask. Bring to volume with distilled
deionized water containing 1.5 ml concentrated HN03/liter
Concentration: 100 mg/liter.
b. Transfer 0, 0.5, 1.0, 2.0, 4.0, and 5.0 ml
solution from a) to separate 100 ml volumetric
flasks. Bring to volume with distilled deionized
water containing 5 ml HNOg/liter. The concentrations
of these standards will be 0, 0.5, 1, 2, 4 and
5 mg Ni/liter .
Standard Addition
1. a. Take the 2, 4 and 5 mg/liter standards and pipet 5 ml
from each into separate 10 ml volumetric flasks.
Add to each 2 ml of the prepared sample. Bring
to volume with distilled deionized water containing
-------
8.58-4
5 ml HN03 per liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Bring to volume with distilled deionlzed
water containing 5 ml HN03/liter. This is the
blank.
Note: The absorbance from the blank will be 1/5 that produced
by the prepared sample. The absorbance from the spiked
standard will be 1/2 that produced by the standard plus the
contribution from 1/5 of the prepared sample. Keeping these
in mind, the correct dilutions to produce optimum absorbance
can be judged.
Instrument Operation
Wavelength: 232.0 nanometers (352.4 nm may be used. It is
less susceptible to interference, but is somewhat
less sensitive.)
Optimum Concentration Range: 0.3-5 mg/liter
Lower Detection Limit: 0.04 mg/liter
Fuel: Acetylene
Oxidant: Air
Type of Flame: Oxidizing
A general outline for instrument operation is given in
section 8.49. Follow the manufacturer's instructions
for the Spectrophotometer being used.
Quantification
The absorbance of spiked samples and blank vs. the
concentrations are plotted accordng to the Method of Standard
-------
8.58-5
Additions as defined in section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line, a non-
linear interference is present. This can sometimes be over-
come by dilution, or addition of other reagents, if there is
some knowledge about the waste.
-------
8.58-6
Graphite Furnace
Reagents
Prepare working standards and blank to bracket the range
5-100 mg Ni/liter by making appropriate dilutions of the
standards described in the direct aspiration method.
Procedure
The Sample is digested, standard additions are prepared,
and the analysis conducted as described under the direct
aspiration method.
Instrument Operation
Wave length: 232.0 nanometers
Optimum Concentration Range: 5-100 ug/liter
Lower Detection Limit: 1.0 ug/liter
Purge Gas: Argon or nitrogen
Drying Time and Temp: 30 sec at 125° C
Ashing Time and Temp: 30 sec at 900° C
Atomizing Time and Temp: 10 sec at 2700° C
These conditions are based on a 20 ul injection, con-
tinuous flow purge gas, and non-pyrolytic graphite on a
Perkin Elmer model HGA 2100 furnace. Other equipment
will have different requirements. Follow the manufacturer's
manual.
-------
8.59-1
Method 8.59
SELENIUM
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of selenium in a
waste or an Extraction Procedure Extract. While both methods
are generally aceptable for RCRA type samples, the graphite
furnace method is simpler to use than the hydride. However
it is sensitive to chloride interference when the concentration
is greater than 800 mg/liter. The presence of sulfate in
concentrations greater than 200 mg/liter will also interfere.
If it does not exceed 2000 mg/liter the sulfate can be suppressed.
The hydride method requires greater preparation and is sensitive
to high concentrations of chromium, copper, mercury, silver
cobalt and molybdenum.
Summary of Methods
In the furnace method the sample is digested using
nitric acid and hydrogen peroxide. The method is most
accurate for solutions containing 5 - 100 ug Selenium/liter.
In the gaseous hydride method the sample is digested
using nitric and sulfuric acids. The selenium is then reduced
from the +4 to the +2 oxidation state with stannous chloride,
and converted to the hydride SeHg, using zinc metal. The
hydride is swept into an argon entrained hydrogen flame for
analysis.
-------
8.59-2
It is most accurate when employed for solutions containing
2 - 20 ug Selenium/liter. However high concentrations of
chromium, copper, mercury, silver, cobalt and molybdenum can
interfere in the analysis.
Graphite Furnace Method
Reagents
1. Standard Stock Solution
1000 ug/liter solution may be purchased, or prepared
as follows:
Dissolve 0.3453g of selenous acid (assay 94.6% ^8003)
in distilled deionized water. Add to a 200 ml volumetric
flask and bring to volume (1 ml » 1 mg Se).
2. Nickel Nitrate Solution, 5%
Dissolve 24.780g of ACS reagent grade Ni(N03)2 * 6H20
in distilled deionized water and make up to 100 ml.
3. Nickel Nitrate Solution, 1%
Dilute 20 ml of the 5% solution to 100 ml with distilled
deionized water.
4. 30% Hydrogen Peroxide
5. Sodium Hydroxide
6. Concentrated Nitric Acid
Procedure
1. Sample Preparation
a. Transfer 100 gms(or in the case of EP extracts
100 ml) of well-mixed sample to a 250 ml beaker
-------
8.59-3
and add 2 ml of 30% H202 and 1 ml of concentrated
HNOj. Heat on a hot plate for 1 hour at 95°C or
until the volume is less than 50 ml.
b. If particulate material remains in the sample after oxida-
tion, add additional HN03 and 1^02 and digest until no
changes are noted in the amount of particulate matter
remaining.
c. At this point cool and bring to 50 ml with distilled
deionized water. The mixture should be centrifuged
and only the supernatant used for the remaining
steps of the analysis.
d. Transfer quantitatively 25 ml of this solution
into a 50 ml volumetric flask. Add 5 ml of 1% nickel
nitrate solution and bring to 50 ml with distilled
deionized water.
No_te; If sulfate is present at concentrations of more
than 200 mg/1 add 10 ml of 5% nickel nitrate solution
instead of 5 ml of 1% solution.
2. Prepare standards from stock solution. The following
provides standards in the optimum working range:
a. Pipet 1 ml stock solution into a 1 liter volumetric
flask. Bring to volume with distilled deionized
water containing 1.5 ml concentrated HN03 Per
liter. The concentration of this solution is 1000
ug Se/liter (1 ml - 1 ug Se)
b. Prepare 6 working standards by transferring 0, 1.0,
2.5, 5.0, 7.5 and 10.0 ml from a. into 100 ml volumetric
flasks. Add 1 ml concentrated HN03, 2 ml 30%
-------
8.59-4
and 2 ml 5% nickel nitrate solution. Bring to volume
with distilled deionized water. The concentrations
of these working standards are 0, 10, 25, 50, 75,
and 100 ug Se/liter. They are ready for injection.
Note! If sulfate is present in the sample at concen-
trations of more than 200 mg/1 add 20 ml of 5% nickel
nitrate solution instead of 2 ml before bringing to
volume.
3. Standard Addition
a. Take the 50, 75 and 100 ug/liter standards and
pipet 5 ml from each into separate 10 ml volumetric
flasks. Add to each 2 ml of the prepared sample.
Add 0.3 ml of 1% nickel nitrate solution and 0.3
ml H202 solution. Bring to volume with distilled
deionized water.
b. Add 2 ml of prepared sample to a 10 ml volumetric
flask. Add 0.3 ml of 1% nickel nitrate solution
and 0.3 ml 1^2®?. solution. Bring to volume
with distilled deionized water. This is the
blank.
Note; that the absorbance from the blank will
be 1/5 that producd by the prepared sample. The
absorbance from the spiked standards will be 1/2
that produced by the standards alone plus the
contribution from 1/5 of the prepared sample.
Keeping these in mind, the correct dilutions to
produce optimum absorbance can be judged.
-------
8.59-5
Instrument Operation
Wavelength: 196.0 nanometers
Optimum Concentration Range: 5 - 100 ug/liter
Lower Detection Limit: 2 ug/liter
Drying Time and Temperature: 30 seconds at 125°C
Ashing Time and Temperature: 30 seconds at 1,200°C
Atomizing Time and Temperature: 10 seconds at 2,700°C
Pure Gas: Argon
These conditions are based on a 20 ul injection,
continuous flow purge gas, and non-pyrolytic graphite in a
Perkin Elmer Model HGA 2100 furnace. Other equipment will
have different requirements. Follow the manufacturer's
manual.
Quantification
The absorbance of spiked samples and blank vs.
the concentrations are plotted according to the Method
of Standard Additions as defined in section 8.49. The
extrapolated value will be 1/5 the concentration of the
original sample.
If the plot does not result in a straight line,
non-linear interference is present. This can sometimes be
overcome by dilution, or additon of other reagents if there
is some knowledge about the waste.
-------
^.59-6
Gaseous Hydride Method
Reagents
1. Stannous Chloride Solution: Dissolve lOOg SnCl2 in
100 ml concentrated HC1.
2. Zinc slurry: Add 50g zinc metal dust (200 mesh)
to to 100 ml deionized distilled water.
3. Diluent: Add 100 ml 18N H2S04 and 400 ml concentrated
HC1 to 400 ml deionized distilled water In a 1-liter
volumetric flask and bring to volume with deionized
distilled water.
4. Standard Stock Solution: 1000 mg/liter solution may be
purchased, or prepared as follows: Dissolve 0.3453g of
selenious acid (assay 94.6% of H2Se03) in distilled
deionized water. Add to a 200 ml volumetric flask and
bring to volume (1 ml » 1 mg Se).
Apparatus
1. Atomic Absorption Spectrophotometer with Boling Burner.
2. Flow Meter, capable of measuring 1 1/min, such as that
used for auxiliary argon.*•
3. Medicine Dropper, capable of delivering 1.5 ml,
fitted into a size "0" rubber stopper.
4. Reaction Flask, a pear-shaped vessel with side arm and
50 ml capacity, both arms having 1 14/20 joint.2
5. Special Gas Inlet-Outlet Tube, constructed from a micro
cold finger condenser-* by cutting off the portion
below the S 14/20 ground glass joint.
1 Gilmont No. 12 or equivalent
2 Scientific Glass JM-5835 or equivalent
3 Scientific Glass JM-5325 or equivalent
-------
8.59-7
6. Magnetic Stirrer, strong enough to homogenize the zinc
slurry.
7. Drying Tube, 100-mm-long polyethylene tube filled with
glass wool to keep particulate matter out of the burner
Apparatus Set-Up
JM-3325
Medicine
Dropper in
Size "0"
Rubber
Stopper
Argon
Flow
Meter
•JM-5835
(Auxiliary Air)
Argon
(Nebulizer
Air)
Connect the apparatus with the burner as shown above.
Connect the outlet of the reaction vessel to the
auxiliary oxidant input of the burner with tygon
tubing. Connect the inlet of the reaction vessel
to the outlet side of the auxiliary oxident (argon
supply) control valve of the instrument.
-------
8.59-8
Procedure
1. Sample Preparation
To a 50 gm sample (or in the case of EP extracts a
50 ml sample) add 10 ml concentrated HN03 and 12
ml of 18 N 112804. Evaporate the sample on a hot plate
until white 803 fumes are observed (a volume of about
20 ml). Do not let it char. If it chars, stop the
digestion, cool and add additional HNOg. Maintain an
excess of HN03 (evidence of brown fumes) and do not
let the solution darken, because selenium may be reduced
and lost. When the sample remains colorless or straw
yellow during evolution of 803 fumes, the digestion is
complete.
Cool the sample, add about 25 ml distilled deionized
water and again evaporate to 803 fumes just to expel
oxides of nitrogen. Cool. Add 40 ml of concentrated
HCl and bring to a volume of 100 ml with distilled
deionized water.
2. Prepare working standards from the standard stock solution.
The following provides standards in the optimum working
range:
a. Pipet 1 ml stock solution into a 1 liter volumetric
flask. Bring to volume with distilled deionized
water containing 1.5 ml concentrated HN03/liter.
The concentration of this solution is 1 mg Se/liter.
(1 ml = 1 ug Se)
b. Prepare 6 working standards by transferring 0, 0.5,
1.0, 1.5, 2.0 and 2.5 ml from a. into 100 ml volumetric
-------
8.59-9
flasks. Bring to volume with diluent. The
concentrations of these working standards are 0, 5,
10, 15, 20 and 25 ug Se/liter.
3. Standard Additions
a. Take the 15, 20, and 25 ug standards and transfer
quantitatively 25 ml from each into separate 50 ml
volumetric flasks. Add to each 10 ml of the prepared
sample. Bring to volume with distilled deionized
water containing 1.5 ml HN03/liter.
b. Add 10 ml of prepared sample to a 50 ml volumetric flask.
Bring to volume with distilled deionized water containing
1.5 ml HN03 per liter. This is the blank.
Instrument Operation
Fuel: Argon-hydrogen flame
Wavelength: 196.0 nanometers
Optimum Concentration Range: 2-20 ug Se/liter
Lower Detection Limit: 2 ug/liter
Turn on the argon and adjust the flow rate to about 8
liters/minute with auxiliary argon flow at 1 liter/min.
Turn on the hydrogen, adjust flow rate to about 7 liters/
minute and ignite. The flame is colorless. The hand may be
passed 1 ft. above the burner to detect heat, in order to
insure ignition.
Follow the instructions given with the atomic absorption
spectrophotometer .
-------
8.59-10
Treatment of Samples and Standards
Transfer a 25-ml portion of the digested sample or
standard to the reaction vessel. Add 0.5 ml SnCl2 solution.
Allow at least 10 minutes for the metal to be reduced to its
lowest oxidation state. Attach the reaction vessel to the
special gas inlet-outlet glassware. Fill the medicine
dropper with 1.50 ml zinc slurry that has been kept in suspension
with the magnetic stirrer. Firmly insert the stopper containing
the medicine droppper into the side neck of the reaction
vessel. Squeeze the bulb to introduce the zinc slurry into
the sample or standard solution. The metal hydride will
produce a peak almost immediately. When the recorder pen
returns part way to the base line, remove the reaction vessel.
Quantification
The spiked samples and blank vs. concentrations are
plotted according to the Method of Standard Additions, as
defined in section 8.49. The extrapolated value will be
1/10 the concentration of the original sample due to a dilution
during preparation.
If the plot does not result in a straight line, a non-linear
interference is present. This can sometimes be overcome by
dilution, or addition of other reagents if there is some
knowledge about the waste.
-------
8.60-1
Method 8.60
SILVER
Scope and Application
The following atomic absorption procedures are approved
methods for determining the concentration of Silver in a
waste or an Extraction Procedure Extract.
Summary of Method
The sample is digested using nitric and/or hydrochloric
acids and the concentration of silver measured using either a
flame or graphite furnace equipped atomic absorption spectro-
photometer. The flame method is most accurate when employed with
solutions containing 0.1 - A mg Ag/liter, while the graphite
furnace procedure is best suited for solutions containing 1-25
ug Ag/liter.
Precaution
Silver in concentrations of 0.4 to 1 mg/liter can cause
damage to liver, kidneys and spleen. Natural drinking waters
do not generally contain more than 2 ug/liter.
Comments
1. Silver samples and silver nitrate standards should not
be stored. Silver is light-sensitive and has a tendency
to plate out on the container walls. Discard solutions
after use.
2. The use of hydrochloric acid is avoided to prevent precipi-
tation of silver chloride.
3. If plating out or AgCl formation is suspected it can be
-------
8.60-2
redissolved by addition of cyanogen iodide. However,
this can only be done after digestion to prevent formation
of toxic hydrogen cyanide under acid conditions.
Direct Aspiration
Reagents
1. Air, cleaned and dried through a suitable filter to
remove oil, water, and other foreign substances. The
source may be a compressor or a cylinder of industrial
grade compressed air.
2. Acetylene should be of high purity. Acetone, which is
always present in acetylene cylinders, can be prevented
from entering and damaging the burner head by replacing
a cylinder when its pressure has fallen to 7 kg/cm^
(100 pslg) acetylene.
3. Concentrated Nitric Acid (HN03>, Ultrex grade.
A. Concentrated Ammonium hydroxide (NH40H)
5. Silver Standard Stock Solution; (1000 mg/liter):
Dissolve 0.7874 g anhydrous silver nitrate (AgN03>
analytical reagent grade, in deionized distilled water.
Add 5 ml cone HN03 and bring to volume in a 500 ml
volumetric flask. 1 ml = 1 mg Ag.
6. Iodine solution, IN:
Dissolve 20 grams potassium iodide, (KI), analytical
reagent grade, in 50 ml distilled deionized water. Add
12.7 grams iodine (I2)> analytical reagent grade, and
dilute to 100 ml. Place in a brown bottle.
-------
8.60-3
7. Cyanogen Iodide solution:
To 50 ml deionized distilled water add 4.0 ml concentrated
NH4
-------
8.60-4
c. Transfer quantitatively to a 100 ml volumetric flask
and bring to volume with distilled deionized water.
3. If there are particulates such as silicates remaining in
the sample it must be centrifuged and only the supernatant
aspirated.
4. Prepare working standards from stock solution. To bracket
the optimum concentration range the following is suggested:
a. Transfer 10 ml stock solution to a 100 ml volumetric
flask and bring to volume with distilled deionized
water containing 10 ml HN03/liter. Concentration:
100 ing/liter.
Note: If, in the sample preparation, it was necessary
to add cyanogen iodide to insure redissolving of precipi-
tated silver, then the standards must be treated in the
same manner. Do not add acidified water in step 4. a.
Transfer 10 ml of stock solution to a small beaker. Add
distilled deionized water to make about 80 ml. Make the
solution basic (pH above 7) with ammonium hydroxide.
Rinse the pH meter electrodes into the solution with
distilled deionized water. Add 1 ml cyanogen, iodide and
allow to stand 1 hour. Transfer quantitatively to a 100
ml volumetric flask and bring to volume with distilled
deionized water.
-------
8.60-5
b. Transfer 0, 0.5, 1.0, 2.0, 3.0 and 4.0 ml from a. to
separate 100 ml volumetric flasks. Bring to volume
with distilled deionized water containing 10 ml
HN03/liter. The concentrations of these working
standards will be 0, 0.5, 1.0, 2.0, 3.0, and 4.0 mg
Ag/liter.
Note; If a. was treated with cyanogen iodide these
working standards must be brought to volume with distilled
deionized water made basic with ammonium hydroxide and
containing 10 ml cyanogen iodide/liter.
Standard Addition
a. Take the 2, 3, and 4 mg standards and pipet 5 ml from
each into separate 10 ml volumetric flasks. Add to
each 2 ml of the prepared sample. Bring to volume
with distilled deionized water containing 10 ml
HN03/liter.
b. Add 2 ml of prepared sample to a 10 ml volumetric flask.
Bring to volume with distilled deionized water containing
10 ml HN03 per liter. This is the blank.
Note; For a sample treated with cyanogen iodide these
standard additions are brought to volume with distilled
deionized water made basic with ammonium hydroxide
and containing 10 ml cyanogen iodide/liter.
Note; The absorbance from the blank b. will be 1/5
that produced by the prepared sample. The absorption
from the spiked standards will be 1/2 that produced by the
standards plus the contribution from 1/5 of the prepared
-------
8.60-6
sample. Keeping these in mind the correct dilutions
to produce optimum absorbance can be judged.
Instrument Operation
Wavelength: 328.1 nanometers
Optimum Concentration Range: 0.1-4 mg/liter
Lower Detection Limit: 0.01 mg/liter
Fuel: Acetylene
Oxidant: Air
Type of flame: oxidizing
A general outline for instrument operation is given
in section 8.49. Follow the manufacturer's instructions
for the model being used.
Quantification
The absorbances of spiked samples and blank vs. the con-
centrations are plotted according to the Method of Standard
Additions as defined in section 8.49. The extrapolated
value will be 1/5 the concentration of the original sample.
If the plot does not result in a straight line a
nonlinear interference is present. This can sometimes be
overcome by dilution, or addition of other reagents if there
is some knowledge about the waste.
Records
It is recommended that recorder outputs chart be retained
as a permanent record of the results and test conditions.
-------
8.55-1
Method 8.55
CYANIDE
Scope and Application
The following procedure can be used to determine
the concentration of inorganic cyanide in a waste
or leachate.
Inorganic cyanide may be present as either a simple soluble
salt (e.g., NaCN) or complex radical (e.g., K3(Fe(CN)g)).
These metal complexes have varying tendencies to dissociate
and form toxic hydrogen cyanide in the presence of acids.
It is such soluble cyanides that this method will address.
This method shall not be used to determine the "reactive"
cyanide content of wastes containing iron-cyanide complexes.
Summary
The waste is divided into two parts. One is chlorinated
to destroy susceptible complexes. Each part is then distilled
to remove interferences and analyzed for cyanide. The fraction
amenable to chlorination is determined by the difference in
values.
During the distillation, cyanide is converted to hydrogen
cyanide vapor which is trapped in a scrubber containing sodium
hydroxide solution. This solution is then titrated with
standard silver nitrate.
-------
8.55-
Interferences
1. Sulfides interfere with the titration. They may be
precipitated with cadmium.
2. Fatty acids form soaps under alkaline titration
conditions and interfere. They may be extracted with a
suitable solvent.
3. Oxidizing agents may decompose the cyanide. They
may be treated with ascorbic acid.
4. Thiocyanate presence will interfere by distilling
over in the procedure. This can be prevented by addition of
magnesium chloride.
5. Aldehydes and ketones may convert cyanide to
cyanohydrin under the acid distillation conditions.
Reagents
1. Calcium Hypochlorite solution: Dissolve 5 g of
hypochlorite (Ca(OCl)2) in 100 ml of distilled water.
2. Sodium Hydroxide solution (1.25N): Dissolve 50 g of
sodium hydroxide (NaOH) in distilled water and dilute to 1
liter.
3. Ascorbic acid: crystals.
4. Potassium Iodide-starch paper.
5. Lead acetate paper
6. Cadmium carbonate (powdered)
7. Hexane
8. Acetic acid solution (1 + 9)
-------
8.55-3
9. Concentrated H2S04
10. Silver Nitrate Standard solution (0.0192N):
Dry 5g AgN(>3 crystals to constant weight at 40°C.
Weigh out 3.2647 grams and dissolve in distilled
water. Dilute 1000ml. (1ml - Img CN)
11. Rhodanine Indicator solution:
Dissolve 20mg p-dimethyl-amino-benzalrhodanine in
100ml acetone.
12. Magnesium Chloride solution:
Weigh 510g MgC^.ei^O into a 1 liter volumetric
flask. Dissolve and bring to volume with distilled
water.
Apparatus;
1. Microburet, 5.0 ml, for titration.
2. Flasks, condenser, and tubing are needed as shown in
the diagram. The boiling flask should be of 1 liter size with
inlet tube and provision for a condenser. The gas absorber
may be a Fisher-Milligan scrubber. Assemble as shown in the
diagram.
-------
8.55-4
Procedure
1. Test and treat the sample as follows If It is known
or suspected that Interferences are present:
a. Sulfides:
If a drop of the distillate on lead acetate test
paper indicates the presence of sulfides, treat 25 ml more of
the sample than that required for the cyanide determination
with powdered cadmium carbonate. Yellow cadmium sulfide
precipitates if the sample contains sulfide. Repeat this
operation until a drop of the treated sample solution does
not darken the lead acetate test paper. Filter the solution
through a dry filter paper into a dry beaker, and from the
filtrate, measure the sample to use for analysis. Avoid a
large excess of cadmium and a long contact time in order to
minimize a loss by complexation or occlusion of cyanide on
the precipitated material. Sulfides should be removed prior
to preservation with sodium hydroxide.
b. Fatty acids:
1. Acidify the sample with acetic acid (1 + 9)
to pH 6.0 to 7.0.
Caution; This operation must be performed in the hood
and the sample left there until it can be made alkaline
again after the extraction has been performed.
-------
8.55-5
2. Extract with iso-octane, hexane, or chloroform
(preference in order named) with a solvent volume equal to
20% of the sample volume. One extraction is usually adequate
to reduce the fatty acids below the interference level.
Avoid multiple extractions or a long contact time at low pH
in order to keep the loss of HCN at a minimum. When the
extraction is completed, immediately raise the pH of the
sample to above 12 with NaOH solution.
c. Oxidizing agents:
1. Test a drop of the sample with potassium iodide-
starch test paper (Kl-starch paper); a blue color indicates
the need for treatment. Add ascorbic acid, a few crystals at
a time, until a drop of sample produces no color on the
indicator paper. Then add an additional 0.6 g of ascorbic
acid for each liter of sample volume.
2. Take two sample aliquots. To one 500.ml aliquot or
a volume diluted to 500 ml, add calcium hypochlorite solutior
dropwise while agitating and maintaining the pH between 11
and 12 with sodium hydroxide solution (1.25N).
Caution: The initial reaction product of alkaline chlori-
nation is the very toxic gas cyanogen chloride; therefore,
this reaction should be performed in a hood.
For convenience, the sample may be agitated in a 1 liter
beaker by means of a magnetic stirring device.
3. Test for residual chlorine with Kl-starch paper and
maintain this excess for one hour, continuing agitation. A
-------
8.55-6
distinct blue color on the test paper indicates a sufficient
chlorine level. If necessary, add additional hypochlorite
solution.
4. After one hour, add 0.5 g portions of ascorbic acid
until Kl-starch paper shows no residual chlorine. Add
an additional 0.5 g of ascorbic acid to insure the presence
of excess reducing agent.
Both the chlorinated and unchlorinated aliquots are now
separately distilled as follows:
5. Place 500 ml of sample, or an aliquot diluted to 500
ml in the 1 liter boiling flask. Add 50 ml of sodium hydrox-
ide (1.25N) to the absorbing tube and dilute if necessary with
distilled water to obtain an adequate depth of liquid in the
absorber. Connect the boiling flask, condenser, absorber and
trap in the train.
6. Start a slow stream of air entering the boiling
flask by adjusting the vacuum source. Adjust the vacuum so
that approximately one bubble of air per second enters the
boiling flask through the air inlet tube.
Caution; The bubble rate will not remain constant after
the reagents have been added and while heat is being applied
to the flask. It will be necessary to readjust the air rate
occasionally to prevent the solution in the boiling flask
from backing up into the air inlet tube.
-------
8.55-7
7. Slowly add 25 ml cone, sulfuric acid through the air
inlet tube. Rinse the tube with distilled water and allow
the air flow to mix the flask contents for 3 min. Pour 20ml
of magnesium chloride solution into the air inlet and wash
down with a stream of water.
8. Heat the solution to boiling, taking care to prevent
the solution from backing up into and overflowing from the
air inlet tube. Reflux for one hour. Turn off heat and
continue the airflow for at least 15 minutes. After cooling
the boiling flask, disconnect absorber and close off the
vacuum source.
9. Drain the solution from the absorber into a 250 ml
volumetric flask and bring up to volume with distilled water
washings from the absorber tube.
10. Titration:
a. Add the solution or an aliquot diluted to 250 ml
to a 500 ml erlenmeyer flask. Add 10-12 drops
Rhodanine indicator.
Titrate with standard silver nitrate to the first change
in color from yellow to brownish-pink. Titrate a distilled
water blank using the same amount of sodium hydroxide and
indicator as in the sample.
The analyst should familiarize himself with the end
point of the titration and the amount of indicator to be used
before actually titrating the samples. A 5 or 10 ml mlcroburet
-------
8.55-8
may be conveniently used to obtain precise titration.
b. Titrate a blank using distilled water.
Calculation;
(A - B)l,000 x 250
a. CN, mg/1 = ml orig. sample ml of aliquot titrated
where:
A. = volume of AgNC>3 for titration of sample.
B = volume of AgNOg for titration of blank.
b. Cyanide amenable to chlorination:
CN, mg/1 - A - B
A = mg/1 total cyanide in unchlorinated aliquot
B = mg/1 total cyanide in chlorinated aliquot
-------
8.55-9
ALLIHN CONDENSER
AIR INLET
ONE LITER •
BOILING FLASK
— CONNECTING TUBING
GAS ABSORBER
SUCTION
Figure 8.55-1
APPARATUS FOR CYANIDE DISTILLATION
-------
8.80-1
METHOD 8.80
DIRECT INJECTION
Direct injection is the simplest and most precise technique
for introducing a liquid into a measurement instrument since
transfer losses are eliminated. Samples are introduced into
the gas chromatograph by injection through the injection port,
into the liquid chromatograph using either the injection port
or sample loop, or into the atomic absorbtion spectrometer
by aspiration into the flame or injection into the graphite
furnace.
Direct injection should only be used with liquids free of
particulate matter and in the case of the gas chromatograph
also free of non-volatile species (e.g., lipids, polymers)
in order to prevent column degradation.
When using direct injection of wastes caution must be
exercised to prevent column or detector overload. To insure
that valid results are obtained the analyst should initially
conduct range finding experiments to insure that the sample
matrix is free enough from interfering species so that the
contaminant of Interest can be determined if present In the
sample at a levl of 1 ug/gm of sample level. If such sensi-
tivity cannot be obtained, then additional sample cleanup
(e.g., column chromatography, liquid-liquid extraction,
exclusion chromatography) must first be carried out.
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. . o*
(,(, Revision B 4/15/81 8.56-1
Method 8.56
TOTAL ORGANIC HALIDE
1. Scope and Application
1.1 This method is to be used for the determination of Total Organic
Hal ides as Cl" by carbon adsorption, and requires that all
samples be run in duplicate. Under conditions of duplicate
analysis, the reliable limit of sensitivity is 5 ug/L. Organic
halides as used in this method are defined as all organic species
containing chlorine, bromine and iodine that are adsorbed by
granular activated carbon under the conditions of the method.
Fluorine containing species are not determined by this method.
1.2 This is a microcoulometric-titration detection method applicable to
the determination of the compound class listed above in drinking
and ground waters, as provided under 40 CFR 265.92.
1.3 Any modification of this method, beyond those expressly permitted,
shall be considered as major modifications subject to application
and approval of alternate test procedures under 40 CFR 260.21.
1.4 This method is restricted to use by, or under the supervision of,
analysts experienced in the operation of a pyrolysis/microcolumeter
and in the interpretation of the results.
2. Summary of Method
2.1 A sample of water that has been protected against the loss of
volatiles by the elimination of headspace in the sampling
container, and is free of undissolved solids, is passed through a
column containing 40 mg of activated carbon. The column is washed
-------
f-t
Revision B 4/15/81 8.56-2
to remove any trapped inorganic halides, .a/id, is then.D.vrolvzed to
convert the adsorbed organohaTides to a titratable species that can
be measured by a microcoulometric detector.
3. Interferences
3.1 Method interferences may be caused by contaminants, reagents,
glassware, and other sample processing hardware. All of these
materials must be routinely demonstrated to be free from
interferences under the conditions of the analysis by running
method blanks.
3.1.1 Glassware must be scrupulously cleaned. Clean all glassware
as soon as possible after use by treating with chromate
cleaning solution. This should be followed by detergent
washing in hot water. Rinse with tap water and distilled
water, drain dry, and heat in a muffle furnace at 400°C
for 15 to 30 minutes. Volumetric ware should not be heated
in a muffle furnace. Glassware should be sealed and stored
in a clean environment after drying and cooling, to prevent
any accumulation of dust or other contaminants.
3.1.2 The use of high purity reagents and gases help to minimize
interference problems.
3.2 Purity of the activated carbon must be verified t>efore use. Only
carbon samples which register less than 1000 ng/40 mg should be
used. The stock of activated carbon should be stored in its
granular form in a glass container with a Teflon seal. Exposure to
the air must be minimized, especially during and after milling and
sieving the activated carbon. No more than a two-week supply
-------
Revision B 4/15/81 8.56-3
should be prepared in advance. Protect carbon at all times from
all sources of halogenated organic vapors. Store prepared carbon
and packed columns in glass containers with Teflon seals.
3.3 This method is applicable to samples whose inorganic-halide
concentration does not exceed the organic-halide concentration by
more than 20,000 times.
4. Safety
The toxicity or carcinogenicity of each reagent 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-handling data sheets should also be made available to all
personnel involved in the chemical analysis.
5. Apparatus and Materials (All specifications are suggested. Catalog
numbers are included for illustration only).
5.1 Sampling equipment, for discrete or composite sampling
5.1.1 Grab-sample bottle - Amber glass, 250-ml, fitted with
Teflon-lined caps. Foil may be substituted for Teflon if
the sample is not corrosive. If amber bottles are not
available, protect samples from light. The container must
be washed and muffled at 400°C before use, to minimize
contamination.
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Revision B 4/15/81 8.56-4
5.2 Adsorption System
5.2.1 Dohrmann Adsorption Module (AD-2), or equivalent,
pressurized, sample and nitrate-wash reservoirs.
5.2.2 Adsorption columns - pyrex, 5 cm long X 6-mm OD X 2-mm ID.
5.2.3 Granular Activated Carbon (GAC) - Filtrasorb-400,
Calgon-APC, or equivalent, ground or milled, and screened to
a 100/200 mesh range. Upon combustion of 40 mg of GAC, the
apparent-halide background should be 1000-mg Cl~
equivalent or less.
5.2.4 Cerafelt (available from Johns-Manvilie), or equivalent -
Form this material into plugs using a 2-mm ID
stainless-steel borer with ejection rod (available from
Dohrmann) to hold 40 mg of GAC in the adsorption columns.
CAUTION: Do not touch this material with your fingers.
5.2.5 Column holders (available from Dohrman).
5.2.6 Volumetric flasks - 100-mL, 50-mL.
A general schematic of the adsorption system is shown in
Figure 1.
5.3 Dohrmann microcoulometric-titration system (MCTS-20 or DX-20), or
equivalent, containing the following components:
5.3.1 Boat sampler.
5.3.2 Pyrolysis furnace.
5.3.3 Microcoulometer with integrator.
5.3.4 Titration cell.
A general description of the analytical system is shown in
Figure 2.
5.4 Strip-Chart Recorder.
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,
Revision B 4/15/81 8.56^5
6. Reagents
6.1 Sodium sulfite - 0.1 M, ACS reagent grade (12.6 g/L).
6.2 Nitric acid - concentrated.
6.3 Nitrate-Wash Solution (5000 mg NO^/L) - Prepare a nitrate-wash
solution by transferring approximately 8.2 gm of potassium nitrate
into a 1-litre volumetric flask and diluting to volume with reagent
water.
6.4 Carbon dioxide - gas, 99.9% purity.
6.5 Oxygen - 99.955 purity.
6.6 Nitrogen - prepurified.
6.7 70/6 Acetic acid in water - Dilute 7 volumes of acetic acid with 3
volumes of water.
6.8 Trichlorophenol solution, stock (1 uL = 10 yg Cl") - Prepare a
stock solution by weighing accurately 1.856 gm of trichlorophenol
into a 100-mL volumetric flask. Dilute to volume with methanol.
6.9 Trichlorophenol solution, calibration (1 uL = 500 ng Cl") -
Dilute 5 ml of the trichlorophenol stock solution to 100 ml with
methanol.
6.10 Trichlorophenol standard, instrument-calibration - First, nitrate
wash a single column packed with 40 mg of activated carbon as
instructed for sample analysis, and then inject the column with
10 uL of the calibration solution.
6.11 Trichlorophenol standard, adsorption-efficiency (100 ug C1~/L) -
Prepare a adsorption-efficiency standard by injecting 10 vl of
stock solution into 1 liter of reagent water.
6.12 Reagent water - Reagent water is defined as a water in which an
-------
Revision B 4/15/81 8.56-6
interferent is not observed at the method detection limit of each
parameter of interest.
6.13 Blank standard - The reagent water used to prepare the calibration
standard should be used as the blank standard.
7. Calibration
7.1 Check the adsorption efficiency of each newly-prepared batch of
carbon by analyzing 100 mL of the adsorption-efficiency standard,
in duplicate, along with duplicates of the blank standard. The net
recovery should be within 5% of the standard value.
7.2 Nitrate-wash blanks (Method Blanks) - Establish the repeatability
of the method background each day by first analyzing several
nitrate-wash blanks. Monitor this background by spacing nitrate-
wash blanks between each group of eight pyrolysis determinations.
7.2.1 The nitrate-wash blank values are obtained on single columns
packed with 40 mg of activated carbon. Wash with the
nitrate solution as instructed for sample analysis, and then
pyrolyze the carbon.
7.3 Pyrolyze duplicate instrument-calibration standards and the blank
standard each day before beginning sample analysis. The net
response to the calibration-standard should be within 3% of the
calibration-standard value. Repeat analysis of the
instrument-calibration standard after each group of eight pyrolysis
determinations, and before resuming sample analysis after cleaning
or reconditioning the titration cell or pyrolysis system.
8. Sample Preparation
8.1 Special care should be taken in the handling of the sample to
-------
Revision B 4/15/81 8.56-7
minimize the loss of volatile organohalides. The adsorption
procedure should be performed simultaneously on duplicates.
8.2 Reduce residual chlorine by the addition of sulfite (1 ml of 0.1 M
per liter of sample). Addition of sulfite should be done at the
time of sampling 1f the analysis is meant to determine the TOX
concentration at the time of sampling. It shou-ld be recognized
that TOX may increase on storage of the sample. Samples should be
stored at 4°C without headspace.
8.3 Adjust pH of the sample to approximately 2 with concentrated HN03
just prior to adding the sample to the reservoir.
9. Adsorption Procedure
9.1 Connect two columns in series, each containing 40 mg of
100/200-mesh activated carbon.
9.2 Fill the sample reservoir, and pass a metered amount of sample
through the activated-carbon columns at a rate, of approximately
3 mL/min. NOTE: 100 ml of sample is the preferred volume for
concentrations of TOX between 5 and 500 ug/L; 50 ml for 501 to 1000
yg/L, and 25 ml for 1001 to 2000 yg/L.
9.3 Wash the columns-in-series with 2 ml of the 5000-mg/L nitrate
solution at a rate of approximately 2 mL/min to displace inorganic
chloride ions.
10. Pyrolysis Procedure
10.1 The contents of each column is pyrolyzed separately. After rinsing
with the nitrate solution, the columns should be protected from the
atmosphere and other sources of contamination until ready for
further analysis.
-------
Revision B 4/15/81 8.56-8
10.2 Pyrolysis of the sample is accomplished in two stages. The
volatile components are pyrolyzed in a CCL-rich atmosphere at a
low temperature to assure the conversion of brominated
trihalomethanes to a titratable species. The less volatile
components are then pyrolyzed at a high temperature in an (L-rich
atmosphere.
NOTE: The quartz sampling boat should have been previously muffled
at 800°C for at least 2 to 4 minutes as in a previous analysis,
and should be cleaned of any residue by vacuuming.
10.3 Transfer the contents of each column to the quartz boat for
individual analysis.
10.4 If the Dohrmann MC-1 is used for pyrolysis, manual instructions are
followed for gas flow regulation. If the MCT-20 is used, the
information on the diagram in Figure 3 is used for gas flow
regulation.
10.5 Position the sample for 2 minutes in the 200°C zone of the
pyrolysis tube. For the MCTS-20, the boat is positioned just
outside the furnace entrance.
10.6 After 2 minutes, advance the boat into the 800°C zone (center) of
the pyrolysis furnace. This second and final stage of pyrolysis
may require from 6 to 10 minutes to complete.
11. Detection
The effluent gases are directly analyzed in the microcoulometric-titra-
tion cell. Carefully follow manual instructions for optimizing cell
performance.
-------
Revision B 4/15/81 8.56-9
12. Breakthrough
Because the background bias can be of such an unpredictable nature, it
can be especially difficult to recognize the extent of breakthrough of
organohalides from one column to another. All second-column
measurements for a properly operating system should not exceed
10-percent of the two-column total measurement. If the 10-percent
figure is exceeded, one of three events can have happened. Either the
first column was overloaded and a legitimate measure of breakthrough was
obtained - in which case taking a smaller sample may be necessary; or
channeling or some other failure occurred - in which case the sample may
need to be rerun; or a high, random, bias occurred and the result should
be rejected and the sample rerun. Because knowing which event has
occurred may not be possible, a sample analysis should be repeated often
enough to gain confidence in results. As a general rule, any analyses
that is rejected should be repeated whenever sample is available. In
the event that the second-column measurement is equal to or less than
the nitrate-wash blank value, the second-column value should be
disregarded.
13. Quality Control
13.1 Before performing any analyses, the analyst must demonstrate the
ability to generate acceptable accuracy and precision with this
procedure by the analysis of appropriate quality-control check
samples.
13.2 The laboratory must develop and maintain a statement of method
accuracy for their laboratory. The laboratory should update the
accuracy statement regularly as new recovery measurements are made.
-------
Revision B 4/15/81 8.56-10
13.3 It is recommended that the laboratory adopt additional
quality-assurance practices for use with this method. The specific
practices that would be most productive will depend upon the needs
of the laboratory and the nature of the samples. Field duplicates
may be analyzed to monitor the precision of the sampling
technique. Whenever possible, the laboratory should perform
analysis of standard reference materials and participate in
relevant performance-evaluation studies.
14. Calculations
OX as Cl" is calculated using the following formula:
(Cj- c3) * (c2 - c3 ) u yg/L Total Organ1c Halide
where:
C, = ug Cl" on the first column in series
C« = ug Cl" on the second column in series
C3 = predetermined, daily, average, method-blank value
(nitrate-wash blank for a 40-mg carbon column)
V = the sample volume in L
15. Accuracy and Precision
These procedures have been applied to a large number of drinking-water
samples. The results of these analysis are summarized in Tables I and
II.
16. Reference
Oressman, R., Najar, G., Redzikowski, R., paper presented at the
Proceedings of the American Water Works Association Water Quality
Technology Conference, Philadelphia, Dec. 1979.
-------
Revision B 4/15/81 8.56-11
TABLE I
PRECISION AND ACCURACY DATA FOR MODEL COMPOUNDS
Model
Compound
CHCU
0
CHBrCl2
CHBr2Cl
CHBr3
Pentachlorophenol
Dose
ug/L
98
160
155
160
120
Dose
as ug/L Cl
88
106
79
67
80
Average
% Recovery
89
98
86
111
93
Standard
Deviation
14
9
n
8
9
No. of
Replicate;
10
11
13
11
7
TABLE II
PRECISION DATA ON TAP WATER ANALYSIS
Sample
A
B
C
Avg. halide
ug Cl/L
71
94
191
Standard
Deviation
4.3
7.0
6.1
No. of
Replicates
8
6
4
-------
Revision B 4/15/81 8.56-12
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Revision B 4/15/81 8.56-13
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-------
Revision B 4/15/81 8.56-14
-------
Revision B 4/15/81 8.57-1
Method 8.57
Sulfides
1. Scope and Application
1.1 This method is applicable to the measurement of total
and dissolved sulfides in drinking, surface and saline
waters, domestic and industrial wastes.
1.2 Acid insoluble sulfides are not measured by this method.
Copper sulfide is the only common sulfide in this class.
1.3 This method is suitable for the measurement of sulfide
in concentrations above 1 mg/1
2. Summary of Method
2.1 Excess iodine is added to a sample which may or may
not have been treated with zinc acetate to produce zinc
sulfide. The iodine oxidizes the sulfide to sulfur
under acidic conditions. The excess iodine is back
titrated with sodium thiosulfate or phenylarsine oxide.
3. Comments
3.1 Reduced sulfur compounds, such as sulfite, thiosulfate
and hydrosulfite, which decompose in acid may yield erratic
results. Also, volatile iodine-consuming substances
will give high results.
3.2 Samples must be taken with a minimum of aeration.
Sulfide may be volitilized by aeration and any ozygen
inadvertently added to the sample may convert sulfide
to an unmeasurable form.
3.3 If the sample is not preserved with zinc acetate, the
analysis must start immediately. Similarly, the
measurement of dissolved sulfides must also be commenced
immediately.
4. Apparatus: Ordinary laboratory glassware
5. Reagents
5.1 Hydrochloric acid, HC1, 6N
5.2 Standard iodine solution, 0.0250 N: Dissolve 20 to 25 g
KI in a little water in a liter volumetric flask and add
3.2 g iodine. Allow to dissolve. Dilute to 1 liter and
standardize against 0.0250 N sodium thiosulfate or
phenylarsine oxide using a starch indicator.
5.3 Phenylarsine oxide 0.0250 N: commercially available.
5.4 Starch indicator: commercially available.
5.5 Procedure for standardization (see Residual Chlorine-
iodometric titration)
6. Procedure
6.1 Unprecipitated sample
6.1.1 Place a known amount of standard iodine solution
(5.2) into a 500 ml flask. The amount should
be estimated to be in excess of the amount of
sulfide expected.
6.1.2 Add distilled water, if necessary, to bring the
volume to approximately 20 ml.
6.1.3 Add 2 ml of 6N HC1 (5.1)
6.1.4 Pipet 200 ml of sample into the flask, keeping
the tip of the pipet below the surface of the
sample.
6.1.5 if the iodine color disappears, add more iodine
-------
/„ -
Revision B 4/15/81 8.57-2
until the color remains. Record the total
number of milliliters of the standard iodine used
in performing steps 6.1.1 and 6.1.5.
6.1.6 Titrate with reducing solution (0.0250 N sodium
thiosulfate or 0.0250 N phenylarsine oxide
solution (5.3)) using the starch indicator (5.4)
until the blue color disappears. Record the
number of milliliters used.
6.2 Precipitated samples
6.2.1 Add the reagents to the sample in the original
bottle. Perform steps 6.1.1, 6.1.3, 6.1.5, and
6.1.6.
6.3 Dewatered samples
6.3.1 Return the glass fibre filter paper which con-
tains the sample to the, original bottle. Add
200 ml of distilled water. Perform steps 6.1.1,
6.1.3, 6.1.5, and 6.1.6.
6.3.2 The calculations (7) should be based on the
original sample put throug the filter.
7. Calculations
7.1 One ml of 0.0250 N standard iodine solution (5.2)
reacts with 0.4 mg of sulfide present in the titration
vessel.
7.2 Use the formula
mg/1 sulfide = 400(A-B)/ml sample
where:
A=ml of 0.0250 N standard iodine solution (5.2)
B=ml of 0.0250 N standard reducing sodium
thiosulfate or phenylarsine oxide solution (5.3)
8. Precision and Accuracy
8.1 Precision and accuracy for this method have not been
determined.
HI.S. GOVERNMENT PRINTING OFFICE: 1981 341-082/248 1-3
-------
8.82-1
Method 8.82
HEADSPACE METHOD
Scope and Application
This method provides a procedure for the extraction of
volatile organic compounds in pastes and solids. The static
headspace technique is a simple method which allows large
numbers of samples to be analyzed in a relatively short period
of time. Because of the large variability and complicated
matrices of waste samples in the solid and paste forms,
detection limits for this method may vary widely among samples.
The method works best for compounds with boiling points less
than 125°C. Due to their low solubility, low molecular
weight compounds can only be detected at high concentrations
or at reduced pressure.
The sensitivity of this method will depend on the equili-
bria of the various compounds between the vapor and dissolved
phases .
Static Headspace Technique
Summary of Method
The waste is collected in sealed glass containers and
allowed to equilibrate at 90°C. A sample of the headspace
gas is withdrawn with a gas tight syringe for analysis by the
appropriate gas chromatographic method.
Apparatus
1. Gas-tight syringe - 5-cc.
2. Head space standard solutions - Prepare two standard solu-
tions of the compounds being determined at the 50-ng/ul and
-------
8.82-2
250-ng/ul coneentratioos. Standard solutions should be prepared
using methanol, methane, or other appropriate solvent. The
standard solutions should be stored at less than 0°C, then
allowed to warm to room temperature before dosing. Fresh
standards should be prepared weekly. Procedures for preparing
standards are outlined in the Purge and Trap Procedure of this
manual (Method 8.83).
3. Vials, 125 ml "Hypo-Vials" (Pierce Chemical Co., #12995), or
equivalent•
4. Septa, "Tuf-Bond" (Pierce #12720), or equivalent.
5. Seals, aluminum, (Pierce #13214), or equivalent.
6. Crimper, hand, (Pierce #13212), or equivalent.
Procedure
1. Place 10.0-g each of the well-mixed waste sample into five
separate 125-ml septum seal vials.
2. Dose one sample vial through the septum with 200-ul of the
50-ng/ul standard methanol solution. Dose a second vial with
200-ul of the 250-ng/ul standard.
3. Place the two dosed sample vials and one non-dosed sample into
a 90°C water bath for 1 hour. Store the two remaining samples
near 4°C for possible future analyses.
4. While maintaining the sample at 90°C, withdraw 2.0-ml of the
head gas with a gas tight syringe and analyze by injecting
into a GC, operating under the appropriate conditions for
the GC measurement method being used. Analyze all three
samples in exactly the same manner. Subtract the peak
areas of compounds found in the undosed sample from the
corresponding compounds contained in the dosed samples.
-------
8.82-3
If a positive response is noted then the waste has not been
demonstrated to be free of the contaminant of interest and
is thus not fundamentally different than the listed waste.
If no response is noted then the required sensitivity
lug/gm sample) of the procedure must be confirmed using
spiked samples.
Note; Standard quality assurance protocols should be employed,
including blanks, duplicates, and dosed samples, as described
in Section 10.
Bibliography
1. "Interim Methods for the Sampling and Analyses of Priority
Pollutants in Sediments and Fish Tissue," U.S. Environmental
Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268 [1980].
2. "Master Scheme for the Analysis of Organic Compounds in
Water, Part I: State-of-the-Art Review of Analytical
Operations," U.S. Environmental Protection Agency,
Environmental Research Laboratory, Athens, Georgia 30605.
3. "An EPA Manual for Organic Analysis Using Gas Chromatography
Mass Spectrometry," W.L. Budde and J.W. Eichelberger, U.S.
Environmental Protection Agency, Environmental Monitoring
Support Laboratory, Cincinnati, Ohio, 1979, EPA/600/8-79/006,
Order Number PB-297164.
-------
8.83-1
Method 8.83
PURGE AND TRAP METHOD
Scope and Application
This method covers a procedure for the extraction of pur-
geable organic compounds from aqueous liquids and free flowing
paste samples prior to gas chromatographic analysis.
The success of the extraction depends on partitioning
the compounds between the sample phase and gaseous headspace
phases. This partitioning is a function of temperature,
interfacial area, the volatility of the species being analyzed
for, its solubility in the liquid being purged, and the
volatility of the waste matrix. For highly volatile matrices,
direct injection preceded by dilution, if necessary, should be
used. For pastes, dilution of the sample until it becomes free
flowing is used to insure adequate interfacial area. The
success of this method also depends on the level of interferences
in the sample; results may vary due to the large variability
and complicated matrices of solid waste samples.
Summary of Method
An inert gas is bubbled through the sample contained in
a specially-designed purging chamber. This purging transfers
the volatile compounds from the liquid phase to the vapor
phase. The gaseous effluent is then swept through a short
sorbent tube where the organic compounds are trapped. After
purging is completed, the trap is heated and backflushed to
-------
8.83-2
desorb the compounds into a gas chromatograph for subsequent
identification and measurement.
Apparatus
1. Vial, with cap--40 ml capacity screw cap (Pierce #13075
or equivalent). Detergent wash and dry vial at 105°C for one
hour before use.
2. Septum, Teflon faced silicone (Pierce #12722 or equiva-
lent). Detergent wash and dry at 105°C for one hour before
use.
3. Purge and Trap Device; The purge and trap equipment
consists of three separate pieces of apparatus: a purging
device, a trap, and a desorber. The complete device is avail-
able commercially from several vendors or can be constructed
in the laboratory according to the specifications of Bellar
and Lichtenberg (1). The sorbent trap consists of a 1/8 in.
O.D. (0.105 in. I.D.) x 25 cm long stainless steel tube
packed with the appropriate absorbent as described in Table
8.83-1 (See Figures 8.83-1 through 8.83-4). 10-cm traps may
be used providing that the recoveries are demonstrated to be
comparable to the 25-cm traps.
Reagents
1. Trap Materials, see Table 8.83-1.
2. Activated carbon Filtrasorb 200 (Calgon Corp.) or equivalent
3. Organic-free water
-------
8.83-3
Organic-free water Is defined as water free of Inter-
ferences when employed in the purge and trap procedure.
It is generated by passing distilled or deionized water
through a carbon filter bed containing activated carbon.
A water system (Millipore Super-Q or equivalent) may
be used to generate organic-free deionized water. Organic-
free water may also be prepared by boiling deionized distilled
water for 15 minutes. Subsequently, while maintaining the
temperature at 90°C, bubble a contaminant-free inert gas
through the water for one hour. While still hot, transfer
the water to a narrow mouth screw cap bottle equipped
with a Teflon seal.
Procedures
1. Assemble the purge and trap device (see Figures 8.83-1
through 8.83-4). Purge parameters to be used depend on
the compounds being analyzed for; see Table 8.83-1.
Pack the trap as shown in Figure 8.83-2 and condition
overnight at a nominal 180°C by backflushing with an
inert gas flow of at least 20 ml/min. Daily, prior to
use, condition the traps for 10 minutes by backflushing
at 180°C.
2. Remove standards and samples from cold storage (approx-
imately an hour prior to an analysis) and bring to room
temperature by placing In a warm water bath at 20-25°C.
3. Adjust the purge gas (nitrogen or helium) flow rate
according to Table 8.83-1.
-------
8.83-4
4. Attach the trap inlet to the purging device, and
set the device to the purge mode. Open the syringe valve
located on the purging device sample introduction needle.
5. Remove the plunger from a 5 ml syringe and attach a closed
syringe valve. Open the sample bottle (or standard) and
carefully pour the sample into the syringe barrel until
it overflows.
Note; For pastes it may be necessary to dilute the sample
by adding a non-volatile solvent to the sample. In such
cases diluting can be performed in the purging device.
6. 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. Since this process
of taking an aliquot destroys the validity of the sample
for future analysis, the analyst should fill a second
syringe at this time to protect against possible loss of
data.
7. Add 5.0 ml of the spiking solution through the valve
bore, then close the valve.
8. Attach the syringe-valve assembly to the syringe valve
on the purging device. Open the syringe valve and inject
the sample into the purging chamber. Close both valves
and purge the sample for the time specified in Table
8.83-1. If method 8.02 will be used for analysis of the
sample, dry the trap by maintaining a flow rate of 40
ml/minute dry purge for 6 minutes.
-------
8.83-5
9. Attach the trap to the chromatograph, and adjust the
device to the desorb mode. Introduce the trapped materials
to the GC column by rapidly heating the trap to the backflush
temperature indicated in Table 8.83-1, while backflushing
the trap with an inert carrier gas at 20 to 60 ml/minute
for 4 minutes. If rapid heating cannot be achieved, 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 initial program temperature of 45
or 50°C.
10. While the trap is being desorbed into the gas chromato-
graph, clean the purging chamber. After the purging
device has been emptied, continue to allow the purge gas
to vent through the chamber until the frit is dry, and
ready for the next sample. After desorbing the sample
for four 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. Maintain the trap tempera-
ture at 180°C. After approximately seven 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.
Note; If this bake out step is omitted when using GC/MS
as the measurement technique, the amount of water entering
the GC/MS system will progressively increase causing
deterioration and potential shut down of the system.
-------
8.83-6
11. The analysis of blanks is most important in the purge
and trap technique since the purging device and the trap
can become contaminated by residues from very concentrated
samples or by vapors in the laboratory. Prepare blanks by
filling a sample bottle with organic-free water. Blanks
should be sealed, stored at 4°C, and analyzed with each
group of samples.
-------
References
1. "Determining Volatile Organics at Microgram-per-liter
Levels by Gas Chromatography," T.A. Bellar and J.J.
Lichtenberg, Journal. AWWA, 66, 739-744, Dec. 1974.
8.83-7
-------
8.83-8
Table 8.83-1
PURGE AND TRAP PARAMETERS
Analysis
Method*
Purge Gas
Purge Gas
Flow Rate
(ml/mln)
Purge Time
(minutes )
Purge
Temperature
(°C)
Desorbtlon
Gas Flow
Rate (ml/mln)
Sorbants To
8.01
Nitrogen or
Helium
40
11.0
180°
20
A
8.02 8.03
Nitrogen or Helium
Helium
40 20+1
12.0 30.0
180° 170°
20 20
B C
8.24
Nitrogen or
Helium
40
12.0
180°
20
D
Be Used in
Packing Tube
Porous polymer packing, 60/80 mesh,
chromatographic grade Tenax GC
(2,6-Diphenylene Oxide).
Three percent OV-1 on Chromosorb-W,
60/80 mesh.
Silica gel, 35-60 mesh Davison grade-15
or equivalent.
Coconut charcoal, 6/10 mesh, Barnaby
Chaney C.A. - 580-26 lot # M - 2649
or equivalent .
B
Porous polymer packing, 60/80 mesh
chromatographic grade Tenax GC
(2,6-Diphenylene Oxide).
Three percent OV-1 on Chromosorb-W,
60/80 mesh.
Silica gel, 35-60 mesh, Davison grade-15
or equivalent.
Porous polymer packing, 50/80 mesh,
chromatographic grade Tenax GC
(2,6-Diphenylene Oxide).
-------
8.83-9
Three percent OV-1 on Chromosorb-W,
60/80 mesh.
Silica gel, 35-60 mesh, Davison grade-15
or equivalent.
Porous polymer packing, 60/80 mesh,
chromatographic grade Tenax GC
(2,6-Diphenylene Oxide).
Three percent OV-1 on Chromosorb-W,
60/80 mesh.
Silica gel, 35-60 mesh, Davison grade-15
or equivalent.
*Measurement method to be employed for identification and
quantification
-------
8.83-10
OPTIONAL
FOAM
TRAP
0. D.
I—14MM 0. D
INLET « IN.
0.0.
0. D. EXIT
10WIM GLASS FRIT
MEDIUM POROSITY
SAMPLE INLET
2-WAY SYRINGE VALVE
•17CM. 20 GAUGE SYRINGE NEEDLE
. 0. D. RUBBER SEPTUM
—IQlCia. 0. D. I/IS IN. O.D.
fl/STAIWLESSSTB.
13X MOLECULAR
SIEVE PURGE
GAS FILTER
PURGE GAS
ROW
CONTROL
Figure 8.83-1
PURGING DEVICE
PACKING PROCEDURE
GLASS
WOOL
GRADE 15
SILICA GEL8CM
TENAX 15CM
35i OV-1 1CMJ ^
GLASS 5MM &
WOOL
CONSTRUCTION
COMPRESS10W
• FITTWCNL'T
AND FERRULES
RESISTANCE RTRE
WRAPPED SOLID
THERMOCOUPLE/
CONTROLLER
SENSOR
ELECTRONIC
TEMPERATURE
CONTROL
AND
PYROMETER
TUBING 25CU
0.105 IN. I.D.
0.125 IN. O.D.
STAINLESS STE
Figure 8.83-2
TRAP PACKINGS AND CONSTRUCTION
TO INCLUDE DESORB CAPABILITY
-------
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
GULATOR
PURGE GAS ...
FLOW CONTROL]^!,
13X MOLECULAR ^
SIEVE FILTER ~
COLUMN OVEN
, n n n n_L CONFIRMATORY COLUMN
DETECTOR
ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SB "CT10N VALVE
6-PORT ThAP INLET
VALVEJ RESISTANCE WIRE
4w£ THAP {OFF
ROW3 22°C
H£ATER
PURGING
DEVICE
Note:
ALL LINES BET.YEEJV
TRAP AND GC
SHOULD BE HEATED
TO BO°C.
Figure 8.83-3
SCHEMATIC OF PURGE AND TRAP DEVICE-PURGE MODE
8.83-10
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
PURGE GAS
FLOW CONTROL
LIQUID INJECTION PORTS
13X MOLECULAR^
iiEVE FILTER p
^OPTIONAL 4-PORT COLUMN
SaECTION VALVE
6-PORT TRAP INLET
COLUMN OVEN
nnnn I CONFIRMATORY COLUMN
iTn_L>TO DETECTOR
ITT ^-ANALYTICAL COLUUM
PURGING
DEVICE
* CONTROL
Note:
ALL LINES
TRAP AND GC
SHOULD BE HEATED
TO 80°C,
Figure 8.83-4
SCHEMATIC OF PURGE AND TRAP DEVICE DESORB MODE
-------
8.84-1
Method 8.84
SHAKE OUT PROCEDURE
Introduction
This procedure provides a method for treating liquid wastes
not soluble in the extraction solvent in order to extract the
organic species prior to measurement by chromatography or, if
necessary, further clean-up.
Summary of Method
Samples are made acid or alkaline, if necessary, then
extracted three times, with the appropriate solvent using vigo-
rous agitation. After the combined extracts are dried with
anhydrous sodium sulfate, they are concentrated in a Kuderna-
Danish Apparatus.
Apparatus
1. Separatory funnel - 250 ml, with Teflon stopcock.
2. Drying column - 20 mm ID Pyrex chromatographic column
with coarse frit.
3. Kuderna-Danish (K-D) Apparatus. [Kontes K-570000 or
equivalent.]
4. Boiling chips - solvent extracted, approximately 10/40
mesh.
5. Water bath - Heated, with concentric ring cover, capable
of temperature control ( + 2° C). The bath should be used in
a hood.
Reagents
1. Sodium hydroxide, (ACS) 10 N in distilled water.
-------
8.84-2
2. Sulfuric acid, (1+1), Mix equal volumes of concentrated
H2S04 (ACS) with distilled water.
3. Methylene chloride, acetone, 2-propanol, hexane, toluene-
Pesticide quality or equivalent.
4. Sodium sulfate (ACS) Granular, anhydrous (purified by
heating at 400° C for four hours in a shallow tray).
Procedure
1. Transfer 50 gms of sample to the separatory funnel.
2. Adjust the pH of the sample to that indicated in Table
8.84-1.
3. Add 50 ml of the appropriate extraction solvent
(see Table 8.84-1),
4. Seal and shake the separatory funnel for 60 seconds
with periodic venting to release vapor pressure.
5. Allow the phases to separate for a minimum of ten
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,
but may include stirring, filtration of the emulsion through
glass wool, or centrifugation.
6. Collect the extract and then repeat the extraction two more
times using fresh portions of solvent.
7. Combine the three extracts and discard the now extracted
waste.
-------
8.84-3
8. Filter the extract and then dry it by passing it through a
drying column containing 10 cm of anhydrous sodium sulfate.
9. Transfer the dried extract to the K-D apparatus. Add 1-2
clean boiling chips to the flask and attach a three-ball
Snyder column. Prewet the Snyder column by adding about 1
ml extraction solvent to the top. Place the K-D 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 and allow it to drain for at least 10 minutes
while cooling.
10. Transfer to 10 ml volumetric flask and dilute to volume.
-------
Table 8.84-1
EXTRACTION CONDITIONS
8.84-4
Compound
Acetonitrile
Acrolein
Acrylamlde
Acrylonltrlle
Benzene
|B en zo( a) anthracene
Benzo(a) pyrene
Benzotrichloride
Benzyl chloride
Benzo(b)fluoranthene
Bis (2-chloroethoxy-
methane)
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl)
ether
Carbon disulfide
Carbon tetrachloride
Chlordane
Chlorinated dibenzo
dioxins
Chlorinated biphenyls
Chloroacetaldehyde
Chlorobenzene
Extraction pH
NA
NA
NA
NA
NA
5-9
> 11
5-9 or > 11
5-9 or > 11
5-9
NA
NA
NA
NA
NA
5-9 or >11
6-8
5-9 or >11
NA
NA
Extraction
Solvent
NA
NA
NA
NA
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
NA
NA
NA
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
NA
NA = Method not applicable to analysis for this compound.
-------
EXTRACTION CONDITIONS cont . (2)
8.84-5
Compound
2-Chlorophenol
Chrysene
Creosote
Cresol(s)
Cresylic acid(s)
Dichlorobenzene(a)
Dichloroethane(s)
Dichloromethane
Dichlorophenoxy-
acetic acid
Dichloropropanol
2,4-Dimethylphenol
Dinltrobenzene
4,6-Dlnltro-O-cresol
2,4-Dinitro toluene
Endrln
Ethyl Ether
Formaldehyde
Formic Acid
Heptachlor
Hexachlorobenzene
Hexachloro butadiene
Hexachloroe thane
Extraction pH
12
5-9
5-9 or > 11
12
12
5-9 or > 11
NA
NA
> 7 or > 11
5-9 or > 11
12
5-9 or > 11
12
5-9 or > 11
5-9 or > 11
NA
NA
5-9 or > 11
5-9 or > 11
5-9 or > 11
5-9 or > 11
Extraction
Solvent
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
NA
Ethyl Ether or
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
-------
EXTRACTION CONDITIONS cont. (3)
8.84-6
Compound
Hexachlorocyclo-
pentadiene
Lindane
Maleic anhydride
Me t h a n o 1
Methomyl
Methyl ethyl ketone
Methyl isobutyl ketone
Naphthalene
Napthoquinone
Nit robenz ene
4-Nitrophenol
Paraldehyde (trimer of
acetaldehyde
Pentachlorophenol
Phenol
Phorate
Pho spho rodi thioic acid
esters
Phthalic anhydride
2-Picoline
Pyridine
Tetrachlorobenzene(s)
Tetrachloroethane(s)
Extraction pH
5-9 or > 11
5-9 or > 11
5-9 or > 11
NA
6.5 - 7.5
> 11
> 11
5-9 or > 11
5-9 or > 11
5-9
12
NA
12
12
6-8
5-9
5-9 or > 11
5-9 or > 11
5-9 or > 11
5-9 or > 11
'NA
Extrac tion
Solvent
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
-------
EXTRACTION CONDITIONS cont. (4)
8.84-7
Compound
Tetrachloroethene
Tetrachlorophenol
Toluene
Toluenediamlne
Toluene diisocyanate( s)
Toxaphene
Trlchloroethane
Trichloroethene(s)
Trichlorof luorome thane
Trichlorophenol(s)
2,4,5-TP(Silvex)
Trichloropropane
Vinyl chloride
Vlnylldene chloride
Xylene
Extraction pH
NA
12
NA
> 11
59
5-9 or > 11
NA
NA
NA
12
<7 or > 11
.
NA
NA
NA
NA
Extraction
Solvent
NA
Methylene Chloride
NA
Methylene Chloride
Methylene Chloride
Methylene Chloride
NA
NA
NA
Methylene Chloride
Ethyl Ether or
Methylene Chloride
NA
NA
NA
NA
-------
8.85-1
Method 8.85
SONICATION METHOD
Scope and Application
This method covers a procedure for the extraction of non-
volatile and semi-volatile organic compounds from solids. The
sonication produces solid disruption to ensure intimate contact
of the sample matrix with the extraction solvent.1
Summary of Method
A weighed sample of the solid waste is ground, mixed with
the extraction medium, then dispersed into the solvent using
sonication. The resulting solution may then be cleaned up further
or analyzed directly using the appropriate technique (Methods
8 .24 through 8 .25) .
Apparatus
1. Apparatus for Grinding.*t
The necessity for grinding and the choice of grinding
apparatus will depend on the physical and chemical charac-
teristics of the solid waste material in question. Any of
1 The high energy vibrations produced by this method may produce
artifacts and may drive off some semi-volatile compounds*
* Grinding is only necessary if the waste cannot either pass
through a 1-mm standard seive or be extracted through a 1-mm
diameter hole.
t Specific equipment listed in this method are for descriptive
purposes only. Equivalent equipment is available from other
manufactures and laboratory supply companies.
-------
8.85-2
the following grinders, or their equivalent, would be
suitable:
a. Fisher Mortar Model 155 Grinder or equivalent.
Fisher Scientific Co., Catalogue Number 8-323. This
grinder handles all except gummy, fibrous or oily
materials.
Sonication
For purposes of solid waste dispersion, ultra-sonication
must be performed by use of a horn-type sonicator. The
following equipment or its equivalent is suitable.
a. Sonifer/Cell disruptor, model W-350, Ultrasonics Inc.
b. Sonic Dismembrator model 300, Fisher Scientific Co.,
Catalogue Number 15-338-40.
With either apparatus a sonicator probe with a titanium
tip must be used.
Reagents
1. Any solvent appropriate for GC or GC/MS analysis. Choice of
solvent will depend on the substances being analyzed for.
The solvent of choice should be appropriate for the method
of measurement to be used and which will give an analyte to
solvent partition coefficient of at least 1 to 1000.
2. Sodium sulfate, (ACS) Granular anhydrous (purified by heating
at 400° C for 4 hours in a shallow tray).
Procedure
1. Grind, or otherwise subdivide the waste, in a manner
such that it either passes through a 1-mm seive or can be
extruded through a 1-mm hole. Introduce sufficient sample
-------
8.85-3
into the grinding apparatus to yield at least 10 gms after
grinding.
2. Weigh 10.0 gm of suitably dispersed material into a 75
ml glass flask, add 30 ml of an appropriate solvent. Sonicate
with agitation for approximately 15 minutes. Filter the
resulting suspension. Reextract the solid residue with an
additional 30 ml portion of solvent. Repeat the extraction
a third time so as to sonicate for a total of 45 minutes*
Solvent of choice should be one with properties which
allow for efficient GC, LC, or GC/MS analysis, and which
has an analyte to solvent partition coefficient on the
order of 1 to 1000.
3. After the extraction is complete, filter the extract and dry
it by passing it through a 4-inch column of sodium sulfate
which has been washed with the extracting solvent. Collect
the dried extract in a 500-ml Kuderna-Danish (K-D) flask
fitted with a 10 ml graduated concentrator tube and a 3
ball Snyder column. Wash the extractor flask and sodium
sulfate column with 100-125 ml of the extracting solvent.
If the extract is expected to contain no interfering organics
(therefore requiring no clean-up step), it may be concentrated
using the K-D apparatus and analyzed by the appropriate gas
or liquid chromatographic technique. If further clean-up is
required proceed accordingly (see Section 9 of this manual)
after concentration of the extract.
-------
8.85-4
4. Heat the K-D apparatus on a steam bath until the liquid
level in the collection tube is below 5 ml. Turn off the
heat and allow K-D to cool. Transfer to 10 ml volumetric
flask and adjust volume to 10 ml.
-------
8.86-1
Method 8.86
SOXHLET EXTRACTION METHOD
Scope and Application
This method provides a procedure for the extraction of
semi-volatile and nonvolatile organic compounds from waste materials
which are in a "solid" state, prior to analysis by the appropriate
gas chromatographic or liquid chromatographic technique.
Summary of Method
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 concentrated, and may either be
cleaned up further, or analyzed directly by the appropriate
measurement technique (Methods 8.04 through 8.25).
Apparatus
1. Soxhlet extractor - 40 mm ID, with 500-ml round bottom
flask.
2. Kuderna-Danish Apparatus [Kontes K-570000 or equivalent]
with 3-ball Snyder column.
3. Chromatographic column—Pyrex, 20 mm ID, approxi-
mately 400 mm long, with coarse fritted plate on bottom
and an appropriate packing medium.
Reagents
1. An appropriate solvent or solvent mixture yielding no
measurable residue on evaporation.
-------
8.86-2
2. Anhydrous granular sodium sulfate, ACS (purified
by heating at 400° C for 4 hours in a shallow tray) .
Procedure
1. Blend 10 grams of the solid sample with an equal weight of
anhydrous sodium sulfate and place in either a glass or
paper extraction thimble.
2. Place the sample in the extractor thimble. (If any
problems are encountered in using the thimble, e.g., if the
sample clogs the thimble, an alternative to the thimble
would be to place a plug of glass wool in the extraction
chamber, transfer the sample into the chamber, then cover
the sample with another plug of glass wool.)
3. Place 300 ml of the solvent into a 500-ml roundbottom flask
containing a boiling stone; attach the flask to the extractor,
and extract the solids for 16' hours.
4. After the extraction is complete, cool the extract; then
filter the extract and dry it by passing it through a 4-inch
column of sodium sulfate which has been washed with the
extracting solvent. Collect the dried extract in a 500-ml
Kuderna-Danish (K-D) flask fitted with a 10ml graduated
concentrator tube. Wash the extractor flask and sodium
sulfate column with 100-125 ml of the extracting solvent.
If the extract is expected to contain no interfering organics
(therefore requiring no clean-up step), it may be concen-
trated using the K-D apparatus and analyzed by the appropriate
gas or liquid chromatographic technique. If further clean-up
is required proceed accordingly (see Section 9 of this
manual) after concentration of the extract.
-------
8.86-3
5. Add 1-2 clean boiling chips to the flask and attach
a three-ball Snyder column. Prewet the Snyder column by
adding about 1 ml solvent to the top. Place the K-D apparatus
on a steam or hot water bath so that the concentrator tube and
the entire lower rounded surface of the flask is bathed in
hot water or 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 and allow it to
drain for at least 10 minutes while cooling.
6. Transfer to 10 ml volumetric flask and adjust volume to 10 ml.
-------
8.86-4
References
1. "Interim Methods for the Sampling and Analysis of Priority
Pollutants In Sediments and Fish Tissue", U.S. Environmental
Protection Agency; Environmental Monitoring and Support
Laboratory; Cincinnati, Ohio 45268.
2. "Recovery of Organic Compounds from Environmentally Contami-
nated Bottom Materials", T. Bellar, J. Lichtenberg, S. Lonneman;
U.S. Environmental Protection Agency; Environmental Monitoring
and Support Laboratory; Cincinnati, OH 45268.
-------
9.01-1
Method 9.01
LIQUID - LIQUID EXTRACTION
Introduction
The following procedure provides a method of sample
clean-up to be used when interferences prevent direct
chromatographic measurement of the compound being analyzed
for. The method makes use of the differential solubility of
the compounds of interest and interfering species.
Summary of Method
Removal of interferences is accomplished by a series of
liquid-liquid extractions conducted at different pHs and with
a variety of solvents.
Apparatus
1. Separatory funnel with Teflon stopcock.
2. Kuderna-Danish (K-D) Apparatus [Kontes-K-570000 or
equivalent! equipped with a three ball Snyder column.
3. Boiling chips - solvent extracted, approximately 10/40
mesh.
4. Drying column - 20 mm I.D. Pyrex chromatographic column
with coarse frit.
5. Water or steam bath.
Reagents
1. Sodium hydroxide-(ACS) 10 N in distilled water.
2. Sulfuric acid-(l + 1) Mix equal volumes of concentrated
(ACS) with distilled water.
-------
9.01-2
3. Methylene chloride, acetone, 2-propanol, hexane, toluene-
Pesticide quality or equivalent.
4. Sodium sulfate-(ACS) Granular, anhydrous (purified by
heating at 400°C for 4 hrs. in a shallow tray).
5. Distilled Water
Procedure
1. Place 10 gms of the extract or organic liquid waste to
be cleaned up into the separatory funnel.
2. Add 20 ml of the solvent indicated in Table 9.01-1.
3. Add 20 ml of distilled water and adjust the pH to 12-13
with sodium hydroxide. Partition the sample into the solvent
and aqueous phases by shaking the funnel for one minute with
periodic venting to release vapor pressure. Allow the organic
layer to separate from the water phase for a minimum of ten
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,
but may include stirring, filtration of the emulsion through
glass wool, or centrifugation layer.
4« Separate the aqueous phase and transfer to a 125 ml
Erlenmeyer flask.
5. Reextract the solvent layer twice more with 20 ml portions
of distilled water at pH 12-13. Combine aqueous extract.
6. At this point the species being analyzed for will be
in either the organic or aqueous phase (See Table 9.01-1). If
in the aqueous phase discard the organic phase and proceed to
-------
9.01-3
step 7. If in the organic phase discard the aqueous phase
and proceed to step 12.
7. Transfer the aqueous phase to a clean separatory funnel.
8. Adjust the aqueous layer to a pH of 1-2 with sulfuric
acid.
9. Add 20 ml of solvent to the funnel and shake for two
minutes. Allow the solvent to separate from the aqueous
phase and collect the solvent in a 100 ml Erlenmeyer
flask.
10. Add a second 20 ml volume of solvent to the separatory
funnel and reextract at pH 1-2 a second time, combining
the extracts in the Erlenmeyer flask.
11. Perform a third extraction in the same manner.
12. Pour the combined organic extracts through a drying
column containing 10 cm of anhydrous sodium sulfate, and
collect it in a Kuderna-Danish (K-D) flask equipped with
a 10 ml concentrator tube. Rinse the Erlenmeyer flask
and column with 20 ml of solvent to complete the quantitive
transfer.
13. Add 1-2 clean boiling chips to the flask and attach
a three-ball Snyder column. Prewet the Snyder column by
adding about 1 ml solvent to the top. Place the K-D apparatus
on a steam or hot water bath so that the concentrator tube and
the entire lower rounded surface of the flask is bathed in
hot water or vapor. Adjust the vertical position of the
apparatus and the water temperature as required to complete
-------
9.0^-4
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 R-D apparatus and allow it to
drain for at least 10 minutes while cooling.
14. If the appropriate analytical solvent is the same as
that used for the above extraction then transfer extract to
10 ml volumetric flask and adjust volume to 10 ml. If a
different solvent is to be used for sample measurement proceed
as in step 15 where 2 propanol is used for illustrative
purposes. Other solvents should be used as appropriate.
15. Increase the temperature of the hot water bath to 95-100°C.
Remove the Snyder column and rinse the flask and its lower
joint into the concentrator tube with 1-2 ml of 2-propanol.
Note: A 5-ml syringe is recommended for this operation.
Attach a micro-Snyder column to the concentrator tube
and prewet the column by adding about 0.5 ml 2-propanol
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. 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 the liquid reaches 2.5 ml, remove
the K-D apparatus and allow it to drain for at least 10
-------
9.01-5
minutes while cooling. Add an additional 2 ml of 2-
propanol through the top of the mlcro-Snyder column and resume
concentrating as hefore. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus and allow it to
drain for at least 10 minutes while cooling. Remove the
micro-Snyder column and rinse its lower joint into the
concentrator tube with a minimum amount of 2-propanol.
Transfer to a 10 ml volumetric flask and adjust the extract
volume to 10 ml. Store in refrigerator, if further processing
will not be performed immediately. If the sample extract
requires no further cleanup, proceed with gas chromatographic
analysis. If the sample requires further cleanup, proceed
as appropriate.
-------
9.01-6
Table 9.01-1
APPROPRIATE CLEAN-UP SOLVENT AND STEP 6 PHASE
Compound Being Analyzed For
Benzo(a)anthracene
Benzo(a)pyrene
Benzotrichloride
Benzyl chloride
Benzo(b)fluoranthene
Solvent
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Step 6 Phase
Solvent
Solvent
Solvent
Solvent
Solvent
Chlordane
Chlorinated dibenzodioxins
2-Chlorophenol
Chrysene
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Solvent
Solvent
Aqueous
Solvent
Creosote
Cresol(s)
Cresylic acid(s)
Dichlorobenzene(s)
Dichlorophenoxy-
acetic acid
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Ethyl ether
Solvent
Aqueous
Aqueous
Solvent
Aqueous
Dichloropropanol
2,4-Dimethylphenol
Dinitrobenzene
4,6-Dinitro-o-cresol
2,4-Dinitrotoluene
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Solvent
Aqueous
Solvent
Aqueous
Solvent
-------
9.01-7
Table 9.01-1 (cont.)
APPROPRIATE CLEAN-UP SOLVENT AND STEP 6 PHASE
Compound Being Analyzed For
Endrin
Heptachlor
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Hexachlorocyclopentadiene
Lindane
Maleic anhydride
Methomyl
Solvent
Dichloromethane
Dlchloromethane
Dlchloromethane
Dlchloromethane
Dlchloromethane
Dlchloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Step 6 Phase
Solvent
Solvent
Solvent
Solvent
Solvent
Solvent
Solvent
Solvent
Solvent
Napthalene
Napthoquinone
Nitrobenzene
4-Nltrophenol
Pentachlorophenol
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Solvent
Solvent
Solvent
Aqueous
Aqueous
Phenol
Phorate
Phosphorodithlolc acid
esters
Phthallc anhydride
2-Plcollne
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Dichloromethane
Aqueous
Aqueous
Solvent
Solvent
Solvent
-------
9.01-8
Table 9.01-1 (cont.)
APPROPRIATE CLEAN-UP SOLVENT AND STEP 6 PHASE
Compound Being Analyzed For
Pyridine
Tetrachlorobenzene(s)
Tetrachlorophenol
Toluenediamine
Toxaphene
Trlchlorophenol(s)
2,4,5-TP (Silvex)
Solvent
Dlchloromethane
Dichloromethane
Dlchloromethane
Dlchloromethane
Dichloromethane
Dlchloromethane
Ethyl ether
Step 6 Phase
Solvent
Solvent
Aqueous
Solvent
Solvent
Aqueous
Aqueous
-------
10.0-1
Section 10.0
QUALITY CONTROL, QUALIFY ASSURANCE
INTRODUCTION
The procedures and methods that make up the body of this
manual outline how to test for hazardous characteristics in
solid wastes. If performed by qualified analysts the Agency
feels that the procedures and methods will give a true verdict
of the wastes being tested.
Yet, the described techniques are only valuable if they
are performed with proper care. Unless the stated criteria of
the quality controls and assurances called for in every
technique are met the derived data will be of little value to
either the Agency or the generator.
Section 10 is meant to give guidance to qualified analysts
who will be testing solid wastes for hazardous characteristics.
Since it was developed for use in the testing of dilute
aqueous samples, it is not to be followed exactly. Rather it
should be used as a guideline that outlines acceptable quality
control procedures.
In the hands of a qualified analyst Section 10 can be used
as a yardstick to compare the acceptability of data derived
by generators of solid wastes. In future editions of this
manual EPA will provide more specific quality assurance and
control criteria for use in solid waste testing programs.
-------
EPA-600/4-79-019
HANDBOOK FOR ANALYTICAL QUALITY CONTROL
IN WATER AND WASTEWATER LABORATORIES
March 1979
ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
CINCINNATI, OHIO 45268
-------
DISCLAIMER
The mention of trade names or commercial products in this handbook is for illustration
purposes and does not constitute endorsement or recommendation for use by the U.S.
Environmental Protection Agency.
11
-------
ABSTRACT
One of the fundamental responsibilities of water and wastewater management is the
establishment of continuing programs to insure the reliability and validity of analytical
laboratory and field data gathered in water treatment and wastewater pollution control
activities.
This handbook is addressed to laboratory directors, leaders of field investigations, and other
personnel who bear responsibility for water and wastewater data. Subject matter of the
handbook is concerned primarily with quality control (QC) for chemical and biological tests
and measurements. Chapters are also included on QC aspects of sampling, microbiology,
biology, radiochemistry, and safety as they relate to water and wastewater pollution
control. Sufficient information is offered to allow the reader to inaugurate or reinforce
programs of analytical QC that emphasize early recognition, prevention, and correction of
factors leading to breakdowns in the validity of water and wastewater pollution control
data.
in
-------
ACKNOWLEDGMENTS
This handbook was prepared by the Environmental Monitoring and Support Laboratory
(EMSL) of the United States Environmental Protection Agency. The contributions of the
following individuals in preparing the handbook are gratefully acknowledged:
J. B. Anderson
D. G. Ballinger
E. L. Berg
R. L. Booth
R. H. Bordner
P. W. Britton
J. F. Kopp
H. L. Krieger
J. J. Lichtenberg
L. B. Lobring
J. E. Longbottom
C. I. Weber
J. A. Winter
Technical editing and preparation of the final manuscript were performed under contract to
John F. Holman & Co. Inc., 1346 Connecticut Avenue, N.W., Washington, D.C. 20036.
Inquiries regarding material contained in the handbook should be made to Environmental
Protection Agency, Environmental Research Center, Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268.
IV
-------
Chapter
TABLE OF CONTENTS
Page
PREFACE iii
ACKNOWLEDGMENTS v
1 IMPORTANCE OF QUALITY CONTROL 1-1
1.1 General 1-1
1.2 Quality Assurance Programs 1-1
1.3 Analytical Methods 1-2
1.4 Reference 1-4
2 LABORATORY SERVICES 2-1
2.1 General 2-1
2.2 Distilled Water 2-1
2.3 Compressed Air 2-5
2.4 Vacuum 2-5
2.5 Hood System 2-5
2.6 Electrical Services 2-6
2.7 References 2-6
3 INSTRUMENT SELECTION 3-1
3.1 Introduction 3-1
3.2 Analytical Balances 3-1
3.3 pH/Selective-Ion Meters 3-3
3.4 Conductivity Meters 3-6
3.5 Turbidimeters (Nephelometers) 3-7
3.6 Spectrometers 3-8
3.7 Organic Carbon Analyzers 3-13
3.8 Gas Chromatographs 3-14
3.9 References 3-14
4 GLASSWARE 4-1
4.1 General 4-1
4.2 Types of Glassware 4-2
4.3 Volumetric Analyses 4-3
4.4 Federal Specifications for Volumetric Glassware 4-4
4.5 Cleaning of Glass and Porcelain 4-5
4.6 Special Cleaning Requirements 4-6
4.7 Disposable Glassware 4-7
4.8 Specialized Glassware 4-7
4.9 Fritted Ware 4-8
4.10 References 4-9
5 REAGENTS, SOLVENTS, AND GASES 5-1
5.1 Introduction 5-1
5.2 Reagent Quality 5-1
-------
5.3 Elimination of Determinate Errors 5-4
5.4 References 5-4
6 QUALITY CONTROL FOR ANALYTICAL PERFORMANCE 6-1
6.1 Introduction 6-1
6.2 The Industrial Approach to QC 6-1
6.3 Applying Control Charts in Environmental Laboratories 6-2
6.4 Recommended Laboratory Quality Assurance Program 6-9
6.5 Outline of a Comprehensive Quality Assurance Program 6-10
6,6 Related Topics 6-13
6.7 References 6-13
7 DATA HANDLING AND REPORTING 7-1
7.1 Introduction 7-1
7.2 The Analytical Value •. 7-1
7.3 Glossary of Statistical Terms 7-3
7.4 Report Forms 7-5
7.5 References 7-11
8 SPECIAL REQUIREMENTS FOR TRACE ORGANIC ANALYSIS ... 8-1
8.1 Introduction 8-1
8.2 Sampling and Sample Handling 8-1
8.3 Extract Handling 8^
8.4 Supplies and Reagents 8-5
8.5 Quality Assurance 8-7
8.6 References 8-10
9 SKILLS AND TRAINING 9-1
9.1 General 9-1
9.2 Skills 9-2
9.3 Training 9-4
10 WATER AND WASTEWATER SAMPLING 10-1
10.1 Introduction 10-1
10.2 Areas of Sampling 10-2
10.3 References 10-6
11 RADIOCHEMISTRY 11-1
11.1 Introduction 11-1
11.2 Sample Collection 11-1
11.3 Laboratory Practices 11-2
11.4 Quality Control . 11-4
11.5 References 11-5
12 MICROBIOLOGY 12-1
12.1 Background 12-1
12.2 Specific Needs in Microbiology 12-1
12.3 Intralaboratory Quality Control 12-2
12.4 Intel-laboratory Quality Control 12-2
12.5 Development of a Formal Quality Assurance Program 12-3
12.6 Documentation of a Quality Assurance Program 12-3
VI
-------
12.7 Chain-of-Custody Procedures for Microbiological Samples .... 12-3
12.8 References 12-10
13 AQUATIC BIOLOGY 13-1
13.1 Summary of General Guidelines 13-1
13.2 Discussion 13-2
13.3 Reference 13-4
14 LABORATORY SAFETY 14-1
14.1 Law and Authority for Safety and Health 14-1
14.2 EPA Policy on Laboratory Safety 14-5
14.3 Laboratory Safety Practices 14-7
14.4 Report of Unsafe or Unhealthful Condition 14-15
14.5 References 14-15
Appendix A-Suggested Checklist for the Safety Evaluation of EPA Laboratory Areas A-l
VII
-------
LIST OF FIGURES
Figure No. Page
4-1 Example of Markings on Glassware 4-5
6-1 Essentials of a Control Chart 6-2
6-2 Shewhart Control Chart for Percent Recovery Data 6-5
6-3 Procedure for Evaluating QC Data From a Monitoring Study 6-12
7-1 Example of Bench Sheet 7-8
7-2 Example of Summary Data Sheet 7-9
7-3 Example of STORE! Report Form 7-10
7-4 Flow Chart of the Sequence of Events During a Controlled Series of
Laboratory Measurements 7-12
12-1 Example of Chain-of-Custody Sample Tag (a) Front, (b) Back 12-5
12-2 Example of a Sample Log Sheet 12-6
12-3 Example of a Chain-of-Custody Record 12-7
14-1 Health and Safety Inspection Checklist 14-8
14-2 Report of Unhealthful or Unsafe Condition 14-16
14-3 Notice of Unhealthful or Unsafe Condition 14-17
LIST OF TABLES
Table No. Page
2-1 Water Purity 2-1
2-2 Requirements for Reagent Water 2-2
2-3 Comparison of Distillates From Glass and Metal Stills 2-3
3-1 Performance Characteristics of Typical pH/Selective-Ion Meter 3-5
3-2 Electrical Conductivity of Potassium Chloride Reference Solutions . . . 3-7
4-1 Tolerances for Volumetric Glassware 4-4
4-2 Fritted Ware Porosity 4-8
6-1 Analyses of Total Phosphate-Phosphorus Standards, in Milligrams Per
Liter of Total PO4-P 64
6-2 Estimates of the Range (R = \A - B\) and the Industrial Statistic (I=\A-
B\I(A + B) of Three Different Parameters for Various Concentration
Ranges 6-7
6-3 Shewhart Upper Control Limits UCL and Critical Range Rc Values for
the Differences Between Duplicate Analyses Within Specific Con-
centration Ranges for Three Parameters 6-8
6-4 Critical Range Values for Varying Concentration Levels 6-9
9-1 Skill-Time Rating of Standard Analytical Operations 9-3
via
-------
10-1 Guidance for Water/Wastewater Sampling 10-2
11-1 Sample Handling, Preservation, Methodology, and Major Instrumentation
Required 11-2
IX
-------
Chapter 1
IMPORTANCE OF QUALITY CONTROL
1.1 General
The analytical laboratory provides qualitative and quantitative data for use in decision-
making. To be valuable, the data must accurately describe the characteristics and
concentrations of constituents in the samples submitted to the laboratory. In many cases,
because they lead to faulty interpretations, approximate or incorrect results are worse than
no result at all.
Ambient water quality standards for pH, dissolved oxygen, heavy metals, and pesticides are
set to establish satisfactory conditions for drinking water, fishing, irrigation, power
generation, or other water uses. The laboratory data define whether conditions are being
met and whether the water can be used for its intended purposes. In wastewater analyses,
the laboratory data identify the characteristics of the treatment plant influent and the final
load imposed upon receiving water resources, as well as the effectiveness of steps in the
treatment process. Decisions on process changes, plant modifications, or the construction of
new facilities may be based upon the results of water laboratory analyses. The financial
implications of such decisions suggest that extreme care be taken in analysis.
Effective research in water pollution control also depends upon a valid laboratory data base,
which in turn may contribute to sound evaluations of both the progress of the research itself
and the viability of available water pollution-control alternatives.
The analytical data from water and wastewater laboratories may also be used to determine
the extent of compliance of a polluting industry with discharge or surface water standards.
If the laboratory results indicate a violation of a standard, remedial action is required by the
responsible parties. Both .legal and social pressures can be brought to bear to protect the
environment. The analyst should realize not only that he has considerable responsibility for
providing reliable laboratory descriptions of the samples at issue, but also that his profes-
sional competence, the validity of the procedures used, and the resulting values reported
may be challenged (perhaps in court). For the analyst to meet such challenges, he should
support the laboratory data with an adequate documentation program that provides valid
records of the control measures applied to all factors bearing on the final results of investi-
gations.
1.2 Quality Assurance Programs
Because of the importance of laboratory analyses in determining practical courses of action
that may be followed, quality assurance programs to insure the reliability of the water and
wastewater data are essential. Although all analysts practice quality control (QC) in amounts
depending upon their training, professional pride, and the importance of their particular
projects, under actual working conditions sufficiently detailed QC may be neglected. An
established, routine, quality assurance program applied to each analytical test can relieve
analysts of the necessity of originating individual QC efforts.
1-1
-------
Quality assurance programs have two primary functions in the laboratory. First, the
programs should continually monitor the reliability (accuracy and precision) of the results
reported; i.e., they should continually provide answers to the question "How good (accurate
and precise) are the results obtained?" This function is the determination of quality. The
second function is the control of quality (to meet the program requirements for reliability).
As an example of the distinction between the two functions, the processing of spiked
samples may be a determination of measurement quality, but the use of analytical grade
reagents is a control measure.
Each analytical method has a rigid protocol. Similarly, QC associated with a test must
include definite required steps for monitoring the test and insuring that its results are
correct. The steps in QC vary with the type of analysis. For example, in a titration,
standardization of the titrant on a frequent basis is an element of QC. In any instrumental
method, calibration and checking out of instrumental response are also QC functions. All of
the experimental variables that affect the final results should be considered, evaluated, and
controlled.
In summary, laboratory data, in quantitative terms, e.g., in milligrams per liter, are reported
by the analyst. These values are interpreted by industrial plant engineers to show
compliance or noncompliance with permits for discharge, by state pollution control agencies
to define the need for additional sampling and analysis to confirm violations, or by EPA to
demonstrate that prescribed waste treatment was sufficient to protect the surface waters
affected by the discharge.
This handbook discusses the basic factors of water and wastewater measurements that
determine the value of analytical results and provides recommendations for the control of
these factors to insure that analytical results are the best possible. Quality assurance
programs initiated from, and based upon, these recommendations should increase
confidence in the reliability of the reported analytical results.
Because ultimately a laboratory director must assume full responsibility for the reliability of
the analytical results submitted, the laboratory director must also assume full responsibility
in both design and implementation for the corresponding quality assurance program.
1.3 Analytical Methods
Many analytical methods for common water pollutants have been in use for many years and
are used in most environmental laboratories. Examples are tests for chloride, nitrate, pH,
specific conductance, and dissolved oxygen. Widespread use of an analytical method in
water and wastewater testing usually indicates that the method is reliable, and therefore
tends to support the validity of the reported test results. Conversely, the use of little-known
analytical techniques forces the water and wastewater data user to rely on the judgment of
the laboratory analyst, who must then defend his choice of analytical technique as well as
his conclusions. Present Federal regulations, notably section 304(h) of Public Law 92-500
(Federal Water Pollution Control Amendments of 1977) and the Interim Drinking Water
Regulations specifically require the use of EPA-approved methods of analysis.
Uniformity of methodology within a single laboratory as well as among a group of
cooperating laboratories is required to remove methodology as a variable when there are
many data users. Uniformity of methodology is particularly important when several
1-2
-------
laboratories provide data to a common data bank (such as STORET*) or cooperate in joint
field surveys. A lack of uniformity of methodology may raise doubts as to the validity of
the reported results. If the same constituents are measured by different analytical
procedures within a single laboratory, or by a different procedure in different laboratories,
it may be asked which procedure is superior, why the superior method is not used
throughout, and what effects the various methods and procedures have on the data values
and their interpretations.
Physical and chemical measurement methods used in water or wastewater laboratories
should be selected by the following criteria:
a. The selected methods should measure desired constituents of water samples in the
presence of normal interferences with sufficient precision and accuracy to meet the
water data needs.
b. The selected procedures should use equipment and skills ordinarily available in the
average water pollution control laboratory or water supply laboratory.
c. The selected methods should be sufficiently tested to have established their validity.
d. The selected methods should be sufficiently rapid to permit repetitive routine use in
the examination of large numbers of water samples.
The restriction to the use of EPA methods in all laboratories providing data to EPA permits
the combination of data from different EPA programs and supports the validity of decisions
made by EPA.
Regardless of which analytical methods are used in a laboratory, the methodology should be
carefully documented. In some reports it is stated that a standard method from an
authoritative reference (such as ref. 1) was used throughout an investigation, when close
examination has indicated, however, that this was not strictly true. Standard methods may
be modified or entirely replaced because of recent advances in the state of the art or
personal preferences of the laboratory staff. Documentation of measurement procedures
used in arriving at laboratory data should be clear, honest, and adequately referenced; and
the procedures should be applied exactly as documented.
Reviewers can apply the associated precision and accuracy of each specific method when
interpreting the laboratory results. If the accuracy and precision of the analytical
methodology are unknown or uncertain, the data user may have to establish the reliability
of the result he or she is interpreting before proceeding with the interpretation.
The necessarily strict adherence to accepted methods in water and wastewater analyses
should not stifle investigations leading to improvements in analytical procedures. Even with
accepted and documented procedures, occasions arise when the procedures must be
modified; e.g., to eliminate unusual interferences, or to yield increased sensitivities. When a
*STORET is the acronym used to identify the computer-oriented U.S. Environmental
Protection Agency water quality control information system; STORET stands for STOrage
and RETrieval of data and information.
1-3
-------
modification of a procedure is necessary, it should be carefully formulated. Data should
then be assembled using both the original and the modified procedures to show the
superiority of the latter. Such results can be brought to the attention of the organizations
responsible for standardization of procedures. To increase the benefit, the modified
procedures should be written in a standard format for routine use as applicable. The
standard format usually includes scope and application, principle, equipment, reagents,
procedure, calculation of results, and expected precision and accuracy.
Responsibility for the results obtained from use of a nonstandard procedure (i.e., one that
has not become accepted through wide use) rests with the analyst and his supervisor.
In field operations, because it may be difficult to transport samples to the laboratory, or to
examine large numbers of samples (e.g., for gross characteristics), the use of rapid field
methods yielding approximate answers is sometimes required. Such methods should be used
only with a clear understanding that the results obtained.are not as reliable as those
obtained from standard laboratory procedures. The fact that such methods have been used
should be documented, and the results .should not be reported in the same context with
more reliable analytical information. When only approximate values are available, perhaps
obtained for screening purposes in the field only, the data user would then be so informed.
1.4 Reference
1. Standard Methods for the Examination of Water and Wastewater, 14th Edition,
American Public Health Association, New York (1975).
1-4
-------
Chapter 2
LABORATORY SERVICES
2.1 General
Quality control of water and wastewater laboratory analyses involves consideration and
control of the many variables that affect the production of reliable data. The quality of the
laboratory services available to the analyst must be included among these variables. An
abundant supply of distilled water, free from interferences and other undesirable
contaminants, is an absolute necessity. An adequate source of clean, dry, compressed air is
needed. Electrical power for routine laboratory use and voltage-regulated sources for
delicate electronic instrumentation must be provided. This chapter, therefore, will be
devoted to describing methods of maintaining the quality of these services, as used in water
and wastewater laboratory operations.
2.2 Distilled Water
Distilled or demineralized water is used in the laboratory for dilution, preparation of reagent
solutions, and final rinsing of glassware. Ordinary distilled water is usually not pure. It may
be contaminated by dissolved gases and by materials leached from the container in which it
has been stored. Volatile organics distilled over from the feed water may be present, and
nonvolatile impurities may occasionally be carried over by the steam, in the form of a spray.
The concentration of these contaminants is usually quite small, and distilled water is used
for many analyses without further purification. However, it is highly important that the
still, storage tank, and any associated piping be carefully selected, installed, and maintained
in such a way as to insure minimum contamination.
Water purity has been defined in many different ways, but one generally accepted definition
states that high-purity water is water that has been distilled or deionized, or both, so that it
will have a specific resistance of 500,000 n or greater (or a conductivity less than 2.0
//mho/cm). This definition is satisfactory as a base to work from, but for more critical
requirements, the breakdown shown in table 2-1 has been suggested to express degrees of
purity (1).
Table 2-1
WATER PURITY
Degree of Purity
Pure
Very Pure
Ultrapure
Theoretically Pure
Maximum
Conductivity
(/imho/cm)
10
1
0.1
0.055
Approximate
Concentration
of Electrolyte
(mg/1)
2-5
0.2-0.5
0.01-0.02
0.00
2-1
-------
The American Society for Testing and Materials (ASTM) specifies four different grades of
water for use in methods of chemical analysis and physical testing. The method of
preparation of the various grades of reagent water determines the limits of impurities. The
various types of reagent water and ASTM requirements are listed in table 2-2.
Type I grade water is prepared by the distillation of feed water having a maximum
conductivity of 20 jumho/cm at 25°C followed by polishing with a mixed bed of
ion-exchange materials and a 0.2-jum membrane filter.
Type II grade water is prepared by using a still designed to produce a distillate having a
conductivity of less than 1.0 /zmno/cm at 25°C. This may be accomplished by double
distillation or the use of a still incorporating special baffling and degassing features.
Type III grade water is prepared by distillation, ion exchange, or reverse osmosis, followed
by polishing with the 0.45-jum membrane filter.
Type IV grade water is prepared by distillation, ion exchange, reverse osmosis, or
electrodialysis.
Properly designed metal stills from reputable manufacturers offer convenient and reliable
sources of distilled water. These stills are usually constructed of copper, brass, and bronze.
All surfaces that contact the distillate should be heavily coated with pure tin to prevent
metallic contamination. The metal storage tank should be of sturdy construction with a
tight-fitting cover, and have a filter in the air vent to remove airborne dust, gases, and fumes.
For special purposes, an all-glass distillation unit may be preferable to the metal still. These
stills are usually smaller, and of more limited capacity than the metal stills. An actual
comparison in which the distillates from an all-glass still and a metal still were analyzed
spectrographically for certain trace metal contaminants is given in table 2-3. It can be seen
that the all-glass still produced a product that had substantially lower contamination from
zinc, copper, and lead.
All stills require periodic cleaning to remove solids that have been deposited from the feed
water. Hard water and high-dissolved-solids content promote scale formation in the
Table 2-2
REQUIREMENTS FOR REAGENT WATER
Grade
of Water
Type I
Type II
Type III
Type IV
Maximum
Total
Matter
(mg/1)
0.1
0.1
1.0
2.0
Maximum
Electrical
Conductivity
at 25°C
(jumho/cm)
0.06
1.0
1.0
5.0
Minimum
Electrical
Resistivity*
at 25°C
(Mn • cm)
16.67
1.0
1.0
0.2
pH at 25°C
6.2-7.5
5.0-8.0
Minimum Color
Retention Time
ofKMnO4
(min)
60
60
10
10
"See reference 2.
2-2
-------
Table 2-3
COMPARISON OF DISTILLATES FROM GLASS AND
METAL STILLS
Element and Concentration Gug/1)
Source
Zn B Fe Mn Al Cu Ni Pb
All-Glass Still
Metal Still
9
12
13
1 <
2 <
:i
-------
A piping system for delivering distilled water to the area of use within the laboratory is a
convenient feature. In this case, special care should be taken that the quality of the water is
not degraded between the still and the point of use. Piping may be of tin, tin-lined brass,
stainless steel, plastic, or chemically resistant glass, depending on the quality of the water
desired, its intended use, and on available funds. Tin is best, but is also very expensive. As a
compromise, plastic pipe or glass pipe with Teflon* O-rings at all connecting joints is
satisfactory for most purposes. The glass pipe has an obvious advantage when freedom from
trace amounts of organic materials is important.
When there is no piped-in supply, distilled water will probably be transported to the
laboratory and stored in polyethylene or glass bottles of about 5-gal capacity. If stored in
glass containers, distilled water will gradually leach the more soluble materials from the glass
and cause an increase in dissolved solids. On the other hand, polyethylene bottles contain
organic plasticizers, and traces of these materials may be leached from the container walls.
These are of little consequence, except in some organic analyses. Rubber stoppers often
used in storage containers contain leachable materials, including significant quantities of
zinc. This is usually no problem, because the water is not in direct contact with the stopper.
However, the analyst should be aware of the potential for contamination, especially when
the supply is not replenished by frequent use.
The delivery tube may consist of a piece of glass tubing that extends almost to the bottom
of the bottle, and that is bent downward above the bottle neck, with a 3- to 4-ft piece of
flexible tubing attached for mobility. Vinyl tubing is preferable to latex rubber, because it is
less leachable; however, a short piece of latex tubing may be required at the outlet for better
control of the pinchcock. The vent tube in the stopper should be protected against the
entrance of dust.
Ordinary distilled water *is quite adequate for many analyses, including the determination of
major cations and anions. Certain needs may require the use of double- or even
triple-distilled water. Redistillation from an alkaline permanganate solution can be used to
obtain a water with low organic background. When determining trace organics by solvent
extraction and gas chromatography, distilled water with sufficiently low background may be
extremely difficult to obtain. In this case, preextraction of the water with the solvent used
in the respective analysis may be helpful in eliminating undesirable peaks in the blank.
Certain analyses require special treatment or conditioning of the distilled water, and these
will now be discussed.
2.2.1 Ammonia-Free Water
Removal of ammonia can be accomplished by shaking ordinary distilled water with a strong
cation exchanger, or by passing distilled water through a column of such material. For
limited volumes of ammonia-free water, use of the Quikpure (Box 254, Chicago, 111.)
500-ml bottle is highly recommended. The ion-free water described in section 2.2.3 is also
suitable for use in the determination of ammonia.
2.2.2 Carbon-Dioxide-Free Water
Carbon-dioxide-free water may be prepared by boiling distilled water for 15 min and cooling
to room temperature. As an alternative, distilled water may be vigorously aerated with a
Trademark of E. I. duPont de Nemours & Co.
2-4
-------
stream of inert gas for a period sufficient to achieve saturation and CO2 removal. Nitrogen is
most frequently used. The final pH of the water should lie between 6.2 and 7.2. It is not
advisable to store CO2 -free water for extended periods.
2.2.3 Ion-Free Water
A multipurpose high purity water, free from trace amounts of the common ions, may be
conveniently prepared by slowly passing distilled water through an ion-exchange column
containing one part of a strongly acidic cation-exchange resin in the hydroxyl form. Resins
of a quality suitable for analytical work must be used. Ion-exchange cartridges of the
research grade, available from scientific supply houses, have been found satisfactory. By
using a fresh column and high-quality distilled water, a water corresponding to the ASTM
designation for type I reagent water (2) (maximum 0.1 mg/1 of total matter and maximum
conductivity of 0.06 mho/cm) can be obtained. This water is suitable for use in the
determination of ammonia, trace metals, and low concentrations of most cations and
anions. It is not suited to some organic analyses, however, because this treatment adds
organic contaminants to the water by contact with the ion-exchange materials.
2.3 Compressed Air
The quality of compressed air required in the laboratory is usually very high, and special
attention should be given to producing and maintaining clean air until it reaches the outlet.
Oil, water, and dirt are undesirable contaminants in compressed air, and it is important to
install equipment that generates dry, oil-free air. When pressures of less than 50 psi are
required, a rotary-type compressor, using a water seal and no oil, eliminates any addition of
oil that would subsequently have to be removed from the system. Large, horizontal,
water-cooled compressors will usually be used when higher pressures are required.
Compression heats air, thus increasing its tendency to retain moisture. An aftercooler is
therefore necessary to remove water. Absorption filters should be used at the compressor to
prevent moisture from entering the piping system. Galvanized steel pipe with threaded,
malleable-iron fittings, or solder-joint copper tubing should be used for piping the air to the
laboratory.
When the compressed air entering the laboratory is of low quality, an efficient filter should
be installed between the outlet and the point of use to trap oil, moisture, and other
contaminants. As an alternative, high-quality compressed air of the dry grade is
commercially available in cylinders when no other source exists.
2.4 Vacuum
A source of vacuum in the chemical laboratory, while not an absolute necessity, can be a
most useful item. While used primarily as an aid in filtration, it is also sometimes used in
pipetting and in speeding up the drying of pipets.
2.5 Hood System
An efficient hood system is a requirement for all laboratories. In addition to removing the
various toxic and hazardous fumes that may be generated when using organic solvents, or
that may be formed during an acid digestion step, a hood system may also be used to
remove toxic gases that may be formed during atomic absorption analyses or other
2-5
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reactions. A regular fume hood should have a face velocity of 100 ft/min (linear) with the
sash fully open.
2.6 Electrical Services
An adequate electrical system is indispensable to the modern laboratory. This involves
having both 115- and 230-V sources in sufficient capacity for the type of work that must be
done. Requirements for satisfactory lighting, proper functioning of sensitive instruments,
and operation of high-current devices must be considered. Any specialized equipment may
present unusual demands on the electrical supply.
Because of the special type of work, requirements for a laboratory lighting system are quite
different from those in other areas. Accurate readings of glassware graduations, balance
verniers, and other measuring lines must be made. Titration endpoints, sometimes involving
subtle changes in color or shading, must be observed. Levels of illumination, brightness,
glare, and location of light sources should be controlled to facilitate ease in making these
measurements and to'provide maximum comfort for the employees.
Such instruments as spectrophotometers, flame photometers, atomic absorption equipment,
emission spectrographs, and gas chromatographs have complicated electronic circuits that
require relatively constant voltage to maintain stable, drift-free instrument operation. If the
voltage of these circuits varies, there is a resulting change in resistance, temperature, current,
efficiency, light output, and component life. These characteristics are interrelated, and one
cannot be changed without affecting the others. Voltage regulation is therefore necessary to
eliminate these conditions.
Many instruments have built-in voltage regulators that perform this function satisfactorily.
In the absence of these, a small, portable, constant-voltage transformer should be placed in
the circuit between the electrical outlet and the instrument. Such units are available from
Sola Basic Industries, Elk Grove Village, 111., and are capable of supplying a constant output
of 118 V from an input that varies between 95 and 130 V. When requirements are more
stringent, special transformer-regulated circuits can be used to supply constant voltage. Only
the instrument receiving the regulated voltage should be operated from such a circuit at any
given time. These lines are in addition to, and separate from, the ordinary circuits used for
operation of equipment with less critical requirements.
Electrical heating devices provide desirable heat sources, and should offer continuously
variable temperature control. Hot plates and muffle furnaces wired for 230-V current will
probably give better service than those that operate on 115V, especially if the lower voltage
circuit is only marginally adequate. Water baths and laboratory ovens with maximum
operating temperatures of about 200°C perform well at 115 V. Care must be taken to
ground all equipment that could constitute a shock hazard. The three-pronged plugs that
incorporate grounds are best for this purpose.
2.7 References
1. Applebaum, S. B., and Crits, G. J., "Producing High Purity Water," Industrial Water
Engineering (Sept./Oct. 1964).
2. "Water," Part 31 of 1977 Book of ASTM Standards, p. 20, American Society for
Testing and Materials, Philadelphia (1977).
2-6
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Chapter 3
INSTRUMENT SELECTION
3.1 Introduction
The modern analytical laboratory depends very heavily upon instrumentation. This
statement may be completely obvious, but it should be remembered that the exceptional
emphasis on electronic equipment has really begun since the development of the transistor
and the computer. To a certain extent, analytical instrumentation is always in the
development stage, with manufacturers continually redesigning and upgrading their
products, striving for miniaturization, better durability and sensitivity, and improved
automation. For laboratory supervisors and staff members the net result is a bewildering
stream of advertising brochures, announcements, and catalogs of newly available equipment.
Consequently, the selection of analytical equipment is always difficult.
The instruments commonly used in water and wastewater analysis include the following:
Analytical balance
pH/selective-ion meter
Conductivity meter
Turbidimeter
Spectrometers (visible, ultraviolet (UV), infrared (IR), and atomic absorption (AA))
Total carbon analyzer
Gas chromatograph (GC)
Gas chromatograph/mass spectrometer (GC/MS)
Temperature devices (such as ovens and water baths)
Recorders
These devices represent basic equipment used in routine work and should be the subject of
careful consideration before purchase. Further, their operation and maintenance ought to be
primary considerations in sustained production of satisfactory data. Obviously, fundamental
understanding of instrument design will assist the analyst in the correct use of instruments
and in some cases will aid in detecting instrumental failures. Calibration of all laboratory
instruments with primary standards is encouraged whenever practical. This normally
involves a National Bureau of Standards standard reference material or calibration and
certification procedures. Calibration checks with secondary standards, made in each
laboratory or available from private sources, are encouraged on a frequent basis if not
required by the analytical method each time an analysis is made.
In the pages that follow an attempt is made to discuss basic instrument design and to offer
some remarks about desirable instrumental features.
3.2 Analytical Balances
The most important piece of equipment in any analytical laboratory is the analytical
balance. The degree of accuracy of the balance is reflected in the accuracy of all data related
to weight-prepared standards. Although the balance should therefore be the most protected
and cared-for instrument in the laboratory, proper care of the balance is frequently
overlooked.
3-1
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There are many fine balances on the market designed to meet a variety of needs. Types of
balances include top-loading, two-pan, microanalytical, electroanalytical, semianalytical,
analytical, and other special-purpose instruments. Each type of balance has its own place in
the scheme of laboratory operation, but analytical single-pan balances are by far the most
important in the production of reliable data.
Single-pan analytical balances range in capacity from the 20-g to the popular 200-g models
with sensitivities from 0.01 to 1 mg. Features of single-pan balances may include mechanical
and electronic switching of weights, digital readout, automatic zeroing of the empty
balance, and automatic preweighing and taring capabilities. Even with all the design
improvements, however, modern analytical balances are still fragile instruments, the
operation of which is subject to shock, temperature, and humidity changes, to mishandling,
and to various other insults. Some of the precautions to be observed in maintaining and
prolonging the dependable life of a balance are as follows:
a. Analytical balances should be mounted on a heavy, shockproof table, preferably one
with an adequately large working surface and with a suitable drawer for storage of
balance accessories. The balance level should be checked frequently and adjusted
when necessary.
b. Balances should be located away from laboratory traffic and protected from sudden
drafts and humidity changes.
c. Balance temperatures should be equilibrated with room temperature; this is
especially important if building heat is shut off or reduced during nonworking hours.
d. When the balance is not in use, the beam should be raised from the knife edges, the
weights returned to the beam, objects such as the weighing dish removed from the
pan, and the weighing compartment closed.
e. Special precautions should be taken to avoid spillage of corrosive chemicals on the
pan or inside the balance case; the interior of the balance housing should be kept
scrupulously clean.
f. Balances should be checked and adjusted periodically by a company service man or
balance consultant; if service is not available locally, the manufacturer's instructions
should be followed as closely as possible. Service contracts, including an automatic
preventive maintenance schedule, are encouraged.
g. The balance should be operated at all times according to the manufacturer's
instructions.
Standardized weights to be used in checking balance accuracy, traceable to the National
Bureau of Standards, may be purchased from various supply houses. A complete set of
directions for checking the performance of a balance is contained in part 41 of ASTM
Standards (1).
Because all analytical balances of the 200-g capacity suitable for water and wastewater
laboratories have about the same design specifications with reference to sensitivity,
precision, convenience, and price, it is safe to assume that there is no clear preference for a
certain model, and selection can be made on the basis of availability of service.
3-2
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3.3 pH/Selective-lon Meters
The concept of pH as a means of expressing the degree of effective acidity or alkalinity
instead of total acidity or alkalinity was developed in 1909 by Sorenson (2). It was not until
about 1940 that commercial instruments were developed for routine laboratory measure-
ment of pH.
»
A basic meter consists of a voltage source, amplifier, and scale or digital readout device.
Certain additional refinements produce varying performance characteristics between models.
Some models incorporate expanded scales for increased readability, solid state circuitry for
operating stability and extreme accuracy, and temperature and slope adjustment to correct
for asymmetric potential of glass electrodes. Other features are scales that facilitate use of
selective-ion electrodes, recorder output, and interfacing with complex data-handling
systems.
In routine pH measurements the glass electrode is used as the indicator and the calomel
electrode as the reference. Glass electrodes have a very fast response time in highly buffered
solutions. However, accurate readings are obtained slowly in poorly buffered samples, and
particularly so when changing from buffered to unbuffered samples. Electrodes, both glass
and calomel, should be well rinsed with distilled water after each reading, and should be
rinsed with, or dipped several times into, the next test sample before the final reading is
taken. Weakly buffered samples should be stirred during measurement. When not in use,
glass electrodes should not be allowed to become dry, but should be immersed in an
appropriate solution consistent with the manufacturer's instructions. The first steps in
calibrating an instrument are to immerse the glass and calomel electrodes into a buffer of
known pH, set the meter to the pH of the buffer, and adjust the proper controls to bring the
circuit into balance. The temperature-compensating dial should be set at the temperature of
the buffer solution. For best accuracy, the instrument should be calibrated against two
buffers that bracket the expected pH of the samples.
The presence of a faulty electrode is indicated by failure to obtain a reasonably correct
value for the pH of the second reference buffer solution after the meter has been
standardized with the first reference buffer solution. A cracked glass electrode will often
yield pH readings that are essentially the same for both standards. The response of
electrodes may also be impaired by failure to maintain the KC1 level in the calomel
electrode, by improper electrode maintenance, or by certain materials such as oily
substances and precipitates that may coat the electrode surface. Faulty electrodes can often
be restored to normal by an appropriate cleaning procedure. Complete and detailed cleaning
methods are given in part 31 of ASTM Standards (3), and are also usually supplied by the
electrode manufacturer.
Because of the asymmetric potential of the glass electrode, most pH meters are built with a
slope adjustment that enables the analyst to correct for slight electrode errors observed
during calibration with two different pH buffers. Exact details of slope adjustment and
slope check may vary with different models of instruments. The slope adjustment must be
made whenever electrodes are changed, subjected to vigorous cleaning, or refilled with fresh
electrolyte. The slope adjustment feature is highly desirable and recommended for
consideration when purchasing a new meter.
Most pH meters now available are built with transistorized circuits rather than vacuum
tubes, which greatly reduces the warmup time and increases the stability of the meters.
3-3
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Also, many instruments are designed with a switching circuit so that the entire conventional
0 to 14 scale of pH may be used to read a single pH unit with a corresponding increase in
accuracy.
This expanded-scale feature is of definite value when the meter is used for potentiometric
titrations and selective-ion work. It is of dubious value, however, in routine analyses,
because pH readings more precise than ±0.1 are seldom required. Primary standard buffer
salts are available from the National Bureau of Standards* and should be used in situations
where extreme accuracy is necessary. Preparation of reference solutions from these salts
requires some special precautions and handling (3,4) such as the use of low-conductivity
dilution water, drying ovens, and carbon-dioxide-free purge gas. These solutions should be
replaced at least once each month.
Secondary standard buffers may be prepared from NBS salts or purchased as solutions from
commercial vendors. Routine use of these commercially available solutions, which have been
validated by comparison to NBS standards, is recommended.
The electrometric measurement of pH varies with temperature because of two effects. The
first effect is the change in electrode output with temperature. This interference can be
controlled by use of instruments having temperature compensation or by calibrating the
instrument system including the electrode at the temperature of the samples. The second
effect is the change of pH of the sample with temperature. 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.
Typical characteristics of a conventional expanded-scale meter are shown in table 3-1.
3.3.1 pH Electrodes
A wide variety of special- and general-purpose pH electrodes are now available to meet all
applications in the general analytical laboratory. A survey through any laboratory supply
catalog may confuse more than clarify the selection process. A rugged, full-range, glass- or
plastic-bodied combination electrode is a good choice for routine use. An added
convenience is an electrode that contains solid geltype filling materials not requiring the
normal maintenance of an electrode containing liquid filling solutions.
3.3.2 Selective-Ion Electrodes
Electrodes have been developed to measure almost every common inorganic ion normally
measured in the water and wastewater laboratory. Application of these electrodes has
progressed at a much slower pace and currently only three are approved for EPA monitoring
applications.
Reference 5 includes methods for use of fluoride, ammonia, and dissolved oxygen
electrodes. Various techniques for use of these and other electrodes are reviewed in
references 6 through 9. A major problem in measuring the total parameter with electrodes is
that of relating the ion activity to ion concentration. Because the electrodes only measure
*NBS, Office of Standard Reference Materials, Institute for Materials Research, Washington,
D.C. 20234.
3-4
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activity, the challenge is to put all of the parameters of interest into the same measurable
ionic form and then to modify the activity to be proportional to the concentration. The
technique of known addition (spiking of samples) is recommended when unproven electrode
methods are being used or when sample matrix problems are suspected or not controlled by
prior distillation or separation techniques.
3.4 Conductivity Meters
Solutions of electrolytes conduct an electric current by the migration of ions under the
influence of an electric field. For a constant applied EMF, the current flowing between
opposing electrodes immersed in the electrolyte will vary inversely with the resistance of the
solution. The reciprocal of the resistance is called conductance and is expressed in reciprocal
ohms (mhos). For natural water samples where the resistance is high, the usual reporting
unit is micromhos.
Most conductivity meters on the market today use a cathode-ray tube, commonly known as
the "magic eye," for indicating solution conductivity. A stepping switch for varying
resistances in steps of 10X facilitates reading conductivities from about 0.1 to about
250,000 nmho. The sensing element for a conductivity measurement is the conductivity
cell, which normally consists of two thin plates of platinized metal, rigidly supported with a
very precise parallel spacing. For protection, the plates are mounted inside a glass tube with
openings in the side walls and submersible end for access of sample. Variations in designs
have included use of hard rubber and plastics for protection of the cell plates. Glass may be
preferable, in that the plates may be visually observed for cleanliness and possible damage,
but the more durable encasements have the advantage of greater protection and reduced cell
breakage. Selection of various cell designs is normally based on personal preference with
consideration of sample type and durability requirements.
In routine use, cells should be frequently examined to insure that (a) the platinized coating
of plates is intact; (b) plates are not coated with suspended matter; (c) plates are not bent,
distorted, or misalined; and (d) lead wires are properly spaced.
Temperature has a pronounced effect on the conductance of solutions, and must be
corrected for when results are reported. The specified temperature for reporting data used
by most analytical groups (and all EPA laboratories) is 25°C. Data correction may be
accomplished by adjusting sample temperatures to 25°C, or by use of mathematical or
electronic adjustment.
Instrumental troubles are seldom encountered with conductivity meters because of the
design simplicity. When troubles occur, they are usually in the cell, and for most accurate
work the following procedures should be used:
a. Standardize the cell and establish a cell factor by measuring the conductivity of a
standard potassium chloride solution (standard conductivity tables may be found in
various handbooks).
b. Rinse the cell by repeated immersion in distilled water.
c. Again, immerse the cell in the sample several times before obtaining a reading.
3-6
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d. If the meter is equipped with a magic eye, determine the maximum width of the
shadow at least twice, once by approaching the endpoint from a low reading
upward, and once from a high reading downward.
Because the cell constants are subject to slow change even under ideal conditions, and
sometimes to more rapid change under adverse conditions, it is recommended that the cell
constant be periodically established. Table 3-2 can be used for this operation.
For instruments reading in mhos, the cell constant is calculated as follows:
.I\ -i ' ./Y f\
L =
where
106 Kx
L = cell constant
ATj = conductivity, in micromhos per centimeter, of the KC1 solution at the
temperature of measurement
K2 = conductivity, in micromhos per centimeter, of the KC1 solution at the same
temperature as the distilled water used to prepare the reference solution
KX = measured conductance, in mhos
Many different manufacturers produce conductivity meters that perform well on water and
wastewater samples. Selection should be made consistent with sampling requirements,
availability of service and sales, and individual personal preference.
3.5 Turbidimeters (Nephelometers)
Many different instrument designs have been used for the optical measurement of turbidity
by measurement of either transmission or reflection of light. An equal or even greater
Table 3-2
ELECTRICAL CONDUCTIVITY OF POTASSIUM CHLORIDE REFERENCE SOLUTIONS
Solution
A
B
C
Normality
0.1
0.01
0.001
Method of Preparation
7.4365 g/1 KC1 at 20°C
0.7440 g/1 KC1 at 20°C
Dilute 1 00 ml of B to 1 .0 1 at 20°C
Temperature
(°C)
0
18
25
0
18
25
25
Conductivity
(jimho/cm)
7,138
11,167
12,856
773
1,220
1,408
147
3-7
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number of materials have been used or proposed as calibration standards. As described in
reference 10, EPA has standardized on the instrument design and the standard turbidity
suspension of Formazin for instrument calibration.
Both the analyst and the user of turbidity data should keep in mind that a turbidity
measurement is not a substitute for particle weight or residue analysis. Turbidity
instruments can be calibrated to give gravimetric data on specific sample types, but the
influence of particle geometry, specific gravity, refractive index, and color make estimates
of total weight impractical on a variety of sample types.
For production of data with maximum accuracy and precision, the following precautions
should be observed:
a. Protect the sample cuvette from scratches and fingerprints.
b. Use a constant orientation of the sample cuvette while calibrating the instrument
and analyzing samples.
c. Use a well-mixed sample in the sample cuvette; do not take readings until finely
dispersed bubbles have disappeared.
d. Dilute samples containing excess turbidity to some value below 40 nephelometric
turbidity units (NTU); take reading; and multiply results by correct dilution factor.
3.6 Spectrometers
Because a large portion of routine quantitative measurements are performed colorimetri-
cally, the spectrometer or filter photometer is usually the workhorse of any analytical
laboratory. Indeed, the versatility of such instruments and the number of demands imposed
upon them have resulted in a variety of designs and price ranges. Systematic listing and
detailed discussion of all instrumental types are beyond the scope of this chapter; however,
ultraviolet, visible, infrared, and atomic absorption instruments will be discussed.
A spectrometer is an instrument for measuring the amount of light or radiant energy
transmitted through a solution or solid material as a function of wavelength. A spectrometer
differs from a filter photometer in that it uses continuously variable, and more nearly
monochromatic, bands of light. Because filter photometers lack the versatility of
spectrometers, they are used most profitably where standard methodologies are used for
routine analysis.
The essential parts of a spectrometer include the following:
a. A source of radiant energy
b. Monochromator or other device for isolating narrow spectral bands of light
c. Cells (cuvettes) or sample holders for containing samples under investigation
d. A photodetector (a device to detect and measure the radiant energy passing through
the sample)
3-8
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Each of the essential features listed, especially the monochromator and the photodetector
system, varies in design principles from one instrument to another. Some of the
characteristics of the commonly used Perkin-Elmer model 124 double-beam grating
spectrometer are the following:
Light source
Visible region Tungsten
Lamp UV region Deuterium lamp
Wavelength accuracy ±0.5 nm
Spectral bandwidth 0.5, 1.0, and 2.0 nm
Photometric presentation
Linear transmittance 0 to 100 percent
Linear absorbance 0 to \A or 0 to 2A
Photodetector R-136 190 to 800 nm
Sample cells 1.0 to 10 cm
3.6.1 Visible Range
Desirable features on a visible-range spectrometer are determined by the anticipated use of
the instrument. Simple, limited programs requiring use of only a few parameters can
probably be supported by inexpensive but reliable filter photometers. On the other hand, if
the laboratory programs require a wide variety of measurements on diverse samples at low
concentrations, more versatile instruments may be needed. One of the prime considerations
is adaptability to various sample cell sizes from 1.0 to 10.0 cm.
3.6.2 Ultraviolet Range
A UV spectrometer is similar in design to a visible-range instrument except for differences in
the light source and in the optics. The UV light source is a hydrogen- or deuterium-discharge
lamp, which emits radiation in the UV portion of the spectrum, generally from about 200
nm to the low-visible-wavelength region. The optical system and sample cells must be
constructed of UV-transparent material, which is usually quartz. A grating used in a UV
system may be specially cut (blazed) in the UV region for greater sensitivity.
3.6.3 Infrared Range
A number of instrumental modifications are required in the construction of spectrometers
for measurements in the IR region because materials such as glass and quartz absorb IR
radiant energy, and ordinary photocells do not respond to it. Most IR spectrometers use
front-surfaced mirrors to eliminate the necessity for the transmission of radiant energy
through quartz, glass, or other lens materials. These mirrors are usually parabolic to focus
the diffuse IR energy. Such instruments must be protected from high humidities and water
vapor to avoid deterioration of the optical system and the presence of extraneous
absorption bands in the IR.
3-9
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The energy or light source for an IR instrument may be a Nernst glower or a Globar. Each
of these sources has certain characteristics that recommend it for use, but the more rugged
Globar is more commonly used because it also has a more stable emission. The receiving or
detection unit may be a thermocouple, bolometer, thermistor, or photoconductor cell. The
type of analysis being performed dictates the degree of sophistication required in the IR
instrumentation selected to acquire usable data.
3.6.4 Proper Use of Spectrometers
The spectrometer manufacturer's instructions for proper use should be followed in all cases.
Several safeguards against misuse of the instruments, however, are mandatory.
Instruments should be checked for wavelength alinement. If a particular colored solution is
to be used at a closely specified wavelength, considerable loss of sensitivity can be
encountered if the wavelength control is misalined. In visible-range instruments, an excellent
reference point is the maximum absorption for a diluted solution of potassium
permanganate, which has dual peaks at 526 and 546 nm. On inexpensive instruments with
less resolution the permanganate peak appears at 525 to 550 nm as a single, flat-topped
peak.
For both UV and IR instruments, standard absorption curves for many organic materials
have been published so that reference material for standard peaks is easily available.
Standard films of styrene and other transparent plastics are available for IR wavelength
checks. A very good discussion of factors that affect the proper performance of
spectrometers and standard reference materials available to calibrate them can be found in
publications of the National Bureau of Standards.* Use of certified standards is encouraged
whenever practical.
Too much emphasis cannot be placed on care of absorption cells. All absorption cells should
be kept scrupulously clean, free of scratches, fingerprints, smudges, and evaporated film
residues. Matched cells should be checked to see that they are equivalent, and any
differences should be .accounted for during use or in the final data. Directions for cleaning
cells are given in chapter 4.
Generally speaking, trained technicians may operate any of the spectrometers successfully;
however, because interpretation of data from both the UV and IR instruments is becoming
increasingly complex, mere compliance with the operations manual may not be sufficient
for completely accomplishing the special techniques of sample preparation, instrument
operation, and interpretation of absorption curves.
3.6.5 Atomic Absorption
There are a number of differences in the basic design and accessories for atomic absorption
(AA) equipment that require consideration before purchase and during subsequent use.
These choices concern the light sources, nebulizer burners, optical systems, readout devices,
and mode conversions. Because some of these choices are not readily obvious, the purchaser
or user must be familiar with the types and numbers of samples to be analyzed and the
*NBS, Office of Standard Reference Materials, Institute for Materials Research, Washington,
D.C. 20234.
3-10
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specific elements to be measured before making the choice. For a program analyzing a wide
variety of samples for a number of elements at varying concentrations, an instrument of
maximum versatility would be best. Most of the discussion that follows applies to use of
instrumentation in absorption, emission, or fluorescence modes.
3.6.5.1 Lamps
Hollow-cathode (HC) lamps or electrodeless-discharge lamps (EDL) are available for over 70
elements with single-element or multielement capability. Multielement lamps are consid-
erably cheaper per element than single-element lamps, but the savings may not be realized if
the lamps are not used strategically, because all the elements in the cathode deteriorate
when the lamp is used, regardless of which element is measured. The deterioration
phenomena result from the different volatilities of metals used in the cathode. One metal
volatilizes (sputters) more rapidly than the others and redeposits upon an area of the other
cathode metals. Thus, with continual use, a drift in signal will be noted with at least one
metal increasing and the other (or others) decreasing. If one can ignore the dubious cost
savings of multielement lamps, use of single-element lamps could result in more precise and
accurate data.
The line intensities of one element in a multielement HC lamp will usually be less than those
of a lamp containing a pure cathode of the same element because this element must share
the discharge energy with the other elements present. However, this reduction should not
affect the output by a factor of more than one-sixth to one-half, depending on the
particular combination and the number of elements combined. The output can be even
greater in some multielement lamps because alloying may permit a higher operating current
than for the case of the pure cathode. All HC lamps have life expectancies that are related to
the volatility of the cathode metal, and. for this reason the manufacturer's recommendations
for operating should be closely followed.
Recent improvements in design and manufacture of HC lamps and EDLs have resulted in
lamps with more constant output and longer life. Under normal conditions an HC lamp may
be expected to operate satisfactorily for several years. HC lamps used to be guaranteed for a
certain minimum ampere-hour period, but this has been changed to a 90-day warranty. It is
good practice to date newly purchased lamps and to inspect them immediately upon receipt.
The operating current and voltage indicated on the lamp should not be exceeded during use.
An increase in background noise or a loss of sensitivity are signs of lamp deterioration.
A basic design feature of AA spectrometers is the convenience of the HC lamp changeover
system. Some instruments provide for as many as six lamps in a rotating turret, all
electronically stabilized and ready for use by simply rotating the lamp turret. Other
instruments provide for use of only one lamp at a time in the lamp housing, and require
manual removal and replacement whenever more than one element is to be measured. A
quick-changeover system enables frequent lamp changes during the period of operation. If
necessary lamp changes are infrequent, however, multilamp mounts do not represent a great
convenience.
After the proper lamp has been selected, the HC current should be adjusted according to the
manufacturer's recommendations and allowed to electronically stabilize (warm up) before
use. During this 15-min period, the monochromator may be positioned at the correct
wavelength, and the proper slit width may be selected. For those instruments employing a
multilamp turret, a warmup current is provided to those lamps not in use, thereby
3-11
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minimizing the warmup period after the turret is rotated. In a single-lamp instrument, the
instability exhibited during warmup is minimized by the use of a double-beam optical
system.
3.6.5.2 Burner Types
The most difficult and inefficient step in the AA process is converting the metal in the
sample from an ion or a molecule to the neutral atomic state. It is the function of the
atomizer/burner to produce the desired neutral atomic condition of the elements. With
minor modifications burners are the same as those used for flame photometry.
Basically there are two different types of burners. They are the total-consumption, or
surface-mix burner, and the laminar-flow or premix burner. There are many variations of
these two basic types, such as the Doling, the high-solids, the turbulent-flow, the triflame,
the nitrous oxide burners, and many others. As one might expect, there are many
similarities among the various burners, the different names resulting from the different
manufacturers. The element being determined and the type of sample solution dictate the
type of burner to be used. Generally, all types and makes of burner can be adjusted
laterally, rotationally, and vertically for selection of the most sensitive area of the flame for
the specific element sought.
Nonflame techniques have gained wide acceptance in AA analysis because of the extreme
sensitivity and the capability to directly introduce very diverse sample matrices. These
systems, which replace the conventional flame burner, come in various designs using
electrical resistance to produce temperatures as high as 3,500°C.
3.6.5.3 Single- and Double- (Split-) Beam Instruments
There is a great deal of existing uncertainty among instrument users about the relative
merits of single- and double-beam instruments. Neither system is appropriate for all cases.
With a single-beam instrument the light beam from the source passes directly through the
flame to the detector. In a double-beam system the light from the source is divided by a
beam splitter into two paths. One path, the reference beam, goes directly to the detector.
The second path, the sample beam, goes through the flame to the detector. The chopper
alternately reflects and passes each beam, creating two equal beams falling alternately upon
the detector. If the beams are equal, they cancel the alternate impulses reaching the
detector, and no signal is generated. If the beams are different, the resulting imbalance
causes the detector to generate a signal that is amplified and measured. The difference
between the reference and sample beams is then determined as a direct function of absorbed
light. The advantages of the double-beam design are that any variations in the source are of
reduced importance, and smaller dependence is placed upon the stability of the power
supply. Conversely, stabilization of the power supply can eliminate the apparent need for
the split-beam system. Furthermore, the beam splitter requires additional mirrors or optical
accessories that cause some loss of radiant energy.
A single-beam system does not monitor source variations, but offers certain other
advantages. It allows use of low-intensity lamps, smaller slit settings, and smaller gain. As a
consequence, the single-beam instrument, properly designed, is capable of operating with
lower noise and better signal-to-noise ratio, and therefore with better precision and
improved sensitivity. Because the simplified optical system conserves radiant energy,
3-12
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especially in the shorter wavelengths, it facilitates operation in the low-wavelength range.
With this advantage, it should be possible to obtain better sensitivity for those elements with
a strong resonance line below 350 nm and particularly those below 300 nm. Background
correction techniques are also available in single-beam systems.
Double-beam instruments, however, offer the opportunity to perform more sophisticated
techniques like background correction, two-channel multielement analysis, use of internal
standards, and element rationing. If one of these techniques is necessary, a double-beam
instrument must be considered.
3.6.5.4 Readout Devices
Readout devices in even the lower cost AA instruments include digital concentration
display. Using high-speed digital electronics, data-handling techniques encompass multiple
calibration standards, regression analysis to characterize the calibration curves, mean
variance and standard deviation statistical programs for sample calculations, and various
forms of printout reports in addition to recorder output. Choice of a readout system is
predicated largely upon laboratory needs and availability of budget. In general, any step
toward automation is desirable, but the degree of automation should be compatible with the
laboratory program.
3.6.5.5 Miscellaneous Accessories
A number of instruments contain a mode selector, making an instrument usable for either
absorption or emission. The conversion to emission may be a desirable feature because
certain elements are more amenable to analysis by this method. Some models offer an
option of atomic fluorescence and can also be used as a UV/visible spectrometer.
Automatic sample changers are offered for almost all instruments on the market, and as has
been previously stated, any automation feature is desirable. However, unless a laboratory
program performs a large number of repetitious measurements daily, an automatic sample
changer is not required. As a practical measure, other commonly used sample-changing
devices, although not expressly designed for AA use, can be interfaced with most AA
instruments.
3.7 Organic Carbon Analyzers
A number of instruments designed to measure total organic carbon (TOC) in waters and
wastes have appeared on the market in the past several years. The first of these units
involved pyrolysis followed by IR measurement of the carbon dioxide formed. Sample
injection of 20 to 200 n\ in a carrier gas of air or oxygen was performed with a syringe.
Combustion at 800°C to 900°C followed by IR analysis was performed automatically with
final output on an analog recorder. Systems using these principles are still produced and
represent a large part of the TOC market.
Other .techniques of TOC analysis that modify every phase of the original TOC instruments
have been introduced. Sample presentation in small metallic boats and purging of CO2 from
solution are two new techniques. Wet chemical oxidation, either external to the instrument
or within the instrument, using various oxidants including ultraviolet irradiation is now in
wide use. Measurement of the CO2 by reduction to methane (CH4) and quantitation with a
flame ionization detector are also available. Various methods of data handling are now used,
3-13
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ranging from recorder output to direct readings and printouts of concentration. Techniques
are also available for measuring materials like soil and sludges, and also the volatile
component of the TOC. Sensitivity on some systems has been extended down to the
microgram per liter level. The major problems associated with TOC measurements are
interference from forms of inorganic carbon and the difficulty of obtaining a representative
sample in the presence of particulate matter. Each system has its own procedure for sample
pretreatment or for accounting for these problems. When choosing a TOC instrument,
consideration should be given to the types of samples to be analyzed, the expected
concentration range, and the forms of carbon to be measured.
3.8 Gas Chromatographs
Because GC's are available from a large number of manufacturers, selection of a particular
manufacturer may be based on convenience. No single multipurpose GC instrument permits
analysis of a wide range of compounds. In this case, a GC/MS could be considered (11). If,
however, relatively few types of environmentally significant compounds are being surveyed,
an inexpensive system equipped with a glass-lined injection port, electrolytic conductivity
detector, and analog recorder is a good choice. A review of the organic methods (12) to be
used will give the analyst all the necessary information on the specific instrument,
apparatus, and materials necessary for each type or class of compounds. A discussion of
various quality control considerations in GC analysis is given in chapter 8 of this manual.
Data handling requirements vary widely, and the need to automate GC data collection is
determined by the extent of the sample load. In a routine monitoring laboratory, GC
systems incorporating their own microprocessers and report generating capabilities would be
useful in solving this problem. Because such systems greatly increase the cost, the overall
economy of this choice must be considered.
3.9 References
1. "Single Arm Balances Testing," Part 41 of 1976 Book of ASTM Standards, American
Society for Testing and Materials, Philadelphia (1976).
2. Sorenson, S. P. L., Biochem. Z., 21, 201 (1909).
3. "pH of Water and Wastewater," from Part 31 of 1976 Book of ASTM Standards,
American Society for Testing and Materials, Philadelphia (1976).
4. Catalog of NBS Standard Reference Materials, NBS Special Publication 260, National
Bureau of Standards (1975-76).
5. Methods for Chemical Analysis of Water and Wastes, U.S. EPA, Office of Research and
Development, EMSL, Cincinnati (1978).
6. Rechnitz, G. A., "Ion-Selective Electrodes," Chem. Eng. News (June 12, 1967), p. 146.
7. Riseman, Jean M., "Measurement of Inorganic Water Pollutants by Specific Ion
Electrode," American Laboratory (July 1969), p. 32.
8. Koryta, Jiri, "Theory and Application of Ion-Selective Electrodes," Anal. Chim. Acta,
61, 329-411 (1972).
9. Covington, A. L, "Ion-Selective Electrodes," CRC Critical Review in Anal. Chem. (Jan.
1974), pp. 355-406.
10. Black, A. P., and Hannah, S. A., "Measurement of Low Turbidities," J. Am. Water
Works Assoc.,57, 901 (1965).
11. EPA GC/MS Procedural Manual, Budde, W. L., and Eichelberger, J. W., Editors, 1st
Edition, Vol. 1, U.S. EPA, Office of Research and Development, EMSL, Cincinnati (in
press).
3-14
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12. Methods for Organic Analysis of Water and Wastes, U.S. EPA, EMSL, Environmental
Research Center, Cincinnati (in preparation).
3-15
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Chapter 4
GLASSWARE
4.1 General
The measurement of trace constituents in water demands methods capable of maximum
sensitivity. This is especially true for metals and trace organics such as pesticides, as well as
for the determination of ammonia and phosphorus. In addition to sensitive methods,
however, there are other areas that require special consideration. One such area is that of the
cleanliness of laboratory glassware. Obviously, the very sensitive analytical systems are more
sensitive to errors resulting from the improper use or choice of apparatus, as well as to
contamination effects due to an improper method of cleaning the apparatus. The purpose of
this chapter is to discuss the kinds of glassware available, the use of volumetric ware, and
various cleaning requirements.
4.2 Types of Glassware
Laboratory vessels serve three functions: storage of reagents, measurement of solution
volumes, and confinement of reactions. For special purposes, vessels made from materials
such as porcelain, nickel, iron, aluminum, platinum, stainless steel, and plastic may be
employed to advantage. Glass, however, is the most widely used material of construction.
There are many grades and types of glassware from which to choose, ranging from student
grade to others possessing specific properties such as super strength, low boron content, and
resistance to thermal shock or alkali. Soft glass containers are not recommended for general
use, especially for storage of reagents because of the possibility of dissolving of the glass (or
of some of the constituents of the glass). The mainstay of the modern analytical laboratory
is a highly resistant borosilicate glass, such as that manufactured by Corning Glass Works
under the name "Pyrex" or by Kimble Glass Co. as "Kimax." This glassware is satisfactory
for all analyses included in reference 1.
Depending on the particular manufacturer, various trade names are used for specific brands
possessing special properties such as resistance to heat, shock, and alkalies. Examples of
some of these special brands follow:
a. Kimax- or Pyrex-brand glass is a relatively inert all-purpose borosilicate glass.
b. Vycor-brand glass is a silica glass (96 percent) made to withstand continuous
temperatures up to 900° C and can be down-shocked in ice water without breakage.
c. Coming-brand glass is claimed to be 50 times more resistant to alkalies than
conventional ware and practically boron-free (maximum 0.2 percent).
d. Ray-Sorb- or Low-Actinic-brand glass is used when the reagents or materials are light
sensitive.
e. Corex-brand labware is harder than conventional borosilicates and therefore better
able to resist clouding and scratching.
The use of plastic vessels, containers, and other apparatus made of Teflon, polyethylene,
polystyrene, and polypropylene has increased markedly over recent years. Some of these
4-1
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materials, such as Teflon, are quite expensive; however, Teflon stopcock plugs have
practically replaced glass plugs in burets, separatory funnels, etc., because lubrication to
avoid sticking or "freezing" is not required. Polypropylene, a methylpentene polymer, is
available as laboratory bottles, graduates, beakers, and even volumetric flasks. It is crystal
clear, shatterproof, autoclavable, and chemically resistant.
The following are some points to consider in choosing glassware or plasticware:
a. The special types of glass listed above, other than Pyrex or Kimax, generally are not
required to perform the analyses given in "Methods for Chemical Analysis of Water
and Wastes" (1).
b. Unless instructed otherwise, borosilicate or polyethylene bottles may be used for the
storage of reagents and standard solutions.
c. Dilute metal solutions are prone to plate out on container walls over long periods of
storage. Thus, dilute metal standard solutions must be prepared fresh at the time of
analysis.
d. For some operations, disposable glassware is entirely satisfactory. One example is
the use of disposable test tubes as sample containers for use with the Technicon
automatic sampler.
e. Plastic bottles of polyethylene and Teflon have been found satisfactory for the
shipment of water samples. Strong mineral acids (such as sulfuric acid) and organic
solvents will readily attack polyethylene and are to be avoided.
f. Borosilicate glassware is not completely inert, particularly to alkalies; therefore,
standard solutions of silica, boron, and the alkali metals are usually stored in
polyethylene bottles.
For additional information the reader is referred to the catalogs of the various glass and
plastic manufacturers. These catalogs contain a wealth of information such as specific
properties, uses, and sizes.
4.3 Volumetric Analyses
By common usage, accurately calibrated glassware for precise measurements of volume has
become known as volumetric glassware. This group includes volumetric flasks, volumetric
pipets, and accurately calibrated burets. Less accurate types of glassware including
graduated cylinders and serological and measuring pipets also have specific uses in the
analytical laboratory when exact volumes are unnecessary.
The precision of volumetric work depends in part upon the accuracy with which volumes of
solutions can be measured. There are certain sources of error that must be carefully
considered. The volumetric apparatus must be read correctly; that is, the bottom of the
meniscus should be tangent to the calibration mark. There are other sources of error,
however, such as changes in temperature, which result in changes in the actual capacity of
glass apparatus and in the volume of the solutions. The capacity of an ordinary glass flask of
1000-ml volume increases 0.025 ml/deg with rise in temperature, but if the flask is made of
borosilicate glass, the increase is much less. One thousand milliliters of water or of most
O.IN solutions increases in volume by approximately 0.20 ml/deg increase at room
4-2
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temperature. Thus solutions must be measured at the temperature at which the apparatus
was calibrated. This temperature (usually 20°C) will be indicated on all volumetric ware.
There may also be errors of calibration of the apparatus; that is, the volume marked on the
apparatus may not be the true volume. Such errors can be eliminated only by recalibrating
the apparatus or by replacing it.
Volumetric apparatus is calibrated to contain or to deliver a definite volume of liquid. This
will be indicated on the apparatus with the letters "TC" (to contain) or "TD" (to deliver).
Volumetric flasks are calibrated to contain a given volume and are available in various shapes
and sizes.
Volumetric pipets are calibrated to deliver a fixed volume. The usual capacities are 1
through 100 ml although micropipets are also available. Micropipets are most useful in
furnace work and are available in sizes ranging from 1 to 100 n\.
In emptying volumetric pipets, they should be held in a vertical position and the outflow
should be unrestricted. The tip of the pipet is kept in contact with the wall of the receiving
vessel for a second or two after the free flow has stopped. The liquid remaining in the tip is
not removed; this is most important.
Measuring and serological pipets should also be held in a vertical position for dispensing
liquids; however, the tip of the pipet is only touched to the wet surface of the receiving
vessel after the outflow has ceased. For those pipets where the small amount of liquid
remaining in the tip is to be blown out and added, indication is made by a frosted band near
the top.
Burets are used to deliver definite volumes. The more common types are usually of 25- or
50-ml capacity, graduated to tenths of a milliliter, and are provided with stopcocks. For
precise analytical methods in microchemistry, microburets are also used. Microburets
generally are of 5- or 10-ml capacity, graduated in divisions of hundredths of a milliliter.
Automatic burets with reservoirs are also available ranging in capacity from 10 to 100 ml.
Reservoir capacity ranges from 100 to 4,000 ml.
General rules in regard to the manipulation of a buret are as follows: Do not attempt to dry
a buret that has been cleaned for use, but rinse it two or three times with a small volume of
the solution with which it is to be filled. Do not allow alkaline solutions to stand in a buret
because the glass will be attacked, and the stopcock, unless made of Teflon, will tend to
freeze. A 50-ml buret should not be emptied faster than 0.7 ml/s, otherwise too much liquid
will adhere to the walls and as the solution drains down, the meniscus will gradually rise,
giving a high false reading. It should be emphasized that improper use or reading of burets
can result in serious calculation errors.
In the case of all apparatus for delivering liquids, the glass must be absolutely clean so that
the film of liquid never breaks at any point. Careful attention must be paid to this fact or
the required amount of solution will not be delivered. The various cleaning agents and their
use are described later.
4.4 Federal Specifications for Volumetric Glassware
Reference 2 contains a description of Federal specifications for volumetric glassware. The
National Bureau of Standards no longer accepts stock quantities of volumetric apparatus
4-3
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from manufacturers or dealers for certification and return for future sale to consumers. This
certification service is still available, but apparatus will be tested only when submitted by
the ultimate user, and then only after an agreement has been reached with the Bureau
concerning the work to be done.
Consequently, the various glass manufacturers have discontinued the listing of NBS-certified
ware. In its place catalog listings of volumetric glass apparatus that meet the Federal
specifications are designated as class A and all such glassware is permanently marked with a
large "A." These NBS specifications are listed in table 4-1. The glassware in question
includes the usual burets, volumetric flasks, and volumetric pipets.
In addition to the "A" marking found on calibrated glassware and the temperature at which
the calibration was made, other markings also appear. These include the type of glass, such
Table 4-1
TOLERANCES FOR VOLUMETRIC
GLASSWARE*
Type
of
Glassware
Graduated flask
Transfer pipet
Buret3
Capacity2
(ml)
25
50
100
200
250
300
500
1,000
2,000
2
5
10
25
30
50
100
200
5
10
30
50
100
Limit of Error
(ml)
0.03
0.05
0.08
0.10
0.11
0.12
0.15
0.30
0.50
0.006
0.01
0.02
0.025
0.03
0.05
0.08
0.10
0.01
0.02
0.03
0.05
0.10
1 Abridged from reference 3.
2 Less than and including.
3 Limits of error are of total or partial capacity.
Customary practice is to test the capacity at five
intervals.
4-4
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as Pyrex, Corex, or Kimax; the stock number of the particular item; and the capacity of the
vessel. If the vessel contains a ground-glass connection, this will also be included along with
the TD or TC symbol. An example of the markings usually found on volumetric ware is
shown in figure 4-1. Class A glassware need not be recalibrated before use. However, if it
should become necessary to calibrate a particular piece of glassware, directions may be
found in texts (4) on quantitative analysis.
4.5 Cleaning of Glass and Porcelain
The method of cleaning should be adapted to both the substances that are to be removed,
and the determination to be performed. Water-soluble substances are simply washed out
with hot or cold water, and the vessel is finally rinsed with successive small amounts of
distilled water. Other substances more difficult to remove may require the use of a
detergent, organic solvent, dichromate cleaning solution, nitric acid, or aqua regia (25
percent by volume concentrated HNO3 in concentrated HC1). In all cases it is good practice
to rinse a vessel with tap water as soon as possible after use. Material allowed to dry on
glassware is much more difficult to remove.
Volumetric glassware, especially burets, may be thoroughly cleaned by a mixture containing
the following: 30 g of sodium hydroxide, 4 g of sodium hexametaphosphate (trade name,
Calgon), 8 g of trisodium phosphate, and 1 1 of water. A gram or two of sodium lauryl
sulfate or other surfactant will improve its action in some cases. This solution should be
used with a buret brush.
Dichromate cleaning solution (chromic acid) is a powerful cleaning agent; however, because
of its destructive nature upon clothing and upon laboratory furniture, extreme care must be
taken when using this mixture. If any of the solution is spilled, it must be cleaned up
immediately. Chromic acid solution may be prepared in the laboratory by adding 1 1 of
concentrated sulfuric acid slowly, with stirring, to a 35-ml saturated sodium dichromate
solution. This mixture must be allowed to stand for approximately 15 min in the vessel that
is being cleaned and may then be returned to a storage bottle. Following the chromic acid
wash, the vessels are rinsed thoroughly with tap water, then with small successive portions
of distilled water. The analyst should be cautioned that when chromium is included in the
PYREX fl
USA
y*-
J 1 0
-*•
500 ml ± 0.20 ml <*-
TP ?n°r ^
NO 5680 *
GLASS CO.
TYPE
STANDARD j
TAPER » 5 ^
SIZE 19
TO
CONTAIN
, ., STOCK
NO.
KIMAX
* USA
s
V
,. *. TP 9n°r
^ NO 28013
Figure 4-1. Example of markings on glassware.
4-5
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scheme of analysis, it is imperative that the last traces of dichromate be removed from the
apparatus. To this end, a substitute for dichromate cleaning solution, called Nochromix,* is
available and may be used to advantage. Fuming nitric acid is another powerful cleaning
agent, but is disagreeable to handle. As with dichromate, when the acid becomes dilute, the
cleaning mixture is no longer effective. A mixture of concentrated sulfuric and fuming nitric
acids is even more efficient but is also hazardous to use. A persistent greasy layer or spot
may be removed by acetone or by allowing a warm solution of sodium hydroxide, about 1 g
per 50 ml of water, to stand in the vessel for 10 to 15 min; after rinsing with water, dilute
hydrochloric acid, and water again, the vessel is usually clean. Alcoholic potassium
hydroxide is also effective in removing grease. To dry glass apparatus, rinse with acetone and
blow or draw air through it.
4.6 Special Cleaning Requirements
Absorption cells, used in spectrophotometers, should be kept scrupulously clean, free of
scratches, fingerprints, smudges, and evaporated film residues. The cells may be cleaned with
detergent solutions for removal of organic residues, but should not be soaked for prolonged
periods in caustic solutions because of the possibility of etching. Organic solvents may be
used to rinse cells in which organic materials have been used. Nitric acid rinses are
permissible, but dichromate solutions are not recommended because of the adsorptive
properties of dichromate on glass. Rinsing and drying of cells with alcohol or acetone before
storage is a preferred practice. Matched cells should be checked to see that they are
equivalent by placing portions of the same solution in both cells and taking several readings
of the transmittance (T, percent) or optical density (OD) values.
For certain determinations-, especially trace metals, the glassware should also be rinsed with
a 1:1 nitric acid-water mixture. This operation is followed by thoroughly rinsing with tap
water and successive portions of distilled water. This may require as many as 12 to 15 rinses,
especially if chromium is being determined. The nitric acid rinse is also especially important
if lead is being determined.
Glassware to be used for phosphate determinations should not be washed with detergents
containing phosphates. This glassware must be thoroughly rinsed with tap water and
distilled water. For ammonia and Kjeldahl nitrogen, the glassware must be rinsed with
ammonia-free water. (See ch. 2.)
Glassware to be used in the determination of trace organic constituents in water, such as
chlorinated pesticides, should be as free as possible of organic contaminants. A chromic acid
wash of at least 15 min is necessary to destroy these organic residues. Rinse thoroughly with
tap water and, finally, with distilled water. Glassware may be dried for immediate use by
rinsing with redistilled acetone. Otherwise glassware may be oven dried or drip dried.
Glassware should be stored immediately after drying to prevent any accumulation of dust
and stored inverted or with mouth of glassware covered with foil.
Bottles to be used for the collection of samples for organic analyses should be rinsed
successively with chromic acid cleaning solution, tap water, distilled water, and, finally,
several times with a redistilled solvent such as acetone, hexane, petroleum ether, or
chloroform. Caps are washed with detergent, rinsed with tap water, distilled water, and
*Available from Godax Laboratories, 6 Varick Street, New York, N.Y. 10013.
4-6
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solvent. Liners are treated in the same way as the bottles and are stored in a sealed
container.
4.7 Disposable Glassware
When the risk of washing a pipet for reuse becomes too great, as in the case of use with
toxic materials, or when the cost of washing glassware becomes prohibitive, disposable
vessels may be the answer, provided they meet the necessary specification. Various types are
available including bacteriological, serological, and microdilution pipets. Disposable glass-
ware generally is made of soft glass although plastic vessels and pipets are also available.
4.8 Specialized Glassware
The use of vessels and glassware fitted with standard-taper, ground-glass, and ball-and-socket
joints has increased because of certain advantages such as less leakage and fewer freezeups.
Standard-taper, interchangeable ground joints save time and trouble in assembling apparatus.
They are precision-ground with tested abrasives to insure an accurate fit and freedom from
leakage. Ball and socket joints increase flexibility of operation and eliminate the need for
exact alinements of apparatus. Symbols and their meaning as applied to standard joints,
stoppers, and stopcocks are shown below.
4.8.1 Standard Taper (1)
The symbol J is used to designate interchangeable joints, stoppers, and stopcocks that
comply with the requirements of reference 5. All mating parts are finished to a 1:10 taper.
The size of a particular piece appears after the appropriate symbol. Primarily because of
greater variety of apparatus equipped with 5 fittings, a number of different types of
identifications are used:
a. For joints-a two-part number as 5 24/40, with 24 being the approximate diameter
in millimeters at the large end of the taper and 40 the axial length of taper, also in
millimeters
b. For stopcocks-a single number, as J 2, with 2 mm being the approximate diameter
of the hole or holes through the plug
c. For bottles-a single number, as J 19, with 19 mm being the appropriate diameter at
top of neck. However, there are differences in dimensions between the bottle and
flask stoppers
d. For flasks and similar containers—a single number, as J 19, with 19 mm being the
appropriate diameter of the opening at top of neck
4.8.2 Spherical Joints (?)
The designation f is for spherical (semiball) joints complying with reference 5. The complete
designation of a spherical joint also consists of a two-part number, as 12/2, with 12 being
the approximate diameter of the ball and 2 the bore of the ball and the socket, also in
millimeters.
4-7
-------
4.8.3 Product Standard (§)
The symbol £ is used for stopcocks with Teflon plugs, the mating surfaces being finished to
a 1:5 taper. As with £ stopcocks, a single number is used. Thus, | 2 means a Teflon
stopcock with a hole of approximately 2-mm diameter in the plug.
4.9 Fritted Ware
For certain laboratory operations the use of fritted ware for filtration, gas dispersion,
absorption, or extractions may be advantageous.
There are six different porosities of fritted ware available, depending on its intended use.
Porosity is controlled in manufacture, and disks are individually tested and graded into these
classifications. The extra-coarse and coarse porosities are held toward the maximum pore
diameter as listed. The medium, fine, very fine, and ultrafme are held toward the minimum
pore diameter as listed in table 4-2.
Pore sizes are determined by the method specified in reference 6.
4.9.1 Recommended Procedures for Maximum Filter Life
a. New Filters. Wash new filters by suction with hot hydrochloric acid, followed by a
water rinse.
b. Pressure Limits. The maximum, safe, differential pressure on a disk is 15 lb/in2.
c. Thermal Shock. Fritted ware has less resistance to thermal shock than nonporous
glassware. Hence, excessive, rapid temperature changes and direct exposure to a
flame should be avoided. Heating in a furnace to 500° C may be done safely,
provided the heating and cooling are gradual. Dry ware may be brought to constant
weight by heating at 105°C to 110°C.
Never subject a damp filter of ultrafine porosity to a sudden temperature change. Steam
produced in the interior may cause cracking.
Table 4-2
FRITTED-WARE POROSITY
Porosity
Grade
Extra Coarse
Coarse
Medium
Fine
Very Fine
Ultrafine
Designation
EC
C
M
F
VF
UF
Pore Size
(Mm)
170-220
40-60
10-15
4-5.5
2-2.5
0.9-1.4
Principal Uses
Coarse filtration ; gas dispersion,
absorption
Coarse filtration; gas dispersion,
absorption
Filtration and extraction
Filtration and extraction
General bacterial filtration
General bacterial filtration
washing, and
washing, and
4-8
-------
4.9.2 Cleaning of Used Filters
In many cases, precipitates can be removed by rinsing with water, passed through from the
underside, with the pressure not exceeding 15 lb/in2. The suggestions that follow will be
helpful in dealing with material that will not be removed by such a reverse water-wash:
Material Removal Agent
Albumen Hot ammonia or hydrochloric acid
Aluminous and siliceous residues Hydrofluoric acid (2 percent) followed by concen-
trated sdlfuric acid; rinse immediately with
water until no trace of acid can be detected.
Copper or iron oxides Hot hydrochloric acid plus potassium chlorate
Fatty materials Carbon tetrachloride
Mercuric sulfide Hot aqua regia
Organic matter Hot, concentrated cleaning solution, or hot, con-
centrated sulfuric acid with a few drops of sodi-
ium nitrite
Silver chloride Ammonium or sodium hyposulfite
4.10 References
1. Methods for Chemical Analysis of Water and Wastes, U.S. EPA, Office of Research and
Development, EMSL (1978).
2. Hughes, J. C., Testing of Glass Volumetric Apparatus, NBS Circular 602, National
Bureau of Standards (1959).
3. Peffer, E. L., and Mulligan, G. C., Testing of Glass Volumetric Apparatus, NBS Circular
434, National Bureau of Standards (1941).
4. Willard, H. H., and Furman, N. H., Elementary Quantitative Analysis—Theory and
Practice, D. Van Nostrand Co., Inc., New York (1947).
5. Interchangeable Ground Glass Joints, Commercial Standard CS-21-30, National Bureau
of Standards (Aug. 25, 1930).
6. Maximum Pore Diameter and Permeability of Rigid Porous Filters for Laboratory Use,
E-128, American Society for Testing and Materials, Philadelphia (1968).
4-9
-------
Chapter 5
REAGENTS, SOLVENTS, AND GASES
5.1 Introduction
The objective of this chapter is to provide general information and suggestions that will
serve to keep the analyst conscious of his responsibilities in analytical quality control, as
they relate to reagents, solvents, and gases. While the material presented here will assist the
analyst in producing high-quality data, it is by no means complete. It is incumbent on the
analyst to obtain details of special precautions required to insure proper selection,
preparation, and storage of reagents, solvents, and gases from the descriptions of individual
methods.
5.2 Reagent Quality
Chemical reagents, solvents, and gases are available in a wide variety of grades of purity,
ranging from technical grade to various ultrapure grades. The purity of these materials
required in analytical chemistry varies with the type of analysis. The parameter being
measured and the sensitivity and specificity of the detection system are important factors in
determining the purity of the reagents required. For many analyses, including most
inorganic analyses, analytical reagent grade is satisfactory. Other analyses, such as trace
organic and radiological, frequently require special ultrapure reagents, solvents, and gases. In
methods where the purity of reagents is not specified it is intended that analytical reagent
grade be used. Reagents of lesser purity than that specified by the method should not be
used. The labels on the container should be checked and the contents examined to verify
that the purity of the reagents meets the needs of the particular method involved. The
quality of reagents, solvents, and gases required for the various classes of analyses—inorganic,
metals, radiological, and organic—are discussed in this section.
Reagents must always be prepared and standardized with the utmost of care and technique
against reliable primary standards. They must be restandardized or prepared fresh as often as
required by their stability. Stock and working standard solutions must be checked regularly
for signs of deterioration; e.g., discoloration, formation of precipitates, and change of
concentration. Standard solutions should be properly labeled as to compound, concentra-
tion, solvent, date, and preparer.
Primary standards must be obtained from a reliable source, pretreated (e.g., dried, under
specified conditions), accurately prepared in calibrated volumetric glassware, and stored in
containers that will not alter the reagent. A large number of primary standards are available
from the National Bureau of Standards (NBS). A complete listing of available standards is
given in reference 1. Primary standards may also be obtained from many chemical supply
companies. Suppliers for special quality reagents, solvents, and gases are noted in later
discussions of the various classes of analyses. Reagents and solvents of all grades are available
from many chemical supply houses.
There is some confusion among chemists as to the definition of the terms "Analytical
Reagent Grade," "Reagent Grade," and "ACS Analytical Reagent Grade." A review of the
literature and chemical supply catalogs indicates that the three terms are synonymous.
Hereafter, in this document, the term "Analytical Reagent Grade" (AR) will be used. It is
5-1
-------
intended that AR-grade chemicals and solvents shall conform to the current specifications of
the Committee on Analytical Reagents of the American Chemical Society (2).
References 3 through 5 devote several chapters to problems related to preparation,
standardization, and storage of reagents. The information provided therein is particularly
appropriate to inorganic determinations. The type of volumetric glassware to be used, the
effect of certain reagents on glassware, the effect of temperature on volumetric
measurements, purity of reagents, absorption of gases and water vapor from the air,
standardization of solutions, instability, and the need for frequent standardization of certain
reagents are among the topics discussed. It is recommended that the analyst become
thoroughly familiar with these publications.
5.2.1 General Inorganic Analyses
In general, AR-grade reagents and solvents are satisfactory for inorganic analyses. Primary
standard reagents must, of course, be used for standardizing all volumetric solutions.
Commercially prepared reagents and standard solutions are very convenient and may be
used when it is demonstrated that they meet the method requirements. All prepared
reagents must be checked for accuracy.
The individual methods specify the reagents that require frequent standardization, or other
special treatment, and the analyst must follow through with these essential operations. To
avoid waste, the analyst should prepare a limited volume of such reagents, depending on the
quantity required over a given period of time. Examples and brief discussions of the kind of
problems that occur are given in section 5.3.
As far as possible, distilled water used for preparation of reagent solutions must be free of
measurable amounts of the constituent to be determined. Special requirements for distilled
water are given in chapter 3 of this manual and in individual method descriptions.
Compressed gases, such as oxygen and nitrogen, used for total organic carbon determination
may be of commercial grade.
5.2.2 Metals Analyses
All standards used for atomic absorption and emission spectroscopy should be of
spectroquality. It is recommended that other reagents and solvents also be of spectroquality,
although AR grade is sometimes satisfactory. Standards may be prepared by the analyst in
the laboratory, or spectrographically standardized materials may be purchased commer-
cially. Standards required for determination of metals in water are not generally available
from the National Bureau of Standards.
Analytical-reagent-grade nitric and hydrochloric acids must be specially prepared by
distillation in borosilicate glass and diluted with deionized distilled water. All other reagents
and standards are also prepared in deionized water.
In general, fuel and oxidant gases used for atomic absorption can be of commercial grade.
Air supplied by an ordinary laboratory compressor is quite satisfactory, if adequate pressure
is maintained and necessary precautions are taken to filter oil, water, and possible trace
metals from the line. For certain determinations such as aluminum, AR-grade nitrous oxide
is required.
5-2
-------
5.2.3 Radiological Analyses
The great sensitivity of radioactive counting instruments requires that scintillation-grade
reagents and solvents, or equivalent, be used for all radioactivity determinations. Some of
the reagents, for example, strontium carbonate and yttrium oxide carriers used for the
determination of strontium-90 and yttrium-90, must be stable, that is, free of radioactivity.
Barium sulfate, used for coprecipitation of radium, must be free from all traces of radium.
These reagents and solvents are commercially available from chemical supply houses.
Calibrated standard sources of specific radioactive materials with known count and date of
counting are available from various suppliers. No single company supplies all standards.
Gases used for radioactive counting must be of high purity and extra dry. Gases such as
helium and air are aged for about 30 days to allow radioactive background to decay. All
gases are checked for background before use. Some cylinders contain inherent radioactivity
that is imparted to the gas. When this background is above normal, the gas should not be
used for radioactivity determinations.
5.2.4 Organic Chemical Analyses
The minimum purity of reagents that can be used for organic analyses is AR grade.
Reference-grade standards should be used whenever available. Special note should be taken
of the assay of standard materials. Owing to the great sensitivity (nanogram and
subnanogram quantities) of gas chromatography (GC), which is often used to quantitate
organic results, much greater purity is frequently required (6). The specificity of some GC
detectors requires that reagents and solvents be free of certain classes of compounds. For
example, analyses by electron capture require that reagents and solvents be free of
electronegative materials that would interfere with the determination of specific compounds
in the sample. Similarly, use of the flame photometric detector requires that reagents and
solvents be free from sulfur and phosphorus interference. Pesticide-quality solvents,
available from several sources, are required when doing low-level work. AR-grade solvents
are permissible when analyzing industrial waste samples.
However, the contents of each container must be checked to assure its suitability for the
analyses. Similarly, all analytical reagents and other chemicals must also be checked
routinely.
The quality of gases required for GC determinations varies somewhat with the type of
detector. In general, the compressed gases are a prepurified dry grade. Grade A helium from
the U.S. Bureau of Mines has always been satisfactory. The Dohrmann nitrogen detection
system requires the use of ultrapure hydrogen for satisfactory results. Argon-methane used
for electron-capture work should be oxygen free and should have an oxygen trap in the
supply line. The use of molecular-sieve, carrier-gas filters and drying tubes is required on
combustion gases and is recommended for use on all other gases. It is recommended that the
analyst familiarize himself with an article by Burke (7) on practical aspects of GC.
All reagents, solvents, and adsorbents used for thin-layer chromatography must be checked
to be certain that there are no impurities present that will react with the chromogenic
reagent or otherwise interfere with subsequent qualitative or quantitative determinations.
Glass-backed layers prepared in the laboratory or precoated layers supplied by a
manufacturer may be used; however, precoated layers are more difficult to scrape. It is
5-3
-------
recommended, therefore, that layers prepared in the laboratory be used when zones are to
be scraped to recover isolated compounds. Plastic-backed layers are generally unsatisfactory
for this type of analysis.
Adsorbents most commonly used for column chromatographic cleanup of sample extracts
are Florisil,* silica gel, and alumina. These must be preactivated according to the method
specifications and checked for interfering constituents.
5.3 Elimination of Determinate Errors
To produce high-quality analytical data, determinate errors must be eliminated or at least
minimized. For purposes of this discussion, we assume that a competent analyst and reliable
equipment in optimum operating condition are available. Thus, determinate errors that
might result from an inexperienced or careless analyst and poor equipment are eliminated.
The remaining sources of error are the reagents, solvents, and gases that are used throughout
the analyses. The quality of these materials, even though they are AR grade or better, may
vary from one source to another, from one lot to another, and even within the same lot.
Therefore, the analyst must predetermine that all of these materials are free of interfering
substances under the conditions of the analyses. To do this he must have a regular check
program. Materials that do not meet requirements are replaced or purified so that they can
be used.
5.3.1 Reagent Blank
The first step the analyst must take is to determine the background or blank of each of the
reagents and solvents used in a given method of analysis. The conditions for determining the
blank must be identical to those used throughout the analysis, including the detection
system. If the reagents and solvents contain substances that interfere with a particular
analysis, they should be treated so that they can be used, or satisfactory reagents and
solvents must be found.
5.3.2 Method Blank
After determining the individual reagent or solvent blanks, the analyst must determine the
method blank to see if the cumulative blank interferes with the analyses. The method blank
is determined by following the procedure step by step, including all of the reagents and
solvents, in the quantity required by the method. If the cumulative blank interferes with the
determination, steps must be taken to eliminate or reduce the interference to a level that
will permit this combination of solvents and reagents to be used. If the blank cannot be
eliminated, the magnitude of the interference must be considered when calculating the
concentration of specific constituents in the samples being analyzed.
A method blank should be determined whenever an analysis is made. The number of blanks
to be run is determined by the method of analysis and the number of samples being
analyzed at a given time. In some methods, such as the AutoAnalyzer procedures, the
method blank is automatically and continuously compensated for because a continuous
flow of the reagents passes through the detector. In other procedures, such as the gas
chromatographic determination of pesticides, a method blank is run with each series of
samples analyzed. Usually this is one blank for every nine samples.
"Trademark of Floridin Co.
5-4
-------
5.3.3 Elimination of Interferences and Other Sources of Error
Procedures for eliminating or at least minimizing impurities that produce specific
interferences or high general background vary with the reagent and method involved. These
procedures may include the following: recrystallization, precipitation, distillation, washing
with an appropriate solvent, or a combination of these. Examples of procedures used for
various types of analyses are given below. For complete information, the analyst should
consult the individual methods.
5.3.3.1 General Inorganic Analyses
Analytical-reagent-grade chemicals and solvents usually present no interference problems in
inorganic analyses. However, some reagents do not always meet methods requirements. An
example is potassium persulfate used in phosphorus and nitrogen determinations. This
reagent is frequently contaminated with ammonia. Therefore, it is routinely purified by
passing air through a heated water solution of the reagent. The purified potassium persulfate
is recovered by recrystallization.
A problem more commonly encountered in inorganic analyses is the rapid deterioration of
the standard reagents and other ingredients. To minimize or eliminate this problem, some
reagents, for example, ferrous ammonium sulfate, must be standardized daily. Others, such
as sodium thiosulfate used for dissolved oxygen determination, may require a substitute
reagent such as phenylarsine oxide. Solid phenol, which readily oxidizes and acquires a
reddish color, can be purified by distillation. Starch indicator used for idiometric titrations
may be prepared for each use or preserved by refrigeration or by addition of zinc chloride or
other suitable compounds.
5.3.3.2 Metals Analyses
In general, spectrograde chemicals, solvents, and gases present no interference problems in
atomic absorption or emission spectrographic determinations. However, standards that do
not meet the requirements of the method are sometimes obtained. Ordinarily, no effort is
made to purify them. They are simply replaced by new reagents of sufficient purity. Some
reagents may form precipitates on standing. Such reagents will reduce the accuracy of
quantitative analyses and should not be used.
5.3.3.3 Radiological Analyses
In general, reagents that do not meet the purity requirements for radiological determina-
tions are replaced with reagents that are satisfactory. However, in some instances (for
example, barium sulfate used for coprecipitation of radium) it may be necessary to perform
repeated recrystallization to remove all forms of radium, and reduce the background count
to a usable level. In some instances, solvents that do not meet requirements may be distilled
to produce adequate purity. In some cases, gases having background counts may be usable
after aging as described earlier. If not, they should be replaced with gases that are
satisfactory.
5.3.3.4 Organic Analyses
Many AR-grade chemicals and solvents, and at times pesticide-quality solvents, do not meet
the specifications required for the determination of specific organic compounds. Impurities
5-5
-------
that are considered trace, or insignificant, for many analytical uses are often present in
greater quantities than the organic constituents being measured. Coupled with the
several-hundred-fold concentration of the sample extract that is usually required, such
impurities can cause very significant interferences in trace organic analyses. Reagents and
solvents found to be unsatisfactory, under the conditions of the analyses, must be replaced
or cleaned up so that they are usable. Some useful cleanup procedures are—
a. Washing the inorganic reagents with each solvent that the reagent contacts during
the analysis
b. Washing the adsorbents, such as silica gel G and Florisil, with the solvents that are
used for a specified column or thin-layer chromatographic procedure, or reactivating
the Florisil by firing to 630°C
c. Preextracting distilled water with solvents used for the particular analysis involved
d. Preextracting aqueous reagent solutions with the solvents involved
e. Redistilling solvents in all-glass systems using an efficient fractionating column
f. Recrystallizing reagents and dyes used in colorimetric or thin-layer determinations
If the reagents and solvents thus produced are not of sufficient purity, they should be
replaced.
Dirty gases (quality less than specified) are particularly troublesome in gas chromatographic
analyses. They may reduce the sensitivity of the detector, and produce a high or noisy
baseline. If this occurs, the cylinder should be replaced immediately. Similarly, if cylinders
of compressed gases are completely emptied in use, the end volumes of the gas may produce
a similar and often more severe effect. Oils and water may get into the system and foul the
detector. When this occurs the system must be dismantled and cleaned. Overhaul of the
detector may be required. To reduce chances of this, it is recommended that all gas
cylinders be replaced when the pressure falls to 100 to 200 psi. Filter driers are of little help
in coping with this type of contamination.
5.3.4 Storing and Maintaining Quality of Reagents and Solvents
Having performed the tasks of selecting, preparing, and verifying the suitability of reagents,
solvents, and gases, the analyst must properly store them to prevent contamination and
deterioration prior to their use. Borosilicate glass bottles with ground-glass stoppers are
recommended for most standard solutions and solvents. Plastic containers such as
polyethylene are recommended for alkaline solutions. Plastic containers must not be used
for reagents or solvents intended for organic analyses. However, plastic containers may be
used for reagents not involved with organic analyses if they maintain a constant volume, and
if it is demonstrated that they do not produce interferences and do not absorb constituents
of interest. It is important that all containers be properly cleaned and stored prior to use.
(Refer to ch. 4 for details.)
Standard reagents and solvents must always be stored according to the manufacturer's
directions. Reagents or solvents that are sensitive to the light should be stored in dark
bottles and in a cool, dark place. It is particularly important to store materials used for
5-6
-------
radiological determinations in dark bottles, because photoluminescence will produce high
background if light-sensitive detectors are used for counting. Some reagents require
refrigeration.
Adsorbents for thin-layer and column chromatography are stored in the containers that they
are supplied in, or according to the requirements of individual methods. When new stock
solutions are necessary, dilutions of the old and new standard should be compared to
determine their accuracy.
The analyst should pay particular attention to the stability of the standard reagents.
Standards should not be kept longer than recommended by the manufacturer or in the
method. Some standards are susceptible to changes in normality because of absorption of
gases or water vapor from the air. Provisions for minimizing this effect are given in
reference 4.
The concentration of the standards will change as a result of evaporation of solvent. This is
especially true of standards prepared in volatile organic solvents. Therefore, the reagent
bottles should be kept stoppered, except when actually in use. The chemical composition of
certain standards may change on standing. Certain pesticides, for instance, will degrade if
prepared in acetone that contains small quantities of water. Thus, it is essential that working
standards be frequently checked to determine changes in concentration or composition.
Stock solutions should be checked before preparing new working standards from them.
5.4 References
1. Catalog of Standard Reference Materials, NBS Special Publication 260, National Bureau
of Standards (June 1975).
2. "Reagent Chemicals," American Chemical Society Specifications, 5th Edition, American
Chemical Society, Washington, D.C. (1974).
3. Manual on Industrial Water and Industrial Waste Water, 2nd Edition, ASTM Special
Publication 148-H, American Society for Testing and Materials (1965), p. 869.
4. "Standard Methods for Preparation, Standardization, and Storage of Standard Solutions
for Chemical Analysis," from Part 31 of 1976 Book of ASTM Standards, American
Society for Testing and Materials, Philadelphia (1977).
5. Standard Methods for the Examination of Water and Wastewater, 13th Edition,
American Public Health Association, New York (1971).
6. Methods for Organic Pesticides in Water and Wastewater, U.S. EPA, Environmental
Research Center, Cincinnati (1971).
7. Burke, J., "Gas Chromatography for Pesticide Residue Analysis; Some Practical
Aspects," J. Assoc. Off. Anal. Chem., 48, 1037 (1965).
5-7
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Chapter 6
QUALITY CONTROL FOR ANALYTICAL PERFORMANCE
6.1 Introduction
Previous chapters discussed basic elements of quality control (QC) pertaining to laboratory
services, instrumentation, glassware, reagents, solvents, and gases; the reader should refer to
the appropriate sections to determine necessary specifications and requirements for QC.
Assuming that these basic variables are under QC, that approved methods are being used,
and that the complete system is initially under QC, valid precision and accuracy data must
initially be developed for each method and analyst. Then, to insure that valid data continue
to be produced, systematic daily checks must show that the test results remain reproducible,
and that the methodology is actually measuring the quantity in each sample. In addition,
QC must begin with sample collection and must not end until the resulting data have been
reported. QC of analytical performance within the laboratory is thus but one vital link in
the dissemination of valid data to the public. Understanding and conscientious use of QC
among all field sampling personnel, analytical personnel, and management personnel is
imperative. Technical approaches are discussed in the following sections.
6.2 The Industrial Approach to QC
In the 1920's, Dr. Walter A. Shewhart of Bell Telephone Laboratories, Inc. developed the
theory of control charts as a basic method for evaluating the quality of products from
manufacturing processes. His book (1) on statistical QC grew out of this original work.
Later, acceptance of his concepts and related statistical techniques within industry led to
refined, quantitative evaluations of product quality in manufacturing. Dr. Shewhart's work
on production processes assumed a uniform product manufactured in large numbers and
inspected on a continuous basis through the periodic analysis of samples of « production
units. The resulting data, Xj, x2, . . .xn, were then used to estimate _precision, as the
standard deviation S or range .R, and accuracy, as the arithmetic mean X. These statistics
were calculated as follows:
S =
n- 1
R = the largest of the Xi - the smallest of the X.
_
V —
These statistics were evaluated by plotting them on control charts developed from similar
statistics taken while the process was under properly controlled operation. The elements
common to such control charts are represented in figure 6-1. They include an expected
value (the central line) and an acceptable range of occurrence (the region between upper and
lower control limits).
6-1
-------
CO
<
u.
O
UJ
UPPER CONTROL LIMIT
CENTRAL LINE
LOWER CONTROL LIMIT
I
10 15
SAMPLE NUMBER
20
Figure 6-1. Essentials of a control chart.
There are many reference sources available that discuss in great detail the classic Shewhart
control charts and related statistics that have since been developed for specific industrial
applications (2-4). In addition, many authors have discussed applications of a related type of
control chart called a cumulative-summation (cusum) chart (2,4). Rather than evaluating
each sample independently, the cusum chart evaluates the cumulative trend of the statistics
from a series of samples. Because each successive point is based upon a cumulative data
trend, cusum charts are often considered more effective than control charts in recognizing
process changes and, therefore, may minimize losses from production of unacceptable units;
however, cusum charts require the more difficult calculations, and optimally designed
Shewhart techniques have been found to be almost as effective (2,4), so there is no universal
agreement on the choice between them.
6.3 Applying Control Charts in Environmental Laboratories
In industrial applications, separate control charts are recommended for each product, each
machine, and each operator. Analogous system variables in an environmental laboratory are
the parameter, the instrument, and the analyst. However, environmental laboratories
routinely have to contend with a variable that has no industrial counterpart—the true
concentration level of the investigated parameter, which may vary considerably among
samples. Unfortunately, the statistics that work well for industry are sensitive to the
variability in true concentration that is common in environmental analysis; e.g., the classic X
and R statistic values increase substantially as concentration increases. This variability in
true concentration means there are no expected values for randomly selected samples, so
that the accuracy of testing methodology must be evaluated indirectly through the recovery
of standards and spikes. As a result, it has been difficult for environmental laboratories to
satisfactorily apply industrial QC techniques.
There are two possible approaches to the solution of the problem of variation in the true
concentration level; either use of a statistic that is not sensitive to this variation or
application of the industrial techniques within restricted concentration ranges. Obviously,
the former should be preferred because it actually solves the problem and does not require
the development and maintenance of a series of charts for each parameter.
6.3.1 Quality Control Charts for Accuracy
Two replacements for the Shewhart X control chart have been suggested for evaluating the
recovery of a series of different standards or spikes. One of these, a cusum chart using the
6-2
-------
square of the difference between the observed and true values, is described in an EPA
Region VI QC manual (5). The other alternative uses the classic Shewhart technique to
evaluate the percent recovery instead of X. It is recommended that the percent recovery be
calculated as
observed
P= 100
known
for standards, or
observed - background
spike
for recovery of spikes into natural water backgrounds. An example of the linear relationship
between percent recovery and the known concentration of standards and spikes is demon-
strated in the accuracy plots of a recent EPA method study report on analysis of mercury
(6). Both approaches are being used on a daily basis by various environmental laboratories.
The data in table 6-1 were used in the EPA Region VI manual (5) to illustrate the
development of a cusum chart. The actual data have been reordered here to appear in
ascending order of the known values. Note that the mean and the range of the df2 values
increase with increasing concentration level, and this violates a basic premise for acceptable
control chart statistics. Because the percent recovery data do not show any such trend, it is
the recommended control chart statistic for controlling accuracy.
From the data in table 6-1, a Shewhart control chart for percent recovery can be calculated
in the following way:
Average percent recovery
The standard deviation for percent recovery
22
6-3
-------
'234,074- (2,310)2/23
22
= 9.70
Therefore, the upper control limit becomes the following:
UCL = P + 3Sp
= 100.4 + 3(9.70)
= 129.5
Table 6-1
ANALYSIS1 OF TOTAL PHOSPHATE-PHOSPHORUS STANDARDS, IN mg/1
TOTAL PO4-P
Point
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
Known
0.34
0.34
0.40
0.49
0.49
0.49
0.50
0.50
0.50
0.52
0.66
0.66
0.67
0.68
0.83
0.98
1.3
1.3
1.6
2.3
2.3
3.3
4.9
Obtained
0.33
0.34
0.40
0.49
0.49
0.63
0.47
0.53
0.56
0.59
0.70
0.60
0.65
0.65
0.80
0.75
1.2
1.3
1.7
2.3
2.4
3.3
4.6
Difference
0.01
0.00
0.00
0.00
0.00
-0.14
0.03
-0.03
-0.06
-0.07
-0.04
0.06
0.02
0.03
0.03
0.23
0.10
0.00
-0.10
0.00
-0.10
0.00
0.30
«>
0.0001
0.0000
0.0000
0.0000
0.0000
0.0196
0.0009
0.0009
0.0036
0.0049
0.0016
0.0036
0.0004
0.0009
0.0009
0.0529
0.0100
0.0000
0.0100
0.0000
0.0100
0.0000
0.0900
Percent
Recovery
Pi
97
100
100
100
100
129
94
106
112
113
106
91
97
96
96
77
92
100
106
100
104
100
94
Pi2
9,409
10,000
10,000
10,000
10,000
16,641
8,836
11,236
12,544
12,769
11,236
8,281
9,409
9,216
9,216
5,929
8,464
10,000
11,236
10,000
10,816
10,000
8,836
Totals
2,310
234,074
1 Using a colorimetric method with persulfate digestion.
6-4
-------
and the lower control limit becomes
LCL= 100.4- 29.1
= 71.3
The completed control chart is shown in figure 6-2.
Following normal procedures, the control chart must indicate the conditions under which it
was developed; i.e., laboratory name, parameter, method of analysis, date of preparation,
and any other information unique to the initializing data, such as range of concentration
and identification of analyst(s). A control chart is not generally applicable under other
conditions.
To verify the control chart, the initializing data should be checked to be sure that none of
the values exceeds these new control limits. In addition, if its distribution is proper, about
68 percent of the initializing data should fall within the interval P ± SP. It has been
suggested that the control chart is not valid if less than 50 percent of the initializing data
falls within this interval.
In applying the control chart, either of the following two conditions would indicate an
out-of-control situation:
a. Any point beyond the control limits
b. Seven successive points on the same side of the value P of the central line
When an out-of-control situation occurs, analyses must be stopped until the problem has
been identified and resolved, after which the frequency should be increased for the next few
percent-recovery QC checks. The problem and its solution must be documented, and all
analyses since the last in-control point must be repeated or discarded.
140r-
£ 120
O
O
!r 100
H-
ULl
£ 80
LU
Q.
60
UCL
.- P
LCL
10 15
SAMPLE NUMBER
20
25
Figure 6-2. Shewhart control chart for percent recovery data.
6-5
-------
A final note of caution regarding use of a single percent-recovery P control chart over a
broad concentration range is necessary. As noted earlier for the analysis of mercury, a good
linear relationship of the form
X = P (known concentration) + K
where K is a constant, seems appropriate for many parameters. However, to justify use of a
single percent-recovery control chart, K must be small enough relative to the /"(known
concentration) term that it has little or no practical effect upon the value of X. This will
usually be true for moderate or high concentration levels, but may not be true at very low
concentration levels. As a result, for some parameters it may be necessary to develop a
separate percent-recovery or Shewhart X chart for each standardized low concentration level
sample.
6.3.2 Quality Control Charts for Precision
Because the characteristics of the range statistic change as concentration changes, two
alternatives to Shewhart's R chart have been used in environmental laboratories to evaluate
the precision of routine sample analyses.
One alternative is a cusum chart using the sum of the squared difference between duplicate
determinations on randomly selected routine samples (5). Because the range R for duplicate
analyses is equal to the difference between them, the cusum statistic equals the sum of
squared ranees^""!/?2. However, if R changes significantly as concentration level changes,
then R2 is affected even more and, therefore, is not as good a criterion for judging whether
precision of the system is within acceptable limits.
The other alternative uses a chart similar tq_the R chart, but the chart statistic is either the
percent relative standard deviation (lOOS/AT), the coefficient of variation (CV or S/X), or
the industrial statistic /. For the duplicate determinations A and B, I equals the absolute
value of their difference divided by their sum, or \A - B\/(A +B), and can be shown to be
equivalent to the other two statistics:
lOO(CV)=100-=
= ,oo
= 100
200 [A - B\
A+B
(A+B)/2
1 R
A+B
2007
For the sake of computational ease, / seems to be a logical alternative to R.
6-6
-------
The next concern is whether / is independent of changes in concentration level. Based upon
experience with duplicates on routine samples taken during the last 2 years by EPA Region
VII, / appears to decrease substantially as concentration increases. In recognition of this
possible dependency, control charts for / should only be developed from and applied to
results within a limited concentration range. Note that control charts for R could be applied
under similar limitations.
As an illustration of the concentration dependency of these precision statistics, table 6-2
provides estimates of R and / for different concentration ranges of three parameters. These
parameters were selected because approximately 100 sets of duplicates were available that
were well distributed over a reasonably broad concentration range. The ranges for the sum
of duplicates A + B used in table 6-2 were selected because they were convenient and the
data tended to be well distributed among them. Data judged to be out of control were
discarded before any calculations were made.
Table 6-2 indicates the concentration dependence of both the range R and the industrial
statistic / for these three parameters. Because / is not independent of concentration and is
Table 6-2
ESTIMATES OF THE RANGE (R = \A-B\) AND THE INDUSTRIAL STATISTIC
[/= \A-B\I(A+B)] OF THREE DIFFERENT PARAMETERS FOR VARIOUS CON-
CENTRATION RANGES1
Parameter
BOD, 5-day (mg/1)
Chromium 0/g/l)
Copper (ptg/1)
Range of
A+B
2 to <20
20 to <50
50 to <1 00
100to<300
300 to <600
600 to <2,000
2,000 up
10 to <20
20 to <50
50 to <1 00
100 to <300
300 to <1, 000
1 ,000 up
10to<30
30 to <50
50 to <100
100to<200
200 to <400
400 up
No. of
Sets of
Duplicates
21
30
27
29
17
12
3
32
15
16
15
8
5
16
23
21
26
10
3
*A+B
11.7
35.2
72.2
204.1
394.4
1,041
6,683
12.3
33.4
72.4
170.3
480.3
6,340
22.2
38.2
70.8
131.9
268.0
702.0
2fl
1.04
1.94
3.33
6.52
11.1
12.1
177
0.32
0.57
1.12
3.80
5.25
76.0
0.93
1.35
1.14
2.33
2.81
4.56
27
0.0888
0.0552
0.0462
0.0319
0.0282
0.0116
0.0264
0.0306
0.0170
0.0155
0.0223
0.0109
0.0120
0.0617
0.0368
0.0169
0.0177
0.0105
0.0065
1 From EPA Surveillance and Analysis Laboratory, Region VII.
2 Average values.
6-7
-------
Table 6-3
SHEWHART UPPER CONTROL LIMITS (UCL) AND CRIT-
ICAL RANGE Rc VALUES FOR THE DIFFERENCES BE-
TWEEN DUPLICATE ANALYSES WITHIN SPECIFIC CON-
CENTRATION RANGES FOR THREE PARAMETERS1
Parameter
BOD, 5-day (mg/1)
Chromium (Mg/1)
Copper (jug/1)
Concentration
Range2
1 to<10
10to<25
25 to <50
50 to <1 50
I50to<300
300 to <1, 000
1,000 up
5to<10
10to<25
25 to <50
50to<150
1 50 to <500
500 up
5 to <1 5
1 5 to <25
25 to <50
50 to <1 00
100to<200
200 up
UCL
3.40
6.34
10.9
21.3
36.3
39.6
579
1.05
1.86
3.66
12.4
17.2
249
3.04
4.41
3.73
7.62
9.19
14.9
Rc
3.5
6
11
21
36
3 40
3579
1
2
4
12
317
3249
3
4
5
8
39
315
1 From EPA Surveillance and Analysis Laboratory, Region VII.
2 Equal to half of the range of A + B given in table 6-2.
3 Based on fewer than 1 5 sets of duplicate analyses.
more difficult to calculate and develop control charts for, the use of R charts for a series of
sequential concentration ranges for each parameter seems practical. However, because the
primary concern when using any range chart is whether the upper control limit has been
exceeded, an even more practical approach would be to develop a table of these limits for all
concentration levels of each parameter. As an example, table 6-3 contains the calculated
Shewhart upper control limits for the range R from duplicate analyses within the various
concentration levels for the three parameters in table 6-2. These limits were calculated, as
usual, from the Shewhart factor D4 for ranges based upon duplicate analyses and the
appropriate average value of the range R given in table 6-2. For example, the UCL for 25 to
50 mg/1 of BOD was calculated as follows:
UCL =
= 3.27(3.33)
= 10.9
6-8
-------
Table 6-3 also contains a critical range Rc column. Because the data from EPA Region VII
were almost always whole units with only a very occasional half unit reported, the Rc value
is the UCL value rounded to the nearest whole unit at higher concentration levels and to the
nearest half unit for the lowest concentration level. However, there is an exception to this
rule among the low-concentration Rc values for copper that demonstrates an advantage
beyond the simplicity of using such tables. The UCL value for copper at 25 to 50 Mg/1 is
inconsistent with the UCL values for adjacent concentration levels, and the Rc value has
been adjusted to resolve this inconsistency. Without the table, such inconsistencies could
very easily go unnoticed.
The examples in table 6-4 illustrate how to use the Rc values in table 6-3. This technique,
consisting of the development and use of a table of critical-range Rc values at different
concentration levels, is recommended to control precision. Normal control chart procedures
should be followed as in section 6.3.1 regarding identification and verification of the table.
The table should be updated periodically as additional, or more current, data become
available, or whenever the basic analytical system undergoes a major change. If any
difference between duplicate analyses exceeds the critical-range value for the appropriate
concentration level, then analyses must be stopped until the problem is identified and
resolved, and the frequency should be increased for the next few precision checks. After
resolution, the problem and its solution must be documented, and all analyses since the last
in-control check must be repeated or discarded.
6.4 Recommended Laboratory Quality Assurance Program
A minimum laboratory quality assurance program should include control procedures for
each parameter as described in the following sections.
6.4.1 Standard Curves
A new standard curve should be established with each new batch of reagents, using at least
seven concentration levels.
6.4.2 Quality Control Checks for Each Analytical Run
With each batch of analyses, the following tests should be run:
a. One blank on water and reagents
Table 6-4
CRITICAL RANGE VALUES FOR VARYING CONCENTRATION
LEVELS
Parameter
BOD (mg/1)
Chromium (Mg/1)
Copper (Mg/1)
Duplicates
20 and 24
60 and 75
46 and 51
R
4
15
5
RC
6
12
15
R
-------
b. One midpoint standard
c. One spike to determine recovery
d. One set of duplicate analysis
The results from b through d should be compared with previous in-control data by using the
appropriate technique recommended in section 6.3.
6.4.3 Intel-laboratory QC
An interlaboratory QC program would require each laboratory to do the following:
a. Analyze reference-type samples to provide independent checks on the analytical
system. These may be available from EPA as QC samples, from the National Bureau
of Standards as standard reference materials, or from commercial sources. If
performance limits are not provided, the results should fall within the routine limits
of each laboratory for a standard at a level comparable to the specified true value.
b. Participate in performance evaluation and method studies as available from EPA,
from the American Society for Testing and Materials, and from other agencies.
6.5 Outline of a Comprehensive Quality Assurance Program
In the following discussion the symbols used represent the results of analysis according to
the scheme:
A j = first replicate of sample A
AI = second replicate of sample A
B = sample taken simultaneously with sample A
B$ F = field spike into sample B
BSL - laboratory spike into sample B
DF = field spike into distilled water
DL = laboratory spike into distilled water
T = true value for all spikes
The laboratory spikes BSL and DL are the only analyses that may not be necessary. All
other analyses must be done simultaneously.
6.5.1 Steps for the Field Personnel
A comprehensive quality assurance program would include the following steps for each
parameter in the monitoring study:
a. Take independent simultaneous samples A and B at the same sampling point.
Depending on the parameter, this might involve side-by-side grab samples or
composite samplers mounted in parallel.
6-10
-------
b. Split sample A into the equal-volume samples A t and A 2.
c. Split sample B into equal volumes and add a spike T to one of them; the latter
sample becomes sample 5SF. As with all spikes, the addition of T should
approximately double the anticipated concentration level.
d. Add the same spike T to a distilled water sample furnished by the laboratory and
designate this sample as DF.
These QC samples must be treated in the same way as routine samples; i.e., the volume, type
of container, preservation, labeling, and transportation must be the same for all.
6.5.2 Steps for the Laboratory
The laboratory personnel should perform the following steps for quality assurance:
a. Analyze the blank and midpoint standard recommended in section 6.4. If results are
unsatisfactory, resolve problems before continuing.
b. Analyze sample DF. If the percent recovery of T is unsatisfactory (see section
6.3.1), create a similarly spiked, distilled-water sample DL and analyze to test for a
systematic error in the laboratory or fundamental problems with the spike. If the
percent recovery of T from DL is satisfactory, any systematic error occurred before
the samples reached the laboratory.
c. Analyze samples B and Bs F. If B is below the detection limit, or if B is greater than
1071 or less than 0.1 T, disregard the remainder of this step and proceed to step d. If
the percent recovery of T from 5SF is unsatisfactory (see section 6.3.1), spike an
aliquot of sample B the same way in the laboratory so that a similar recovery can be
anticipated. Analyze this sample 5SL to test for immediate sample interferences or a
bad background result B. If the percent recovery from Bs L is satisfactory, then the
interference must require a longer delay before analysis, or other special conditions
not present in the laboratory, in order to have a noticeable effect upon recovery of
the spike.
d. Analyze A^ and A2. If the absolute (unsigned) difference between these results
exceeds the critical value (see section 6.3.2), then precision is out of control.
e. Calculate the absolute difference between A^ and B. If it is unsatisfactory (see
section 6.3.2), the field sampling procedure did not provide representative samples.
If initial results at each of the laboratory steps were satisfactory, then the validity of the
related data has been indisputably established. If results at any step are unsatisfactory,
resolution depends upon the problem identified. Laboratory problems may just require that
the analyses be repeated, but field problems will usually require new samples. Figure 6-3 is
intended to clarify the interdependence of the preceding laboratory steps b through e.
In figure 6-3 it must be noted that there is no way to identify additive sample interferences;
i.e., those that have an equal effect upon the background-plus-spike results (5SF or 5SL)
and the background result B. Recovery of a spike will not show such interferences.
6-11
-------
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Problems causing systematic errors that may occur in the field include the following:
a. Contaminated preservative, distilled water, or containers
b. Contamination by sampling personnel
c. Deterioration through excessive holding time or use of an ineffectual preservation
technique
d. Use of a bad field-spiking procedure
6.6 Related Topics
6.6.1 Advanced Laboratory Automation and Its Effect on QC
Advanced laboratory automation systems under development analyze samples automatically
and use a control computer to interpret the resulting data and produce an analytical report.
The primary benefits of such a system are not only that the data-recording and calculation
errors common to manual analyses have been inherently eliminated, but also that extensive
QC can be accomplished quite easily and cheaply. The computer can be programed to
automatically recognize different kinds of QC samples and to establish or recall appropriate
control limits. Thus the QC overhead is reduced considerably and QC procedures previously
too costly or complex become practical.
As an example of a QC procedure that is considered impractical for manual use, regression
could be used to determine the relationship between concentration change and the accuracy
and precision statistics discussed earlier. Using these relationships, very responsive, single
accuracy and precision charts could be developed for each parameter. As computer-assisted
analysis becomes common, automated laboratories will very likely replace the manual
procedures recommended earlier in this chapter with evaluation criteria based upon
regressions.
6.6.2 Method Comparability (Equivalency)
Requirements for method comparability are under development for proposed alternatives to
the methodology specified in Public Law 92-500, section 304(g). A final version of these
requirements should be available from EPA at a later date.
6.7 References
1. Shewhart, W. A., Economic Control of Quality of Manufactured Product, Van Nostrand
Reinhold Co., Princeton, N.J. (1931).
2. Duncan, A. J., Quality Control and Industrial Statistics, 3d Edition, Ch. 18, R. D. Irwin,
Inc., Homewood, 111. (1965).
3. Grant, E. L., and Leavenworth, R. S., Statistical Quality Control, 4th Edition,
McGraw-Hill Book Co., Inc., New York (1972).
4. Davies, O. L., and Goldsmith, P. L., Statistical Methods in Research and Production, 4th
Edition, Hafner Publishing Co., New York (1972).
5. Laboratory Quality Control Manual, 2d Edition, U.S. EPA, Region VI, Surveillance and
Analysis Division, Ada, Okla. (1972).
6-13
-------
6. Winter, J., Britton, P., Clements, H., and Kroner, R., "Total Mercury in Water," EPA
Method Study 8, pp. 35-36, U.S. EPA, Office of Research and Development, EMSL, Cin-
cinnati (Feb. 1977).
6-14
-------
Chapter 7
DATA HANDLING AND REPORTING
7.1 Introduction
To obtain meaningful data on water quality, the sample collector must obtain a
representative sample and then deliver it unchanged for analysis. The analyst must perform
the proper analysis in the prescribed fashion, complete calculations, and convert results to
final form for permanent recording of the analytical data in meaningfu.1, exact terms. These
results are transferred to a storage facility for future interpretation and use.
The following sections discuss processing of actual values, recording and reporting of data in
the proper way, some means of quality control of data, and the storage and retrieval of data.
7.2 The Analytical Value
7.2.1 Significant Figures
The term "significant figure" is used, sometimes rather loosely, to describe a judgment of
the reportable digits in a result. When the judgment is not soundly based, meaningful digits
are lost or meaningless digits are reported. On the other hand, proper use of significant
figures gives an indication of the reliability of the analytical method used.
The following discussion describes the process of retention of significant figures.
A number is an expression of quantity. A figure or digit is any of the characters 0, 1,2, 3, 4,
5, 6, 7, 8, 9, which, alone or in combination, serve to express a number. A significant figure
is a digit that denotes the amount of the quantity in the particular decimal place in which it
stands. Reported analytical values should contain only significant figures. A value is made
up of significant figures when it contains all digits known to be true and one last digit in
doubt. For example, if a value is reported as 18.8 mg/1, the 18 must be firm while the 0.8 is
somewhat uncertain, but presumably better than one of the values 0.7 or 0.9 would be.
The number zero may or may not be a significant figure depending on the situation.
Final zeros after a decimal point are always meant to be significant figures. For example, to
the nearest milligram, 9.8 g is reported as 9.800 g.
Zeros before a decimal point with nonzero digits preceding them are significant. With no
preceding nonzero digit, a zero before the decimal point is not significant.
If there are no nonzero digits preceding a decimal point, the zeros after the decimal point
but preceding other nonzero digits are not significant. These zeros only indicate the position
of the decimal point.
Final zeros in a whole number may or may not be significant. In a conductivity
measurement of 1,000 /zmho/cm, there is no implication by convention that the conductiv-
ity is 1,000 ± 1 jumho. Rather, the zeros only indicate the magnitude of the number.
7-1
-------
A good measure of the significance of one or more zeros interspersed in a number is to
determine whether the zeros can be dropped by expressing the number in exponential form.
If they can, the zeros may not be significant. For example, no zeros can be dropped when
expressing a weight of 100.08 g in exponential form; therefore the zeros are significant.
However, a weight of 0.0008 g can be expressed in exponential form as 8 X 10~4 g, so the
zeros are not significant. Significant figures reflect the limits in accuracy of the particular
method of analysis. It must be decided whether the number of significant digits obtained for
resulting values is sufficient for interpretation purposes. If not, there is little that can be
done within the limits of the given laboratory operations to improve these values. If more
significant figures are needed, a further improvement in method or selection of another
method will be required.
Once the number of significant figures obtainable from a type of analysis is established, data
resulting from such analyses are reduced according to set rules for rounding off.
7.2.2 Rounding Off Numbers
Rounding off of numbers is a necessary operation in all analytical areas. It is automatically
applied by the limits of measurement of every instrument and all glassware. However, when
it is applied in chemical calculations incorrectly or prematurely, it can adversely affect the
final results. Rounding off should be applied only as described in the following sections.
7.2.2.1 Rounding-Off Rules
If the figure following those to be retained is less than 5, the figure is dropped, and the
retained figures are .kept unchanged. As an example, 11.443 is rounded off to 11.44.
If the figure following those to be retained is greater than 5, the figure is dropped, and the
last retained figure is raised by 1. As an example, 11.446 is rounded off to 11.45.
If the figure following those to be retained is 5, and if there are no figures other than zeros
beyond the five, the figure 5 is dropped, and the last-place figure retained is increased by
one if it is an odd number or it is kept unchanged if an even number. As an example, 11.435
is rounded off to 11.44, while 11.425 is rounded off to 11.42.
7.2.2.2 Rounding Off Arithmetic Operations
When a series of numbers is added, the sum should be rounded off to the same number of
decimal places as the addend with the smallest number of places. However, the operation is
completed with all decimal places intact, and rounding off is done afterward. As an
example,
The sum must be rounded off to 33.4.
7-2
-------
When one number is subtracted from another, rounding off should be completed after the
subtraction operation, to avoid possible invalidation of the operation.
When two numbers are to be multiplied, all digits are carried through the operation, then
the product is rounded off to the number of significant digits of the multiplier with the
fewer significant digits.
When two numbers are to be divided, the division is carried out on the two numbers using
all digits. Then the quotient is rounded off to the number of significant digits of the divisor
or dividend, whichever has the fewer.
When a number contains n significant digits, its root can be relied on for n digits, but its
power can rarely be relied on for n digits.
7.2.2.3 Rounding Off the Results of a Series of Arithmetic Operations
The preceding rules for rounding off are reasonable for most calculations; however, when
dealing with two nearly equal numbers, there is a danger of loss of all significance when
applied to a series of computations that rely on a relatively small difference in two values.
Examples are calculation of variance and standard deviation. The recommended procedure is
to carry several extra figures through the calculations and then to round off the final answer
to the proper number of significant figures.
7.3 Glossary of Statistical Terms
To clarify the meanings of statistical reports and evaluations of water quality data, the
following statistical terms are introduced. They are derived in part from usage (1,2) of the
American Society for Quality Control.
Accuracy— The difference between an average value and the true value when the latter is
known or assumed.
Arithmetic mean— The arithmetic mean (or average) of a set of n values is the sum of the
values divided by n :
y — _,_-- ___________
n
Bias— A systematic error due to the experimental method that causes the measured values to
deviate from the true value.
Confidence limit, 95 percent— The limits of the range of analytical values within which a
single analysis will be included 95 percent of the time,
95 percent CL = X ± 1 .965
where CL is the confidence level and S is the estimate of the standard deviation.
7-3
-------
Constant—A nonvarying qualitative or quantitative characteristic of the population.
Geometric mean—A measure of central tendency for data from a positively skewed
distribution (log normal),
_ -
Xa = antilog
g n
Interference-A biological or chemical attribute of a test sample that positively or negatively
offsets the measured result from the true value. If interference that is not segregated and
identified is present, it enlarges or reduces the method bias.
Median—Middle value of all data ranked in ascending order. If there are two middle values,
the median is the mean of these values.
n—The number of values Xt reported for a sample.
N—The total number of values X{ of the entire population or universal set of data.
Population—The total set of units, items, or measurements under consideration.
Precision-Relative to the data from a single test procedure, the degree of mutual agreement
among individual measurements made under prescribed conditions.
Precision data—Factors that relate to the variations among the test results themselves; i.e.,
the scatter or dispersion of a series of test results, without assumption of any prior
information.
Range—The difference between the highest and lowest values reported for a sample.
Relative deviation (coefficient of variation)—The ratio of the standard deviation S of a set of
numbers to their mean X expressed as percent. It relates standard deviation (or precision) of
a set of data to the size of the numbers:
£
CV = RD (percent) =100—
X
Relative error—The mean error of a series of measured data values as a percentage of the
true value Xt,
\X- X\
RE (percent) = 100
%t
Sample-Groups of units or portions of material, taken from a larger collection of units or
quantity of material, that provide information to be used for judging the quality of the total
collection or entire material as a basis for action on them or on their production processes.
7-4
-------
Series—A number of test results with common properties that identify them uniquely.
Skewness—A measure of the asymmetry of a frequency distribution,
(Xt ~ X)3
K =
no*
This measure is a pure signed number. If the data are perfectly symmetrical, the skewness is
zero. If K is negative, the long tail of the distribution is to the left. If K is positive, the long
tail extends to the right.
Standard deviation—The square root of the variance of the universe,
a =
\ 2
•(, /N
Standard deviation estimate-The most widely used measure to describe the dispersion of a
set of data. Normally X ± S will include 68 percent, and X ± 2S will include about 95
percent of the data from a study:
S =
n- 1
Standard deviation, single analyst—A measure of dispersion for data from a single analyst
that is calculated here using an equation developed by Youden for his nonreplicate study
design (3),
where D = the difference in paired values obtained from a single analyst.
Universe—The total set of items or measurements.
Variable—An experimentally determined estimate denoted X for a particular quality or trait
of the population.
7.4 Report Forms
The analytical information reported should include the measured parameters; the details of
the analysis such as burette readings, absorbance, wavelength, normalities of reagents,
correction factors, blanks; and the reported data values.
7-5
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To reduce errors in manipulation of numbers a general rule is to reduce handling and
transposition of data to an absolute minimum. Ideally, a report form includes preliminary
information about the sample and its analysis, and the same form is used for the final
entering of data into a computer; however, such report forms are not yet common. Rather,
a variety of methods is used to record data.
7.4.1 Loose Sheets
Reporting of data onto loose or ring-binder forms is a means of recording data that allows
easy addition of new sheets, removal of older data, or collection of specific data segments.
However, the easy facility for addition or removal also permits loss or misplacement of
sheets, mixups in date sequence, and ultimately questionable status of the data for formal
display or presentation as courtroom evidence.
7.4.2 Bound Books
The use of bound books is an improvement in data recording that tends to result in a
chronological sequence of data insertion. Modification beyond a simple lined book improves
its effectiveness with little additional effort. Numbering of pages encourages use of data in
sequence and also aids in referencing data through a table of contents ordered according to
time, type of analysis, kind of sample, and identity of analyst.
Validation can be easily accomplished by requiring the analyst to date and sign each analysis
on the day completed. This validation can be strengthened further by providing space for
the laboratory supervisor to witness the date and the completion of the analyses.
A further development of the bound notebook is the commercially available version
designed for research-type work. These notebooks are preprinted with book and page
numbers, and spaces for title of project, project number, analyst signature, witness
signature, and dates. Each report sheet has a detachable duplicate sheet that allows
up-to-date review by management without disruption of the notebook in the laboratory.
The cost of research-type bound notebooks is about four times that of ordinary note-
books.*
Use of bound notebooks has been limited to research and development work where an
analysis is part of a relatively long-term project, and where the recording in the notebook is
the prime disposition of the data until an intermediate or final report is written.
However, bound notebooks can and should be used in routine analytical laboratories such as
those concerned with water quality. The need for repeated information on sampling and
analyses can be answered by use of preprinted pages in the bound notebooks.
7.4.3 Preprinted Report Forms
Most field laboratories and installations repetitively analyzing fixed parameters develop their
own system of compiling laboratory data that may include bound notebooks, but a means
of forwarding data is also required. Usually, laboratories design forms to fit a related group
*Scientific Notebook Co., 719 Wisconsin Ave., Oak Park, 111. 60304.
7-6
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of analyses or to report a single type of analysis for a series of samples. As much
information as possible is preprinted to simplify use of the form. With loose-sheet,
multicopy forms (using carbon or NCR paper) information can be forwarded on the desired
schedule while also allowing retention of data in the laboratory. Still, the most common
means for recording data in rough form are internal bench sheets or bound books. The
bench sheet or book never leaves the laboratory but serves as the source of information for
transfer of data to appropriate report forms. (See fig. 7-1.)
In most instances the supervisor and analyst wish to look at the data from a sampling point
or station in relation to other sampling points or stations on a particular river or lake. This
review of data by the supervisor prior to release is a very important part of the QC program
of the laboratory; however, such reviews are not easily accomplished with bench sheets. For
review purposes, a summary sheet can be prepared that displays a related group of analyses
from a number of stations such as shown in figure 7-2. Because the form contains space for
all of the information necessary for reporting data, the completed form can also be, used to
complete the data forms forwarded to the computer storage and retrieval system.
The forms used to report data to storage systems provide spaces for identification of the
sampling point, the parameter code, the type of analysis used, and the reporting
terminology. Failure to provide the correct information can result in rejection of the data,
or insertion of the data into incorrect parameter fields. As sample analyses are completed,
the data values are usually reported in floating decimal form along with the code numbers
for identifying the parameter data fields and the sampling point data fields. Figure 7-3
shows an example of a preprinted report form used for forwarding data to keypunch.
7.4.4 Plastic-Coated Labels and Forms
A recent addition to good sample handling and data management is the availability of
plastic-coated (blank or preprinted) labels, report forms, and bound report books. These
materials are waterproof, do not disintegrate when wet or handled, can be written on while
wet, and retain pencil or waterproof ink markings though handled when wet.
7.4.5 Digital Readout
Instrumental analyses, including automated, wet-chemistry instruments such as the Tech-
nicon AutoAnalyzer, the atomic absorption spectrophotometer, the pH meter, and the
selective electrode meter, provide digital readout of concentrations, which can be recorded
directly onto report sheets without further calculation. Programmed calculators can be used
to construct best-fit curves, to perform regression analyses, and to perform series of
calculations leading to final reported values.
7.4.6 Keypunch Cards and Paper Tape
Because much of the analytical data generated in laboratories is first recorded on bench
sheets, then transferred to data report forms, keypunched, and manipulated on small
terminal computers (or manipulated and stored in a larger data storage system), there is a
danger of transfer error that increases with each data copy. The analyst can reduce this error
by recording data directly from bench sheets onto punch cards that can be retained or
forwarded immediately to the data storage system. Small hand-operated keypunch machines
are available.
7-7
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LABORATORY BENCH DATA
*— y. rr
DATE OP COMPOSITE SAMPLE
LL
M 1 1 1 1 1 1 II
1 1
i i
Fecal Cohform llkll^
MF/100
1 1 1 1 1 M 1 1 1 1 1 1 1
i i
1 1
NHj-N + OrgN
1 1 1 1 1 0 0 0 0 0
1 II II 1 1 1 II
NH3-N
1111100000
mg/1
000
1 1
4 S <
000
1 1 M 1 1 1 1 1 1 MM
i i
LL
N02-N + N03-N
1 I 1 1 1 0 0 0 0 0
1 II 1 II 1 1 II
mg/1
000
1 1
ITEM _
I I
P, Total
I 1 1 I 1 0 0 0 0 0
mg/1
M II 1 1 II 1
i i
1 1
ITEM _
1 1
LL
P, Soluble
1 1 1 1 1 0 0 0 0 0
II 1 1 1 1 1 1 II
TOC
1111100000
II 1 II 1 1 1 II
mg/1
000
1 1
4 5 6
mg/1
000
1 1
i i
1 1
8 7
1 1
| |
8 1
Phenol
1 til 100000
1 M 1 II 1 1 II
6S43210133
Cyanide
till 100000
II 1 1 1 II 1 II
6543210123
Mg/1
000
1 1
456
mg/1
000
1 1
456
COMPUTER CODED DATA
II 1 1 1 M M M 1
1 i M M 1
13-18
N'N'M 1 1 1 1 1 IIJI 1
19-23 24-27 28 29 30
MM M- 1 1 1 1 Ml II 1
31-35 36-39 40 41 42
l°|0|6|3|5 Ml ||
43-47 48-51 52 53 54
|0|0|6|1|0| | (I 1 Mi II 1
55-59 60-63 64 65 66
COLUMN 80 (BLANK)
CHG
|0|0|6|3|0 | || 1 1 II II II 1
67-71 72-75 76 77 78 79
NEXT CARD. REPEAT COLUMNS 1 80 ABOVE
|0|0|6J6|5| | || | Ml || |
19-23 24-27 28 29 30
|0|0|6|6|6| | | II III II 1
31-35 36-39 40 41 42
|0|0|6|8|0 I I || 1 II II 1
43-47 48-51 52 53 54
|3M7]3|0| | || | Mill 1
55-59 60-63 64 65 66
COLUMN 80 (BLANK)
CMS.
|OJO|7|2|0|| I I I I 1 II II II 1
67-71 72-75 76 77 78 79
Figure 7-3. Example of STORET report form.
7.4.7 STORET-Computerized Storage and Retrieval of Water Quality Data
Because of their ability to record, store, retrieve, and manipulate huge amounts of data, the
use of computers is a natural outgrowth of demands for meaningful interpretation of the
great masses of data generated in almost all technical activities.
In August 1961, numerous ideas were brought together concerning the basic design of a
system called STORET for storage and retrieval of water pollution control' data. A
7-10
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refinement of this system is now operated by the Technical Data and Information Branch,
Division of Applied Technology, Office of Water Programs, EPA.
This is a State/Federal cooperative activity that provides State water pollution control
agencies with direct, rapid access into a central computer system for the storage, retrieval,
and analysis of water quality-control information.
If properly stored, the data can be retrieved according to such descriptors as the point of
sampling, the date, and the specific parameters stored, or all data at a sample point or series
of points can be extracted as a unit.
Full details on use of the STORET system are given in the recently revised STORET
handbook (4).
7.4.8 Automated Laboratory Systems
The use of digital readout, keypunch cards, and paper tape have been overshadowed by the
development of customized, fully automated online computer systems that make measure-
ments, calculate results, perform quality control, and report analytical data simultaneously
from a full range of laboratory instruments. (See fig. 7-4.) Such systems can contain the
following functions:
a. Manual or automatic sampling and testing of a series of samples, standards,
replicates, and check samples
b. Detection of the measurement signals from the series of samples
c. Conversion of signals to concentrations, generation of a standard curve, and
calculation of sample values in final units
d. Calculation of the deviation and recovery values of the results and indication of
acceptance or nonacceptance based on limits established by the analyst
e. Provision of the output in a form designated by the analyst: dial, paper recording
chart, digital readout, cathode ray tube, or printed report form
The degree of hands-on operation required in the system is specified by the analyst.
If an automated system is properly designed and operated, most calculation and transposi-
tion errors are avoided and the proper level of quality control is automatically exerted.
Laboratory automation systems for water analyses are being developed and coordinated by
EMSL-Cincinnati for use in a number of EPA laboratories (5).
7.5 References
1. "Guide for Measure of Precision and Accuracy," Anal. Chem. 33, 480 (1961).
2. "Glossary of General Terms Used in Quality Control," Quality Progress, Standard Group
of the Standards Committee, American Society for Quality Control, 11(1), 21-2 (1969).
3. Youden, W. J., Statistical Techniques for Collaborative Tests, Association of Official
Analytical Chemists, Washington, D.C. (1967).
7-11
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MEASURE TIMING,
BASELINE, AND
CALIBRATION STANDARDS
MEASURE SAMPLES,
CHECK STANDARDS,
REPLICATES, SPIKES,
AND BLANKS
COMPUTE AND DISPLAY
INTERIM RESULTS AND
STATISTICS
ALERT
OPERATOR
TO
ACTION
Figure 7-4. Flow chart of the sequence of events
during a controlled series of laboratory
measurements.
7-12
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4. "STORET, EPA's Computerized Water Quality Data Base, the Right Answer," U.S. EPA,
Office of Water and Hazardous Materials, Washington, D.C. (1977).
5. Budde, W. L., Almich, B. P., and Teuschler, J. M., The Status of the EPA Laboratory
Automation Project, EPA-600/4-77-025, U.S. EPA, Office of Research and Develop-
ment, EMSL, Cincinnati (1977).
7-13
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Chapter 8
SPECIAL REQUIREMENTS FOR TRACE ORGANIC ANALYSIS
8.1 Introduction
The high sensitivity of the instrumentation used in trace organic chemical analysis and the
low concentration of organic compounds being investigated dictate that special attention
must be given to this analytical field. Contamination of the sample from any possible source
must be diligently guarded against, and interferences in the sample must be carefully
controlled. Strict attention to method and highly refined technique are required to produce
valid quantitative and qualitative results.
Two manuals have been developed by the Environmental Monitoring and Support Labora-
tory (EMSL) in Cincinnati covering the two broad areas of quantitative and qualitative trace
organic analysis. Reference 1 contains both general and specific quality assurance programs
designed to insure the production of acceptable measurements when using the methods
contained in that manual. Reference 2 contains specific, detailed quality assurance programs
covering both the performance of the instrument and the interpretation of the resulting
spectra.
This chapter is organized with three sections devoted to general material applicable to all
organic analysis, and one section abstracting highly specialized materials from each of the
two manuals cited.
8.2 Sampling and Sample Handling
Regardless of the intent, all numbers generated by a water quality laboratory are ultimately
represented as the concentration levels in the sample matrix at the time of collection. Such
numbers tend, automatically, to endorse the sample collection, preservation, and shipment
procedures. Thus, quality assurance programs limited to the care of the sample beginning
with its receipt by the laboratory are inadequate. The laboratory must share responsibility
for the preservation and shipment of all samples that it will accredit with concentration
values. Two approaches are available that will generally protect the laboratory from
generating numbers that may not reflect actual conditions of the sample at the time of
collection. The best, but usually least practical solution, is for the laboratory personnel to
collect all samples. The alternative is for the laboratory to adopt a policy of sample rejection
based on minimum standards of sample identification and age. Guidelines for establishing
these standards are discussed in this section. It is recommended that copies of this material
be supplied to all sample collectors along with an understanding of the specific policy of the
laboratory toward rejecting samples that do not meet these criteria.
In all of the cases to be discussed, it is the responsibility of the project director to (a)
coordinate his sampling, preservation, and shipment with the laboratory, (b) obtain clean
sample containers from the laboratory, (c) provide adequate sample identification and
compositing instructions, and (d) provide duplicates and blanks as required by the
laboratory. Additional prearrangements should be made with the laboratory if sample
splitting is desired, to create separate supernatant and settleable matter samples, or if
calculations on a wet-weight basis in addition to the standard dry-weight calculations are
desired.
8-1
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Bottles and caps are to be supplied by the laboratory because rigorous cleaning is required
even for new bottles. New and recycled bottles should be washed as described in reference
1. Samples received in bottles of unknown origin or questionable cleanliness should be
rejected by the laboratory. For water samples that are to be analyzed by solvent extraction
methods, narrow-mouthed, screw-cap bottles such as Boston rounds are preferred because
they have less tendency to leak and are easy to handle in the laboratory. One-liter bottles
are generally more expensive than quart bottles, thus most laboratories use the clear 32-oz
containers. The bottle should be sealed with Teflon (du Pont) lined caps. For water samples
of intermediate pH, aluminum-foil-lined caps may be used when Teflon (du Pont) is not
available. Oil and grease samples can be collected with Polyseal caps-the conical liner pro-
vides an excellent seal against pressure during shipment and sample extraction. Screw-cap,
widemouthed glass bottles are preferred for sediment samples because they are easier to
handle in tlie field and in the laboratory. Precleaned, 16- or 32-oz bottles with Teflon (du
Pont) or foil-lined screw-cap closures should be provided by the laboratory. Masking tape or
other suitable labels should be applied to the dry bottles.
Sampling for purgeable organics requires special consideration and equipment. The sample
container should consist of a 45-ml, screw-cap vial fitted with a Teflon (du Pont) faced, sili-
cone septum.* The vials, septa, and caps should be washed in hot detergent water and thor-
oughly rinsed with tapwater and organic-free water, then dried at 105°C for 1 h. The vials
should then be cooled to room temperature in a contaminant-free area. When cool, the vials
should be sealed with the septa, Teflon (du Pont) side down, and screw cap and maintained
in this sealed condition until filled with sample.
The bottles used for collecting water samples for solvent extraction should not be overfilled
or prerinsed with sample before filling because oil and other material that can cause
erroneously high results may remain in the sample bottle after rinsing. Bottles should be
filled with sample to about 90 percent of capacity, and the level should be marked to
determine if leakage occurs. Because full sample bottles are difficult to pour from during
extractions, complete filling should be avoided. In the collection of sediment samples,
nonrepresentative debris such as large stones or wood should be discarded.
Multiple samples are usually required for purgeable organics analysis because of leakage and
because the measurement process is destructive to the sample. All vials should be identified
with waterproof labels. The water sample vials are filled to overflowing from a bubble-free
source so that a convex meniscus is formed at the top. They are sealed by carefully placing
the septum, Teflon (du Pont) side down on the opening of the vial and screwing the cap
firmly in place.
Shipment and receipt of samples must be coordinated with the laboratory to minimize time
in transit because it is the prerogative of the laboratory to reject samples where delays in
shipment have caused them to age beyond acceptable holding times. To avoid the need to
resample, the sampler should determine in advance the most efficient and reliable form of
transportation for the samples. All samples for organic analysis should arrive at the
laboratory on the same day collected, or should be shipped and maintained at less than 4°C
for arrival by the next day. The samples are usually shipped in insulated ice chests. Water
samples should be prechilled before packing to reduce the ice requirements during shipment.
*Vial and septum are available from Pierce Chemical Co., P.O. Box 117, Rockford, 111.
61105. Vial: No. 13074; septum: No. 12722.
8-2
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The bottles should be stabilized in the container with styrofoam and covered with ice. The
information needed to identify the samples should be attached to the outside of the ice
chest.
A part of the quality assurance program for a laboratory must be the development of a clear
policy for accepting or rejecting samples. Because organic analyses are expensive in terms of
manpower and supplies, it is poor management to commit such resources just to obtain data
of questionable validity. Water samples that clearly have not met the preservation criteria
during shipment (e.g., in the case of a spill) should be accepted only if resampling is
impossible. Results from such samples must be qualified in the laboratory report.
Upon receipt, the samples should be checked for adequate identification, sample tempera-
ture or presence of ice in the chest, and leakage. Samples for volatile organics should be
checked for air bubbles, although it is extremely difficult to avoid the development of very
small bubbles regardless of the type of sample bottle employed. The samples are logged in
with the receipt time noted. Unless the condition of the samples fails to meet the criteria for
acceptance by the laboratory, required preservatives are added immediately and the samples
are all refrigerated.
The laboratory staff should be alerted to the arrival of the samples, so that the required
analysis can begin as soon as possible. When sediment samples are to be reported on a
wet-weight basis, or when a water sample is to be filtered or divided into supernatant and
settleable subsamples for separate analysis, the sample processing should begin promptly.
When analysis of water samples is to be restricted to the water phase only, filtration is
required. This may be accomplished with 4.7-cm, glass-fiber filter disks that have been
preextracted with acetone and allowed to dry. The filter disks must not contain organic
binder. The disks are placed on a membrane filter holder and up to a liter of water is
filtered. The filtrate is transferred without rinsing to a clean sample container and treated as
a normal, whole-water sample.
When separate reporting is required for the settleable and supernatant phases, the water
sample (or a large portion of it) is allowed to settle overnight in a closed glass container at
4°C. If phase separation occurs, the supernatant phase is carefully decanted or siphoned into
a graduated cylinder without disturbing the surface of the settled material. After the volume
of supernatant is noted, the supernatant is filtered as just described into a clean glass
container and stored at 4°C. The volume of settled material is determined either by marking
the slurry level on the side of the sample container for later calibration, or by transferring
the slurry without washing into a calibrated vessel. A portion of well-mixed slurry is
removed for a determination of percent solids. The remaining slurry is stored in a sealed
glass container at 4°C.
When both dry-weight an'd wet-weight results are required for sediments and sludges, a
percent-solids determination should be performed soon after receipt of the sample. A
representative portion (ideally 10 to 25 g) of well-mixed sample is weighed into a tared
Erlenmeyer flask and dried at 105°C to a constant weight. Then percent solids are
calculated for the sample.
Routine laboratory management involves detailed recordkeeping beginning with the initial
contact with the sample collector. A master flowsheet should be prepared for each sample,
listing parameters to be determined and pretreatment operations to be performed. The
8-3
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master flowsheet is kept at a fixed location and is designed to handle dated entries
indicating when operations are completed. Methods requiring extraction permit double
entry on the master flowsheet so laboratory throughput times can be calculated in terms of
receipt-to-extraction and receipt-to-completion times. A separate set of forms follows the
sample through the laboratory. Each sample form records information for a class of
parameters and details cleanup operations and other method options. Calculations and cross
references to chromatograph files are entered on the sample form also.
A chain-of-custody program lends significant legal support to the results generated. The
program begins when the laboratory dispatches the sample collectors. Each time a sample is
collected, a form is initiated stating where and when the sample was collected, with cross
reference to a number on the sample container. The sample collectors return to the
laboratory with the samples in their guarded possession and deliver them personally to the
responsible person in the laboratory. The person (custodian) receiving the samples in turn
signs each ledger to acknowledge receipt and locks the sample in a refrigerator. The analyst
then comes to the custodian and signs for the sample that he is taking for analysis. In this
way full documentation of the sample handling is maintained from sample collection
through completion of the analysis.
8.3 Extract Handling
Each method in reference 1 is prepared in sections identified by titles. This style of
presentation is aimed at presenting the analyst logical places to interrupt his analysis. Often,
because of the length of the method, the analyst is unable to complete an analysis in a single
day. When planning a partial analysis, certain factors must be considered. Because the
organics are generally more stable in solvent than in water, it is always preferable to extract
a sample and hold the extract rather than to hold the sample. Extracts to be held overnight
or longer should first be dried by treatment with sodium sulfate.
The methods include several transfers of the solvent extract from one piece of glassware to
another. These quantitative transfers are made using several small portions of solvent to
wash the walls of the previous container. A 5-ml, luer-lock glass syringe with a 2-in., 20-gage
needle is convenient for this purpose. Solvents tend to creep up the outside of a container,
such as an ampoule, while pouring. To minimize contamination during transfer, solvent ex-
tracts are poured as rapidly as possible. Extracts can become contaminated not only from
oils from the skin of the handler but also from other extracts handled at the same time
where deposits on the outside of the container are unintentionally transferred from one con-
tainer to another. Instead of using labels, contamination problems from this source canjbe
reduced by etching permanent numbers on ampoules. Sample log sheets should be used1 to
index the extract with a numbered ampoule, eliminating the need for tape or wax pencils.
Of the several ways to dry a solvent extract with sodium sulfate, passing it through a
chromatographic column packed with 2 to 3 in. of anhydrous crystals is the most
convenient and quantitative. When a wet extract is being transferred from a separatory
funnel to a Kuderna-Danish (K-D) concentrator, it should be drained directly through the
column to eliminate the need for an intermediate piece of glassware and the resulting
transfer step. Prewashing the sodium sulfate column with extracting solvent is recom-
mended, although interferences can be controlled by preheating the salt in a shallow tray at
400°C for 30 min. After the extract has passed through the column, 20 to 30 ml of
extracting solvent are used to wash the residual extract from the column.
8-4
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The standard K-D concentration apparatus consists of a three-ball Snyder column, a 500-ml
flask, and a calibrated ampoule. It is designed to concentrate 100- to 300-ml volumes of
extract to a final volume of 5 to 10 ml. To use the apparatus, one 10/20 mesh boiling chip
(previously rinsed with solvent and heated for 1/2 h) is added, and the assembly is supported
above a concentric-ring water bath with the tip of the ampoule below the surface of the
water. The lower ground-glass joint must be kept above the water. The water temperature
should be adjusted for mild distillation, with no chamber flooding or splashing (about 10°C
to 20° C above the boiling point of the solvent). When the volume of liquid reaches 1 to 2 ml
(checked frequently), the assembly should be removed and allowed to cool. The chambers
and flask will drain to a final volume near 10 ml. The flask and the lower ground-glass joint
are rinsed with a minimum of solvent. The evaporation can be continued with the
microscale K-D concentrator if further concentration is required.
The microscale K-D concentrator is designed to concentrate extracts from 5 to 10 ml to 1.0
ml. A fresh boiling chip must be added, and a two-ball micro-Snyder column is attached to
the ampoule. The ampoule is supported above the water bath, and the extract is
concentrated to about 0.7 ml. The column and ground-glass joint are rinsed with a minimum
of solvent, and the volume is adjusted to 1.0 ml.
The K-D concentrator can be used to exchange solvents. When the sample is dissolved in a
solvent unsuitable for a cleanup operation or for gas chromatography (GC), it can be
displaced by a suitable higher boiling solvent. The actual volumes that should be used to
effect the exchange vary with the solvent pairs depending upon the difference in boiling
points and azeotrope formations, but a general procedure is to concentrate the extract to 10
ml, add 20 ml of the higher boiling solvent, and reconcentrate to 10 ml.
After extractions and subsequent K-D concentrations, solubilities of some materials may be
exceeded. High sulfur levels are a particular problem encountered in sediment extractions.
Extracts should be decanted from ampoules where sulfur has crystallized. In some samples
the extractable organic levels are so high the extract tends to solidify and will not
concentrate further. When this occurs, a small aliquot of the extract should be taken and
diluted as appropriate for final analysis.
A significant source of the variation in GC analysis can be attributed to the injection of a
portion of the extract into the analytical system. Manual injections of 2 to 3 n\ with the use
of a 10-/il syringe will introduce variance even when the injection volumes are determined to
the nearest 0.05 pi. Of a variety of injection techniques in use, the solvent flush technique
h^s been found to be acceptable for quantitative work. This technique is described in detail
in reference 1.
8.4 Supplies and Reagents
Reference compounds of materials should be assayed and of 98 percent purity or higher. If
the purity is less than 98 percent, the appropriate correction factor must be included in all
calculations of standard concentration. The reference materials should be cataloged, dated,
and stored in a refrigerator.
Stock solutions of these reference materials should be prepared in a high-boiling, inert
solvent, if possible, to minimize errors due to evaporation or solvent-induced decomposi-
8-5
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tion. The laboratory should have an accurate six-place balance for preparing small quantities
of reference standards. The following method of preparing stock solutions is recommended:
Weigh 10 mg (to the nearest 0.01 mg) of reference standard into a small aluminum weighing
pan.* Drop the entire pan into a 10-ml volumetric flask. Dissolve the reference material in
about 5 ml of solvent, then dilute to volume. Label the solution with compound name,
concentration, solvent used, date prepared, and initials of preparer. Store the volumetric in a
refrigerator, except when preparing dilutions. When the standard is a replacement for an
existing stock solution, the two solutions should be compared, and the results as well as the
suspected reason for any variation should be permanently recorded in the laboratory files.
Working standards are prepared from one or more stock solutions after they have warmed to
room temperature. As these working standards usually represent three to six orders of
magnitude of dilution of the stock standards, it is obviously necessary to take great care in
preparing them. Serial dilutions are recommended with a maximum of 1:100 dilution for
each step. Although 10-jul volumes can be read within 1 percent with a 10-jul syringe, the
inherent problems with the dead volume in the syringe make the use of such equipment less
desirable for preparing working standards than volumetric pipets. New working standards
should be prepared frequently unless long-term stability has been demonstrated. When
several compounds are combined into a single standard for simultaneous GC, they must be
closely monitored for chemical interactions.
Pesticide-quality solvents are usually required, and each new lot should be checked for
interferences prior to use. The solvent check, representing approximately 10 percent more
solvent than required for any method, should be concentrated and analyzed for method
interferences under all GC conditions applicable to that solvent. If interfering peaks or a
broad solvent front are observed, the solvent should be redistilled in an all-glass distillation
system, with a distillation column.** If interferences persist, the solvent lot should be
discarded. This preliminary lot check does not eliminate the need for routine solvent blanks
to monitor for purity changes over a period of time.
Diethyl ether must be shown to be free of peroxides before use. Peroxide test stripst can be
used for a quick, convenient test. The alumina column procedure for removing peroxides,
described in literature supplied with the test strips, has been used successfully to remove all
peroxides from the solvent. The solvent should always be stabilized with 2 percent
volume/volume ethyl alcohol. Chromatographic elution patterns are based on ether
containing this alcohol.
Granular sodium sulfate should be purchased in glass containers. If purchased in a large
container (5-lb bottles or larger), it should be transferred to smaller bottles for daily use.
Before sodium sulfate is used for chromatographic work, it should be heated to 400°C for
30 min and shown not to be contaminated. When the sodium sulfate is used to dry extracts
before concentration, the heating is usually unnecessary because impurities will be removed
by preelution of the drying column with solvent.
*Available from The Perkin-Elmer Corp., Norwalk, Conn.; No. 219-0041.
**Available from Lab Glass, Inc., North West Blvd., Vineland, N.J. 08360; Widner No.
LG-5930.
tAvailable from Scientific Products, 1430 Waukegan Rd., McGaw Park, 111. 60085; Quant
peroxide test strip, No. PI 126-8.
8-6
-------
Florisil (Floridin Co.) is purchased from the manufacturer preactivated at 630°C and trans-
ferred to glass containers with Teflon (du Pont) or foil-lined lids. Prior to use the Florisil
(Floridin Co.) is heated in a shallow open dish for 5 h in a 130°C oven. The Florisil (Floridin
Co.) can then be transferred into a sealed, glass bottle and stored indefinitely at 130°C until
needed. Cleanup methods using Florisil (Floridin Co.) require that a lauric acid value be
determined for each lot before use. In addition, the determination of a pesticide elution pat-
tern is recommended. These procedures are described in detail in reference 1.
Carrier gases are a very important part of the chromatographic system; therefore, special
care should be taken in their selection and handling. Only high-purity or equivalent carrier
gases should be purchased, and they should be filtered, online, through a 5-A molecular
sieve. Porous polymer and other column packings degrade at elevated temperatures in the
presence of trace quantities of oxygen. Oxygen in the carrier gas also adversely affects the
performance of electron-capture detectors. Therefore, some type of online oxygen-removal
system is recommended for these applications. The chemical traps should be changed or
regenerated with each new cylinder of carrier gas. One purifier* has been found to last
through many cylinders without replacement. A better grade of gas is required for
temperature programing than for isothermal operation. Combustion gases may be of lower
quality but should be at least equivalent to dry air or purified hydrogen. Regulators should
have stainless steel internal parts and be of two-stage design. External tubing should be of
good quality, such as refrigeration tubing. Such tubing should be rinsed with solvent and
heated at 200°C under gas flow before use in the analytical system.
The purchase of precoated GC column packings is strongly recommended over the
preparation of coated materials in the laboratory. The commercial products are, generally,
of higher quality and consistency than those prepared by the average analyst. All tubing
should be cleaned before packing by passing a series of solvents (e.g., hexane, chloroform,
and acetone) through it. Glass columns should also be silylated. (Instructions are included
with the purchase of silylating agents.) Dry the tubing thoroughly before packing it.
A vibrator should never be used to settle the packing material in the column. Such vibration
may fracture the solid support material, expose uncoated active sites, and produce inferior
chromatographic separations. When vacuum alone is inadequate and further settling is
required, the column may be tapped with a pencil or similar object while the vacuum is
being applied. Unless otherwise stated, stainless-steel tubing should be packed before coiling
it to fit the GC apparatus.
8.5 Quality Assurance
8.5.1 Measurements
Most of the quality assurance programs suggested in chapter 6 of this manual cannot easily
be adapted to the methods for organic compounds. The reasons for this, and the suggested
approach for a suitable program for the organic analytical laboratory are discussed in detail
in reference 1 and only summarized here.
Reference 1 suggests that quality assurance for organic analysis be divided into three
separate categories. The first category represents the determination of purgeable com-
*Available from Matheson Gas Products, P.O. Box E, Lyndhurst, N.J. 07071; Hydrox
Purifier model No. 8301.
8-7
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pounds. This determination is performed in a closed analytical system; the complete analysis
can be performed in 1 h; and the number of theoretically possible interferences is somewhat
limited. The second category represents liquid/liquid partition methods in a regulatory
situation. Here a very limited number of compounds are being measured; there is a high
occurrence of positive results; and it is important to establish that the method works
satisfactorily on the particular sample matrix. The third category represents liquid/liquid
partition methods in a monitoring situation. Here a large number of compounds are often
being measured simultaneously; there is a low occurrence of positive results; and each
sample matrix may be different. Quality assurance is aimed at establishing that the
laboratory is using the method correctly.
The purgeable methods are unique among organic methods because the standards are treated
in exactly the same way as the samples, and there is no inherent method bias. The methods
are amenable to a variety of quality assurance programs. The approach that has been found
applicable to all types of samples and provides the maximum data for the expended effort
consists of the addition of one or more internal standards to the matrix before purging. Data
generated in this program provide a continuous monitoring of the equipment and establishes
matrix applicability for the test.
For liquid/liquid extraction methods in a regulatory situation, the emphasis is placed on
duplicates and dosed samples. Both field duplicates and laboratory duplicates are used in the
program to establish sampling and subsampling validity. Trie dosing of samples to establish
method accuracy for the matrix is an integral part of this program. Where the analytical
program will extend over a long period of time the construction of control charts is
recommended.
When the liquid/liquid extraction methods are used for monitoring, the emphasis is placed
on an external control series. A standard laboratory matrix is developed. With each series of
samples the matrix is dosed and analyzed with the samples. Data generated over a period of
time can be used to monitor the performance of the equipment and the analyst, with
relatively tight specifications to define problems that arise. Control charts can be con-
structed to alert the analyst to problems, but there is no provision for rejection of results for
samples of this type.
8.5.2 Identifications
The combined gas chromatography/mass spectrometer (GC/MS) has emerged as the most
important single instrument at the disposal of the environmental analytical chemist. It alone
can provide both the sensitivity and the high degree of certainty necessary for an
identification culled from a complex environmental matrix. The instrument has generated
an aura of well-deserved respect, and its results are seldom questioned. For these reasons it is
mandatory that strict quality assurance programs be followed in both the generation and the
interpretation of mass spectra. The EMSL, with the cooperation of many other EPA GC/MS
users, has produced a procedural manual (2) generally for use with a Finnigan quadrupole
instrument. A detailed quality assurance program constitutes an integral part of the manual.
The actual detailed program is beyond the scope of this manual but has been summarized in
the following paragraphs.
To insure that a quadrupole mass spectrometer generates quality spectra, the program
provides for at least daily performance evaluation with a reference compound, and
-------
readjustment of the instrument as necessary. The operator prepares a solution of decafluoro-
triphenylphosphine (DFTPP).* When this compound is injected into the GC/MS using any
of several compatible GC columns, the resulting elaborate spectrum can be evaluated using
criteria developed at EMSL-Cincinnati (3).
Decafluorotriphenylphosphine key ions and the ion-abundance criteria that are used for
determining whether the mass spectrometer is generating high-quality spectra are as follows:
Abundance Criteria
Mass (amu) 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 ! 98
197 Less than 1 percent of mass 198
198 . Base peak, ] 00 percent relative abundance
199 5 to 9 percent of mass 198
275 10 to 30 percent of mass 198
365 1 percent (or greater) of mass 198
441 Less than mass 443
442 Greater than 40 percent of mass ] 98
443 17 to 23 percent of mass 442
If these specifications are not met, the guidelines provided for adjusting the instrument (2,3)
must be considered.
Having produced quality spectra, the analyst must always ascertain, by analyzing a method
blank under exactly the same analytical and instrumental conditions, that the spectra are
relevant. Most commercial systems have a software program, extracted ion current profile
(EICP), that permits the sample and the blank to be overlayed on a graphic display device
when each is scanned for a particular mass. When large numbers of spectra result ffrom a
sample and the blank must be checked for a match for each one, this technique simplifies
the screening process.
If the spectra are found to be unique to the sample, it is normally processed through a
mass-spectral search-and-match system. The computerized version of such a system consists
*Available from PCR Research Chemicals, Inc., P.O. Box 1778, Gainsville, Fla. 32602; No.
11898-4.
8-9
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of an organized collection of many thousands of compound spectra stored in a large central
computer accessed by the analyst through telephone linkups. On the basis of 8 or 10 major
masses the computer can rapidly search the complete system for similar spectra. These
spectra are ranked for similarity to the unknown using a mathematical algorithm. The
resulting similarity index (SI), which can be further refined to a quality index (2), is a
measure of the degree of confidence the analyst may place in the identification. If a match
is not found, the analyst must revert to manual interpretation of the spectra and deduce the
structure of the compound by its fragmentation patterns.
After a tentative identification is made, several other types of supporting experiments
become possible. Retention-time GC/MS data of a pure compound (standard) may be
compared with analogous data from the sample component. Similarly, the mass spectrum of
the standard, obtained under the same conditions that were used for the sample, may be
compared with the sample component spectrum. The standard may be dissolved in water at
an appropriate concentration, isolated, and measured. The recovery of this spike in the same
fraction in which the suspected component appeared and the observation of equivalent mass
spectra for the spike and the sample component constitute strong evidence for confirmation
of the identification.
8.6 References
1. Methods for Organic Analysis of Water and pastes, U.S. EPA, EMSL, Environmental
Research Center, Cincinnati (in preparation).
2. EPA GC/MS Procedural Manual, Budde, W. L., and Eichelberger, J. W., editors, 1st
Edition, Vol. 1, U.S. EPA, Office of Research and Development, EMSL, Cincinnati (in
press).
3. Eichelberger, J. W., Harris, L. E., and Budde, W. L, Anal. Chem., 47, 995 (1975).
8-10
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Chapter 9
SKILLS AND TRAINING
9.1 General
Analytical operations in the laboratory can be graded according to the degree of
complexity. Some analyses require no sample treatment, and the measurement can be
performed in minutes on a simple instrument. Other determinations require extensive
sample preparation prior to complex instrumental examination. Consequently, work
assignments in the laboratory should be clearly defined. Each analyst should be completely
trained and should fully understand all the assignments of his job before being given new
responsibilities. In this regard, all analysts, subprofessional or professional, should be
thoroughly instructed in basic laboratory operations, according to the degree of professional
maturity. Some of the basic operations that should be reviewed periodically with laboratory
personnel follow.
9.1.1 Sample Logging
Routine procedure for recording of samples entering the laboratory and assigning primary
responsibility should be emphasized. The information that is required and the routing of the
sample to the analyst is then established. The stability, preservation, and storage of samples
prior to analyses are then discussed.
9.1.2 Sample Handling
The analyst should understand thoroughly at which points in his procedures the sample is to
be settled, agitated, pipetted, etc., before he removes it from the original container.
9.1.3 Measuring
The analysts, especially new employees and subprofessionals, should be instructed in the use
of volumetric glassware. The correct use of pipettes and graduates should be emphasized as
discussed in chapter 4 of this manual.
9.1.4 Weighing
Because almost every measuring operation in the analytical laboratory is ultimately related
to a weighing operation, the proper use of the analytical balance should be strongly
emphasized. Maintenance of the balance, including periodic standardization, should be
repeatedly emphasized to all personnel. The correct use and maintenance of balances is
discussed in chapter 3 of this manual.
9.1.5 Glassware
All glassware should be washed and rinsed according to the requirements of the analysis to
be performed. Not only must the personnel assigned to these tasks be instructed, but also all
lab personnel should know the routine for washing and special requirements for particular
uses of glassware. In addition, the precision tools of the laboratory such as pipets, burets,
9-1
-------
graduates, and Nessler tubes should be inspected before use for cleanliness, broken delivery
tips, and clarity of marking. Defective glassware should be discarded or segregated.
9.1.6 Instrumentation
Operation and maintenance of analytical instrumentation is of primary consideration in the
production of valid data. All instruments must be properly calibrated, quality-control
checks documented, and standard curves verified on a routine basis. Details on instrumental
quality control are presented in chapter 3 of this manual.
9.1.7 Data Handling and Reporting
As with sample logging, the routine procedure for recording results of analyses and pertinent
observations, including quality control checks, should be emphasized. Analytical data
should be permanently recorded in meaningful, exact terms and reported in a form that
permits future interpretation and unlimited use. Details are discussed in chapter 7 of this
manual.
9.1.8 Quality Control
The need to continuously assess precision and recovery values of methodology is a prime
responsibility of the analyst. Self-evaluation through the analyses of replicates and recovery
of spikes from samples representative of the daily workload provides confidence and
documentation of the quality of the reported data.
9.1.9 Safety
Laboratory safety should be discussed on a continuing basis with all employees, but it
should be emphasized when an employee is assigned to perform new duties.
9.1.10 Improvement
In summary, quality control begins with basic laboratory techniques. Individual operator
error and laboratory error can be minimized if approved techniques are consistently
practiced. To insure the continued use of good technique, laboratory supervisors should
periodically review the basic techniques and point out areas of needed improvement with
each analyst.
Continuing improvement of technical competence by all laboratory personnel is, of course,
the final responsibility of the laboratory supervisor. In a well-organized laboratory, however,
a big-brother attitude of higher ranking to lower grade personnel should be encouraged; each
person should be eager to share experience, tricks of the trade, special skills, and special
knowledge with subordinates. Obviously, efficiency and results will improve.
9.2 Skills
The cost of data production in the analytical laboratory is based largely upon two factors:
the pay scale of the analyst, and the number of data units produced per unit of time.
However, because of the large variety of factors involved, estimates of the number of
measurements that can be made per unit of time are difficult. If the analyst is pushed to
9-2
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Table 9-1
SKILL-TIME RATING OF STANDARD ANALYTICAL OPERATIONS
Measurement
Simple Instrumental:
pH
Conductivity
Turbidity
Color
Dissolved Oxygen (Probe)
Fluoride (Probe)
Simple Volumetric:
Alkalinity (Potentiometric)
Acidity (Potentiometric)
Chloride
Hardness
Dissolved Oxygen (Winkler)
Simple Gravimetric:
Solids, Suspended
Solids, Dissolved
Solids, Total
Solids, Volatile
Simple Colorimetric:
Nitrite N (Manual)
Nitrate N (Manual)
Sulfate (Turbidimetric)
Silica
Arsenic
Complex, Volumetric, or Colorimetric:
BOD
COD
TKN
Ammonia
Phosphorus, Total
Phenol (Distillation Included)
Oil and Grease
Fluoride (Distillation Included)
Cyanide
Special Instrumental:
TOC
Metals (by AA), No Preliminary Treatment
Metals (by AA), With Preliminary Treatment
Organics (by GC), Pesticides, Without Cleanup
Organics (by GC), Pesticides, With Cleanup
Skill Required
(Rating No.)1
1
1
1
1
1,2
1,2
1
1
1
1
1,2
1,2
1,2
1,2
1,2
2
2
2
2
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
2,3
3,4
3,4
Number
Per Day
100-125
100-125
75-100
60-75
100-125
100-125
50-75
50-75
100-125
100-125
75-100
20-25
20-25
25-30
25-30
75-100
40-50
70-80
70-80
20-30
2 15-20
25-30
25-30
25-30
50-60
20-30
15-20
25-30
8-10
75-100
150
60-80
3-5
2-4
1 Skill-required rating numbers are defined as follows:
1 -aide who is a semiskilled subprofessional with minimum background or
training, comparable to GS-3 through GS-5. (Continued)
9-3
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produce data at a rate beyond his capabilities, unreliable results may be produced. On the
other hand, the analyst should be under some compulsion to produce a minimum number of
measurements per unit of time, lest the cost of data production become prohibitive. In table
9-1, estimates are given for the number of determinations that an analyst should be
expected to perform on a routine basis. The degree of skill required for reliable performance
is also indicated.
The time limits presented in the table are based on use of approved methodology. A tacit
assumption has been made that multiple analytical units are available for measurements re-
quiring special equipment, as for cyanides, phenols, ammonia, nitrogen, and COD. For some
of the simple instrumental or simple volumetric measurements, it is assumed that other
operations such as filtration, dilution, or duplicate readings are required; in such cases the
number of measurements performed per day may appear to be fewer than one would nor-
mally anticipate.
9.3 Training
For more experienced, higher grade personnel, formal training in special fields, possibly
leading to specialization, should be almost mandatory. Such training can be fostered
through local institutions and through the training courses provided by the EPA. Regional
policies on after-hours, Government-supported training should be properly publicized.
Formalized training for lower grade personnel, comparable to GS-3 to GS-5, is relatively
scarce. However, skills can be most efficiently improved at the bench level on a personal,
informal basis by more experienced analysts working in the same area. Exposure of
personnel to pertinent literature should also be a definite program policy.
(Continued)
2—aide with special training or professional with minimum training with
background in general laboratory techniques and some knowledge of
chemistry, comparable to GS-5 through GS-7.
3-experienced analyst capable of following complex procedures with good
background in analytical techniques, professional, comparable to GS-9
through GS-12.
4-experienced analyst specialized in highly complex procedures, profes-
sional, comparable to GS-11 through GS-13.
2 Rate depends on type of samples.
9-4
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Chapter 10
WATER AND WASTEWATER SAMPLING
10.1 Introduction
The quality of data resulting from water and wastewater sampling surveys is dependent
upon the following six major activities: (a) formulating the particular objectives of the water
sampling program, (b) collecting representative water samples, (c) maintaining the integrity
of the water samples through proper handling and preservation, (d) adhering to adequate
chain-of-custody and sample identification procedures, (e) practicing quality assurance in
the field, and (f) properly analyzing the pollutants in the water samples. These areas are
equally important for insuring that environmental data are of the highest validity and
quality.
The present section addresses aspects of quality control (QC) concerned with the collection
of environmental samples and data in the field. It includes a capsule summary of the specific
areas mentioned previously and a list of references (table 10-1) that provide specific
guidance in these areas, rather than a collection of guidelines on sampling procedures.
10.2 Areas of Sampling
The specific areas that comprise an overall water sampling program are as follows.
10.2.1 Objectives of the Particular Sampling Program
The objectives of the sampling program affect all the other aspects of the sampling program.
Sampling program objectives are determined by the following activities: (a) planning
(areawide or basin), (b) permitting, (c) compliance, (d) enforcement, (e) design, (f) process
control, and (g) research and development. The types of water sampling programs to be
employed, depending on suitability to program objectives, include reconnaissance surveys;
point-source characterization; intensive surveys; fixed-station-network monitoring; ground-
water monitoring; and special surveys involving chemical, biological, microbiological, and
radiological monitoring.
Factors that must be considered in meeting the objectives of the sampling program are the
extent of the manpower resources, the complexity of the parameters of interest, the
duration of the survey, the number of samples, the frequency of sampling, the type of
samples (grab or composite), and the method of sample collection (manual or automatic).
10.2.2 Collection of Representative Samples
The objective of all water and wastewater sampling is to obtain a representative portion of
the total environment under investigation. The techniques for obtaining representative water
samples may vary with the length, width, and depth of a body of water, its physical and
chemical parameters, and its type to be sampled (such as municipal or industrial effluents,
surface waters and bottom sediments, agricultural runoff, and sludges). In collecting
representative samples, the following factors should be considered.
10-1
-------
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10.2.2.1 Site Selection
The location of the sampling site is critical in obtaining representative data. Preferably,
water sampling sites for point sources of pollution from municipal and industrial effluents
are located at points of highly turbulent flow to insure good mixing; however, inaccessi-
bility, lack of site security, or power unavailability may preclude use of the best sites, but
these impediments should not be used as reasons for collecting samples at unacceptable
locations. Locations of sampling sites for streams, lakes, impoundments, estuaries, and
coastal areas vary, but in general occur in the following bodies: (a) in water bodies for
sensitive uses (swimming and drinking water supply), (b) in major impoundments or
reservoirs near the mouths of major tributaries and in the rivers entering and leaving the
impoundments, (c) in water bodies polluted by man's activities, (d) in rivers upstream and
downstream from tributaries, and (e) where hydrological conditions change significantly.
10.2.2.2 Types
The basic types of water and wastewater sampling methods are grab sampling and composite
sampling. Composite sampling may be conducted manually or automatically. The six
methods for forming composite samples, all of which depend on either a continuous or
periodic sampling mode, are the following: (a) constant sample pumping rates, (b) sample
pumping rates proportional to stream flow rates, (c) constant sample volumes and constant
time intervals between samples, (d) constant sample volumes and time intervals between
samples proportional to stream flow rates, (e) constant time intervals between samples and
sample volumes proportional to total stream flow volumes since last sample, and (f)
constant time intervals between samples, and sample volumes proportional to total stream
flow rates at time of sampling. The choice of using the grab sampling method or one of the
six compositing sampling methods is determined by program objectives and the parameters
to be sampled.
10.2.2.3 Automatic Samplers
The use of automatic samplers eliminates errors caused by the human element in manual
sampling, reduces personnel cost, provides more frequent sampling than practical for manual
sampling, and eliminates the performance of routine tasks by personnel. Criteria for brand
selection of automatic samplers include evaluations of the intake device, intake pumping
rates, sample transport lines, sample gathering systems (including pumps and scoops), power
supplies and power controls, sample storage systems, and additional desirable features to fit
particular sampling conditions. There are many commercially available automatic samplers;
however, because no single automatic sampler is ideally suited for all situations, the user
carefully selects the automatic sampler most suited for the particular water or wastewater to
be characterized. Precautions must be taken in regard to using certain types of samples in
potentially explosive atmospheres.
10.2.2.4 Flow Measurement
An essential part of any water or wastewater sampling survey as well as a necessary
requirement of the National Pollution Discharge Elimination System (NPDES) permit
program is accurate flow measurement, which can be divided into four categories:
a. Flow measurement in completely filled pipes under pressure—common devices
employed are orifices, Venturi tubes, flow nozzles, Pitot tubes, magnetic flow
meters, ultrasonic flow meters, and elbow meters.
10-3
-------
b. Trajectory methods, either full or partially full, measured at the end of the
pipe--common flow measurement methods are the California and Purdue pipe
methods. These methods are normally considered as estimates rather than accurate
measurements.
c. Flow measurement in open channels and sewers—common methods are the velocity-
area measurement, time-of-passage measurement, and level measurement methods
using weirs and flumes.
d. Miscellaneous flow measurement methods-common methods include use of Man-
ning formula, tracer and salt dilution-techniques, water meters, pump rates, and
measurements of level changes in tanks and calibrated vessels.
Flow measurement data may be instantaneous or continuous. For continuous measure-
ments, a typical system consists of primary devices such as weirs and flumes and secondary
devices such as flow sensors, transmitting equipment, recorders, and totalizers. The
improper installation or design of a primary device or malfunction of any part of a
secondary device results in erroneous flow data. The accuracy of flow measurement data
also varies widely, depending principally on the accuracy of the primary device and the
particular flow measurement method used. In any case, an experienced investigator should
be able to measure flow rates within ±10 percent of the true values.
10.2.2.5 Statistical Approach to Sampling
Four factors must be established for every sampling program: (a) number of samples, (b)
frequency of sampling, (c) parameters to be measured, and (d) sampling locations. These
factors are usually determined in varying degrees by details of the pertinent discharge
permits or are more arbitrarily set by the program resource limitations. Nevertheless, ,the
nature of the statistical methods selected and scientific judgment should be used to establish
the best procedures.
10.2.2.6 Special Sampling Procedures
Special sampling procedures should be employed for municipal, industrial, and agricultural
waters, surface waters as well as bottom sediments and sludges, and for biological,
microbiological, and radiological studies.
10.2.3 Sample Preservation and Handling
During and after collection, if immediate analysis is not possible, the sample must be
preserved to maintain its integrity. The only legally binding reference EPA has for sample
preservation methods is the NPDES permit program specified in reference 13. However,
these sample preservation procedures serve as a guide for other program objectives.
Proper handling of the samples helps insure valid data; consideration must also be given to
care of the field container material and cap material, cleaning, structure of containers,
container preparation for determination of specific parameters, container identification, and
volumes of samples.
10-4
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10.2.4 Chain-of-Custody Procedures
All programs involved in water and wastewater surveys should document and implement a
chain of possession and custody of any sample collected, whether or not the resulting data
are to be used in enforcement cases. Such procedures insure that the samples are collected,
transferred, stored, analyzed, and destroyed only by authorized personnel. See section 12.7
for detailed procedures that can be used on all sample types.
10.2.5 Quality Assurance in the Field
Quality assurance programs for sampling equipment and for field measurement procedures
(of such parameters as temperature, dissolved oxygen, pH, and conductance) are necessary
to insure data of the highest quality. A field quality assurance program administered by a
quality assurance coordinator should contain the following documented elements:
a. The analytical methodology; the special sample handling procedures; and the preci-
sion, accuracy, and detection limits of all analytical methods used.
b. The basis for selection of analytical and sampling methodology. For example, all
analytical methodology for NPDES permits shall be that specified in reference 13, or
shall consist of approved alternative test procedures. Where methodology does not
exist, the quality assurance plan should state how the new method will be
documented, justified, and approved for use.
c. The amount of analyses for quality control (QC), expressed as a percentage of
overall analyses, to assess the validity of data. Generally, the complete quality
assurance program should approximate 15 percent of the overall program with 10
and 5 percent assigned to laboratory QC and field QC, respectively. The plan should
include a shifting of these allocations or a decrease in the allocations depending
upon the degree of confidence established for collected data.
d. Procedures for the recording, processing, and reporting of data; procedures for
review of data and invalidation of data based upon QC results.
e. Procedures for calibration and maintenance of field instruments and automatic
samplers.
f. A performance evaluation system, administered through the quality assurance
coordinator, allowing field sampling personnel to cover the following areas:
(1) Qualifications of field personnel for a particular sampling situation.
(2) Determination of the best representative sampling site.
(3) Sampling technique including location of the points of sampling within the body
of water, the choice of grab or composite sampling, the type of automatic
sampler, special handling procedures, sample preservation, and sample identifi-
cation.
(4) Flow measurement, where applicable.
10-5
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(5) Completeness of data, data recording, processing, and reporting.
(6) Calibration and maintenance of field instruments and equipment.
(7) The use of QC samples such as duplicate, split, or spiked samples to assess the
validity of data.
g. Training of all personnel involved in any function affecting the data quality.
Quality assurance in sample collection should be implemented to minimize such common
errors as improper sampling methodology, poor sample preservation, and lack of adequate
mixing during compositing and testing. The checks listed in the following sections will help
the quality assurance coordinator to determine when the sample collection system is out of
control.
10.2.5.1 Duplicate Samples
At selected stations on a random time frame duplicate samples are collected from two sets
of field equipment installed at the site, or duplicate grab samples are collected. This provides
a check of sampling equipment and technique for precision.
10.2.5.2 Split Samples
A representative subsample from the collected sample is removed and both are analyzed for
the pollutants of interest. The samples may be reanalyzed by the same laboratory or
analyzed by two different laboratories for a check of the analytical procedures.
10.2.5.3 Spiked Samples
Known amounts of a particular constituent are added to an actual sample or to blanks of
deionized water at concentrations at which the accuracy of the test method is satisfactory.
The amount added should be coordinated with the laboratory. This method provides a
proficiency check for accuracy of the analytical procedures.
10.2.5.4 Sample Preservative Blanks
Acids and chemical preservatives can become contaminated after a period of use in the field.
The sampler should add the same quantity of preservative to some distilled water as
normally would be added to a wastewater sample. This preservative blank is sent to the
laboratory for analysis of the same parameters that are measured in the sample and values
for the blank are then subtracted from the sample values. Liquid chemical preservatives
should be changed every 2 weeks—or sooner, if contamination increases above predeter-
mined levels.
10.2.5.5 Precision, Accuracy, and Control Charts
A minimum of seven sets each of comparative data for duplicates, spikes, split samples, and
blanks should be collected to define acceptable estimates of precision and accuracy criteria
for data validation.
10-6
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10.2.5.6 Calibration of Field Equipment
Plans should be developed and implemented for calibrating all field analysis test equipment
and calibration standards to include the following: (a) calibration and maintenance intervals,
(b) listing of required calibration standards, (c) environmental conditions requiring calibra-
tion, and (d) a documented record system. Written calibration procedures should be
documented and should include mention of the following:
a. To what tests the procedure is applicable.
b. A brief description of the calibration procedure. (A copy of the manufacturer's
instructions is usually adequate.)
c. A listing of the calibration standard, the reagents, and any accessory equipment
required.
d. Provisions for indicating that the field equipment is labeled and contains the
calibration expiration date.
10.3 References
1. Huibregtse, K. R., and Moser, J. H., Handbook for Sampling and Sample Preservation of
Water and Waste water, EPA-600/4-76-049 (Sept. 1976).
2. Olson, D. M., Berg, E. L., Christensen, R., Otto, H., Ciancia J., Bryant, G., Lair, M. D.,
Birch, M., Keffer, W., Dahl, T., and Wehner, T., Compliance Sampling Manual,
Enforcement Division, Office of Water Enforcement, Compliance Branch (June 1977).
3. Weber, C. I., Biological Field and Laboratory Methods for Measuring the Quality of
Surface Waters and Effluents, EPA-670/4-73-001, U.S. EPA (July 1973).
4. Lauch, R. P., A Survey of Commercially Available Automatic Wastewater Samplers,
EPA-600/4-76-051, U.S. EPA (Sept. 1976).
5. Shelly, P. E., and Kirkpatrick, G. A., An Assessment of Automatic Sewer Flow
Samplers, EPA-600/2-75-065, U.S. EPA (Dec. 1975).
6. Handbook for Monitoring Industrial Wastewater, U.S. EPA, Technology Transfer (Aug.
1973).
7. Areawide Assessment Procedures Manual, Vol. II, App. D, EPA-600/9-76-014, U.S.
EPA (July 1976).
8. Winter, J. A., Bordner, R., and Scarpino, P., Microbiological Methods for Monitoring
the Environment, Part I-"Water and Wastes" U.S. EPA, EMSL, Cincinnati (1977).
9. Water Measurement Manual, 2d Edition (Revised), U.S. Department of the Interior,
Bureau of Reclamation (1974).
10. Kulin, G., and Compton, P., A Guide to Methods and Standards for Measurement of
Water Flow, Special Publication 421, National Bureau of Standards (May 1975).
11. Methods for Chemical Analysis of Water and Wastewater, U.S. EPA Technology
Transfer (1974).
12. Harris, D. J., and Keffer, W. J., Wastewater Sampling Methodologies and Flow
Measurement Techniques, EPA-907/9-74-005 (Sept. 1974).
13. Guidelines Establishing Test Procedures for the Analysis of Pollutants, Federal Register,
41, No. 232 (Dec. 1976).
10-7
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Chapter 11
RADIOCHEMISTRY
11.1 Introduction
The objective of this chapter is to provide general information and suggestions that enable
the analyst to execute his responsibilities in analytical quality control (QC) as they relate to
radiochemical analyses. Because chemical and radiochemical responsibilities should be
considered together, the following requirements could be included with the items specified
in the preceding chapters as tasks for the radiochemist: verifying the validity of laboratory
data, recommending methodology, interpreting the results, and examining the need for
standard procedures.
Environmental radiation measurements are made daily by Federal, State, local, and private
agencies. The data obtained from these measurements are used by the U.S. EPA and other
agencies for such purposes as estimating doses, describing health effects, establishing
standards and guides, and conducting regulatory activities. It is imperative to insure the
precision and accuracy of the data, so that policy decisions concerning environmental
quality are based on valid and comparable data.
A radiation QC program should be designed to encourage the development and implementa-
tion of QC procedures at all levels of sample collection, analysis, data processing, and
reporting. It should enable the analyst to verify his analyses and document the validity of
the data. In addition, such a program allows the determination of the precision and accuracy
of environmental radiochemical analyses.
11.2 Sample Collection
Analytical results can be no more meaningful than the integrity of the samples that are
analyzed. Representative samples must be collected so the data for any aliquot can be
related to a well-defined pollution source. For most analyses (table 11-1) the sample should
be preserved at the sampling site to maintain its integrity and to minimize activity losses
from absorption on container walls. If at all possible, analyses should be performed soon
after receipt of the samples at the laboratory.
Sample container types and descriptions have already been discussed. Both plastic and glass
containers have been recommended, and each has its particular merit. Cost most likely
determines which of the many types of plastic containers will be used; in general, those
more expensive should be more resistant to adsorption losses. Because sample analyses are
recommended soon after collection, less expensive plastic ware can be tolerated. In any
event, containers should be discarded after use to prevent contamination of subsequent
samples.
Glass bottles are popular items, readily available in all sizes. Although the possibility of
breakage either in handling or shipping is obviously great, the very small sizes for radioactive
tracer standards and calibration sources can withstand breakage. Use of the larger size
bottles for these purposes should therefore be avoided. It is poor economics to ship
radioactive samples in fragile glass containers when unbreakable types serve the purpose
much better.
11-1
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11.3 Laboratory Practices
11.3.1 Laboratory Safety
The general principles of laboratory safety are covered in chapter 14. The hazards to be
avoided are listed, and the importance of good housekeeping practices is stressed. These
practices reduce the potential hazard of the many chemical operations being performed. In
the radiochemical laboratory they eliminate the probability of radiological cross-
contamination from sample to sample and from sample to glassware. Many of the materials
used in the laboratory are potentially hazardous because of their chemical properties or
their radioactivity and therefore should be handled with the utmost care and respect.
In the radiochemical laboratory, the prevention of contamination by radioactive materials
requires attention to radiation-protection practices, an ongoing personnel-monitoring pro-
gram, and the designation of a segregated storage area for radioactive sources following use
in radiochemical analyses. Adequate labeling of work areas, of samples for analysis, of
aliquots, and of separated fractions helps to control radiation hazards and to insure
personnel safety.
The handling of radioactive materials involves safety hazards of a type not usually associated
with a chemical laboratory. Special precautions and instruments should be used to insure
the greatest personal safety. It is imperative to wear monitoring devices (personal film
badges) at all times.
A number of health and contamination hazards are to be considered. Many radioactive
materials are dangerous even in extremely minute quantities if inhaled or ingested. All
radioactive materials are capable of contaminating laboratories, instruments, and clothing.
All radioactive materials in large concentrations are dangerous because of the effects of their
radiation external to their containers. To control these hazards, the following rules should
be in effect at all times.
*
a. For the case of radioactive materials that are capable of being volatilized or airborne,
perform all work in a closed area or hood. Perform distillations, evaporations, and
other such processes in a well-ventilated hood.
b. Do not bring food or liquid refreshments into a laboratory engaged in work with
radioactive materials. The same applies to the counting rooms.
c. Do not smoke when handling radioactive materials.
d. After working with radioactive materials, wash hands thoroughly before eating or
handling uncontaminated materials.
11.3.2 Laboratory Analyses
Standard radiochemical procedures or their equivalent are needed to comply with sensitivity
detection limits for each nuclide as designated by the quality assurance program (2).
Radiochemical procedures have been compiled for a multitude of nuclides in a multitude of
media (3-9) and descriptions of specific separations are to be found in the scientific
literature. As laboratory techniques become more sophisticated and as more sensitive
instrumentation is developed, these procedures Will be improved.
11-3
-------
Depending on the media, the radioactivity levels, and the nuclide composition, there are
several approaches that can be made when a sample is received for QC determination. The
requirements for an acceptable QC program are described in section 11.4.
Before starting such a program, the laboratory should be already set up for radiochemical
analyses and the analysts should have the prescribed education and experience.
11.3.3 Laboratory Radiation Instruments
The types of radiation counting systems (described in ref. 2) needed to comply with the
requirements are set forth in the following paragraphs. Only those instruments needed for
analyzing specific radionuclides are required. Such instruments should meet the specifica-
tions discussed in the next sections.
11.3.3.1 Liquid Scintillation System
A liquid scintillation system must measure tritium with the sensitivity required by the
National Interim Primary Drinking Water Regulations. Efficiency of the system should be
greater than 57 percent for tritium. The tritium figure of merit (ET)i/B should be greater
than 100.
11.3.3.2 Gas-Flow Proportional Counting System or Alternative
A gas-flow proportional counter, or the alternative described later, is required for
measurement of gross alpha- and gross beta-particle activities, «radium-228, strontium-89,
cesium-134, and iodine-131. The detector may be either windowless (internal proportional
counter) or of the thin-window type. A minimum shielding equivalent of 5 cm of lead must
surround the detector. A cosmic (guard) detector should be operated in anticoincidence
with the main detector. The main detector should have an efficiency greater than 20 percent
for polonium-210 and carbon-14 and greater than 40 percent for strontium-90. The detector
background should be less than 1.3 counts per minute.
The detector plateau should be less than 1.5 percent per 100 V and should be at least 100 V
wide for carbon-14 and less than 2 percent per 100 V for strontium-90.
A scintillation system designed for alpha- and beta-particle counting may be substituted for
the gas-flow proportional counter described. In such a system a Mylar disk coated with a
phosphor (silver-activated zinc sulfide) is either placed directly on the sample (for alpha
measurements) or on the face of a photomultiplier tube, enclosed within a light-tight
container along with the appropriate electronics (high-voltage supply, amplifier(s), timer,
and sealer). Radiation shielding, although desirable, is not required for this system.
11.3.3.3 Gamma Spectrometer System
A sodium iodide (Nal) detector connected to a multichannel analyzer is required for
determination of manmade photon emitters. A 7.5- by 7.5-cm Nal crystal is satisfactory;
however, a 10- by 10-cm crystal is recommended. The crystal detector must be shielded
with a minimum of 10 cm of iron or equivalent.
It is recommended but not required that the distance from the center of the crystal detector
to any part on the shield should not be less than 30 cm. The multichannel analyzer, in
11-4
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addition to the appropriate electronics (high-voltage supply, preamplifier, and linear
amplifier), must contain a memory of not less than 200 channels and at least one readout
device.
11.3.3.4 Scintillation Cell System
A scintillation system must be designed to accept scintillation flasks (Lucas cells) for
measurement of radium-226 by the radon-emanation method. The system consists of a
light-tight enclosure for the scintillation flasks, a detector (phototube), and the appropriate
electronics (high-voltage supply, amplifier, timers, and sealers). The scintillation flasks
required for this measurement may either be purchased from commercial suppliers or
constructed according to published specifications.
11.4 Quality Control
The following requirements are recommended for all laboratories:
a. All QC data should be available for inspection to determine validity of laboratory
results.
b. Each laboratory should participate at least twice each year in EPA laboratory
intercomparison studies (10).
c. Each laboratory should participate once each year in an appropriate EPA-
administered parformance study on unknowns. Results must be within the control
limits established by EPA for each analysis.
d. Counting-instrument operating manuals and calibration protocols should be available
to analysts and technicians.
e. Calibration data and maintenance records on all radiation instruments and analytical
balances must be maintained in a permanent record.
f. Minimum daily QC
(1) To verify precision of methods, a minimum of 10 percent of the samples shall be
duplicates. Checks must be within ±2 standard deviations of the mean range.
(2) If less than 20 samples per day are analyzed, a performance standard and a
background sample must be measured. If 20 or more samples are analyzed per
day, a performance standard and a background sample imast be measured with
each 20 samples. Checks must be within ±2 standard deviations of the mean
range.
(3) Quality control performance charts or performance records must be maintained.
11.5 References
1. Federal Register^, No. 133 (July 9, 1976).
2. Jarvis, A. N., et al., The Status and Quality of Radiation Measurements of Water,
EPA-600/4-76-017(Apr. 1976).
11-5
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3. Krieger, H. L., Interim Radiochemical Methodology for Drinking Water, EPA-600/4-
75-008 (Revised), U.S. EPA, Office of Research and Development, Cincinnati (Mar.
1976).
4. Doublas, G. S., Editor, Radioassay Procedures for Environmental Samples, Environ-
mental Health Series, Publication 999-RH-27, U.S. DHEW, Public Health Service,
National Center for Radiological Health (Jan. 1967).
5. Harley, J. H., HASL Procedures Manual, HASL-300, U.S. ERDA, Health and Safety
Laboratory, New York (1972).
6. Kleinberg, J., Editor, Collected Radiochemical Procedures, LA-1721 (Revised), Los
Alamos Scientific Laboratory, Los Alamos (Nov. 1955).
7. Krieger, H. L., et al., Radionuclide Analysis of Environmental Samples, Technical
Report R59-6 (Revised), U.S. DHEW, Public Health Service (Feb. 1966).
8. Nuclear Science Series, Reports NAS-NS-3001 to NAS-NS-3115, National Academy of
Sciences, National Research Council Technical Information 'Center, Office of Informa-
tion Services (1960).
9. "Water," Part 31 of 1977 Annual Bo'ok of ASTM Standards, American Society for
Testing and Materials, Philadelphia (1977).
10. Jarvis, A. N., et al., Environmental Radioactivity Laboratory Intercomparison Studies
Program 1975, EPA-680/4-75-002b, U.S. EPA, Office of Research and Development,
Las Vegas (1975).
11. Ziegler, L. H., and Hunt, H. M., Quality Control for Environmental Measurements Using
Gamma-Ray Spectrometry, U.S. EPA, EPA-600/7-77-144 (Dec. 1977).
12. Kanipe, L. G., Handbook for Analytical Quality Control in Radioanalytical Labora-
tories, U.S. EPA, EPA-600/7-77-088 (Aug. 1977).
11-6
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Chapter 12
MICROBIOLOGY
12.1 Background
The quality assurance program described in this chapter is a synopsis of a detailed program
prescribed in parts IV and V of the EPA microbiological methods manual (1). Quality
assurance is a program of integration of all intralaboratory and interlaboratory quality
control (QC), methods standardization, and QC management practices into a formal,
coordinated, continuing effort.
12.2 Specific Needs in Microbiology
A quality assurance program for microbiological analyses must emphasize the control of
laboratory operations and analytical procedures because the tests measure living organisms
that continually change in response to their environment. Further, because true values
cannot be provided for the microbial parameters, microbiologists do not yet have the
advantages of analytical standards, QC charts, and spiked samples available to other
disciplines for measurement of accuracy. Because known values cannot be applied, it is im-
portant that careful and continuous control be exerted over sampling, personnel, analytical
methodology, materials, supplies, and equipment.
12.3 Intralaboratory Quality Control
Intralaboratory QC is the orderly application within a single laboratory of laboratory
practices necessary to eliminate or reduce systematic error and to control random error.
This within-laboratory program must be practical, integrated, and time-efficient or it will be
bypassed. When properly administered, such a program helps to insure high-quality data
without interfering with the primary analytical functions of the laboratory. This within-
laboratory program should be supplemented by participation of the laboratory in an
interlaboratory quality assurance program such as that conducted by EPA.
Intralaboratory QC for microbiology should cover the following areas:
a. Laboratory operations
(1) Sample collection and handling
(2) Laboratory facilities
(3) Laboratory personnel
(4) Laboratory equipment and instrumentation
(5) Laboratory supplies
(6) Culture media
(7) Analytical methodology
b. Analytical QC
(1) Sterility checks
(2) Positive and negative controls
12-1
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(3) Duplicate analyses
(4) Single-analyst precision
(5) Comparison of results between analysts
(6) Verification of membrane filter analyses
(7) Completion of most probable number analyses,
(8) Data handling
12.4 Interlaboratory Quality Control
An interlaboratory quality -assurance program is an agreed-upon system of minimal
requirements necessary to maintain a quality standard among a group of laboratories. Such a
program may be voluntary or compulsory for participants. It may include the following
activities:
a. Selection and approval of uniform sampling methodology and analytical methodol-
ogy
b. Collaborative studies to establish the precision and accuracy of selected methodol-
ogy
c. Preparation of guidelines to set minimal group standards for personnel, equipment,
instrumentation, facilities, and intralaboratory QC programs
d. Onsite inspection of laboratory capabilities
e. Periodic evaluation of laboratory performance on unknown samples
f. Followup on problems identified in onsite inspections and performance evaluations
As a part of its interlaboratory quality assurance program, EPA has selected microbiological
methodology and standards for laboratory operations (1). EMSL-Cincinnati is currently
conducting research on the development of QC samples for use in performance testing and
method-validation studies.
12.5 Development of a Formal Quality Assurance Program
Unless records are kept of the QC checks and procedures, there is no proof of performance,
no value in future reference, and for practical purposes, no quality assurance program in
operation. To insure a viable quality assurance program, management must first recognize
the need and require the development of a formal program, and then commit 15 percent of
the laboratory man-years to QC activities. The laboratory manager holds meetings with
supervisors and staff workers to establish levels of responsibility and functions of
management, supervisor, and analyst in the quality assurance program. Laboratory person-
nel participate in planning and structuring the program.
Once the quality assurance program is functioning, supervisors review laboratory operations
and QC with analysts on a frequent (weekly) basis. Supervisors use the results of the regular
meetings with laboratory personnel to inform management of the status of the program on a
regular (monthly) basis. These meetings identify problems through participation of labora-
tory personnel and provide the backing of management for actions required to correct
problems.
12-2
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12.6 Documentation of a Quality Assurance Program
A laboratory operating manual should be prepared that describes operation, maintenance,
and QC of laboratory operations and analyses as practiced. The review mechanisms and the
frequency of review in the quality-assurance program are included.
12.6.1 Sampling
A sample log is used to record information on samples received in the laboratory including
details of sample identification and origin, the necessary chain-of-custody information,
analyses performed, and final results.
12.6.2 Laboratory Operations
A QC record is maintained on media preparation, instrument calibration, purchase of
supplies, QC checks on materials, supplies, equipment, instrumentation, facilities, and
analyses.
12.6.3 Analytical Quality Control
A record of analytical QC checks is maintained on positive and negative controls, sterility
checks, single-analyst precision, precision between analysts, and use-test results from
comparison of lots of media, membrane filters, and other supplies.
12.7 CKain-of-Custody Procedures for Microbiological Samples
12.7.1 General
A regulatory agency must demonstrate the reliability of its evidence by proving the chain of
possession and custody of any samples that are offered for evidence or that form the basis
of analytical test results introduced into evidence in any water pollution case. It is
imperative that the office and the laboratory prepare procedures to be followed whenever
evidence samples are collected, transferred, stored, analyzed, or destroyed.
The primary objective of these procedures is to create an accurate written record that can be
used to trace the possession of the sample from the moment of its collection through its
introduction into evidence. A sample is in custody if it is in any one of the following states:
a. In actual physical possession
b. In view, after being in physical possession
c. In physical possession and locked up so that no one can tamper with it
d. In a secured area, restricted to authorized personnel
Personnel should receive copies of study plans prior to the study of a water pollution case.
Prestudy briefings should then be held to apprise participants of the objectives, sample
locations, and chain-of-custody procedures to be followed. After the chain-of-custody
samples are collected, a debriefing is held in the field to verify the adherence to the
chain-of-custody procedures and to determine whether additional samples are required.
12-3
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12.7.2 Rules for Sample Collection
An agency or laboratory engaged in sample study activities should follow these rules:
a. Involve a minimum number of trained persons in sample collection and handling.
b. Establish guidelines for particular procedures to be used for each type of sample
collection, preservation, and handling.
c. Handle samples as little as possible.
d. Obtain stream and effluent samples using the appropriate sampling techniques.
e. Attach sample tag or label securely (see fig. 12-1) to the sample container at the
time the sample is collected. The tag should contain the following items as a
minimum: the serial number of the tag, the station number and location, the date
and time taken, the type of sample, the sequence number (e.g., first sample of the
day—sequence No. 1), the preservative used, the analyses required, and the name of
the sample collector. Tags should be completed legibly in waterproof ink.
f. Use bound field notebooks to record field measurements and other pertinent
information necessary to reconstruct the sample collection processes in the event of
a later enforcement proceeding. Maintain a separate set of field notebooks for each
study and store them in a safe place where they can be protected and accounted for
at all times. Establish a sample log sheet with a standard format to minimize field
entries and include the serial number of the sheet, the date, time, survey, type of
samples taken, volume of each sample, type of analyses, (unique) sample numbers,
sampling location, field measurements (such as temperature, conductivity, dissolved
oxygen (DO), and pH), and any other pertinent information or observation. (See fig.
12-2.) The entries should be signed by the sample collector. The responsibility for
preparing and retaining field notebooks during and after a study should be assigned
to a study coordinator or his designated representative.
g. The sample collector is responsible for the care and custody of the samples until the
samples are properly dispatched to the receiving laboratory or given to an assigned
custodian. The sample collector must insure that each container is in his physical
possession or in his view at all times, or stored in a locked place where no one can
tamper with it.
h. Take color slides or photographs of the sample locations and any visible water
pollution. Sign and indicate time, date, and site location on the back of the photo.
To prevent alteration, handle such photographs according to the established
chain-of-custody procedures.
12.7.3 Transfer of Custody and Shipment
In transfer-of-custody procedures, each custodian or sampler must sign, record, and date the
transfer. Most environmental regulatory agencies develop chain-of-custody procedures
tailored to their needs. These procedures may vary in format and language but contain the
same essential elements. Historically, sample transfer under chain of custody has been on a
12-4
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UVS. ENVIRONMENTAL PROTECTION AGENCY
Station No. Date Time Sequence No.
Station Location
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ENVIRONMENTAL PROTECTION AGENCY
(Local Address)
(b)
Figure 12-1. Example of chain-of-custody sample tag. (a) Front, (b) Back.
sample-by-sample basis, which is awkward and time consuming. However, the EPA National
Enforcement Investigation Center (NEIC) at Denver has set precedent with its bulk transfer
of samples. Bulk transfer is speedier and reduces paperwork and the number of sample
custodians. The following description of bulk transfer of custody is essentially that of the
Office of Enforcement (2).
12-5
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a. Samples must be accompanied by a chain-of-custody record that includes the name
of the study, collectors' signatures, station number, station location, date, time, type
of sample, sequence number, number of containers, and analyses required. (See fig.
12-3.) When turning over possession of samples, the transferor and transferee sign,
date, and time the record sheet. This record sheet allows transfer of custody of a
CHAIN OF CUSTODY RECORD
STATION
NUMBER
STATION LOCATION
DATE
Relinquished by:^™^™/
Relinquished by: is,gn,,u,,/
Relinquished by: ts,ir,,w,,i
Received by: »»„«•,«/
Dispatched by: Date/Time
Received by Mobile Laboratory for field Date/Time
analysis: (Sw,u,,i
Date/Time
Received for Laboratory by: Date/Time
Distribution: Ong.— Accompany Shipment
1 Copy— Survey Coordinator Field Files
Figure 12-3. Example of chain-of-custody record.
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group of samples in the field to the mobile laboratory or to the central laboratory.
When a custodian transfers a portion of the samples identified on the sheet to the
mobile laboratory, the individual samples must be noted in the column with the
signature of the person relinquishing the samples. The laboratory person receiving
the samples acknowledges receipt by signing in the appropriate column.
b. If the custodian has not been assigned, the field custodian or field sampler has the
responsibility of packaging and dispatching samples to the laboratory for analysis.
The dispatch portion of the chain-of-custody record must be filled out, dated, and
signed.
c. To avoid breakage, samples must be carefully packed in shipment containers such as
ice chests. The shipping containers are padlocked for shipment to the receiving
laboratory.
d. Packages must be accompanied by the chain-of-custody record showing identifica-
tion of the contents. The original must accompany the shipment. A copy is retained
by the survey coordinator.
e. If sent by mail, register the package with return receipt requested. If sent by
common carrier, a Government bill of lading should be obtained. Receipts from post
offices and bills of lading will be retained as part of the permanent chain-of-custody
documentation.
f. If delivered to the laboratory when appropriate personnel are not there to receive
them, the samples must be locked in a designated area within the laboratory, so that
no one can tamper with them or must be placed in a secure area. The recipient must
return to the laboratory, unlock the samples, and deliver custody to the appropriate
custodian.
12.7.4 Laboratory Custody Procedures
Suitable laboratory procedures during custody of samples include the following:
a. The laboratory shall designate a sample custodian and an alternate custodian to act
in his absence. In addition, the laboratory shall set aside a sample storage security
area. This should be a clean, dry, isolated room with sufficient refrigerator space
that can be securely locked from the outside.
b. Samples should be handled by the minimum possible number of persons.
c. Incoming samples shall be received only by the custodian who will indicate receipt
by signing the chain-of-custody record sheet accompanying the samples and
retaining the sheet as a permanent record. Couriers picking up samples at the airport
or post office shall sign jointly with the laboratory custodian.
d. Immediately upon receipt, the custodian places the samples in the sample room,
which will be locked at all times except when samples are removed or replaced by
the custodian. To the maximum extent possible, only the custodian shall be
permitted in the sample room.
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e. The custodian shall insure that microbiological samples are properly stored and
maintained at 4°C.
f. Only the custodian will distribute samples to personnel who are to perform tests.
g. The analyst records in his laboratory notebook or analytical worksheet, identifying
information describing the sample, the procedures performed, and the results of the
testing. The notes shall be dated, shall indicate who performed the tests, and should
include any abnormalities that occurred during the testing procedure. The notes
shall be retained as a permanent record in the laboratory. In the event that the
person who performed the tests is not available as a witness at the time of a trial, the
Government may be able to introduce the notes in evidence under the Federal
Business Records Act.
h. Approved methods of laboratory analyses shall be used as required by Public Laws
92-500, 93-523, 92-532, and amendments.
i. Laboratory personnel are responsible for the care and custody of a sample once it is
handed to them and should be prepared to testify that the sample was in their
possession and view or secured in the laboratory at all times from the moment it was
received from the custodian until the tests were run.
j. The laboratory area shall be maintained as a secured area and shall be restricted to
authorized personnel.
k. Once the sample analyses are completed, the unused portion of the sample, together
with identifying labels and other documentation, must be returned to the custodian.
The returned, tagged sample should be retained in the custody room until
permission to destroy the sample is received by the custodian.
1. Samples should be destroyed only upon the order of the laboratory director, in
consultation with previously designated enforcement officials, or when it is certain
that the information is no longer required, or that the samples have deteriorated.
The same destruction procedure is true for tags and laboratory records.
12.7.5 Evidentiary Considerations
Reducing chain-of-custody procedures and promulgated analytical procedures to writing will
facilitate the admission of evidence under Rule 803 (6) of the Federal Rules of Evidence
(Public Law 93-575). Under this statute, written records of regularly conducted business
activities may be introduced into evidence as an exception to the hearsay rule without the
testimony of the person(s) who made the record. Although it would be preferable, it is not
always possible for the individuals who collected, kept, and analyzed samples to testify in
court. In addition, if the opposing party does not intend to contest the integrity of the
sample or testing evidence, admission under Rule 803(6) can save a great deal of trial time.
For these reasons, it is important that the procedures followed in the collection and analyses
of evidentiary samples be standardized and described in an instruction manual, which, if
need be, can be offered as evidence of the regularly conducted business activity followed by
the laboratory or office in generating any given record.
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In criminal cases, however, records and reports of matter observed by police officers and
other law enforcement personnel are not included under the business record exceptions to
the hearsay rule previously cited. (See Rule 803(8), Public Law 93-595.) It is arguable that
those portions of the compliance inspection report dealing with matters other than sampling
and analysis results come within this exception. For this reason, in criminal actions, records
and reports of matter observed by field investigators may not be admissible, and the
evidence may still have to be presented in the form of oral testimony by the person(s) who
made the record or report, even though the materials come within the definition of business
records. In a criminal proceeding, the opposing counsel may be able to obtain copies of
reports prepared by witnesses (even if the witness does not refer to the records while
testifying), which may be used for cross-examination purposes.
Admission of records is not automatic under either of these sections. The business records
section authorizes admission "unless the source of information or the method or circum-
stances of preparation indicate lack of trustworthiness," and the caveat under the public
records exception reads "unless the sources of information or other circumstances indicate
lack of trustworthiness."
Thus, whether or not the inspector anticipates that a report will be introduced as evidence,
the inspector should make certain that the report is as accurate and objective as possible.
12.8 References
1. Winter, J. A., Bordner, R., and Scarpino, P., Microbiological Methods for Monitoring the
Environment, Part I-"Water and Wastes," U.S. EPA, EMSL, Cincinnati (1978).
2. NPDES Compliance Sampling Manual, U.S. EPA, Office of Water Enforcement (June
1977).
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Chapter 13
AQUATIC BIOLOGY
Quality assurance guidelines for aquatic biology programs (fully described in ref. 1) are
summarized in the following section.
13.1 Summary of General Guidelines
Successful quality assurance programs in aquatic biology are based on the following essential
elements:
a. An understanding and acceptance of the importance of quality control (QC) and a
commitment on the part of the biology staff to fully integrate QC practices into
field and laboratory operations
b. A staff with adequate formal training and experience and proper specialization to
meet program needs
c. Adequate field equipment, storage and laboratory space, instrumentation, and
taxonomic references
d. Careful advance preparation and design of field and laboratory studies
e. Strict adherence to approved methodology, where available, and careful consider-
ation of the technical defensibility of the methods and their application
f. Use of replication in sample collection and analysis where feasible, and determina-
tion of the accuracy and precision of the data
g. Frequent calibration of field and laboratory instruments
h. Proper sample identification and handling to prevent misidentification or inter-
mixing of samples
i. Use of blind, split, or other control samples to evaluate performance
j. Development and regular use of in-house reference specimen collections, and use of
outside taxonomic experts to confirm or provide identifications for problem
specimens
k. Meticulous, dual-level review of the results of manual arithmetical data manipula-
tions and transcriptions before the data are used in reports or placed in BIO-
STORET (2)
1. Participation in EPA formal interlaboratory aquatic biology methods studies, and
use of EPA biological reference materials
m. Documentation of methodology and QC practices employed in the program
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13.2 Discussion
A brief description of each of the areas mentioned in section 13.1 is provided here to
indicate the scope of the quality assurance program.
13.2.1 Staff Commitment
To establish and maintain an effective quality assurance program in aquatic biology, the
supervisor must actively support and frequently monitor the use of QC practices in all
aquatic biology activities. This will require the commitment of 10 to 15 percent of the total
manpower resources. The supervisor must review field and laboratory operations with his
staff frequently (weekly) to insure that QC practices are followed and properly docu-
mented.
13.2.2 Staff
The quality and reliability of the data rest heavily on the competence of the staff. The range
of aquatic organisms studied by biological programs is very broad, and each community
requires unique skills in sample collection and analysis, and in data interpretation. Several
disciplines, therefore, must be represented on the staff to deal effectively with the
taxonomy and ecology of the major groups of aquatic organisms, which include the
phytoplankton, zooplankton, periphyton, macrophyton, macroinvertebrates, and fish.
13.2.3 Facilities
The quality of the data also depends upon the availability and performance of laboratory
equipment. Such items as sampling gear, current meters, spectrophotometers, and micro-
scopes must be available and must meet performance standards related to the biological
parameters measured. Laboratory instrumentation must provide the sensitivity and accuracy
required by the state of the art in sample analysis. Adequate laboratory and storage space
must also be provided.
13.2.4 Advance Planning
Thorough advance planning of field and laboratory projects is necessary to maintain the
required control over the technical aspects of the project and to insure the collection of
meaningful data. Factors taken into consideration include the objectives of the study, the
parameters to be measured, station selection, the sampling frequency and replication,
seasonal cycles in the properties of communities of aquatic organisms, and QC measures to
be incorporated into the various phases of the project.
13.2.5 Use of Approved Methodology
Methods in the EPA manual "Biological Field and Laboratory Methods for Measuring the
Quality of Surface Waters and Effluents" (1) should be employed where applicable. The
manual contains concensus methods selected by a committee of senior Agency aquatic
biologists as the preferred methods for use within EPA. If program activities require
methods for parameters not covered by reference 1, methods may be selected from other
sources if their application is technically defensible.
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13.2.6 Replication
Comparisons of biological parameters measured in control and affected regions of water
bodies, and in laboratory experiments, may be meaningless if the precision of the results is
not known. Preliminary measurements must be made in each study to determine the scatter
in the data and to establish the number of replicate samples required to achieve the level of
precision required to detect differences in the responses measured.
13.2.7 Instrument Calibration and Maintenance
Sampling equipment and laboratory instrumentation with mechanical metering devices and
electronic components are calibrated on a regularly scheduled basis to insure the accuracy of
the data. Standards are obtained, such as NBS-certified thermometers for temperature-
measuring devices, class-S weights for balances, and absorption filters for spectrophotom-
eters. Records of calibrations, regular performance checks, and service for each device are
maintained in bound log books in such a manner that the history of performance of the
instruments may be easily reviewed. Analytical reagents are labeled and dated when
received, and are protected from deterioration if labile. The expected shelf life of each
reagent is recorded on the label, and the material is not used after the expiration date.
13.2.8 Sample Labeling
Samples are securely labeled in the field and recorded in a bound log. Information on the
label should include the station, date, time of day, depth, and other relevant information. A
unique lot number is assigned to the sample and recorded on the label. Waterproof paper
and ink must be used for the labels, and are recommended for the field logs. Depending on
requirements, labels are placed inside samples such as macroinvertebrates.
13.2.9 Quality Control Samples
The accuracy of the data from routine analyses such as counts and identification of
organisms and chlorophyll and biomass measurments is determined by introducing blind,
split, or reference samples in the sample processing stream. These samples are either
prepared by the supervisory aquatic biologist, laboratory quality control officer, or
analytical QC coordinator, or are obtained from EMSL-Cincinnati. The results are discussed
at regularly scheduled staff meetings and any problems identified are discussed and
corrected.
13.2.10 Organism Identification and Reference Specimens
Accurate identification of aquatic organisms to the species level is essential to the
interpretation of biological data. A set of reference specimens is established within each
laboratory, to be used as taxonomic (identification) standards in processing samples and in
training new personnel. The set is representative of the aquatic organisms collected by the
program, and each specimen embodies the morphological characteristics essential to the
identification of that taxon. The identity of these specimens is verified by outside
taxonomic experts, who also examine organisms that pose unusually difficult identification
problems in routine sample analysis.
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13.2.11 Data Records, Editing (Proofing), Review
Data collected manually are entered in a bound log or on specialized bench sheets that fully
describe the origin and nature of the sample and that are maintained in a binder or file.
Source data such as organism abundance, metabolic rates, and chlorophyll, which are
manually or electronically manipulated or transcribed from one record to another, are
doublechecked by a second person. All manual calculations and all electronic calculations,
where data are manually keyboarded, are performed twice, except where the source (input)
data are included in the output and can be proofed. Keyboarded data are carefully proofed
before they are submitted for computer manipulation.
13.2.12 Interlaboratory Methods Studies
The aquatic biology programs participate in formal interlaboratory biological methods
studies performed by EMSL-Cincinnati. Studies on chlorophyll and macroinvertebrate
identification methods have been completed, and additional studies on phytoplankton and
periphyton identification methods are planned in the future.
13.2.13 Documentation for the Quality Assurance Program
A laboratory operations manual is available (1) that describes the scope of the program,
organizational structure, qualifications of the staff, available space and equipment,
methodology employed for sample collection and analysis, and QC procedures.
13.3 References
1. Weber, C. I., Editor, Biological Field and Laboratory Methods for Measuring the Quality
of Surface Waters and Effluents, 2d Edition, EPA-670/4-73-001, U.S. EPA (July 1973).
2. Nacht, L., and Weber, C. I., BIO-STORET Final Design Specification, U.S. EPA (1976).
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Chapter 14
LABORATORY SAFETY
14.1 Law and Authority for Safety and Health
Public Law 91-596 is the Occupational Safety and Health Act of 1970 (1). The purpose of
this act is the following:
To assure safe and healthful working conditions for working men and women; by
authorizing enforcement of the standards developed under the act; by assisting and
encouraging the States in their efforts to assure safe and healthful working
conditions; by providing for research, information, education, and training in the
field of occupational safety and health; and for other purposes.
The intent of the act (1) is "to assure so far as possible every working man and woman in
the Nation safe and healthful working conditions and to preserve our human resources."
The responsibility for promulgating and enforcing occupational safety and health standards
rests with the Department of Labor. The Department of Health, Education, and Welfare is
responsible for conducting research on which new standards can be based, and for
implementing education and training programs for producing an adequate supply of
manpower to implement the purposes of the act. These responsibilities are performed by the
National Institute for Occupational Safety and Health (NIOSH).
Section 19(a) of Public Law 91-596 states the following (1):
It shall be the responsibility of the head of each Federal agency to establish and
maintain an effective and comprehensive occupational safety and health program
that is consistent with the standards promulgated under section 6. The head of each
agency shall (after consultation with representatives of the employees thereof)-
(1) provide safe and healthful places and conditions of employment, consistent
with the standards set under section 6;
(2) acquire, maintain, and require the use of safety equipment, personal
protective equipment, and devices reasonably necessary to protect employ-
ees;
(3) keep adequate records of all occupational accidents and illnesses for proper
evaluation and necessary corrective action;
(4) consult with the Secretary with regard to the adequacy as to form and
content of records kept pursuant to subsection (a)(3) of this section; and
(5) make an annual report to the Secretary of Labor with respect to occupa-
tional accidents and injuries and the agency's program under this section.
Such report shall include any report submitted under section 7902(e)(2) of
title 5, United States Code.
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Executive Order 11807 was issued in the fall of 1974 to provide general direction to the
heads of Federal agencies and to the Secretary of Labor in establishing occupational safety
and health programs in the Federal Government. The executive order details the following
duties:
a. Appointment of a Safety and Health Official.
b. Establishment of a management information system.
c. Establishment of an occupational safety and health program.
(1) Adoption of safety and health standards as effective as the Secretary of Labor's
standards.
(2) Institution of procedures for. processing reports from employees on hazardous
conditions.
(3) Institution of periodic inspection of facilities.
(4) Provision for abatement of hazards in facilities.
d. Provision for training of agency personnel.
(1) Training of supervisors at all levels.
(2) Training of those responsible for conducting inspections of facilities.
(3) Training of other employees. Attention is called to the list of OSHA training
requirements.
e. Assistance of the Secretary of Labor.
(1) Compliance with the recordkeeping and reporting requirements.
(2) Observation of the guidelines issued in 28 CFR 1960.
(3) Cooperation with the Secretary of Labor in the performance of his responsi-
bilities.
f. Issuance of guidelines for a safety and health program.
g. Prescription of recordkeeping and reporting requirements.
h. Provision of consulting services to Federal agencies in adoption of standards, training
agency personnel, and in other matters.
i. Upon request and subject to reimbursement, performance of such services as
evaluation of safety and health conditions in the agency, recommendation on the
adoption of standards, inspection of facilities for safety and health hazards, and
training of agency personnel in safety and health matters.
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j. Evaluation of agency programs and report of the condition to the President. This
item in the executive order gives to the Secretary of Labor a degree of enforcement
of safety and health rules and regulations within Federal agencies.
k. Section 4 continues the Federal Advisory Council on Occupational Safety and
Health. Note that at least one-third of the members are to be labor representatives.
14.1.1 Occupational Safety and Health Administration Regulations
The Occupational Safety and Health Act of 1970 covers laboratory workplaces as well as
industrial and manufacturing workplaces (1). Many large laboratories have already come
under the scrutiny of the Occupational Safety and Health Administration (OSHA); small
and medium-sized laboratories can expect direct involvement in the future.
The 1970 act set up NIOSH within the U.S. Department of Health, Education, and Welfare.
NIOSH provides OSHA with the scientific information it needs to effectively perform its
regulatory function.
The initial OSHA regulations were, and in some measure still are, a collection of many
well-established standards taken from industry and from standards-making groups. The
American National Standards Institute, the National Safety Council, the National Fire
Protection Association, and the American Society for Testing and Materials were some of
the prime sources for these OSHA standards. Certain existing State health and safety
regulations that predated OSHA were used to develop the Federal OSHA regulations.
Efforts are continually underway to refine and quantify current OHSA regulations through
court decisions resulting from appeals of compliance citations, through continuing reviews
of standards by OSHA safety compliance officers, through the work of NIOSH, and through
internal reviews.
Because of the continual publication of revisions of the regulations consisting of refinements
and clarifications, as well as dissemination of newly issued regulations, laboratory operators
must expend great efforts to keep strictly up to date. What was permitted yesterday may
not be permitted today.
The first place to start in keeping up to date on OSHA regulations is to obtain a copy of the
Federal or State regulations that apply. In States not operating their own OSHA function,
copies of applicable regulations are usually available without charge from the local office of
the U.S. Department of Labor. In other States, copies of their regulations are available from
similar State offices and boards either free or at a nominal charge. In either case, appropriate
steps must be taken to keep up to date on revisions and reissues. Usually this means getting
on a mailing list for automatic receipt of this material as it is published. Because entire
sections of the regulations may be reissued to incorporate only a few changes, it is usually
necessary to completely study these reissues to determine the actual change that has been
made.
As OSHA moves more into the complicated areas of chemical and biological laboratory
workplaces, the level of the compliance inspector's knowledge must be increased
accordingly.
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14.1.2 Federal and State OSHA
There is no completely identical OSHA standard in force throughout the United States.
A provision of the act of 1970 permits Federal OSHA to certify individual State OSHA
operations when they are satisfied that applicable State regulations and compliance
enforcement methods fulfill the provisions of the Federal act (1).
While many State OSHA applications are in process, the following States and territories are
certified or at least provisionally certified to permit their own OSHA operation:
Alaska
Arizona
California
Colorado
Connecticut
Hawaii
Indiana
Iowa
Kentucky
Maryland
Michigan
Minnesota
Nevada
North Carolina
Oregon
South Carolina
Tennessee
Utah
Vermont
Washington
Wyoming
Virgin Islands
All other states and territories are under Federal OSHA jurisdiction.
Even though the Federal Government supports 50 percent of the cost of State OSHA
programs, these States apparently would prefer that the Federal Government take on the
entire cost burden for them.,
An important difference between Federal and State OSHA operations lies in their
jurisdictions. While State OSHAs are responsible for State-operated workplaces, Federal
OSHA can only inspect Federal workplaces. It cannot enforce compliance. There is,
however, a current effort by the General Accounting Office to urge Congress to remove this
restriction on Federal OSHA's effectiveness in dealing with Federal workplaces.
While Federal OSHA is not permitted to offer a consulting service, most State OSHAs can
and do. Some who were unaware of this difference have called local Federal OSHA offices
for advice. All they succeeded in doing was to invite an inspection. Normally, however,
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inspections of facilities, be they manufacturing or laboratories, are triggered by an
employee's complaint or an accident report, either as required by law or through police,
fire, or hospital channels.
Reactions to OSHA operations vary widely. Many laboratories have been cited for violations
and have been heavily fined. Yet Dr. David Pettit, Technical Director of Kelco's operations
in San Diego, says, "We feel that OSHA standards are not unrealistic, just good laboratory
practice."
14.2 EPA Policy on Laboratory Safety
EPA has issued an Occupational Safety and Health Manual (3) in which policy, responsi-
bilities, accident reporting, inspections, standards, and training are described. The Agency
has also issued a Safety Management Manual (4) that contains information on safety
protection plans at EPA facilities and air, water, and road safety.
The EPA Occupational Safety and Health Manual (3) establishes policy, responsibilities, and
procedures for the conduct of the EPA safety and health program.
The policy of EPA is to administer its programs in a manner that assures adequate
protection of its own employees and property, and that for which it has a responsibility.
Every manager, supervisor, and employee is responsible for identifying risks, hazards, and
unsafe situations or practices and for taking steps to insure adequate safety in the activities
under his supervison.
A facility safety officer designated for each unit must be responsible for assisting the
officer-in-charge in developing, organizing, directing, and evaluating the safety and health
program and coordinating illness and inquiry reporting and recordkeeping requirements;
analyzing accidents and injuries for prevention and control; and providing technical advice
in the implementation of program standards and policy.
Safety in any laboratory requires continuing attention. Use of new or different techniques,
chemicals, and equipment requires careful reading, instruction, and supervision, and may
require consultation with other people with special knowledge or experience.
Prevention of laboratory accidents requires positive attitudes toward safety and training,
and suitable information for understanding laboratory and chemical hazards and their
consequences.
Responsibility for safety within the laboratories for an organization may be considered to
exist at three different levels—individual, supervisory (or instructional), and organizational
(or institutional).
The division of responsibility needs to be clearly assigned and accepted, steps need to be
taken to see that the responsibilities are exercised, and the assignments need to be reassessed
if unexpected problems develop.
Each individual who works in a laboratory is responsible for learning the health and safety
hazards of the chemicals he will be using or producing, and the hazards that may occur from
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the equipment and techniques he will employ, so that he can design his setup and
procedures to limit the effects of any accident. The individual should investigate any
accident that occurs, and record and report the apparent causes and the preventive measures
that may be needed to prevent similar accidents.
The supervisor has the responsibility for giving all the necessary directions, including the
safety measures to be used, and the responsibility of seeing that employees carry out their
individual responsibilities. Whoever directs the activities of others has a concurrent
responsibility to prevent accidental injuries from occurring as a result of the activities.
The organization of which the laboratories are part has a fundamental responsibility to
provide the facilities, equipment, and maintenance for a safe working environment and to
provide an organized program to make the improvements necessary for a safe working
environment. Unless the organization fulfills its responsibilities, it cannot expect its
supervisors, employees, or students to fulfill their responsibilities for laboratory safety.
The Environmental Protection Agency has a" designated safety and health official who is
responsible for assuring that formal safety and health inspections are conducted at all EPA
workplaces. He will notify the Administrator regarding uncorrected safety and health
deficiencies.
14.2.1 Formal Safety Inspections
Formal safety and health inspections at workplaces where there is an increased risk of
accident, injury, or illness because of the nature of the work performed, as in the case of
chemical operations and material-handling or material-loading operations, must be made by
a safety and health specialist. A "Safety and Health Specialist" is defined in 29 CFR
1960.2(h) as a person who meets the Civil Service standards for the position of Safety
Manager/Specialist GS-018, Safety Engineer GS-803, Fire Protection Engineer GS-804,
Industrial Hygienist GS-690, Fire Protection Specialist/Marshall GS-081, or Health Physicist
GS-1306, or who is an employee of an equally qualified military agency or nongovernment
organization.
Formal safety and health inspections need not be made by a safety and health specialist at
workplaces where there is little risk involved, but should be conducted by a person having
sufficient training and experience in the safety and health needs of the workplaces involved
to adequately perform the duties of an inspector as set forth in Executive Order 11807.
Inspectors should be accompanied on formal safety and health inspections by representa-
tives of the officer in charge of the reporting unit being inspected and representatives of the
employees of such establishments. Management and employee representatives should be
familiar with and maintain OSHA standards.
To insure safe and healthful working conditions for EPA employees, safety and health
inspectors are authorized to enter without delay and at reasonable times, any building,
installation, facility, construction site, or other area, workplace, or environment where work
is performed by employees of the Agency, to inspect and investigate during regular working
hours and at other reasonable times, and within reasonable limits and in a reasonable
manner, any such place of employment and all pertinent conditions, structures, machines,
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apparatus, devices, equipment, and materials therein; and to question privately any
employee, or any supervisory employee, or any officer in charge of a reporting unit.
14.2.2 Informal Safety Inspections
An informal safety and health inspection is performed on either a scheduled or unscheduled
basis by the facility safety officer, regional safety officer, supervisory management, or
members of a safety and health committee. EPA Form 1440-2, Health and Safety
Inspection Checklist (fig. 14-1), shall be used by the inspector or spokesman of a safety and
health committee conducting the inspection to note safety and health deficiencies identified
during the inspection process.
A more detailed checklist for safety evaluation of the laboratory is given in appendix A.
This inspection list contains many specific recommendations and guidelines for laboratory
safety.
14.3 Laboratory Safety Practices*
14.3.1 Introduction
14.3.1.1 Safe Use, Handling, and Storage of Chemicals
Chemicals in any form can be safely stored, handled, and used if their hazardous physical
and chemical properties are fully understood and the necessary precautions, including the
use of proper safeguards and personal protective equipment are observed.
The management of every unit within a manufacturing establishment must give whole-
hearted support to a well-integrated safety policy.
14.3.1.2 General Rules for Laboratory Safety
Supervisory personnel should think "safety." Their attitude toward fire and safety standard
practices is reflected in the behavior of their entire staff.
A safety program is only as strong as the worker's will to do the correct things at the right
time.
The fundamental weakness of most safety programs lies in too much lip service to safety
rules and not enough action in putting them into practice.
Safety practices should be practical and enforceable.
Accident prevention is based on certain common standards of education and training of
personnel, and provision of safeguards against accidents.
This description was prepared by Paul F. Hallbach, Chemist, National Training and
Operational Technology Center, U.S. EPA, Cincinnati, Ohio 45268.
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HEALTH AND SAFETY INSPECTION CHECKLIST
1 TITLE
NAME 'NUMBER OF BUILDING INSPECTED (Use separate form for each huiltiingl [ REPORTING UNIT
DATE
PART 1 PHYSICAL CONDITIONS (Check each applicable Item)
ITEM
8 EGRESS
9 LIGHTING
10 VENTILATION
7 1 FLAMMABLE OR NOXIOUS OUST OR VAPORS
IS FIRE SUPRESSiON (Including extinguishers)
SAT
UNSAT
ITEM
19 WATER (anri-svphi>ne and cross connections)
20 ELECTRICAL (fuses, grounding, etc ;
23 PARKING AREA
27 HAZARDOUS WARNING SIGNS
28 EMISSION OF POLLUTANTS lair fluid, solids)
29 OCCUPATIONAL NOISE EXPOSURE
31 PROVISIONS FOR HANDICAPPED
32 OTHER
SAT
UNSAT
PART ll PROCEDURES AND INSTRUCTIONS {Check each applicable item}
33 MATERIALS HANDLING
35 BUILDING MAINTENANCE
EQUIPMENT
38 HA2ABO MONITORING EQUIPMENT
/carton monoxide radiation, etc /
41 BOATING OPERATIONS
AND HEALTH
53 OTHER
REMARKS fConrinueon back, if necessary) (NOTE Use EPA Form )44Cf> '
9
EPA Form 1440-2 (Rev 5-771
PREVIOUS EDITION MAV BE USED UNTIL SUPPLY IS EXHAUSTED
Figure 14-1. Health and safety inspection checklist.
14.3.2 Laboratory Design and Equipment
14.3.2.1 Type of Construction
The construction of the laboratory should generally be fire resistant or noncombustible.
14-8
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Multiple story buildings should have adequate means of exit.
Stairways should be enclosed with brick or concrete walls.
Laboratories should have adequate exit doors to permit quick, safe escape in an emergency
and to protect the occupants from fires or accidents in adjoining rooms. Each room should
be checked to make sure there is.no chance of a person being trapped by fire, explosions, or
release of dangerous gases.
Laboratory rooms in which most of the work is performed with flammable liquids or gases
should be provided with explosion-venting windows.
14.3.2.2 Arrangement of Furniture and Equipment
Furniture should be arranged for maximum use of available space and should provide
working conditions that are efficient and safe.
Aisles between benches should be at least 4 ft wide to provide adequate room for passage of
personnel and equipment.
Desks should be isolated from benches or adequately protected.
Every laboratory should have an eyewash station and a safety shower.
14.3.2.3 Hoods and Ventilation
Adequate hood facilities should be installed where highly toxic or highly flammable
materials are used.
Hoods should be ventilated separately and the exhaust should be terminated at a safe
distance from the building.
Makeup air should be supplied to rooms or to hoods to replace the quantity of air exhausted
through the hoods.
Hood ventilation systems are best designed to have an airflow of not less than 60 ft/min
(linear) across the face of the hood with all doors open, and 150 ft/min (linear) if toxic
materials are involved.
Exhaust fans should be sparkproof if exhausting flammable vapors and corrosive resistant if
handling corrosive fumes.
Controls for all services should be located at the front of the hood and should be operable
when the hood door is closed.
All laboratory rooms should have the air changed continuously at a rate depending on the
materials being handled.
Recent California OSHA regulations require the presence of a means of visual indication of
the existence of the airflow in the hood and specify the height and type of hood exhaust
permitted.
14-9
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14.3.2.4 Electrical Services
Electrical outlets should be placed outside of hoods to afford easy access and thus protect
them from spills and corrosion by gases.
Noninterchangeable plugs should be provided for multiple electrical services.
*
Adequate outlets should be provided and should be of the three-pole-type to provide for
adequate grounding.
Rubber or nonconductive composition shoe soles should be required (except when
flammable vapors are present). Shoe soles should not be of a type that readily absorbs water
or other liquids.
14.3.2.5 Storage
Laboratories should provide for adequate storage space for mechanical equipment and
glassware that will be used regularly.
Flammable solvents should not be stored in glass bottles over 1 1 in size. Large quantities
should be stored in metal safety cans. Quantities requiring containers larger than 1 gal
should be stored outside the laboratory.
Explosionproof refrigerators should be used for the storage of highly volatile and flammable
solvents.
Cylinders of compressed or liquefied gases should not be stored in the laboratory.
Alphabetized storage of chemicals should be avoided to prevent the unintentional mixing of
two incompatible chemicals in an accident situation.
An appropriate antidote must be readily available for every stored chemical compound for
which an antidote is specified.
U.3.2.6 Housekeeping
Housekeeping plays an important role in reducing the frequency of laboratory accidents.
Rooms should be kept in a neat and orderly condition. Floors, shelves, and tables should be
kept free from dirt and from all apparatus and chemicals not in use.
A cluttered laboratory is a dangerous place to work. Maintenance of a clean and orderly
work space is indicative of interest, personal pride, and safetymindedness.
Passageways should be kept clear to all building exits and stairways.
Metal containers should be provided for the disposal of broken glassware and should be
properly labeled.
Separate approved waste disposal cans should be provided for the disposal of waste
chemicals.
14-10
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Flammable liquids not miscible with water and corrosive materials or compounds that are
likely to give off toxic vapors should never be poured into the sink.
Laboratory operators must be sure that unguarded rotating equipment such as belt-driven
vacuum pumps is provided with guards all the way around and that the guards are always in
place.
Whenever heavy laboratory equipment must be moved frequently, rollers should be
provided. In other cases, proper lifting equipment should be available.
14.3.2.7 Fire Protection
Laboratory personnel should be adequately trained regarding pertinent fire hazards
associated with their work.
Personnel should know rules of fire prevention and methods of combating fires.
Fire extinguishers (CO2 type) should be provided at convenient locations and personnel
should be instructed in their use.
Automatic sprinkler systems are effective for the control of fires in chemical laboratories.
14.3.2.8 Alarms
An approved fire-alarm system should be provided.
Wherever a hazard of accidental release of toxic gases exists, a gas alarm system to warn
occupants to evacuate the building should be provided.
Gas masks of oxygen or compressed-air-type should be located near exits and selected
personnel trained to use them.
14.3.3 Handling Glassware
14.3.3.1 Receiving, Inspection, and Storage
Packages containing glassware should be opened and inspected for cracked or nicked pieces,
pieces with flaws that may become cracked in use, and badly shaped pieces.
Glassware should be stored on well-lighted stockroom shelves designed and having a coping
of sufficient height around the edges to prevent the pieces from falling off.
14.3.3.2 Laboratory Practice
Select glassware that is designed for the type of work planned.
To cut glass tubing or a rod, make a straight, clean cut with a cutter or file at the point
where the piece is to be severed. Place a towel over the piece to protect the hands and
fingers, then break away from the body.
14-11
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Large size tubing is cut by means of a heated nichrome wire looped around the piece at the
point of severance.
When it is necessary to insert a piece of glass tubing or a rod through a perforated rubber or
cork stopper, select the correct bore so that the insertion can be made without excessive
strain.
Use electric mantels for heating distillation apparatus, etc.
To remove glass splinters, use a whisk broom and a dustpan. Very small pieces can be picked
up with a large piece of wet cotton.
14.3.4 Gases and Flammable Solvents
14.3.4.1 Gas Cylinders
Large cylinders must be securely fastened so that they cannot be dislodged or tipped in any
direction.
Connections, gages, regulators, or fittings used with other cylinders must not be inter-
changed with oxygen cylinder fittings because of the possibility of fire or explosion from a
reaction between oxygen and residual oil in the fitting.
Return empty cylinders promptly with protective caps replaced.
14.3.4.2 Flammable Solvents
Store in well-ventilated designated areas.
Flash point: the temperature at which a liquid gives off vapor sufficient to form an ignitible
mixture with the air near the surface of the liquid or within the vessel used.
Ignition temperature: the minimum temperature required to initiate or cause self-sustained
combustion independently of the heating or heated element.
Explosive or flammable limits: for most flammable liquids, gases, and solids there is a
minimum concentration of vapor in air or oxygen below which propagation of flame does
not occur on contact with a source of ignition. There is also a maximum proportion of
vapor or gas in air above which propagation of flame does not occur. These limit mixtures of
vapor or gas with air, which if ignited will just propagate flame, are known as the "lower and
higher explosive or flammable limits."
Explosive range: the difference between the lower and higher explosive or flammable limits,
expressed in terms of percentage of vapor or gas in air by volume.
Vapor density: the relative density of the vapor as compared with air.
14-12
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Underwriter's Laboratories classification: a standard classification for grading the relative
hazard of the various flammable liquids. This classification is based on the following scale:
Ether class 100
Gasoline class 90 to 100
Alcohol (ethyl) class 60 to 70
Kerosene class 30 to 40
Paraffin oil class 10 to 20
Extinguishing agents that are appropriate for each of the four classes of fires are required.
14.3.5 Chemical Hazards
14.3.5.1 Acids and Alkalies
Some of the most hazardous chemicals are the strong or mineral acids such as hydrochloric,
hydrofluoric, sulfuric, and nitric.
Organic acids are less hazardous because of their comparatively low ionization potentials;
however, such acids as phenol (carbolic acid), hydrocyanic, and oxalic are extremely
hazardous because of their toxic properties.
Classification of acids is according to mineral or organic composition. Acids should be
stored together, except that perchloric acid should not be placed next to glacial acetic acid.
Picric acid should be stored separately.
14.3.5.2 Oxidizing Materials
Oxidizing agents, in contact with organic matter, can cause explosions and fire. They are
exothermic and decompose rapidly, liberating oxygen, which reacts with organic com-
pounds.
Typical hazardous oxidizing agents are-
Chlorine dioxide
Sodium chlorate; chlorates
Potassium chromate
Chromium trioxide
Perchloric acid; perchlorates
14.3.5.3 Explosive Power
Many chemicals are explosive or form compounds that are explosive and should be treated
accordingly.
A few of the more common examples of this class of hazardous materials are-
Acetylides
Silver fulminate
14-13
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Peroxides
Peracetic acid
Nitroglycerine
Picric acid
Chlorine and ethylene
Sodium metal
Calcium carbide
14.3.5.4 Toxicity
Laboratory chemicals improperly stored or handled can cause injury to personnel by virtue
of their toxicity.
There are four types of exposure to chemicals:
a. Contact with the skin and eyes
b. Inhalation
c. Swallowing
d. Injection
Special classes of toxic agents:
a. Carcinogens-laboratory operators should recheck the OSHA 1974 regulations on
carcinogens.
b. Mercury—complete cleaning of spills is essential for compliance with OSHA limits.
14.3.6 Precautionary Measures
14.3.6.1 Clothing and Personal Protective Equipment
Chemical laboratories should have special protective clothing and equipment readily
available for emergency use and for secondary protection of personnel working with
hazardous materials.
Equipment should be provided for adequate—
a. Eye protection
b. Body protection
c. Respiratory protection
d. Foot protection
e. Hand protection
14-14
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14.3.6.2 Bodily Injury
Burns, eye injuries, and poisoning are the injuries with which laboratory people must be
most concerned.
First emphasis in the laboratory should be on preventing accidents. This means observing all
recognized safe practices using necessary personal protective equipment and exercising
proper control over poisonous substances at the source of exposure.
So that a physician can be summoned promptly, every laboratory should post the names,
telephone numbers, and addresses of doctors to be called in an emergency requiring medical
care.
A consulting physician should specify the type and extent of first aid materials required for
the laboratory.
14.4 Report of Unsafe or Unhealthful Condition
In EPA a procedure has been established for reporting an unsafe or unhealthful condition by
the employee or supervisor. The procedure also provides for communication between the
employees; supervisors; safety official; head of the unit; and, in the case of an unresolved
report, the Department of Labor.
A sample of an unsafe or unhealthful condition reporting form is shown in figure 14-2, and
a sample notice of an unhealthful or unsafe condition is shown in figure 14-3.
14.5 References
1. Public Law 91-596, Occupational Safety and Health Act of 1970 (Dec. 29, 1970).
2. Occupational Safety and Health Manual, U.S. EPA (Jan. 8, 1976).
3. Safety Management Manual, U.S. EPA, TN 1440.1 (Dec. 4, 1972).
14-15
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RNN1440.011
REPORT OF UNHEALTHFUL OR UNSAFE CONDITION
BRIEF DESCRIPTION OF UNHEALTHFUL OR UNSAFE CONDITION
OCCUPATIONAL SAFETY AND HEALTH STANDARD VIOLATEO (If LOCATION (Include organization. Facility and Building/
ACYlON TAKEN BY SUPERVISOR
•IONATURI
EMPLOYING ORGANIZATION
DATE
EPA Perm 14404
-------
NOTICE OF UNHEALTHFUL OR
TO (Name of Offtcer-tn-Chargt of Reporting Unit)
REPORTING UNIT
An inipection conducted by me or
following violationd) of EPA Occ
with the Occupational Safety and t
ITEM
MO.
STANDARD, REGULATION
OR SECTION VIOLATED
UNSAFE WORKING CONDITION
FROM (Name of Inspector)
Occupational Health and Safety Office
Washington. IXC 20460
upational Health and Safety Standards. These Standards have been adopted in compliance
he Health Act of 1970, PL 91-956, Section 19.
DESCRIPTION OF
VIOLATION
LOCATION OF
VIOLATION
NO. OF WORKING DAVS
BY WHICH VIOLATION
MUST BE CORRECTED
AND DATE
Subpart 0 of 29CFR 1960.33, Safety and Health Provisions for Federal Employees, requires that a copy of this Notice shall
be posted immediately in a prominent ptace at or near each place that the violation(s) referred to in the Notice occurred. The
Notice must remained posted until all violations cited therein are corrected, or for three (3) working* days, whichever period is
longer. A copy of this Notice shall be sent to the Health and Safety Committee of the establishment or Reporting Unit and to
any person(s) who made a report of the unhealthful or unsafe condition which precipitated this inspection pursuant to the
provisions of 29CFR 1960.31.
Subpart D of 29CFR 1960.34 requires the Officer-m-Charge of the Reporting Unit to immediately submit an abatement plan
to the Designated Agency Safety and Health Official, if, in his judgment, the correction of the violation will not be possible
within thirty (30) working days*. Such plan shall contain an explanation of the circumstances of the delay in abatement; a
proposed timetable for the abatement, and a summary of steps being taken in the interim to protect employees. A copy of
the plan shall be sent to the Health and Safety Committee of the establishment or Reporting Unit and to any person(s) who
made a report of the unhealthful or unsafe condition which precipitated this inspection pursuant to the provisions of 29CFR
1960.31.
"Under the Occupational Safety and Health Act, the term "Working Day" means Monday through Friday but does not
inc ude Saturday, Sunday, or Federal Holidays.
SIGNATURE OF HEALTH AND SAFETY OFFICER
DATE
EPA Hq Form 14408 (8 771
Figure 14-3. Notice of unhealthful or unsafe condition.
14-17
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APPENDIX A
SUGGESTED CHECKLIST FOR THE SAFETY EVALUATION OF EPA LABORATORY AREAS
A-1
-------
Organization:
Location:
Room(s):
Date:
Building:
By:
Item Inspected
Yes
No
N.A.
Comments
Fire Prevention
I. Is fire alarm facility available? [.36(b)(7)]
2. Are all exits maintained to provide free and unob-
structed egress from all parts of building? [.36(b)(4)]
3. Are all exits free of locks or fastening devices that
could prevent free escape? l.36(b)(4)]
4. a. Is the fire detection system in working order?
b. Is the sprinkler system in working order?
c. Are fire doors in working order? [.36(d)(2>]
5. Are corridors and hallways at least 44 in wide?
6. Do all exits discharge directly to a street, yard,
court, or other open space? [.37(h)(l)]
7. Are all exits marked by proper sign and illuminated?
Are letters in sign not less than 6 in high, 3/4 in
wide? [.37(q)(8)]
8. Is access to exits marked in all cases where the exit or
the way to reach it is not immediately visible?
[.37(q)(5)]
9. Is care taken to insure that no exit signs are obscured
by decorations, furniture, or equipment? [.37(q)(3)J
1 0. Is exit access arranged so that it is not necessary to
travel toward any high hazard area to escape?
t.37(f)(5)J
Note: Adapted from a safety inspection work sheet developed by the Center for Disease Control (CDC).
The pertinent section'of the Code of Federal Regulations, title 29, part 1910 is given within brackets at the
end of each item.
A-2
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Listed below are explanations given by OSHA to questions that are not immediately clear on the safety
checklist for laboratory areas. Each explanation is numbered according to the corresponding question on
the facing page.
1. Self-explanatory.
2. Self-explanatory.
3. At no time should an exit door be locked or fastened in a manner that prevents it from being
immediately opened from the inside of the building in the event of emergency. Safety inspectors
should check all doors marked "EXIT" to insure that they can be readily opened.
4a,b. The building manager should conduct periodic tests (as recommended by the manufacturer or as
required by local code) to insure proper working order of the fire detection system and the
automatic sprinkler system.
4c. Fire doors are designed to be closed in the event of fire. Automatic fire doors normally remain
open; however, the heat produced by a fire will cause them to close. Regular fire doors are designed
to stay closed at all times, except for the passage of personnel. No fire door should ever be blocked
open as this will interfere with its function. Fire doors are typically used to enclose stairways and to
separate buildings and corridors.
5. Self-explanatory.
6. An exit should never discharge into a location that could potentially trap personnel. For example,
an exit should not discharge into a closed courtyard.
7. Self-explanatory.
8. A sign reading "EXIT" or similar designation, with an arrow indicating the direction, shall be placed
in every location where the direction of travel to reach the nearest exit is not immediately apparent.
9. Self-explanatory.
10. In designing and maintaining exit routes from a building, care should be taken to avoid routing
people through or near a high hazard area. An example of a high hazard area is a corridor or room
where flammable liquids are stored.
A-3
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Item Inspected
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
Are aisles maintained clear and unobstructed for
movement of personnel and fire-fighting equipment?
[.22(b)(l)l
Is all fire protection equipment and apparatus identi-
fied with the color red? [.144(a)(l)l
Are portable fire extinguishers maintained fully
charged and operable and kept in designated places at
all times? [.157(a)(l)J
Are fire extinguishers conspicuously located, readily
accessible, and available along normal paths of travel?
[.157(a)(2)]
Are extinguishers and locations conspicuously
marked to indicate intended usage? [.157(a)(4)J
Are extinguishers mounted so that the top is not
more than 5 ft above floor; not more than 3Vi ft if
weight equals more than 40 Ib? [.157(a)(6)l
Are all extinguishers mounted in cabinets placed so
that the instructions face outward? [.157(a)(7)l
Are extinguishers available suited to the class of fire
anticipated in each area? [.157(b)(l)l
Are extinguishers placed according to distances for
proper coverage?
Within 75 ft-class A.
Within 50 in-class B. [.157(c)J
Are extinguishers inspected, maintained, and re-
placed by spares when they are discharged or miss-
ing? [.157(d)]
Are laboratory rooms with potential fire hazards
equipped with proper extinguishers for emergency
situations? [.157(b)l
If flammable liquids are used in a laboratory, is the
mechanical ventilation sufficient to remove vapors
before they reach a hazardous concentration?
[.106(e)(2)(iii)]
Yes
No
N.A.
Comments
A-4
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11. Self-explanatory.
12. Self-explanatory.
13. Self-explanatory. During a safety inspection all fire extinguishers should be checked aginst the
requirements of this question.
14. Fire extinguishers should never be located in places where they are concealed from general view.
15. Fire extinguishers must be clearly labeled to indicate the type of fire they are capable of fighting.
The following code is used to classify types of fires:
Class A -fires in ordinary combustible materials such as wood, cloth, paper, and rubber
Class fi-fires in flammable liquids, gases, and greases
Class C-fires that involve energized electrical equipment where the electrical nonconductivity
of the extinguishing medium is of importance; when electrical equipment is deenergized,
extinguishers for class A or B fires may be safely used
Class D~fires in combustible metals such as magnesium, titanium, zirconium, sodium, and
potassium
16. Inspector should make certain that no fire extinguishers (other than wheeled-type extinguishers) are
placed on the floor.
17. Self-explanatory.
18. See explanation to question No. 15 for classification of fire hazards. Fire extinguishers (of more
than one type, if necessary) should be selected and located according to anticipated fire hazards.
19. Self-explanatory.
20. Self-explanatory.
21. In certain instances, laboratories can be fire traps. When this is the case, fire extinguishers should be
provided inside the laboratory so that occupants can fight their way out of a fire emergency. Safety
inspectors should check for escape routes to the nearest corridor when considering this question.
22. Self-explanatory.
A-5
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Item Inspected
23. Are hose outlets within easy reach of persons stand-
ing on floor and not over 6 ft from floor?
24. Do all connections on dry standpipes have a sign
reading "DRY STANDPIPE-FOR FIRE DEPART-
MENT USE ONLY"? [.158(b)(7)]
25. Are automatic sprinkler systems provided with at
least one fire department connection with at least
4-in pipe size? (.159(b)(l)]
26. Are fire department connections designated "AUTO
SPKR" or "OPEN SPKR"? [.159(b)(4)(v)J
27. Is there a water flow detection device on the sprin-
kler system that will activate the fire alarm?
28. Are fire alarm boxes readily accessible and within
normal path distance of 200 ft? [.163(b)(3)]
29. Are all fire alarm systems inspected and tested at
weekly intervals? [.163(c>]
30. Are "NO SMOKING" signs posted in prohibited
areas? [.106(d)(7)(iii)]
Yes
No
N.A.
Comments
Flammable Liquid Storage
3 1 . Are drums which contain flammable liquids con-
structed of noncombustible materials?
32. Are storage drums vented? [.106(b)(2)(iv)l
33. Are flammable liquids stored in proper containers?
34. Are safety cans and portable containers of flammable
liquids painted red with yellow band or name of con-
tents? [.144(a)(l)(ii)J
35. Are storage cabinets being used for storing flammable
liquids? [.106(d)(3)l
36. Are storage cabinets labeled "FLAMMABLE-KEEP
FIRE AWAY"? (.106(d,)(3)(ii)]
A-6
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23. This question applies to buildings equipped with water and hose standpipe systems. Hose outlets are
normally located in stairways or in the corridor immediately outside the stairway.
24. Self-explanatory. The building manager should be able to answer this question.
25. Fire department connections to automatic sprinkler systems are normally located on the outside of
the building being protected. The size of the connection is usually embossed on a metal plate or on
the connection itself.
26. The designations "AUTO-SPKR" or "OPEN-SPKR" are found embossed on a plate at the fire
department connection. Failure to display one of the designations is a violation of OSHA.
27. This question is self-explanatory. The building manager should know if the facility is equipped with
a sprinkler waterflow detection device.
28. Self-explanatory.
29. This procedure should be part of a regular maintenance program.
30. The laboratory supervisor shall designate areas where smoking is prohibited and "NO SMOKING"
signs shall be placed in these areas.
31. Flammable liquid storage drums should be constructed of steel or some other noncombustible
material.
32. Self-explanatory.
33. Flammable liquids may be safely stored in glass containers if the total capacity of the container is 1
gal or less. Larger quantities of flammable liquids should be stored in safety cans approved by a
recognized testing laboratory.
34. Self-explanatory.
35. Self-explanatory.
36. Self-explanatory.
A-7
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Item Inspected
37. Is the storage area provided with either a gravity or
mechanical exhaust ventilation system?
M06(d)(4)(iv)]
38. Are extinguishers available where flammable or com-
bustible liquids are stored? [.106(d)(7>]
39. Is at least one portable fire extinguisher of rating not
less than 12-B units located outside of, but not more
than 10 ft from, door opening into room used for
storage? [.106(d)(7)(i)(a)]
40. Is at least one portable fire extinguisher with rating
not less than 12-B units located not less than 10 ft
nor more than 25 ft from flammable liquid storage
located outside of a storage room but inside of a
building? [.106(d)(7)(i)(b)]
41. Are "NO SMOKING" signs posted in the flammable
or combustible liquid storage areas? [.106(d)(7)(iii)]
Yes
*
No
N.A.
Comments
Electrical Hazards
42. Are all new electrical installations and all replace-
ments, modifications, or repairs made and being
maintained in accordance with the National Electri-
cal Code? [.309]
43. Does the interior wiring system have a grounded con-
ductor? i.e., three-wire system? [.309-NEC(200-2)J
44. Do all electrical appliances have Underwriter's Labo-
ratories Inc. approval, or that of some other na-
tionally recognized testing laboratory?
[.309-NEC(90-8>]
45. Are the cords of all electrical equipment in good
condition, not frayed or spliced, etc.?
[.309-NEC(400-5)J
46. Are cords used properly (not run under rugs)?
[.309-NEC(400-4)J
47. Is there only one plug-in per socket outlet; i.e., no
multiple plug-ins to one socket? f.309-NEC(240-2)]
48. Are the lighting levels such that good illumination is
provided in all walking, working, and service areas to
insure safety? [.309-NEC(110-16(e))]
A-8
-------
37. The building manager or engineering department should know the answer to this question.
38. Self-explanatory. Fire extinguishers approved for fighting class B fires should be available in
flammable liquids storage areas.
39. This question is self-explanatory but should be reread several times for full comprehension. Fire
extinguisher ratings are often stamped on the title plate along with other specifications. Note: This
question is to be used only when inspecting flammable-liquid storage rooms.
40. This question is self-explanatory, but should be reread several times for full comprehension. Note:
This question is to be used only when inspecting flammable liquid storage outside of a special desig-
nated room, but inside of a building.
41. Self-explanatory.
42. Prior to adding any new electrical facilities to a laboratory, it is the responsibility of the laboratory
supervisor or building manager to check with the electrical contractor to insure that the National
Electrical Code is observed. This rule is retroactive to Mar. 15, 1972.
43. All convenience outlets shall be of the three-pronged type. These outlets should be checked with a
grounding tester to insure proper wiring.
44. Approved appliances will bear the label of a recognized testing laboratory.
45. Self-explanatory.
46. Self-explanatory.
47. Multiple outlet (cube) adaptors or extension cords with multiple outlet ends should not be used to
plug more than one appliance into a single socket.
48. Self-explanatory.
A-9
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Item Inspected
49. Are circuit breaker panels and cutoff switches
located so as to be readily accessible?
[.309-NEC(240-25(b)) and (1926.400(e))]
50. Are all circuit breaker switches marked or labeled?
[.309-NEC(240-25(e)>]
Yes
No
N.A.
Comments
A isles, Exits, Floors, and Stairways
51. Are all floors kept clean and dry? [.22(a)(2)J
52. Are all floors, areas, and passageways free from pro-
truding nails, splinters, holes, and loose boards?
[.22(a)(3)J
53. Are aisles and passageways clear of all obstructions
and in good repair? [.22(b)(l)]
54. Are aisles and passageways wide enough to operate
equipment safely? [.22(b)(l)]
55. Is floor loading within posted maximum limits?
l.22(d)]
56. Does every floor opening having standard guarded
railings? [.23(a)(l)]
57. Do fixed stairs make an angle to the horizontal be-
tween 30° to 50°? [.24(e)J
58. Are all opensided floors, platforms, ramps, 4 ft above
adjacent floor or ground, guarded with railing?
t.23(c)(l)]
A-10
-------
49. Circuit breaker cabinets should never be locked.
50. Each circuit breaker switch shall be labeled as to the lights, outlets, or appliances it controls.
51. Self-explanatory.
52. Self-explanatory.
53. Self-explanatory.
54. Self-explanatory.
55. Safe floor loadings must be posted and observed in all buildings and on all floors.
56. Specifications for standard railing: A standard railing shall consist of a top rail, intermediate rail,
and posts. It shall have a vertical height of 42 in nominal from upper surface top rail to floor,
platform, runway, or ramp level. The top rail shall be smooth surfaced throughout its length. The
intermediate rail shall be approximately halfway between the top rail and the floor, platform,
runway, or ramp.
Specifications for standard railing to be used on stairways: A stair railing shall be of construction
similar to a standard railing (for a floor opening), but the vertical height shall not be more than 34
in nor less than 30 in from upper surface of top rail to surface of tread in line with face of visor at
forward edge of tread.
57. All fixed stairways must rise at an angle within the limits shown:
ACCEPTABLE SAFE ZONE
_L
50°
30° HORIZONTAL
58. Self-explanatory.
A-ll
-------
Item Inspected
59. Are flights of stairs having four or more risers
equipped with railings or handrails? [.23(d)(l )]
60. Are stair treads reasonably uniform and slip resist-
ant? f.24(f)]
61. Are all places kept clean and orderly and in a sanitary
condition? [.141(a)(lXi)]
62. Are trash receptacles of approved use? [.141(a)(3)]
63. Is every enclosed work place so constructed and
maintained to prevent entrance and harborage of
rodents, insects, and vermin? [.141(a)(4)J
64. Is drinking water potable and available within 200 ft
of location at which employees work?
65. Are toilet facilities adequate for both sexes and in
accordance with regulations listed? [.141(c)(l)]
66. Are toilet rooms constructed so that each water
closet occupies a separate compartment equipped
with door, latch, and clothes hanger? {.141(c)(2)(i)]
67. Does door to toilet room have a self-closing device,
and is entrance screened so interior is not visible
from outside? [.141(e)(2)(iii)]
68. Does every water closet have a hinged open-front seat
made of nonabsorbent material? [.141(c)(3)(ii))
69. Are suitable washing facilities available and main-
tained in a sanitary condition? [.141(d)(l)]
70. If employees are allowed to lunch on the premises, is
an adequate space provided for that purpose?
7 1 . Are adequate waste disposal containers provided?
72. Are change rooms provided for each sex where it is
necessary to change clothes? [.141(e)(l)]
73. Are change rooms provided with separate storage
facilities for street clothes and protective clothing?
74. Are noise levels acceptable? [.95]
Yes
No
N.A.
Comments
A-12
-------
59. Self-explanatory.
60. Stair treads should be free of cracks and other uneven areas.
61. Self-explanatory.
62. Trash receptacles used for garbage disposal should be equipped with tight-fitting lids.
63. Self-explanatory.
64. Self-explanatory.
65. Self-explanatory.
66. Self-explanatory.
67. Self-explanatory.
68. Self-explanatory.
69. Suitable washing facilities consist of a lavatory with hot and cold water, a suitable cleansing agent,
and individual hand towels.
70. Self-explanatory. Employees are not permitted to eat in any laboratory that uses agents that are
dangerous to health if ingested. It is the responsibility of the laboratory supervisor to enforce this
rule.
71. Self-explanatory.
72. Self-explanatory.
73. Self-explanatory.
74. If any employees are exposed to noise levels greater than 90 dB at any time during the day, a safety
expert should be consulted to determine if corrective measures need to be taken.
A-13
-------
Item Inspected
75. Is necessary protective equipment provided, used, and
maintained in a sanitary, safe, and reliable condition?
[.132(a)J
76. Are eye protectors provided where machines or
operations present the hazard of flying objects, glare,
liquids, radiation? [.133(a)J
77. Are sufficient washing facilities (including eye washes
and deluge showers) available for all persons required
to handle liquids that may burn, irritate, etc.?
I.lSl(c)]
78. Are employees in the area exposed to air contami-
nants only in accordance with proper limits?
79. Is a respiratory protection program used where
needed? [.134(a)(2)l
80. Are written standard operating procedures governing
the selection and use of respirators established?
8 1 . Has the user of the respirator been instructed and
trained in the proper use and limitations of the respi-
rator? [.134(b)(3)l
82. Are respirators regularly cleaned, disinfected, in-
spected, and stored in a convenient, clean, and sani-
tary location? [.134(b)(5)l
83. Has the person assigned the task requiring a respira-
tor been determined physically able to perform the
work and use the equipment by the local physician?
[.134(b)(10)]
84. Are breathing gas containers marked in accordance
with the American National Standards identifying
contents? [.134(d)(4)]
85. Can gas mask canisters be identified by properly
worded labels and color code or atmospheric con-
taminant? [.134(g)(l)]
86. Is the compressor for supplying air to respirators
equipped with necessary safety and standby equip-
ment? [.134(d)(2)(ii)l
Yes
No
N.A.
Comments
A-14
-------
75. Protective equipment, including personal protective equipment, shall be provided, used, and
maintained in a sanitary and reliable condition wherever work-associated hazards may cause injury
or impairment in function of any part of the body through absorption, inhalation, or physical
contact. Laboratory supervisors should recognize such hazards and take the action necessary to
insure that employees use adequate personal protection to avoid injury.
76. Safety glasses or goggles may be obtained through the CDC Safety Office.
77. All laboratories using liquids that may burn or irritate must be equipped with some type of
emergency eye wash equipment.
78. If laboratory supervisors have any questions concerning safety concentrations of air contaminants,
they should contact the CDC Office of Biosafety.
79. Respirators that are applicable and suitable for the purpose intended shall be provided when such
equipment is necessary to protect the health of employees. The CDC Office of Biosafety will
provide the proper respirators for a given hazard.
80. Self-explanatory.
81. Self-explanatory.
82. Self-explanatory. Respirators should be cleaned and disinfected after each use.
83. Self-explanatory.
84. Self-explanatory.
85. Self-explanatory.
86. Compressors for breathing air should be equipped with receivers of sufficient capacity to enable the
respirator wearer to escape from a contaminated atmosphere in the event of compressor failure;
alarms to indicate compressor failure and overheating shall be installed in the system.
A-15
-------
Item Inspected
87. Is each employee who works in an area where radio-
active material is used furnished and wearing film
badge? [.96(d)(2)l
88. Is radiation exposure of individuals to the body
limited to 1% rems per calendar quarter? [.96(o)j
89. Is each radiation area posted with the proper radia-
tion caution sign? [.96(e)(2)]
90. Are all radiation area employees instructed in the
safety problems, precautions, and devices to mini-
mize exposure? [.96(i)l
91. Are records maintained of the radiation exposure of
all employees who are monitored? [.96(n)]
92. Are radioactive materials stored and disposed prop-
erly? Are storage containers labeled? [.96(j,k)]
Yes
No
N.A.
Comments
98. Is each mobile hydrogen supply unit secured to pre-
vent movement? [.103(b)(l)(iv)(e)]
99. Is the hydrogen storage area permanently marked
"HYDROGEN-FLAMMABLE GAS-NO SMOKING
OR OPEN FLAMES"? [.103(b)(l)(v)J
Compressed Gases
93. Is each portable gas container for gases such as hy-
drogen legibly marked with the name of contents?
[.252(aX2)(i)(a)l
94. Are compressed gas cylinders determined in safe con-
dition by visual and other inspection required in
regulations? [.101(a)l
95. Does the compressed gas cylinder or tank have an
installed pressure relief device? [.101(c)]
96. Are all compressed gas cylinders stored and secured
so they cannot fall? [.252(a)(2)(ii)(b)]
97. Are protection caps in place on compressed gas cylin-
ders except when in use? [.2S2(a)(2)(ii)(d)l
A-16
-------
87. Self-explanatory.
88. Self-explanatory.
89. Self-explanatory.
90. Self-explanatory.
91. Self-explanatory.
92. Self-explanatory.
93. Self-explanatory.
94. Deep dents or heavy corrosion of a gas cylinder might indicate a dangerous situation.
95. Self-explanatory.
96. When either storing or moving a compressed gas cylinder, care must be taken to prevent the cylinder
from falling. OSHA requires that compressed gas cylinders either be chained or strapped in an
upright position at all times.
97. Self-explanatory.
Preface to Questions 98 through 107
A gaseous hydrogen system is one in which the hydrogen is delivered, stored, and discharged in the gaseous
form to the consumer's piping. The system includes stationary or movable containers, pressure regulators,
safety relief devices, manifolds, interconnecting piping, and controls. The system terminates at the point
where hydrogen at service pressure first enters the consumer's distribution piping. Systems having a total
hydrogen content of less than 400 ft^ are not covered by the following questions.
98. Mobile supply units should be strapped or chained to prohibit movement.
99. Self-explanatory.
A-17
-------
Item Inspected
100. Is the hydrogen system in an adequately ventilated
area? [.103(b)(2)(ii)(d)(l)]
101. Is the hydrogen system 20 ft from stored flammable
materials or oxidizing gases? [ . 1 03(b)(2Xii)(d)(2)]
102. Is it 25 ft from open flames, electrical equipment, or
other sources of ignition? [.103(b)(2)(iiXd)(3)]
103. Is it 25 ft from concentrations of people?
104. Is it 50 ft from intakes of ventilation or air-
conditioning equipment and air compressors?
[ . 1 03(b)(2Xii)(d)(5)l
105. Is it 50 ft from other flammable gas storage?
106. Is it protected from damage due to falling objects or
working activity in the area? [.103(b)(2Xii)(d)(7)]
107. Are safety relief devices arranged to discharge up-
ward and unobstructed to the open air and prevent
impingement of gas upon container, adjacent struc-
ture, or personnel? [.109(b)(0(u)(b)I
Yes
No
N.A.
Comments
Storage
108. Is storage of material stable and secure against slid-
ing, collapse, falls, or spills? [.176(b)]
109. Are storage areas kept free from accumulation of
materials that constitute hazards from tripping, fire,
explosion, or pest harborage? (.176(e)J
110. Are dangerous parts of machines or energized equip-
ment, which may injure, colored orange where ex-
posed? [.144(a)(2)J
111. Is yellow the color used for designating physical
hazards and caution in the particular environment?
[.144(a)(3)J
112. Is the color green used to designate "SAFETY" and
location of first aid equipment? t-144(a)(4)]
113. Is purple the color used to designate radiation
hazards? [.144(a)(6)l
114. Are the colors black and white used for traffic and
housekeeping markings? [.144(a)(7)l
A-18
-------
100. Self-explanatory.
101. Self-explanatory.
102. Self-explanatory.
103. Self-explanatory.
104. Self-explanatory.
105. Self-explanatory.
106. Self-explanatory.
107. Self-explanatory.
108. Self-explanatory but very important.
109. Self-explanatory.
110. Self-explanatory.
111. Self-explanatory.
112. Self-explanatory.
113. Self-explanatory.
114. Self-explanatory.
A-19
-------
Item Inspected
115.
116.
117.
118.
Are all accident prevention signs used to minimize
workplace hazards? [.145(e)(e)j
Are all exposed steam and hot water pipes within 7 ft
of floor or working platform covered with an insulat-
ing material or guarded? [.264(e)(4)(iii)]
At all loading docks employing powered industrial
trucks, are wheel chocks used to prevent trailers or
railroad cars from rolling? [.178(k)(l)]
Is medical help readily available through personnel
and first aid supplies approved by consulting physi-
cian? [.151(b)]
Yes
*
No
N.A.
Comments
A-20
-------
115. Self-explanatory.
116. Self-explanatory.
117. Self-explanatory.
118. In the absence of an infirmary, clinic, or hospital in near proximity to the workplace that is used for
treating all injured employees, a person or persons shall be adequately trained to render first aid.
First aid supplies approved by the consulting physician shall be readily available.
A-21
-------
1 REPORT NO. 2.
EPA-600A-79-OJ9
. TITLE AND SUBTITLE
Handbook' for Analytical Quality Control in Water
and Wastewater Laboratories
7. AUTHOR(S)
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
3. RECIPIENT'S ACCESSION«NO.
5. REPORT DATE
March 1979 issuing date
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
Editor: Robert L. Booth, EMSL-Cincinnati
9. PERFORMING ORGANIZATION NAME AND ADDRESS
SAME AS BELOW
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Monitoring and Support Lab. - Cinn, OH
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
15. SUPPLEMENTARY NOTES
10. PROGRAM ELEMENT NO.
1BD801
11. CONTRACT/GRANT NO.
In house
13. TYPE OF REPORT AND PERIOD COVERED
Final
14. SPONSORING AGENCY CODE
EPA/600/06
Project Officer: Robert L. Booth, EMSL-Cincinnati
16. ABSTRACT
This handbook is addressed to laboratory directors, leaders of field investigations,
and other personnel who bear responsibility for water and wastewater data. Subject
matter of the handbook is concerned primarily with quality control (QC) for chemical
and biological tests and measurements. Chapters are also included on QC aspects of
sampling, microbiology, biology, radiochemistry, and safety as they relate to water
and wastewater pollution control. Sufficient information is offered to allow the
reader to inaugurate or reinforce programs of analytical QC that emphasize early
recognition, prevention, and correction of factors leading to breakdowns in the
validity of water and wastewater pollution control data.
17.
a.
DESCRIPTORS
Quality Assurance
Quality Control
Water Analysis
Laboratory Performance
Valid Data
Analytical Quality Control
KEY WORDS AND DOCUMENT ANALYSIS
b.IDENTIFIERS/OPEN ENDED TERMS C. COS AT I Field/Group
Quality Assurance Techniq es
Analytical Measurements
Laboratory Services
Instrument Selection 07/B
Data Handling
Skills and Training
18. DISTRIBUTION STATEMENT
Release to Public
EPA Form 2220-1 (9-73)
19. SECURITY CLASS (This Report)
Unclassified
20. SECURITY CLASS (Thispage)
Unclassified
21. NO. OF PAGES
164
22. PRICE
-------
11.0-1
Section 1LO
SUPPLIERS
Ace Glass Co.
Vineland, NJ 08360
Acurex Corporation
485 Clyde Avenue
Mountain View, CA 94042
Aldrich Chemical Co.
940 W. St. Paul Avenue
Milwaukee, WI 53233
Associated Design and Manufacturing Co
814 North Henry St.
Alexandria, VA 22314
Bethlehem Apparatus Co .
Front and Depot Streets
Hellertown, PA 18055
Calgon Corporation
Pittsburg, PA 15230
Calspan Corporation
Buffalo, NY 14225
Davison Chemical Co.
Div. W.R. Grace
Charles & Baltimore Sts.
Baltimore, MD 21202
(609) 692-3333
(415) 964-3200
(414) 273-3850
(703) 549-5999
(215) 838-7034
(412) 923-2345
(716) 632-7500
(301) 727-3900
-------
ii. o-:
EM Laboratories
Elmsford, NY
Fisher Scientific Co.
711 Forbes Avenue
Pittsburg, PA 15219
Gelman Instrument Co .
600 S. Wagner Road
Ann Arbor, MI 48106
Goldsmith Division
National Lead Co.
Ill Broadway
New York, NY 10006
J.T. Baker Chemical Co.
222 Red School Lane
Phillipsburg, NJ 08865
John Manville Co.
P.O. Box 5108
Denver, CO 80217
Kontes Glass Co.
Spruce Street
Vineland, NJ 08360
Micro Filtration Systems
8 Stransbury Court
Fredericksburg, VA 22401
(914) 592-4660
(412) 562-8300
(800) 521-5120
(212) 732-9400
(201) 859-2151
(303) 979-1000
(609) 692-8500
(703) 786-7315
-------
11.0-3
Mlllipore Corporation
Bedford, MA 01730
Nalge Co.
75 Panorama Creek Dr.
Rochester, NY 14602
Nuclepore Corporation
7035 Commerce Circle
Pleasanton, CA 94566
Pall Trincor Corporation
2395 Springfield Avenue
Vauxhall, NJ 07088
Quicksilver Products, Inc.
350 Brannan
San Francisco, CA 94107
Scientific Glass & Instruments, Inc.
Houston, TX 77001
Selas Corporation of America
1957 Pioneer Road
Huntington Valley, PA 19006
Supelco, Inc .
Supelco Park
Bellefonte, PA 16823
(800) 225-1380
(716) 586-8800
(415) 462-2230
(201) 245-8400
(415) 781-1988
(713) 868-1481
(215) 672-0400
(814) 359-2784
-------
11.0-
Ultrasonics Instruments International Ltd. (516) 333-0213
200 T Shames Drive
Westburg, NY 11580
-------
APPENDIX 1
SAMPLERS AND SAMPLING PROCEDURES FOR HAZARDOUS WASTE STREAMS
by
Emil R. deVera, Bart P. Simmons,
Robert D. Stephens and David L. Storm
California Department of Health Services
Berkeley, California 94704
-------
EPA-600/2-80-018
January 1980
SAMPLERS AND SAMPLING PROCEDURES FOR HAZARDOUS WASTE STREAMS
by
Emil R. deVera, Bart P. Simmons,
Robert D. Stephens and David L. Storm
California Department of Health Services
Berkeley, California 94704
Grant No. R804692010
Project Officer
Richard A. Carnes
Solid and Hazardous Waste Research Division
Municipal Environmental Research Laboratory
Cincinnati, Ohio 45268
MUNICIPAL ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
-------
DISCLAIMER
This report has been reviewed by the Municipal Environmental
Research Laboratory, U.S. Environmental Protection Agency, and approved
for publication. Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products constitute
endorsement or recommendation for use.
ii
-------
FOREWORD
The Environmental Protection Agency was created because of increasing
public and government concern about the dangers of pollution to the
health and welfare of the American people. Noxious air, foul water,
and spoiled land are tragic testimony to the deterioration of our
natural environment. The complexity of that environment and the inter-
play between its components require a concentrated and integrated
attack on the problem.
Research and development is that necessary first step in problem solu-
tion and it involves defining the problem, measuring its impact, and
searching for solutions. The Municipal Environmental Research Laboratory
develops new and improved technology and systems for the prevention,
treatment, and management of wastewater and solid and hazardous waste
pollutant discharges from municipal and community sources, for the
preservation and treatment of public drinking water supplies, and to
minimize the adverse economic, social, health, and aesthetic effects of
pollution. This publication is one of the products of that research;
a most vital communications link between the researcher and the user
community.
This study involved the development of simple but effective sampling
equipment and procedures for collecting, handling, storing, and record-
ing of hazardous wastes. A variety of sampling devices were developed
and/or selected to meet the needs of those who regulate and manage
hazardous wastes. Of particular importance is the development of the
composite liquid waste sampler, the Coliwasa. The sampling procedures
developed were designed to provide maximum protection for the sample
collector, collection of representative samples of the bulk of wastes,
and proper containment, identification, preservation, and handling of
the samples.
Francis T. Mayo, Director
Municipal Environmental Research
Laboratory
iii
-------
ABSTRACT
The goal of this project was to develop simple but effective sampl-
ing equipment and procedures for collecting, handling, storing, and
recording samples of hazardous wastes. The report describes a variety
of sampling devices designed to meet the needs of those who regulate
and manage hazardous wastes. Particular emphasis is given to the
development of a composite liquid waste sampler, the Coliwasa. This
simple device is designed for use on liquid and semi-liquid wastes in
a variety of containers, tanks, and ponds. Devices for sampling solids
and soils are also described.
In addition to the sampling devices, the report describes pro-
cedures for development of a sampling plan, sample handling, safety
precautions, proper recordkeeping and chain of custody, and sample
containment, preservation, and transport. Also discussed are certain
limitations and potential sources of error that exist in the sampling
equipment and the procedures. The statistics of sampling are covered
briefly, and additional references in this area are given.
This report was submitted in partial fulfillment of Research Grant
No. R804692010 by the California Department of Health Services under
sponsorship of the U.S. Environmental Protection Agency.
iv
-------
CONTENTS
Foreword ill
Abstract iv
Figures vi
Tables vit
1. Introduction 1
2. Conclusions ........ 3
3. Recommendations 4
4. Samplers 5
Composite liquid waste sampler 5
Solid waste samplers 9
Soil samplers 13
Procedure for use 14
Pond sampler 20
5. Preparation for Sampling 24
6. Sampling Procedures 26
General considerations 26
Sample handling 39
Field log book 41
Chain of custody record ..... 42
Sample analysis request sheet 42
Sample delivery to the laboratory 42
Shipping of sample 45
7. Receipt and Logging of Sample 45
8. Preservation and Storage of Samples 4g
References 50
Appendices 52
A. Development of the composite liquid 52
waste sampler (Coliwasa) .
B. Parts for constructing the coliwasa 62
C. Checklist of items required in the field 63
sampling of hazardous wastes •
D. Random sampling • 67
E. Systematic errors in using the coliwasa 68
-------
FIGURES
Number
1 Composite liquid waste sampler (Coliwasa) ... 6
2 Grain sampler 10
3 Sampling trier 11
k Trowel or scoop with calibrations 13
5 Soil auger 15
6 Veihmeyer sampler 16
7 Waste pile sampler ..... 20
8 Pond sampler 21
9 Weighted bottle sampler 23
10 Example of waste manifest 28
11 Official sample label 40
12 Example of official sample seal 40
13 Example of chain of custody record 43
14 Example of hazardous waste sample
analysis request sheet 44
A-l Coliwasa, Model 1 53
A-2 Coliwasa, Model 2 55
A-3 Coliwasa, Model 3 56
A-4 Coliwasa, Model 4 58
A-5 Coliwasa, Model 5 58
vi
-------
TABLES
Number Pagj
1 Basic Parts and Costs of a Veihmeyer Soil Sampler ... 17
2 Basic Parts and Approximate Costs of a
Pond Sampler 21
3 Samplers Recommended for Various Types of Waste .... 29
4 Sample Containers and Closures Recommended
for Various Types of Waste 31
5 Sampling Points Recommended for Most Waste
Containers 32
6 Number of Samples to be Collected 34
7 Respiratory Protective Devices Recommended
for Various Hazards 35
8 Methods of Preservation for Hazardous Wastes 49
A-l Relative Volumes of Liquids in the
Two-Phase Mixture 60
A-2 Relative Volumes of Liquids in the
Three-Phase Mixture 60
APPENDIX B. Parts for Constructing the Coliwasa 62
APPENDIX C. Checklist of Items Required in the
Field Sampling of Hazardous Wastes. . . 63, 64, 65, 66
APPENDIX D. Random Sampling ..... 67
APPENDIX E-l Sample Volume 69
vii
-------
SECTION 1
INTRODUCTION
The growth in the size and complexity of industry and the imple-
mentation of air and waste pollution abatement technology has confronted
the nation with the immensely difficult problem of managing large volumes
of waste products that are often toxic, flammable, corrosive, and explo-
sive. The problem is further exacerbated by the complexity of the waste.
This difficult situation is now being addressed at many levels of govern-
ment through a variety of regulatory agencies. Solution is being sought
through a "cradle-to-grave" regulation of waste generation, transport,
reprocessing, and disposal. Significant progress toward a solution is
also being made by the private waste management industry with improved
techniques in handling, resource recovery, and disposal.
The management of hazardous wastes may be addressed primarily as a
chemical problem. With this approach, management decisions must be
founded on proper knowledge of waste chemical compositions. Defining
the information needed on waste composition to support management
decisions presents an additional complication, for such information
varies with waste type and with handling or disposal objectives. Re-
gardless of the details, the required information results from chemical
and physical testing of the waste.
Industrial waste predominantly occurs in volumes that are large
enough to preclude testing or analysis of the entire body of the waste.
Obtaining samples adequate in size for the required testing and repre-
sentative of the bulk volumes of the wastes is therefore necessary.
The obtainment of such representative samples presents special problem,
for many wastes are complex, multiphase mixtures that vary greatly in
viscosity, corrosivity, volatility, flammability, or capability to
generate toxic gases.
This study was conducted to deve?.op specialized equipment and pro-
cedures designed to handle the widest possible variety of waste sampling
situations.
The equipment and procedures that have been developed and described
in this report had their origins in the hazardous waste regulatory pro-
gram of the California Department of Health Services. Early in this
program, the necessity of reliable analytical data on waste composition
became apprent. As a result, the problem of proper sampling of hazardous
-------
wastes was addressed. Review of the wide variety of industrial wastes
produced in California revealed that liquids or liquid-sludge mixtures
accounted for the greatest volume of wastes. Most of these materials
at some point are contained and/or transported in tank (vacuum) trucks
or barrels. A primary concern, therefore, was to develop the capa-
bility for sampling these wastes.
The first prototype of a liquid waste sampler was the tube sampler,
which was designated the composite liquid waste sampler or Coliwasa.1
This sampling device, fabricated from readily available materials, was
taken to the field and tested for its usability and reliability.
The first large-scale sampling of hazardous wastes was conducted
jointly by the California Department of Health Services and the Univer-
sity of Southern California under the sponsorship of the U.S. Environ-
mental Protection Agency (EPA)2 in this sampling program, approxi-
mately 400 waste samples were collected. These samples varied greatly
in composition and in physical characteristics.
Approximately 90% of all wastes sampled were liquids or sludges and
could be sampled with the Coliwasa. The sampling program established the
utility of this sampler. In addition, however, several deficiencies and
needed improvments were demonstrated. Along with the need for liquid
sampling equipment, a need was also demonstrated for simple but effective
equipment for sampling solids, soils, and liquids in large tanks or ponds.
This early study also clearly indicated the need for development of good
safety procedures and sample handling, preservation, and custody procedures.
In November 1976, under a grant from the EPA, the California Depart-
ment of Health Services embarked on a development program to establish
recommended procedures and equipment for the sampling of hazardous wastes.
Commercially available liquid samplers were investigated, but none was
found to be adaptable to sampling hazardous wastes. Equipment development
centered on the Coliwasa, which had been conceived and initially designed
by waste management personnel of the Department. Solid, soil, and pond
sampling equipment was obtained after an extensive review of the litera-
ture and testing of available equipment for efficiency. Criteria used in
choosing candidate procedures were ready availability, reasonable cost,
simplicity of design and operation, and chemical inertness. Candidate
methods and samplers were subjected to laboratory and field tests. Lab-
oratory tests for the liquid samplers consisted of sampling water as well
as multiphase waste mixtures. The samplers were examined for leakage,
ease of use and transfer, and cross contamination. In field tests, the
samplers for liquids and solids were used on actual wastes existing in a
variety of containers, ponds, or soils.
The body of the report gives detailed discussions of recommended
samplers, preparation for sampling, sampling procedures, sample handling,
and recordkeeping. The appendices present a variety of practical support
data for the body of the report.
-------
SECTION 2
CONCLUSIONS
The present study was designed to develop simple but effective
sampling equipment for collecting represenative samples of hazardous
wastes. In addition, recommended procedures for sample collection,
handling, storage, and recording were to be developed. These primary
objectives have been met, and the resulting sampling equipment and
procedures are presented here.
The sampling equipment and procedures were designed to insure the
widest possible applicability in the sampling of various types of
hazardous wastes. The methods, however, are not intended to cover
all possible sampling situations. Professional judgment on applica-
bility must be exercised.
-------
SECTION 3
RECOMMENDATIONS
The next step in the development of standardized sampling methods
should be user verfication. Additional data on applicability, reli-
ability, and other performance characteristics need to be developed
before these recommended methods can become standard methods. This
next phase will require considerable effort by a large number of
collaborators, for the methodology described in this report is
intended to be satisfactory for essentially the entire waste-producing
industry. Significant benefit is to be gained by both industry and
environmental regulatory agencies if efficient, reliable hazardous
waste sampling methods can be established. We therefore strongly recommend
that work on this validation begin immediately.
-------
SECTION 4
SAMPLERS
Sampling of hazardous wastes requires different types of samplers.
Some of these samplers are commercially available, but the others have
to be fabricated. This section lists and describes suitable samplers.
Their uses and commercial availability as well as directions for their
use are reported. Directions for fabricating the commercially unavail-
able samplers are also outlined.
COMPOSITE LIQUID WASTE SAMPLER (COLIWASA)
The Coliwasa is the single most important hazardous waste sampler
discussed in this report. It was chosen from a number of other liquid
samplers, based on laboratory and field tests, as the most practical.
It permits the representative sampling of multiphase wastes of a wide
range of viscosity, corrosivity, volatility, and solids content. Its
simple design makes it easy to use and allow the rapid collection of
samples, thus minimizing the exposure of the sample collector to po-
tential hazards from the wastes. The sampler is not commercially
available, but it is relatively easy and inexpensive to fabricate. The
cost of fabrication is low enough that the contaminated parts may be
discarded after a single use when they cannot be easily cleaned.
The recommended model of the Coliwasa is shown in Figure 1. The
history and development of this sampler is discussed in detail in Appendix
A. The main parts of the Coliwasa consist of the sampling tube, the
closure-locking mechanism, and the closure system.
The sampling tube consists of a 1.52-m(5-ft.) by 4.13-cm(l 5/8-in.)
I.D. translucent plastic pipe, usually polyvinyl chloride (PVC) or boro-
silicate glass plumbing tube. The closure-locking mechanism consists of
a short-length, channeled aluminum bar attached to the sampler's stopper
rod by an adjustable swivel. The aluminum bar serves both as a T-handle
and lock for the sampler's closure system. When the sampler is in
the open position, the handle is place in the T-position and pushed
down against the locking block. This manipulation pushes out the
neoprene stopper and opens the sampling tube. In the close position,
the handle is rotated until one leg of the T is squarely perpendicular
against the locking block. This tightly seats the neoprene stopper
against the bottom opening of the sampling tube and positively locks
the sampler in the close position. The closure tension can be adjusted
by shortening or lengthening the stopper rod by screwing it in or out
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of the T-handle swivel. The closure system of the sampler consists of
a sharply taped neoprene stopper attached to a 0.95-cm (3/8-in.) O.D.
rod, usually PVC. The upper end of the stopper rod is connected to
the swivel of the aluminum T-handle. The sharply tapered neoprene
stopper can be fabricated according to specifications by plastic products
manufacturers at an extremely high price, or it can be made in-house by
grinding down the inexpensive stopper with a shop grinder as described in
Note 1 of Appendix B.
Two types of Coliwasa samplers are made, namely plastic or glass.
The plastic type consists of translucent plastic (usually PVC) sampling
tube. The glass Coliwasa uses borosilicate glass plumbing pipe as the
sampling tube and Teflon plastic stopper rod.
The complete list of parts for constructing the two types of Coliwasa
samplers is given in Appendix B. The suppliers and approximate costs of
the parts as well as the directions for fabricating the commercially
unavailable parts are also given.
The sampler is assembled as shown in Figure 1 and as follows:
1. Attach the swivel to the T-handle with the 3.18 cm(l% in.) long
bolt and secure with the 0.48 cm(3/16 in.) National Coarse(NC)
washer and lock nut.
2. Attach the neoprene stopper to one end of the stopper rod and
secure with the 0.95 cm(3/8 in.) washer and lock nut.
3. Install the stopper and stopper rod assembly in the sampling tube.
4. Secure the locking block sleeve on the block with glue or screws.
This block can also be fashioned by shaping a solid plastic rod
on a lathe to the required dimensions.
5. Position the locking block on top of the sampling tube such that
the sleeveless portion of the block fits inside the tube, the
sleeve sits against the top end of the tube, and the upper end of
the stopper rod slips through the center hole of the block.
6. Attach the upper end of the stopper rod to the swivel of the
T-handle.
7. Place the sampler in the close position and adjust the tension on
the stopper by screwing the T-handle in or out.
Uses
The plastic Coliwasa is used to sample most containerized liquid
wastes except wastes that contain ketones, nitrobenzene, dimethylforamide,
mesityl oxide, and tetrahydrofuran. '
-------
The glass Coliwasa is used to sample all other containerized liquid
wastes that cannot be sampled with the plastic Coliwasa except strong
alkali and hydrofluoric acid solutions.
Procedure for Use
1. Choose the plastic or glass Coliwasa for the liquid waste to be
sampled and assemble the sampler as shown in Figure 1.
2. Make sure that the sampler is clean (see Section 5).
3. Check to make sure the sampler is functioning properly. Adjust
the locking mechanism* if necessary to make sure the neoprene
rubber stopper provides a tight closure.
4. Wear necessary protective clothing and gear and observe required
sampling precautions (see Section 6).
5. Put the sampler in the open position by placing the stopper rod
handle in the T-position and pushing the rod down until the handle
sits against the sampler's locking block.
6. Slowly lower the sampler into the liquid waste. (Lower the sampler
at a rate that permits the levels of the liquid inside and outside
the sampler tube to be about the same. If the level of the liquid
in the sampler tube is lower than that outside the sampler, the
sampling rate is too fast and will result in a nonrepresentative
sample).
7. When the sampler stopper hits the bottom of the waste container,
push the sampler tube downward against the stopper to close the
sampler. Lock the sampler in the close position by turning the T
handle until it is upright and one end rests tightly on the locking
block.
8. Slowly withdraw the sampler from the waste container with one hand
while wiping the sampler tube with a disposable cloth or rag with
the other hand.
9. Carefully discharge the sample into a suitable sample container
(see Section 6) by slowly opening the sampler. This is done by
slowly pulling the lower end of the T handle away from the locking
block while the lower end of the sampler is positioned in a sample
container.
10. Cap the sample container; attach label and seal; record in field
log book; and complete sample analysis request sheet and chain of
custody record.
-------
11. Unscrew the T handle of the sampler and disengage the locking
block. Clean sampler on site (see Section 5) or store the con-
taminated parts of the sampler in a plastic storage tube for
subsequent cleaning. Store used rags in plastic bags for
subsequent disposal.
12. Deliver the sample to the laboratory for analysis (see Section 6).
SOLID WASTE SAMPLERS
A number of tools are available for sampling solid substances. The
most suitable of these for sampling hazardous solid wastes are the grain
sampler, sampling trier, and the trowel or scoop. •
Grain Sampler
The grain sampler (Figure 2) consists of two slotted telescoping
tubes, usually made of brass or stainless steel. The outer tube has a
conical, pointed tip on one end that permits the sampler to penetrate the
material being sampled. The sampler is opened and closed by rotating the
inner tube. Grain samplers are generally 61 to 100 cm (24 to 40 in.)
long by 1.27 to 2.54 cm (% to 1 in.) in diameter, and they are commercially
available at laboratory supply houses.
Uses—
The grain sampler is used for sampling powdered or granular wastes
or materials in bags, fiberdrums, sacks or similar containers. This
sampler is most useful when the solids are no greater than 0.6 cm Os in.)
in diameter.
Procedure for Use—
1. While the sampler is in the close position, insert it into the
granular or powdered material or waste being sampled from a point
near a top edge or corner, through the center, and to a point
diagonally opposite the point of entry.5
2. Rotate the inner tube of the sampler into the open position.
3. Wiggle the sampler a few times to allow materials to enter the
open slots.
4. Place the sampler in the close position and withdraw from the
material being sampled.
5. Place the sampler in a horizontal position with the slots facing
upward.
6. Rotate and slide out the outer tube from the inner tube.
-------
7. Transfer the collected sample in the inner tube into a suitable
sample container (see Section 6).
8. Collect two or more core samples at different points (see Section
6), and combine the samples in the same container.
9. Cap the sample container; attach label and seal; record in field
log book; and complete sample analysis request sheet and chain
of custody record.
10. Clean (see Section 5) or store the sampler in plastic bag for ^
subsequent cleaning.
11. Deliver the sample to the laboratory for analysis (see Section 6).
61-100 cm,
(24-40")
1.27-2.54 cm (%-!")
Figure 2• Grain sampler.
10
-------
\
61-100 cm,
(24-40")
N
\
h
1.27-2.54 cm (%-!")
Figure 3. Sampling trier.
Sampling trier
A typical sampling trier (Figure 3) is a long tube with a slot that
extends almost its entire length. The tip and edges of the tube slot are
sharpened to allow the trier to cut a core of the material to be sampled
when rotated after insertion into the material. Sampling triers are
usually made of stainless steel with wooden handles. They are about 61
to 100 cm (24 to 40 in.) long and 1.27 to 2.54 cm (% to 1 in.) in diameter.
They can be purchased readily from laboratory supply houses.
Uses—
The use of the trier is similar to that of the grain sampler dis-
cussed above. It is preferred over the grain sampler when the powdered or
11
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granular material to be sampled is moist or sticky.
In addition, the sampling trier can be used to obtain soft or loosened
soil samples up to a depth of 61 cm(24 in.) as outlined below.
Procedure for Use—
1. Insert the trier into the waste material at a 0 to 45° angle from
horizontal. This orientation minimizes the spillage of sample
from the sampler. Extraction of samples might require tilting of
the containers.
2. Rotate the trier once or twice to cut a core of material.
3. Slowly withdraw the trier, making sure that the slot is facing
upward.
4. Transfer the sample into a suitable container (see Section 6) with
the aid of a spatula and/or brush.
5. Repeat the sampling at different points (see Section 6). Two or
more times and combine the samples in the same sample container.
6. Cap the sample container; attach the label and seal; record in
field log book; and complete sample analysis request sheet and
chain of custody record.
7. Wipe the sampler clean, or store it in a plastic bag for subsequent
cleaning.
8. Deliver the sample to the laboratory for analysis (see Section 6).
Trowel or Scoop
A garden-variety trowel looks like a small shovel (Figure 4). The
blade is usually about 7 by 13 cm(3 by 5 in.) with a sharp tip. A labor-
atory scoop is similar to the trowel, but the blade is usually more curved
and has a closed upper end to permit the containment of material. Scoops
come in different sizes and makes. Stainless steel or polypropylene scoops
with 7 by 15-cm(2 3/4 by 6-in.) blades are preferred. A trowel can be
bought from hardware stores; the scoop can be bought from laboratory supply
houses.
Uses—
An ordinary zinc-plated garden trowel can be used in some cases for
sampling dry granular or powdered materials in bins or other shallow con-
tainers. The laboratory scoop, however, is a superior choice. It is
usually made of materials less subject to corrosion or chemical reactions,
thus lessening the probability of sample contamination.
12
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The trowel or scoop can also be used in collecting top surface soil
samples.
Procedure for Use—
1. At regular intervals (see Section 6), take small, equal portions of
sample from the surface or near the surface of the material to be
sampled.
2. ,Combine the samples in a suitable container (see Section 6).
3. Cap the container; attach the label and seal; record in field log
book; and complete sample analysis request sheet and chain of
custody record.
i
4. Deliver the sample to the laboratory for analysis (see Section 6).
SOIL SAMPLERS
There is a variety of soil samplers used. For taking soil core
samples, the scoop, sample trier, soil auger, and Veihmeyer sampler can be
used. These samplers are commercially available and relatively inexpensive.
Scoop or Trowel
See the preceding section on solid waste samplers for the description
of a scoop or trowel (Figure 4).
\
Figure 4. Trowel or scoop with calibrations.
13
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Uses—
The scoop is used to collect soil samples up to 8 cm(3 in.) deep.
It is simple to use, but identical mass sample units for a composite sample
are difficult to collect with this sampler. The procedure for use of the
scoop is outlined in the preceding section on solid waste samplers.
Sampling Trier
See the preceding section on solid waste samplers for the description
of a sampling trier (Figure 3).
Uses—
This sampler can be used to collect soil samples at a depth greater
than 8 cm(3 in.)- The sampling depth is determined by the hardness and
types of soil being sampled. This sampler can be difficult to use in
stony, dry, very heavy, or sandy soil. The collected sample tends to be
slightly compacted, but this method permits observation of the core sample
before removal.
Procedure for Use—
Procedure for use of the sampling trier can be found in the section
on solid waste samplers.
Soil Auger
This tool consists of a hard metal central shaft and sharpened spiral
blades (Figure 5). When the tool is rotated clockwise by its wooden
T handle, it cuts the soil as it moves forward and discharges most of the
loose soil upward. The cutting diameter is about 5 cm(2 in.). The length
is about 1 m(40 in.), with graduations every 15.2 cm(6 in.). The length
can be increased up to 2 m(80 in.). This tool can be bought from stores
and, in some cases, from laboratory supply houses.
Uses—
The auger is particularly useful in collecting soil samples at depths
greater than 8 cm(3 in.). This sampler destroys the structure of cohesive
soil and does not distinguish between samples collected near the surface
or toward the bottom. It is not recommended, therefore, when an undis-
turbed soil sample is desired.
Procedure for Use—
1. Select the sampling point (see Section 6) and remove unnecessary
rocks, twigs, and other non-soil materials.
14
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2. Install the sampler's wooden T handle in its socket.
3. Bore a hole through the middle of an aluminum pie pan large enough to
allow the blades of the auger to pass through. The pan will be used
to catch the sample brought to the surface by the auger.
4. Spot the pan against the selected sampling point.
5. Start augering through the hole in the pan until the desired sampling
depth is reached.
6. Back off the auger and transfer the sample collected in the catch pan
and the sample adhering to the auger to a suitable container (see
Section 6). Spoon out the rest of the loosened sample with a sampling
trier.
7. Repeat the sampling at different sampling points (see Section 6), and
combine the samples in the same container as in step 6.
8. Cap the sample container; attach label and seal; record in field log
book; and complete sample analysis request sheet and chain of cus-
tody record.
9. Brush off and wipe the sampler clean, or store it in a plastic bag
for subsequent cleaning.
10. Deliver the sample to the laboratory for analysis (see Section 6).
•—tt—
101.6 cm (40")
5.08 cm (2")
Figure 5. Soil auger.
15
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A. Drive hammer
n
B. Head
C. Tube
D. Point
Standard point
Constricted point
Bulge point
Special point
Point types
Puller jack and grip
Figure 6. Veihmeyer sampler
16
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Veihmeyer Soil Sampler
This sampler was developed by Professor F.J. Veihmeyer of the Univer-
sity of California in Davis.' The parts of a basic sampler and the corres-
ponding costs are given in Table 1, and the basic sampler is shown in
Figure 6.
TABLE 1. BASIC PARTS AND COSTS OF A VEIHMEYER SOIL SAMPLER
Part3 Costb
Tube, 1.5 m (5 ft.) $ 50.40
Tube, 3 m (10 ft.) 84.75
Tip, type A, general use 25.80
Drive head 29.05
Drop hammer, 6.8 kg (15 Ib.) 71.85
Puller jack and gripc 161.90
Total $ 433.75
a Only one of each part is needed. They are manufactured by
Hansen Machine Works, 334 N. 12th Street, Sacramento, CA
95815.
k Based on August 1, 1977, price list.
c Recommended for deep soil sampling.
The tube is chromium-molybdenum steel and comes in various standard
lengths from 0.91 to 4.9 m(3 to 16 ft.) and calibrated every 30.48 cm(12
in.). Longer tubes can be obtained on special order. Different points
(Figure 6) are also available for different types of soil and sampling.
Each point is shaped to penetrate specific types of soil without pushing
the soil ahead of it, thus preventing the core from compacting in the tube.
The standard point is adequate for most general sampling purposes. The
inside taper of each point is designed to keep the sample from being
sucked out of the tube as it is pulled frcm the ground. The drive head
protects the top of the tube from deforming when the tube is driven into
the ground with the drive hammer. The hammer doubles as a drive weight
and handle when pulling the sampler from the ground. When the sampler tube
cannot be pulled easily from the ground, a special puller jack and grip
17
-------
are also available. Specifications for the various parts of the Veihmeyer
sampler are given as follows:
Points Chrome-molly steel, heat-treated. Includes a
standard point for general use, a constricted
point for deep sampling in heavy clay (keeps
core from being sucked out of the tube), a
bulge point for shallow sampling in heavy clay,
and a special point for dry sand. (See Figure
6D).
Drive hammer . . Standard weight is 6.8 kg (15 lb.). (See Figure 6A)
Tubes . , Chrome-molly steel. Maximum length is 4.9 m
(16 ft.). (See Figure 6C).
Head . . Chrome-molly steel, heat-treated. (See Figure 6B).
Puller jack . . Cast aluminum frame with steel roller assembly
and handle.'
Grip . . Chrome-molly steel, heat-treated.
Uses—
The Veihmeyer sampler is recommended for core sampling of most types
of soil. It may not be applicable to sampling stony, rocky, or very wet
soil.
Procedure for Use—
1. Assemble the sampler by screwing in the tip and the drive head on
the sampling tube.
2. Insert the tapered handle (drive guide) of the drive hammer through
the drive head.
3. Place the sampler in a perpendicular position on the soil to be
sampled.
4. With the left hand holding the tube, drive the sampler into the
ground to the desired sampling depth by pounding the drive head
with the drive hammer. Do not drive the tube further than the
tip of the hammer's drive guide.
5. Record the length of the tube that penetrated the ground.
6. Remove the drive hammer and fit the keyhole-like opening on the flat
side of the hammer onto the drive head. In this position, the ham-
mer serves as a handle for the sampler.
18
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7. Rotate the sampler at least two revolutions to shear off the sample
at the bottom.
8. Lower the sampler handle (hammer) until it just clears the two ear-
like protrusions on the drive head and rotate about 90°.
9. Withdraw the sampler from the ground by pulling the handle (hammer)
upwards. When the sampler cannot be withdrawn by hand, as in deep
soil sampling, use the puller jack and grip.
10. Dislodge the hammer from the sampler; turn the sampler tube upside
down; tap the head gently against the hammer; and carefully recover
the sample from the tube. The sample should slip out easily.
11. Store the core sample, preferably, in a rigid, transparent, or trans-
lucent plastic tube when observation of soil layers is to be made.
The use of the tube will keep the sample relatively undisturbed. In
other cases, use a 1000-or 2000-ml (l«-qt. or %-gal) sample container
(see Section 6) to store the sample.
12. Collect additional core samples at different points (see Section 6).
13. Label the samples; affix the seals; record in the field log book;
complete analysis request sheet and chain of custody record; and
deliver the samples to the laboratory for analysis (see Section 6).
Waste Pile Sampler
A waste pile sampler (Figure 7) is essentially a large sampling trier.
It is commercially available, but it can be easily fabricated from sheet
metal plastic pipe. A polyvinyl chloride plumbing pipe 1.52 m(5 ft ) long
by 3.2 cm(l% in.) I.D. by 0.32 cm(1/8 in.) wall thickness is adequate. The
pipe is sawed lengthwise (about 60/40 split) until the last 10 cm(4 in.)
The narrower piece is sawn off and hence forms a slot in the pipe. The
edges of the slot and the tip of the pipe are sharpened to permit the
sampler to cut into the waste material being sampled. The unsplit length
of the pipe serves as the handle. The plastic pipe can be purchased from
hardware stores.
Uses—
The waste pile sampler is used for sampling wastes in large heaps with
cross-sectional diameters greater than 1 m(39. 4 in.). It can also be used
for sampling granular or powdered wastes or materials in large bins, barges,
or silos where the grain sampler or sampling trier is not long enough.
This sampler does not collect representative samples when the diameters of
the solid particles are greater than half the diameter of the tube.
Procedure for Use—
19
-------
1. Insert the sampler into the waste material being sampled at 0 to 45°
from horizontal.
2. Rotate the sampler two or three times in order to cut a core of the
material.
3. Slowly withdraw the sampler, making sure that the slot is facing
upward.
4. Transfer the sample into a suitable container (see Section 6) with
the aid of a spatula and/or brush.
5. Repeat the sampling at different sampling points (see Section 6) two
or more times and combine the samples in the same sample container in
step 4.
6. Cap the container; attach label and seal; record in field log book;
and complete sample analysis request sheet and chain of custody
record.
7. Wipe the sampler clean or store it in a plastic bag for subsequent
cleaning.
8. Deliver the sample to the laboratory for analysis (see Section 6).
i
122-183 cm
(48-72")
5.08-7.62 cm
(2-3") I.D.
Figure 7. Waste pile sampler.
Pond Sampler
The pond sampler (Figure 8) consists of an adjustable clamp attached
to the end of a two or three piece telescoping aluminum tube that serves
as the handle. The clamp is used to secure a sampling beaker. The sampler
is not commercially available, but it is easily and inexpensively fabri-
20
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cated. The tubes can be readily purchased from most hardware or swimming
pool supply stores. The adjustable clamp and sampling beaker can be
obtained from most laboratory supply houses. The materials required to
fabricate the sampler are given in Table 2.
Bolt hole
Beaker, polyprop-
ylene, 250 ml
(1 qt)
Pole, telescoping, aluminum, heavy
duty, 250-450 cm (96-180")
Figure 8. Pond sampler.
TABLE 2. BASIC PARTS AND APPROXIMATE COSTS OF A POND SAMPLER
Quantity
Item
Supplier
Approximate
Cost
Clamp, adjustable, 6.4 to
8.9 cm(2% to 3% in.) for
250-to 600-ml(% to 1%-pt.)
beakers
Tube, aluminum, heavy duty,
telescoping extends 2.5 to
4.5 m(8 to 15 ft.) with
joint cam locking mechanism.
Pole diameters 2.54 cm(l in.)
I.D. and 3.18 011(1% in.) I.D.
Laboratory supply $ 7.00
houses
Olympic Swimming 16.24
Pool Co. 807 Buena
Vista Street, Alameda,
Calif. 94501 or other
general swimming pool
supply houses.
1
4
4
Beaker, polypropylene,
Bolts, 6.35 by 0.64 cm(2k, by
k in.) NC
Nuts, 0.64 cmft in.) NC
Total
Laboratory supply
houses .
Hardware stores
Hardware stores
1.00
.20
.20
$24.64
21
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Uses—
The pond sampler is used to collect liquid waste samples from disposal
ponds, pits, lagoons, and similar reservoirs. Grab samples can be obtained
at distances as far as 3.5 m(ll% ft ) from the edge of the ponds. The
tubular aluminum handle may bow when sampling very viscous liquids if
sampling is not done slowly.
Procedure for Use—
1. Assemble the pond sampler. Make sure that the sampling beaker and
the bolts and nuts that secure the clamp to the pole are tightened
properly.
2. With proper protective garment and gear (see Section 6), take grab
samples from the pond at different distances and depths (see Section
6).
3. Combine the samples in one suitable container (see Section 6).
4. Cap the container; label and affix the seal; record in field log
book; and complete sample analysis request sheet and chain of
custody record.
5. Dismantle the sampler; wipe the parts with terry towels or rags and
store them in plastic bags for subsequent cleaning. Store used
towels or rags in garbage bags for subsequent disposal.
6. Deliver the sample to the laboratory for analysis (see Section 6).
Weighted Bottle Sampler
This sampler (Figure 9) consists of a bottle, usually glass, a weight
sinker, a bottle stopper, and a line that is used to open the bottle and to
lower and raise the sampler during sampling. There are a few variations
of this sampler, as illustrated in the ASTM Methods D 2708 and E 3009.
The ASTM sampler, which uses a metallic bottle basket that also serves as
weight sinker, is preferred. The weighted bottle sampler can either be
fabricated or purchased.
Uses—
The weighted bottle sampler can be used to sample liquids in storage
tanks, wells, sumps, or other containers that cannot be adequately sampled
with a Coliwasa. The sampler cannot be used to collect liquids that are
incompatible or that react chemically with the weight sinker and line.
Procedure for use—
1. Assemble the weighted bottle sampler as shown in Figure 9.
22
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Using protective sampling equipment, in turn, lower the sampler to
proper depths to collect the following samples:
a) upper sample - middle of upper third of tank contents.
b) middle sample - middle of tank contents.
c) lower sample - near bottom of tank contents.
Pull out the bottle stopper with a sharp jerk of the sampler line.
Allow the bottle to fill completely, as evidence by the cessation of
air bubbles.
Raise the sampler and retrieve and cap the bottle. Wipe off the out-
side of the bottle with a terry towel or rag. The bottle can serve
as the sample container.
Label each of the three samples collected; affix seal^ fill out sample
analysis request sheet and chain of custody record; record in the
field log book.
Clean onsite or store contaminated sampler in a plastic bag for sub-
sequent cleaning.
Deliver the sample to the laboratory for analysis (see Section 6).
Instruct the laboratory to perform analysis on each sample or a
composite of the samples.
Washer
Pin
Eyelet
Nut
1000-ml (1-quart) weighted
bottle catcher
Figure 9. Weighted bottle sampler.
23
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SECTION 5
PREPARATION FOR SAMPLING
GENERAL CONSIDERATIONS
Adequate preparation for sampling is necessary to perform proper sam-
pling of any hazardous waste. A checklist of items required for field
sampling helps to ensure proper preparation. Such a checklist is given in
Appendix C. The appendix lists the minimal equipment, accessories, and
supplies necessary to sample any type of solid or liquid waste. When the
type of waste to be sampled is known beforehand, the list can be narrowed
down to the actual pieces of equipment to be used.
When sample analyses are to be performed in the field, such as for pH,
flammability, or explosivity, then the necessary apparatus for such tests
should also be included in the preparation for sampling.
CLEANING AND STORAGE OF SAMPLER
All samplers must be clean before use. Used samplers must be washed
with warm detergent solution (i.e., Liquinox or Alconox), rinsed several
times with tap water, rinsed with distilled water, drained of excess water,
and air dried or dried with a stream of warm, dry air or wiped dry. For
samplers that have been used to sample petroleum products and oil residues,
it may be necessary first to wipe the samplers with absorbent cloth to
eliminate the residues. The equipment is then rinsed with an organic sol-
vent such as petroleum naphtha or trichloroethane, followed by washing with
the detergent solution and rinsing with water. A necessary piece of equip-
ment for cleaning the tube of a Coliwasa is a bottle brush that fits tight-
ly the inside diameter of the tube. The brush is connected to a rod of
sufficient length to allow for reaching the entire length of the sampler
tube. Using this ramrod and fiber-reinforced paper towels, the Coliwasa
tube may be quickly cleaned.
Improper cleaning of sampling equipment will cause cross contamination
of samples. Such contamination is of particular importance in samples
taken for legal or regulatory purposes. Also, contamination becomes im-
portant when sampling wastes from different production sources within the
same time frame. A detailed study of cross contamination as a function of
cleaning procedures has not been carried out. A recommended policy is that
if samples are to be taken for legal or regulatory purposes, or if analysis
is to be performed on samples expected to contain low-level (low ppm range)
24
-------
concentrations of hazardous components, that a fresh, unused sampler be
used. The Coliwasa in particular was designed to be semidisposable. Parts
of the device that become contaminated during sampling (i.e., the tube, the
stopper rod, and the stopper mechanism) may be discarded at little expense.
In addition, or these parts may later be disassembled, secured, and returned
to the laboratory for thorough decontamination and reused.
If the cleaning process has the potential for producing toxic fumes,
ensure adequate ventilation. If the washings are hazardous, store them
in closed waste containers and dispose of them properly in approved dis-
posal sites. Locations of these sites close to one's area may be obtained
by calling the agency in the State responsible for the regulation of hazard-
ous wastes. Store the clean samplers in a clean and protected area. Poly-
ethylene plastic tubes or bags are usually adequate for storing the samplers.
25
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SECTION 6
SAMPLING PROCEDURES
PURPOSES AND GENERAL CONSIDERATIONS
Sampling of hazardous wastes is conducted for different purposes. In
most instances, it is performed to determine compliance with existing regu-
lations promulgated by the different regulatory agencies. In some cases,
it is conducted to obtain data for purposes of classifying, treating, reco-
vering, recycling, or determining compatibility characteristics of the
wastes. Sampling is also conducted as an important part of research activ-
ities.
In general, sampling of hazardous wastes requires the collection of
adequate sized, representative samples of the body of wastes. Sampling
situations vary widely and therefore no universal sampling procedure can be
recommended. Rather, several procedures are outlined for sampling different
types of wastes in various states and containers.
These procedures require a plan of action to maximize safety of sam-
pling personnel, minimize sampling time and cost, reduce errors in sampling,
and protect the integrity of the samples after sampling. The following
steps are essential in this plan of action:
1. Research background information about the waste.
2. Determine what should be sampled.
3. Select the proper sampler.
4. Select the proper sample container and closure.
5. Design an adequate sampling plan that includes the following:
a) Choice of the proper sampling point.
b) Determination of the number of samples to be taken.
c) Determination of the volumes of samples to be taken.
6. Observe proper sampling precautions.
7. Handle samples properly.
8. Identify samples and protect them from tampering.
9. Record all sample information in a field notebook.
10. Fill out chain of custody record.
11. Fill out the sample analysis request sheet.
12. Deliver or ship the samples to the laboratory for analysis.
26
-------
BACKGROUND INFORMATION ABOUT THE WASTE
Accurate background information about the waste to be sampled is very
important in planning any sampling activity. The information is used to
determine the types of protective sampling equipment to be used, sampling
precautions to be observed, as well as the types of samplers, sample con-
tainers, container closures, and preservatives (when needed) required.
Generally, the information about the waste determines the kind of sampling
scheme to be used.
Most often, the information about the waste is incomplete. In these
instances, as much information as possible must be obtained by examining
any documentation pertaining to the wastes, such as the hauler's manifest
(Figure 10). When documentation is not available, information may be
obtained from the generator, hauler, disposer, or processor. The informa-
tion obtained is checked for hazardous properties against references such
as the Dangerous Properties of Industrial Materials,10 the Merck Index,3
the Condensed Chemical Dictionary,H Toxic and Hazardous Industrial Chemi-
cals Safety Manual for Handling and Disposal with Toxicity and Hazardous
Data,l^ or other chemical references.
SELECTION OF SAMPLER
Hazardous wastes are usually complex, multiphase mixtures of liquids,
semisolids, sludges, or solids.1 The liquid and semisolid mixtures vary
greatly in viscosity, corrosivity, volatility, explosivity, and flamma-
bility. The solid wastes can range from powders to granules to big lumps.
The wastes are contained' in drums, barrels, sacks, bins, vacuum trucks,
ponds, and other containers., No single type of sampler can therefore be
used to collect representative samples of all types of wastes. Table 3
lists most waste types and the corresponding recommended samplers to be
used.
SELECTION OF SAMPLE CONTAINER, CONTAINER CLOSURE, AND CLOSURE LINING
Containers
The most important factors to consider when choosing containers for
hazardous waste samples are compatibility, resistance to breakage, and
volume. Containers must not melt, rupture, or leak as a result of chemical
reactions with constituents of waste samples. Thus, it is important to
have some idea of the components of the waste. The containers must have
adequate wall thickness to withstand handling during sample collection and
transport to the laboratory. Containers with wide mouths are desirable to
facilitate transfer of samples from samplers to containers. Also, the
containers must be large enough to contain the required volume of samplers
or the entire volume of a sample contained in samplers.
Plastic and glass containers are generally used for collection and
storage of hazardous waste samples. Commonly available plastic containers
27
-------
are made of high-density or linear polyethylene (LPE), conventional poly-
ethylene, polypropylene, polycarbonate, teflon FEP (fluorinated ethylene
propylene), polyvinyl chloride (PVC), or polymethylpentene. Teflon FEP is
the most inert plastic, but LPE offers the best combination of chemical
resistance and low cost.
Glass containers are relatively inert to most chemicals and can be
used to collect and store almost all hazardous waste samples except those
that contain strong alkali and hydrofluoric acid. Soda glass bottles are
the cheapest and most readily available. Borosilicate such as Pyrex and
Corex glass containers are also commercially available, but they are ex-
pensive and not always readily obtainable. Glass containers are breakable
and much heavier than plastic containers.
CALIFORNIA LIQUID WASTE HAULER RECORD
STATE WATCH RESOURCES CONTROL BOARD
STATS DEPARTMENT OF HEALTH
009-Q00928
mODUCCfl Of WAffTf (MM M fHM br pradwv> ]
Name 1
(**•••* •* TTM) COM N«.
Fkk us Addreac
(MUH«««| (•T»B«T} (CITY)
TatapMone Numfaw. i . _! mll , f> 0 or Canute! No
Or« H*.
wefteiMater treatment, pkkling bath, petroleum refining)
DMcaifTJOW OF WASTE MM* b* fitted by pradWMrlJ
Cheek type of «W«MM:
1 Q Acid solution 6. G Tetreothyl lead (Judge t D Contaminated Mil and atnd
J Q Alkaline tolution 7 D Chemical toilet wwn* 2 Q Cannery warn
3 O Penkldea B. D Tank bottom Mdlfn«ni 3 Q L»t«n wMf«
4 Q P»tot •ludot 9 D Oil 4 Q Mud and wM*r
« DSolvwtt 10 D Drilling «nud B Q Brlr*
G Oth« (SP«ifV) 1 |] 1
ComporMnn. ***" ™"
(ExamplM Hvdroctiforic »CK|, tHrt*. eaufffe tod*, Cone«ntr«tfon.
pn«nolic«. *ol«*mt (list). m«t»lf (Mn), Upow Lowor % ppm
orffjntea Ul«), cv*nld«)
1
a
4.
B
0.
M*ivdou« PropwtiM ol W«t«
DM O non« O to^te D fl«mnMbt« O corrocfev D mMo^w
Bulk Volume Q t*i O ton* C «2 »•* J O otftw ,_
rAn*.iMr.- n ^«.«« 1 1 »»AM n H,^ n -.K—
•hyiieiri Sun G ioi(d G liquid D tfud** G otft*r_Ts.__p_T.
^B«eW Huu4l.ftf In^ni^l^M )!# any^-
Th« w#t« i» d«crlbod to th« b«M of mv •bMltv «nd n w*> dMivorM to • iteonwtf liquid WWM hwl«r (If
•ppliertMl
I eonrfv (or doctwo) und«r ptnotty of portury
ihM tM IwvfOtn* to tru« and comet.
•MMATVN* •» «W«HOllW«0 *•••« AN* TCTVM
HAULER OF WASTE (MM be (Htod by hniM |
LLU
coe> na.
own
Plrb 1.1 p Ttm* Dom
*' a
•*«• Liquid W«t. M*ulM'i RtfiMmten No (tf »pplicM>l«) . , ._ 9.. .. .
VatileM: D vacuum truck _ b*rf *lt. D flMlMri, Doth*/....
(«*«ei^»)
facility nomod baloov «od «*M •ccoptod
1 conifv (or d*cl*r«) undor p*n«lty of porfury
that MM forMDlrw ta tru« and eorract.
«MM«rwN« «* *VT*t0Kt>«» Jt«VMT 4MB TITtB
DISPOSER OF WASTE (Mutt b» HM«1 by dnpOO»r> 1
COOK N*.
Th« haular abova daUvarad tha daacribod watta to thfe dlapOMl facility and it vwa» an •ccaptaWa
matarUI undar tha ttrmt of RWOCS raquirarnann, Stm O#P*rtmant of Haaltfi rwgultttom. and
local raatrtetlona.
Qiuntitv m*Murad at ilta (If aoalicablat: _ Statafaa (if anwl: , . ,
Handing Matt>od(«):
Dracovory r~r~I
n *rMtrMr» iMMelfwI. 1 1 1
Q dtapoort (apOBffy): Q pond O aprMdin* G landftH G Infactfon «rotl , — . — .
coaa t*a.
n*^»*f-' °«»-- .
1 corttfy (or daclara) undar ponarty of partury
that tna foregoing <• true and eorract.
Tha turn 9««nrror tfwll *wbm>t • iaoibia copy of aach completed Record to tha State Department of
FOR INFORMATION RELATED TO SIM L US Oft OTHER EMERGENCIES INVOLVING
HAZARDOUS WA»T» Oft OTHKH MATERIALS CAUL «QOl 434-9300.
Q a.T. *TBBM BMppInf MMM
Figure 10. Example of waste manifest
28
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TABLE 3. SAMPLERS RECOMMENDED FOR VARIOUS TYPES OF WASTE
Waste type
Recommended
sampler
Limitations
Liquids, sludges, Coliwasa
and slurries in
drums, vacuum a) Plastic
trucks, barrels,
and similar
containers , N „,
b) Glass
Liquids and
sludges in ponds,
pits, or lagoons
Powdered or gran-
ular solids in
bags, drums,
barrels, and
similar con-
tainers
Dry wastes in
shallow contain-
ers and surface
soil
Waste piles
Soil deeper
than 8 cm(3 in.)
Pond
Wastes in
storage tanks
a) Grain
sampler
b) Sampling
trier
Trowel or
scoop
Waste pile
sampler
a) Soil auger
b) Veihmeyer
sampler
Weighted
bottle sampler
Not for containers 1.5 m(5 ft ) deep.
Not for wastes containing ketones,
nitrobenzene, dimethylformamide,
mesityl oxide, or tetrahydrofuran^'^.
Not for wastes containing hydro-
fluoric acid and concentrated alkali
solutions.
Cannot be used to collect samples
beyond 3.5 m(11.5 ft ). Dip and
retrieve sampler slowly to avoid
bending the tubular aluminum handle.
Limited application for sampling moist
and sticky solids with a diameter
0.6 cmOs in.).
May incur difficulty in retaining core
sample of very dry granular materials
during sampling.
Not applicable to sampling deeper than
8 cm(3 in.). Difficult to obtain
reproducible mass of samples.
Not applicable to sampling solid
wastes with dimensions greater than
half the diameter of the sampling tube.
Does not collect undisturbed core
sample.
Difficult to use on stony, rocky, or
very wet soil.
May be difficult to use on very
viscous liquids.
29
-------
Wide-mouth 1000-and 2000-ml(l qt-and ^-gal.) glass bottles are recom-
mended for waste samples containing petroleum distillates, chlorinated
hydrocarbons, pesticides, and petroleum residues that are mostly incom-
patible with plastic containers. For all other types of samples, 1000-and
2000-ml(l-qt and %-gal.) wide-mouth LPE bottles are recommended.
Container Closures and Closure Linings
The containers must have tight, screw-type lids. Plastic bottles are
usually provided with screw caps made of the same material as the bottles.
No cap liners are usually required. Glass containers usually come with
glass or rigid plastic screw caps such as Bakelite. The plastic caps are
popularly provided with waxed paper liners. Other liner materials are
polyethylene, polypropylene, neoprene, and Teflon FEP plastics. For con-
taining hazardous waste samples requiring petroleum distillates, chlori-
nated hydrocarbons, pesticides, and petroleum residue analyses. Bakelite
caps with Teflon liners are recommended to be used with glass bottles.
Teflon liners may be purchased from plastic specialty supply houses (e.g.,
Scientific Specialties Service, Inc., P.O. Box 352, Randallstown, Md.
21133).
Table 4 shows most types of wastes and the corresponding sampling
containers and closures recommended.
SAMPLING PLAN
The sampling plan should be well formulated before any actual sampling
is attempted. The plan must be consistent with the objectives of the
sampling. It must include the selected point(s) of sampling and the in-
tended number, volumes, and types (i.e., composite, grab, etc.) of samples
to be taken. These requirements are discussed below.
POINT OF SAMPLING
A representative sample is crucial to the sampling plan. This sample
depends on proper selection of sampling points in the bulk of the waste.
Hazardous wastes are usually multiphase mixtures and are contained and
stored in containers of different sizes and shapes. No single sampling
point can be specified for all types of containers. Table 5 lists most
types of containers used for hazardous wastes and the corresponding
recommended sampling pqints.
NUMBER OF SAMPLES
The number of samples to be taken primarily depends on the information
desired. Table 6 lists the recommended number of samples to be collected
consistent with the information sought and the types of wastes to be
sampled. In hazardous waste management, the properties and the average
concentrations of the hazardous components are usually desired. In this
30
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TABLE 4. SAMPLE CONTAINERS AND CLOSURES RECOMMENDED FOR
VARIOUS TYPES OF WASTE
Waste type
item
Recommended
container
Recommended
closure
Oil wastes except
pesticides, HC,
chlorinated HC, and
photosensitive
wastes
Pesticides, HC,
and chlorinated
HC
Linear polyethylene (LPE)
bottles,3 1000-and 2000-
ml (1-qt. and ^-gal.),
wide mouth
Glass bottles,b wide-
mouth, 1000-and 2000-ml
(1-qt. and ^-
LPE caps
Bakelite caps
with Teflon
liner0
Photosensitive
wastes
Amber LPE or brown
glass** bottles, wide-
mouth, 1000-and 200-ml
(1-qt. and %-gal.)
LPE caps for
the LPE bottles;
Bakelite caps
with Teflon
liner for the
glass bottles
aNalgene, Cat. Nos. 2104-0032 and 2120-0005, or equivalent.
Scientific Products, Cat. Nos. 87519-32 and B7519-64, or equivalent.
Available from Scientific Specialities, P.O. Box 352, Randallstown,
Md.
Scientific Products, Cat. Nos. B7528-050 and 7528-2L, or equivalent.
31
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TABLE 5. SAMPLING POINTS RECOMMENDED FOR MOST .WASTE CONTAINERS
Container type
Sampling point
Drum, bung on one end
Drum, bung on side
Barrel, fiberdrum,
buckets, sacks, bags
Vacuum truck and
similar containers
Pond, pit, lagoons
Waste pile
Storage tank
Soil
Withdraw sample through the bung opening.
Lay drum on side with bung up. Withdraw
sample through the bung opening.
Withdraw samples through the top of barrels,
fiberdrums, buckets, and similar containers.
Withdraw samples through fill openings of
bags and sacks. Withdraw samples through
the center of the containers and to different
points diagonally opposite the point of entry.
Withdraw sample through open hatch. Sample
all other hatches.
Divide surface area into an imaginary grid.3
Take three samples, if possible: one sample
near the surface, one sample at mid-depth or
at center, and one sample at the bottom.
Repeat the sampling at each grid over the
entire pond or site.
Withdraw samples through at least three
different points near the top of pile to
points diagonally opposite the point of
entry.
Sample from the top through the sampling hole.
Divide the surface area into an imaginary
grid.3 Sample each grid.
3The number of grid is determined by the desired number of samples
to be collected, which when combined should give a representative
sample of the wastes.
32
-------
respect, collecting one representative sample of a given waste is usually
adequate. This sample can either be collected from a single sampling point
with a composite sampler, or several samples can be collected from various
sampling points and combine into one composite sample.
When gathering evidence for possible legal actions, multiple samples
of a waste are usually collected. Three identical samples are desirable:
one sample is given to the company or organization responsible for the
waste, the second sample is submitted to the laboratory for analysis, and
the third sample is kept in storage for possible use as a referee sample.
Subdividing a waste sample is not recommended unless it is homogeneous.
VOLUME OF SAMPLES
Sufficient volume of a sample, representative of the main body of the
waste, must be collected. This sample must be adequate in size for all
needs, including laboratory analysis, splitting with other organizations
involved, etc. In collecting liquid waste samples in drums, vacuum trucks,
or similar containers, the volume collected in the Coliwasa usually deter-
mines the volume of the sample. This volume can range from 200 to 1800
(% pt. to 1.9 qt.). In most cases, 1000 ml(l qt.) of a sample is usually
sufficient. Hazardous wastes usually contain high concentrations of the
hazardous components, and only a small aliquot of the sample is used for
analysis.
SAMPLING PRECAUTIONS AND PROTECTIVE GEAR
Proper safety precautions must always be observed when sampling ha-
zardous wastes. In all cases, a person collecting a sample must be aware
that the waste can be a strong sensitizer and can be corrosive, flammable,
explosive, toxic, and capable of releasing extremely poisonous gases.^
The background information obtained about the waste should be helpful in
deciding the extent of sampling safety precautions to be observed and in
choosing protective equipment to be used..
For full protection, the person collecting the sample must use a self-
contained breathing apparatus, protective clothing, hard hat, neoprene
rubber gloves, goggles, and rubber boots.
A self-contained breathing apparatus consists of an air-tight face
mask and a supply of air in a pressure tank equipped with a pressure
regulator. Protective clothing consists of long-sleeved neoprene rubber
coat and pants, or long-sleeved coverall and oil-and-acid proof apron. In
hot weather, the coverall-apron combination might be preferred. Table 7
lists the uses and commercial availability of respiratory protective equip-
ment. All equipment except the respirator must be properly washed and
cleaned between uses (see Section 5).
33
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TABLE 6. NUMBER OF SAMPLES TO BE COLLECTED
Case Information
No. desired
Waste
type
Container type
Number of samples
to be collected
1 Average
concentration
2 Average
concentration
3 Average
concentration
4 Average
concentration
5 Average
concentration
6 Concentration
range
7 Concentration
range
8 Concentration
range
9 Concentration
range
10 Concentration
range
11 Average
concentration
for legal
evidence
12 Average
concentration
13 Average
concentration
Liquid Drum, vacuum truck,
and similar
containers
Liquid Pond, pit, lagoon
Bag, drum, bin
Solid
(powder
or gran
ular)
Waste
pile
Soil
Liquid Drum, vacuum truck,
storage tank
Liquid Ponds, pit, lagoon
Solid Bag, drum, bin
(powder
or gran-
ular)
Waste —
pile
Soil
All All containers
types
Liquid Storage tank
Liquid Storage tank
1 Collected with
Coliwasa
1 Composite sample of
several samples
collected at differ-
ent, sampling points
or levels
Same as Case #2
Same as Case #2
1 Composite sample of
several samples
collected at differ-
ent sampling areas
3 to 10 separate sam-
ples, each from a
different depth of
the liquid
3 to 20 separate sam-
ples from different
sampling points and
depths
3 to 5 samples from
different sampling
points
Same as Case #8
3 to 20 separate sam-
ples from different
sampling areas
3 Identical samples or
1 composite sample
divided into 3
identical samples if
homogeneous
Same as Case #2
Same as Case #6
34
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TABLE 7. RESPIRATORY PROTECTIVE DEVICES RECOMMENDED
FOR VARIOUS HAZARDS
Type of hazard
Recommended respiratory device
Oxygen deficiency
Gaseous contaminant
immediately dangerous
to life
Gaseous contaminant
not immediately
dangerous to life
Particulate contaminant
Combination of gaseous and
particulate contaminants
immediately dangerous to
life
Combination of gaseous
and particulate contaminants
not immediately
dangerous to life
Self-contained breathing apparatus,
hose mask with blower
Self-contained breathing apparatus,
hose mask with blower,
gas mask
Air-line respirator,
hose mask without blower,
chemical-cartridge respirator
Dust, mist, or fume respirator,
air-line respirator,
abrasive-blasting respirator
Self-contained breathing apparatus,
hose mask with blower,
gas mask with special filter
Air-line respirator,
hose mask without blower,
chemical-cartridge respirator
with special filter
Source: Reference 14.
35
-------
The self-contained breathing apparatus may not be required in all
sampling situations. In some cases, gas masks or chemical cartridge-type
respirators with filters will suffice. Table 7 may be used to select the
proper protective respiratory device.
» For added protection in sampling, a second person with a radio-
telephone and first-aid kit must be present to render any necessary
help or call for assistance.
SAMPLING PROCEDURES
The following procedures are recommended for sampling different types
of hazardous wastes in various containers.
Sampling a Drum
Drums containing liquid wastes can be under pressure or vacuum. A
bulging drum usually indicates that it is under high pressure and should
not be sampled until the pressure can be safely relieved. A heavily
corroded or rusted drum can readily rupture and spill its contents when
disturbed; it should only be sampled with extreme caution. Opening the
bung of a drum can produce a spark that might detonate an explosive gas
mixture in the drum. This situation is difficult to predict and must be
taken into consideration every time a drum is opened. The need for full
protective sampling equipment cannot be overemphasized when sampling a
drum.
1. Position the drum so that the bung is up (drums with the bung on the
end should be positioned upright; drums with bungs on the side should
be laid on its side, with the bungs up).
2. Allow the contents of the drum to settle.
3. Slowly loosen the bung with a bung wrench, allowing any gas pressure
to release.
4. Remove the bung and collect a sample through the bung hole with a
Coliwasa, as directed in Section 4.
5. When there is more than one drum of waste at a site, segregate and
sample the drums according to waste types, using a table of random
numbers as outlined in Appendix D.
Sampling a Vacuum Truck
Sampling a vacuum truck requires the person collecting the sample to
climb onto the truck and walk along a narrow catwalk. In some trucks, it
requires climbing access rungs to the tank hatch. These situations pre-
sent accessability problems to the sample collector, who most usually
36
-------
wear full protective sampling gear. Preferably, two persons should
perform the sampling: One person should do the actual sampling and the
other should hand the sampling device, stand ready with the sample con-
tainer, arid help deal with any problems. The sample collector should
position himself to collect samples only after the truck driver has
opened the tank hatch. The tank is usually under pressure or vacuum.
The driver should open the hatch slowly to release pressure or to break
the vacuum.
1. Let the truck driver open the tank hatch.
2. Using protective sampling gear, assume a stable stance on the tank
catwalk or access rung to the hatch.
3. Collect a sample through the hatch opening with a Coliwasa, as
directed in Section 4.
4. If the tank truck is not horizontal, take one additional sample each
from the rear and front clean out hatches and combine all three
samples in one sample container.
5. When necessary, carefully take sediment sample from the tank through
the drain spigot.
Sampling a Barrel, Fiberdrum, Can, Bags, or Sacks Containing Powder
or Granular Waste
The proper protective respirator (see Table 7), in addition to the
other protective gear, must be worn when sampling dry powdered or granu-
lar wastes in these containers. These wastes tend to generate airborne
particles when the containers are disturbed. The containers must be
opened slowly. The barrels, fiberdrums, and cans must be positioned
upright. If possible,, sample sacks or bags in the position you find
them, since standing them upright might rupture the bags or sacks.
1. Collect a composite sample from the container with a grain sampler
or sampler trier, as directed in Section 4.
2. When there is more than one container of waste at a site, segregate
and sample the containers according to a table of random numbers, as
outlined in Appendix D.
Sampling a Pond
Storage or evaporation ponds for hazardous wastes vary greatly in
size from a few to a hundred meters. It is difficult to collect repre-
sentative samples from the large ponds without incurring huge expense
and assuming excessive risks. Any samples desired beyond S.SmCll^ ft)
from the bank may require the use of a boat, which is very risky, or
the use of a crane or a helicopter, which is very expensive. The
37
-------
information sought must be weighed against the risk and expense of col-
lecting the samples. The pond sampler described in Section 4 can be used
to collect samples as far as 3.5 m(ll% ft ) from the bank.
1. Collect a composite sample with pond sampler, as directed in Section
4.
Sampling Soil
The techniques of soil sampling are numerous. The procedures out-
lined below are adopted from ASTM methods.15 The procedures are consist-
ent with the hazardous waste management objective of collecting soil
samples which is usually to determine the amount of hazardous material
deposited on a particular area of land or to determine the leaching rate
of the material and/or determine the residue level on the soil. Elaborate
statistically designed patterns have been designed for sampling soils. If
one of these patterns is to be used, a good statistics book may have to be
consulted. In the following procedures, soil samples are taken in a grid
pattern over the entire site to ensure a uniform coverage.
1. Divide the area into an imaginary grid (see Table 5).
2. Sample each grid and combine the samples into one.
3. To sample up to 8 cm(3 in.) deep, collect samples with a scoop, as
directed in Section 4.
4. To sample beyond 8 cm(3 in.) deep, collect samples with a soil auger
or Veihmeyer soil sampler, as directed in Section 4.
Sampling a Waste Pile
Waste piles can range from small heaps to a large aggregates of
wastes. The wastes are predominantly solid and can be a mixture of powders,
granules, and chunks as large as or greater than 2.54 cm(l in.) average
diameter. A number of core samples have to be taken at different angles
and composited to obtain a sample that, on analysis, will give average
values for the hazardous components in the waste pile.
1. Determine the sampling points (see Table 5).
2. Collect a composite sample with a waste pile sampler according to the
directions in Section 4.
Sampling a Storage Tank
The collection of liquid samples in storage tanks is extremely dis-
cussed in the ASTM methods. The procedure used here is adopted from one
of those methods.16
38
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Sampling a storage tank requires a great deal of manual dexterity.
Usually it requires climbing to the top of the tank through a narrow
vertical or spiral stairway while wearing protective sampling equipment
and carry sampling paraphernalia. At least two persons must always per-
form the sampling: One should collect the actual samples and the other
should stand back, usually at the head of the stairway, and observe, ready
to assist or call for help. The sample collectors must be accompanied by
a representative of the company, who must open the sampling hole, usually
on the tank roof.
1. Collect one sample each from the upper, middle, and lower sections of
the tank contents with a weighted bottle sampler, as outlined in
Section 4.
2. Combine the samples one container and submit it as a composite sample.
SAMPLE HANDLING
After a sample is transferred into the proper sample container, the
container must be tightly capped as quickly as possible to prevent the
loss of volatile components and to exclude possible oxidation from the air.
The use of a preservative or additive is not recommended. However,
if only one or two components of a waste are of interest, and if these
components are known to rapidly degrade or deteriorate chemically or bio-
chemically, the sample may be refrigerated at 4 to 6°C.(39.2 to 42.8°F.)
or treated with preservatives according to Section 8.
To split or withdraw an aliquot of a sample, considerable mixing,
homogenization, or quartering is required to ensure that representative
or identical portions are obtained. When transferring a sample aliquot,
open the container as briefly as possible.
IDENTIFICATION OF SAMPLE
Each sample must be labeled and sealed properly immediately after
collection.
Sample Labels
Sample labels (Figure 11) are necessary to prevent misidentification
of samples. Gummed paper labels or tags are adequate. The label must
include at least the following information:
Name of collector.
Date and time of collection.
Place of collection.
Collector's sample number, which uniquely identifies the sample.
39
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OFFICIAL SAMPLE LABEL
Collector Collector's Sample No.
Place of Collection .
Date Sampled Time Sampled_
Field Information
Figure 11. Example of official sample label.
OFFICIAL SAMPLE SEAL
State of California Public Health Division
Department of Health Services Hazardous Materials Laboratory
Collected by Collector's Sample No.
(signature)
Date Collected Time Collected
Place Collected
Figure 12. Example of official sample seal
40
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Sample Seals
Sample seals are used to preserve the integrity of the sample from the
time it is collected until it is opened in the laboratory. Gummed paper
seals can be used as official sample seals. The paper seal must carry
information such as:
Collector's name
Date and time of sampling
Collector's sample number. (This number must be identical with the
number on the sample label).
The seal must be attached in such a way that it is necessary to break
it in order to open the sample container. An example of a sample seal is
shown in Figure 12.
FIELD LOG BOOK
All information pertinent to a field survey and/or sampling must be
recorded in a log book. This must be a bound book, preferabley with con-
secutively numbered pages that are 21.6 by 27.9 cm(8% by 11 in.). Entries
in the log book must include at least the following:
Purpose of sampling (e.g., surveillance, etc.)
Location of sampling (e.g., hauler, disposal site, etc.) and address
Name and address of field contact
Producer of waste and address
Type of process (if known) producing waste
Type of waste (e.g., sludge, wastewater, etc.)
Declared waste components and concentrations
Number and volume of sample taken
Description of sampling point
Date and time of collection
Collector's sample identification number(s)
Sample distribution (e.g., laboratory, hauler, etc.)
References such as maps or photographs of the sampling site
Field observations
Any field measurements made such as pH, flammability, explositivity,
etc.
Sampling situations vary widely. No general rule can be given as to
the extent of information that must be entered in the log book. A good
41
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rule, however, is to record sufficient information so that someone can
reconstruct the sampling situation without reliance on the collector's
memory.
The log book must be protected and kept in a safe place.
CHAIN OF CUSTODY RECORD
To establish the documentation necessary to trace sample possession
from the time of collection, a chain of custody record must be filled out
and accompany every sample. This record becomes especially important when
the sample is to be introduced as evidence in a court litigation. An
example of a chain of custody record is illustrated in Figure 13.
The record must contain the following minimum information:
Collector's sample number
Signature of collector
Date and time of collection
Place and address of collection
Waste type
Signatures of persons involved in the chain of possession
Inclusive dates of possession
SAMPLE ANALYSIS REQUEST SHEET
The sample analysis request sheet (Figure 14) is intended to accom-
pany the sample on delivery to the laboratory. The field portion of this
form must be completed by the person collecting the sample and should
include most of the pertinent information noted in the log book. The
laboratory portion of this form is intended to be completed by laboratory
personnel and to include:
Name of person receiving the sample
Laboratory sample number
Date of sample receipt
Sample allocation
Analyses to be performed
SAMPLE DELIVERY TO THE LABORATORY
Preferably, the sample must be delivered in person to the laboratory
for analysis as soon as practicable—usually the same day as the sampling.
Consult Section 8 when.sample preservation is required. The sample must
42
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California Department of Health
Hazardous Materials Laboratory
Collector's Sample No,
CHAIN OF CUSTODY RECORD
Hazardous Materials
Location of Sampling: Producer Hauler
Other:
Company's Name
Address
Disposal Site
Telephone ( )_
number street
Co11ector's Name
Date Sampled
city
state zip
Telephone ( )
signature
Time Sampled_
Type of Process Producing Waste_
Waste Type Code Other_
Field Information
hours
Sample Allocation:
1.
2.
3.
1.
2.
3.
name of organization
name of organization
n of Possession
signature
signature
signature
name of organization
title
title
title
inclusive dates
inclusive dates
inclusive dates
Figure 13. Example of chain of custody record
43
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PRIORITY California Department of Health Services
(explain) Hazardous Materials Laboratory
HAZARDOUS MATERIALS SAMPLE ANALYSIS REQUEST
PART I; FIELD SECTION
Collector Date Sampled Time hours
Location of Sampling
name of company, disposal site, etc.
Addres s
number street city state zip
Telephone ( ) Company Contact
HML NO. COLLECTOR'S TYPE OF »
(Lab only) SAMPLE NO. SAMPLE* FIELD INFORMATION**
Analysis Requested_
Special Handling and/or Storage_
PART II: LABORATORY SECTION
Received by Title Date_
Sample Allocation: HML LBL LABL SRL Date_
Analysis Required
*Indicate whether sample is sludge, soil, etc.;**Use back of page for
additional information.
Figure 14. Example of hazardous waste sample analysis request sheet
44
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be accompanied by the chain of custody record and by a sample analysis
request sheet (Figure 14). The sample must be delivered to the person
in the laboratory authorized to receive samples (often referred to as
the sample custodian).
SHIPPING OF SAMPLES
When a sample is shipped to the laboratory, it must be packaged in a
proper shipping container to avoid leakage and/or breakage. A cardboard
box that will provide at least 10 cm(4 in.) of tight packing around the
sample container must be used. Acceptable packing materials include saw-
dust, crumpled newspapers, vermiculite, polyurethane chips, etc. Other
samples that require refrigeration must be packed with reusable plastic
packs or cans of frozen freezing gels in molded polyurethane boxes with
sturdy fiberboard protective case. The boxes must be taped closed with
masking tape or fiber plastic tape.
All packages must be accompanied by a sample analysis sheet and chain
of custody record. Complete address of the sender and the receiving lab-
oratory must legibly appear on each package. When sent by mail, register
the package with return receipt requested. When sent by common carrier,
obtain a copy of the bill of lading. Post office receipts and bill of
lading copies may be used as part of the chain of custody documentation.^
45
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SECTION 7
RECEIPT AND LOGGING OF SAMPLE
Field samples are delivered to the laboratory either personally or
through a public carrier. In the laboratory, a sample custodian should
be assigned to receive the samples. Upon receipt of a sample, the custo-
dian should inspect the condition of the sample and the sample sea],
reconcile the information on the sample label and seal against that on the
chain of custody record, assign a laboratory number, leg in the sample in
the laboratory log book, and store the sample in a secured sample storage
room or cabinet until assigned to an analyst for analysis.
SAMPLE INSPECTION
The sample custodian should inspect the sample for any leakage from
the container. A leaky container containing multiphase sample should
not be accepted for analysis. This sample will no longer be a represen-
tative sample. If the sample is contained in a plastic bottle and the
walls show any bulging or collapsing, the custodian should note that the
sample is under pressure or releasing gases, respectively. A sample
under pressure should be treated with caution. It can be explosive or
release extremely poisonous gases. The custodian should examine whether
the sample seal is intact or broken, since broken seal may mean sample
tampering and would make analysis results inadmissible in court as evi-
dence. Discrepancies between the information on the sample label and seal
and that on the chain of custody record and the sample analysis request
sheet should be resolved before the sample is assigned for analysis. This
effort might require communication with the sample collector. Results of
the inspection should be noted on the sample analysis request sheet and
on the laboratory sample log book.
ASSIGNMENT OF LABORATORY NUMBER
Incoming samples usually carry the inspector's or collector's identi-
fication numbers. To further identify these samples, the laboratory
should assign its own identification numbers, which normally are given
consecutively. Each sample should be marked with the assigned laboratory
number. This number is correspondingly recorded on a laboratory sample
log book along with the information describing the sample. The sample
information is copied from the sample analysis request sheet and cross-
checked against that on the sample label.
46
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ASSIGNMENT OF SAMPLE FOR ANALYSIS
In most cases, the laboratory supervisor assigns the sample for ana-
lysis. The supervisor should review the information on the sample analysis
request sheet, which now includes inspection notes recorded by the labora-
tory sample custodian. The supervisor should then decide what analyses
are to be performed. The sample may have to be split with other labora-
tories to obtain tne necessary information about the sample. The super-
visor should decide on the sample allocation and delineate the types of
analyses to be performed on each allocation. In his own laboratory, the
supervisor should assign the sample analysis to at least one chemist, who
is to be responsible for the care and custody of the sample once it is
handed to him. He should be prepared to testify that the sample was in
his possession or secured in the laboratory at all times from the moment
it was received from the custodian until the analyses were performed.
• The receiving chemist should record in his laboratory notebook the
identifying information about the sample, the date of receipt, and other
pertinent information. This record should also include the subsequent
analytical data and calculations.
47
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SECTION 8
PRESERVATION AND STORAGE OF SAMPLES
Ideally, hazardous waste samples should be analyzed immediately after
collection for maximum reliability of the analytical results. Hazardous
wastes are such complex mixtures that it is difficult to exactly predict
the physical, biological, and chemical changes that occur in the samples
with time. After collection of samples, pH may change significantly in a
matter of minutes; sulfides and cyanides may be oxidized or evolve as
gases; and hexavalent chromium may slowly be reduced to the trivalent
state. Certain cations may be partly lost as a result of adsorption to
the walls of the sample containers. Growth of microorganisms may also
cause changes to certain constituents of the sample. Volatile compounds
may be rapidly lost.
In a number of cases, the above changes may be slowed down or pre-
vented by refrigeration at 4 to 6°C, or by the addition of preservatives.
However, these treatments mostly apply to one or two components or pro-
perties. Refrigeration may deter the evolution of volatile components and
acid gases such as hydrogen sulfides and hydrogen cyanides, but it also
introduces the uncertainty that some salts may precipitate at lower temper-
ature. On warming to room temperature for analysis, the precipitates may
not redissolve, thus incurring error in determining the actual concentra-
tions of dissolved sample constituents. Addition or preservatives may
retard biochemical changes, whereas other additives may convert some
constituents to stable hydroxides, salts, or compounds. Unknown in these
treatments, however, is the possible conversion of other compounds to other
forms (such as the products of nitration, sulfonation, oxidation, etc., of
organic components). In subsequent analyses, the results may not reflect
the original identity of the components.
Thus, both advantages and disadvantages are associated with the refri-
geration and/or addition of preservatives or additives to waste samples.
These methods of preservation or stabilization are not recommended for
hazardous waste samples unless only one or two components or properties
are to be analyzed.
Standard methods books 1"»19 have compilations of useful preservatives
for various constitutents. Table 8 is excerpted from these lists and shows
only the preservation methods that may be used for hazardous wastes.
48
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TABLE 8. METHODS OF PRESERVATION FOR HAZARDOUS WASTES
Waste constituent
to be preserved
Acidity
Alkalinity
Ammonia
Arsenic
Chlorine
Chromium (VI)
Cyanides
Fluoride
Metals:
1) dissolved
2) suspended
3) Total
Mercury
1) dissolved
2) Total
PH
Phenolics
Residue, volatile
Selenium
Specific
conductance
Sulfide
Sulfide
Zinc
Preservation method Storage time
Cool to 4° C
Cool to 4° C
Add 1 ml cone. H2S04/&
Add 6 ml cone. HN03/ &
Cool to 4° C
Add 6 ml cone. H2S04/£
Add 2.5 ml of 50% NaOH/& ;
cool to 4° C
Cool to 4° C
Filter on site; add 5 ml
cone. HN03/&
Filter on site
Add 5 ml cone. HN03/£
Filter; add 5 ml cone. HN03/&
Add 5 ml cone. HN03/£
Determine on site; cool to 4° C
Add H3P04 to pH 4 and 1 g.
CuS04/& ; refrigerate at 4° C
Cool to 4° C
Add 5 ml cone. HN03/£
Cool to 4° C
Add 2 ml of 2N 7
-------
REFERENCES
1. Stephens, R. D. 1976. Hazardous Sampling; Residual Management by Land
Disposal. Proceedings of the Hazardous Waste Research Symposium,
University of Arizona, Tucson, Ariz.
2. Eichenberger, B., J. R. Edwards, K. Y. Chen, and R. D. Stephens. 1977.
A Case Study of Hazardous Wastes Input Into Class I Landfills. U.S.
Environmental Protection Agency, Cincinnati, Ohio.
3. Merck Index, An Encyclopedia of Chemicals and Drugs. 1968. 9th Ed.
Merck & Co., Rahway.
4. Chemical Resistance of Excelon R-4000. 1977. Thermoplastic Process,
Inc., Stirling, N.J.
5. Sampling Procedure for Animal Feed. Official Methods-of Analysis of
the Association of Official Analytical Chemists. 1975. 12th Ed.
Washington, D.C., p. 129.
6. Guidelines on Sampling and Statistical Methodologies for Ambient Pesti-
cide Monitoring. 1974. Federal Working Group on Pest Management, U.S.
Environmental Protection Agency, Washington, D.C., p. III-5.
7. Veihmeyer, F. J. 1929. An Improved Soil-Sampling Tube. Soil Science
27/2): 147-152.
8. American Society for Testing and Materials. 1975. ASTM D270. ASTM
Standards. The Society, Philadelphia, Pa.
9. American Society for Testing and Materials. 1973. ASTM E 300. ASTM
Standards. The Society, Philadelphia, Pa.
10. Sax, I. N. 1968. Dangerous Properties of Industrial Materials. 3rd
Ed. Van Nostrand Reinhold Co. New York, N.Y.
11. Condensed Chemical Dictionary. 1977. 8th Ed. Van Nostrand Reinhold
Co., New York, N.Y.
12. Toxic and Hazardous Industrial Chemicals Safety Manual for Handling
and Disposal With Toxicity and Hazardous Data. 1976. International
Technical Information Institute, Tokyo, Japan.
13. Dunlap, J. 1972. Industrial Waste Law (AB 596), State of California.
50
-------
14. Bureau of Labor Standards. Respiratory Protective Equipment.
Bulletin 226. Safety in Industry; Environmental and Chemical
Hazards, No. 3. U.S. Department of Labor, Washington, D.C.
15. American Society for Testing and Materials. ASTM D1452, D1586 and
D3550. ASTM Standards. The Society, Philadelphia, Pa.
16. American Society for Testing and Materials. ASTM D270 and E 300
ASTM Standards. The Society, Philadelphia, Pa.
17. Compliance Monitoring Procedures. 1974. EPA 330/1-74-/002. U.S.
Environmental Protection Agency, Denver, Colo.
18. Manual of Methods for Chemical Analysis of Water and Wastewater.
1974. EPA-625/6-74-003. U.S. Environmental Protection Agency,
Washington, D.C.
19. Standard Methods for Examination of Water and Wastewater. 1975. 14th
Ed. American Public Health Association, New York, N.Y.
20. Collection, Storage, Transportation, and Pretreatment of Water and
Wastewater Samples. 1971. California Department of Health Services,
Berkeley, Ca.
51
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APPENDICES
APPENDIX A. DEVELOPMENT OF THE COMPOSITE LIQUID WASTE SAMPLER (COLIWASA)
Early in the development of the California waste management program,
the needs were recognized for accurate information about waste composition
and adequate equipment and procedures for sampling and analysis.
In 1975, the California Department of Health Services engaged in a
cooperative study with the University of Southern California (Environmental
Engineering Department) under U.S. Environmental Protection Agency's spon-
sorship to collect and analyze a large number of hazardous waste samples
at a number of Class I disposal sites in Los Angeles County. In the prep-
aration ' of this study, the Department of Health Services personnel designed
and constructed a simple tube sampler suitable for use in liquid and sludge
wastes. The objective of the sampler design was to obtain samples represent-
ative of complex, heterogeneous wastes contained in vacuum trucks and drums.
In preliminary testing, the prototype design shown in Figure A-l appeared to
give good representative samples. At this time, the sampling device was
named the composite liquid waste sampler, or Coliwasa for short.
Requirements for Hazardous Waste Sampling Procedures and Equipment
Approximately 24 of these devices were constructed for use in the Los
Angeles County sampling program. During a 2-week period in 1975, 400 samples
of hazardous wastes were taken from vacuum trucks and drums. The wastes
represented an extremely wide variety of chemical compositions and physical
characteristics. The experience given by this sampling program emphasized
the following important requirements of hazardous waste sampling procedures
and equipment:
1) Sampling equipment must be of simple design to facilitate easy cleaning
or to allow discard if necessary to prevent sample cross contamination.
Many wastes, because of their chemical composition and/or physical nature,
so fouled sampling equipment that it required discarding or extensive
cleaning. Extensive cleaning produces a significant volume of cleaning
waste that must be properly disposed. Equipment that has complicated
valves, levers, and other fittings would never survive many hazardous
wastes.
2) Equipment must be light weight and leak proof. Sampling personnel are
required to climb and move about on tank trucks and other dangerous areas
while holding sampling equipment. Once the sampler is filled with its
52
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charge of hazardous waste, it must not discharge until property
a sample receptacle. Thus, a positive locking mechanism is needed.
3) Several types of sampling equipment must be available, for no one de-
sign or material meets all hazardous waste sampling requirements.
4) Sampling requires a minimum of two persons equipped with the proper
and complete complement of safety equipment. Even at a well-run waste
disposal site using an approved manifest system, surprises occur. A
waste sample collector never knows for sure what he will find when a
vacuum truck or barrel is opened.
183 cm (72")
152 cm (60")
Rod, PVC, 0.95 cm (3/8") O.D.
Pipe, WC, 4.13 cm (1 5/8") I.D.,
A.78 cm (1 7/8") O.D.
Stopper, neoprene, #9
Nut and washer, stainless steel,
0.95 cm (3/8")
Figure 1. Coliwasa, Model 1.
Sampler Selection
A review of the literature was conducted to investigate the availabil-
ity of commercial equipment: that would better suit the sampling require-
ments than the Coliwasa. The guidelines used in the selections were
commercial availability, cost, simplicity in design, chemical inertness,
and adaptability for use in composite sampling of liquid hazardous wastes.
Preliminary tests were performed on a number of candidate liquid
samplers. The tests included physical inspections of the sampling mechan-
isms for ease of operation and applicability. Water was the initial test
liquid. The test water was placed in a fabricated tank made out of a
53
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122-cm(4-ft) tall by 15.2-cm(6-in.) I.D. glass cylinder. Each sampler
under test was lowered slowly into the tank to determine whether the check
valve or closing device or sampling orifice would allow the sample to flow
in the sampler. The sampler was then withdrawn and tested for leakage and
ease of transfer of the collected sample.
None of the commercially available samplers was found to be satis-
factory. The Coliwasa designed by the Department of Health Services
appeared to be the most promising.
First Coliwasa Model
The early design of the Coliwasa (Model 1; Figure A-l) consisted of
1.52-m(5-ft.) by 4.13-cm(l ,5/8-in.) I.D., opaque PVC pipe as the sampling
tube and a neoprene stopper attached to one end of a 0.95-cm(3/8-in.) O.D.
PVC rod as the closing mechanism. To collect a sample with this sampler,
the stopper is pushed out about 5 cm(2 in.) from the bottom end of the
sampling tube. Then the sampler is lowered straight down through the body
of liquid waste to be sampled to the bottom of the waste container. The
liquid in the tube is trapped by plugging the bottom of the tube with the
stopper by pulling up the stopper rod with one hand and holding the tube
with the other hand.
This early design of the Coliwasa, albeit functional, was deficient in
a number of aspects. First, it was difficult to put the sampler in the
close position. The stopper did not easily line up with the bottom opening
of the sampling tube. Several manipulations of the stopper rod were usually
required to effect closure. This difficulty tended to disturb the bottom
layer of the waste being sampled and undoubtedly contributed to the col-
lection of nonrepresentative samples.
Second, the sampler was not equipped with a mechanism that positively
and independently locked it closed. Closure was maintained by using one
hand to hold the sampling tube and the other hand to maintain a constant
upward pressure on the upper end of the stopper rod to keep the stopper
tightly seated against the bottom opening of the sampler. In some samp-
ling instances, this method of closure was not always practical. When
sampling waste containers as deep as the length of the sampler, the opera-
tor could not withdraw the sampler without freeing the hand that maintains
the closing pressure on the stopper rod. Thus, the snug contact between
the neoprene stopper and the inner opening of the sampling tube was the
only force that locked the sampler closed. The weight of the sample con-
tained in the sampling tube has in some cases pushed out the stopper,
resulting in lost samples and exposure of the sample collector to unneces-
sary hazards.
Third, samples contained in the sampler were difficult to transfer
into sample containers at regulated rates, and caused some samples to be
lost from splashings.
-------
Attempts were made to improve the first model of the Coliwasa. These
efforts led to fabrication of the other models shown in Figures A-2 through
A-5, and finally to the recommended version as shown in Figure 1 of the text.
Models 2 and 3
The improvement in the second model of the Coliwasa (Figure A-2) con-
sisted of making diametrical slits, 5 cm(2 in.) deep by 2 cm(0.79 in.) wide,
and indentations 90° from the slits at the top of the sampler tube. The
slits accommodate the T-handle of the stopper rod in the open position, which
allows the stopper to extend down about 5 cm(2 in.) below the bottom of the
sampler. The indentations serve as support for the T-handle when the sam-
pler is placed in the close position. When this improved Coliwasa was tested
the neoprene stopper still did not readily line up with the bottom opening
of the sampling tube. Several twisting manipulations of the stopper rod
were required to bring the sampler into the close position. This problem
was remedied by installing three stainless steel guide wires (18 gauge) on
the stopper, with the upper wire ends secured to the stopper rod, as shown
in Figure A-3. This version (Model 3) of the sampler was again tested. The
sampler was found functional and relatively easy to operate. It can be dis-
assembled and reassembled for cleaning in about 2 minutes. It can be built
for less than $10.00. The closing tension on the stopper of this sampler,
however, is not easily adjusted while sampling. This drawback might incur
some sample loss.
1.91 cm (3/4")
O.D. rod
V
152 cm (60")
155 cm (61")
T~r
t
EXPANDED VIEW OF
TOP OF SAMPLER
0.95 cm (3/8") O.D.
rod, PVC
4.13 cm (1 5/8")
I.D. pipe, PVC
OPEN
CLOSE
S topper, neoprene,
#9, with PVC nut
and washer
Figure A-2. Coliwasa, Model 2.
55
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p
•
o
to
(0
CO
0)
I
56
-------
Model 4
Further investigation into improving the design of the Coliwasa re-
sulted in Model 4(Figure A-4). In this version, the locking mechanism of
the sampler consisted of a threaded PVC plug that rides on a short threaded
0.95-cm(3/8-in.) O.D. metal rod. The metal rod is coupled with the PVC
stopper rod. To sample, the plug is screwed out about 5 cm and then the
stopper rod is pushed downward to open the sampler. The sampler is lowered
slowly into the liquid. Upon reaching the bottom of the container, the
stopper rod is pulled up to close the sampler. The PVC plug is screwed in
until tight to secure the stopper in the close position. This design of
the Coliwasa is simple, functional, and provides the person collecting the
sample with control over the tightness of the stopper against the bottom
of the sampler. It is, likewise, easily disassembled and reassembled for
cleaning. This sampler, however, is slow to operate, the PVC plug does not
screw in and out fast enough. This drawback tends to expose the sample col-
lector to the liquid waste during sampling longer than is perhaps necessary.
Model 5
Another model of the Coliwasa was fabricated using a closing principle
similar to a float valve (Figure A-5). This sampler was fabricated from a
1.52-m(5-ft) by 5.1-cm(2-in.) I.D. plastic, pipe. At the bottom end is a
plastic reducer fitting (5.1-cm(2-in.) to 3.18-cm(1.5-in.) I.D.). A manu-
ally operated neoprene rubber plug attached to a rod is used as the closing
device. When sampling, the rubber plug is raised about 5 cm(2-in.) above
its seat, and the sampler is slowly lowered into the liquid. On reaching
the bottom of the container, the sampler is closed by slowly lowering the
plug back to its seat. The sampler is withdrawn and the sample is dischar-
ged into a sample container. Tests performed on this sampler showed no
leakage of collected samples. This sampler was also found to be the easiest
to disassemble and reassemble for cleaning. However, the annular clearance
between the outside diameter of the stopper and the inside diameter of,, the
sampling tube was too narrow. The sampler tended to stir the liquid mixture
on filling, which could incur the collection of nonrepresentative sample.
In addition, the sampler tended to exclude large particles in the wastes.
Increasing the tube/stopper annular clearance did not seem practical because
it conversely reduced the opening of the reducer fitting of the sampling
tube.
Final Design
A much improved and recommended model of the Coliwasa is shown in Fig-
ure 1 of the text. This model features three main improvements over the
previous models. The first improvement consists of the use of a positive,
quick engaging closing and locking mechanism. This mechanism consists of a
short-length, channeled aluminum bar that is attached to the sampler's
stopper rod by an adjustable swivel. The aluminum bar serves both as a
T-handle and lock for the sampler's closure system. When the sampler is in
the open position, the handle is placed in the T-position and pushed
down agains the locking block. This manipulation pushes out the
57
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neoprene stopper and opens the sampling tube. In the cl ose position, the
handle is rotated until one leg of the T is squarely perpendicular against
the locking block. This tightly seats the neoprene stopper against the
bottom opening of the sampling tube and positively locks the sampler in the
close position. The closure tension can be adjusted by shortening or length-
ening the stopper rod by slightly screwing it in or out of the T handle
swivel. In discharging a collected sample, the T handle is slowly brought
into the T-position. This facilitates the opening of the sampler at a con-
trollable rate and permits the transfer of the sample into a sample container
at a regulated rate, thus minimizing splashing or loss of sample.
The second improvement made is the use of a sharply tapered neoprene
stopper. The sharp taper of the stopper eliminates the use of guide wires
and facilitates the proper seating of the stopper against the opening of the
sampling tube on closure. This stopper can be fabricated to specfications by
plastic products manufacturers at an extremely high price, or it can be made
by simply grinding down the inexpensive and commercially available neoprene
stopper to the desired taper with a shop grinder (Note 1 in Appendix B).
The third improvement is the use of translucent PVC and glass pipes
as the sampling tubes. These tubes permit the observation of the phases of
the liquid waste sample collected in the sampler. The glass sampling tube
is usually used with a Teflon stopper rod. Each tube is used for different
purposes, as described in Section 4 of the text.
The improved model of the Coliwasa was tested in the field and in the
laboratory and found to be the most practical and capable of collecting
representative samples of multiphase liquid wastes samples.
Laboratory Tests
In the laboratory, the testing was conducted using test liquid mixtures
in a 122-cm (4-ft) tall by 15.2-cm(6-in.) I.D. glass cylindrical tank. The
glass tank was ideal for the tests because it permitted observation and
measurement of the relative heights of the liquid phases.
A two-phase liquid mixture consisting of about 13.9 liters of water
and 3.29 liters of waste oil was sampled with the Coliwasa. The sample
was emptied into a 1000-ml (1.056 qt) graduated cylinder. The relative
volumes of the liquids were determined and given in Table A-l.
The results indicate that the Coliwasa is capable of obtaining a repre-
sentative sample of a two-phase liquid mixture.
59
-------
TABLE A-l.
Relative Volumes of Liquids in the Two-Phase Mixture
Item Oil Water
% by height of
phases in tank 19 81
% by volume collected 22 78
A three-phase mixture was sampled next. The mixture was prepared by
combining waste oil, water, and trichloroethylene (TCE) in the test tank.
The TCE extracted some of the waste oil and foamy emulsions formed at the
oil/water and the water/TCE interfaces. However, three distinct phases
were still obtained. Just like the previous experiment, the starting
heights of the liquid phases for each trial were measured. The mixture
was sampled with the ColiVasa. The samples were each discharged into
1000-ml (1.056 qt) graduated cylinders and the relative volumes of the
liquid phases were determined (Table A-2).
TABLE A-2.
Relative Volumes of Liquids in the Three-Phase Mixture
Item Oil Aqueous TCE
Trial I:
% by height of
phases in tank 7.9 77.4 14.7
% by volume collected 9.7 79.6 10.8
Trial II:
% by height of
phases in tank 9.3 75.0 15.0
% by volume collected 10.2 79.0 10.7
Average:
% by height of
phases in tank 8.8 76.0 15.0
% by volume collected 9.8 79.0 11.0
60
-------
The results indicate that the Coliwasa is capable of collecting a
representative sample of a three-phase mixture within 5% accuracy. The
greatest nonrepresentative error, as anticipated, occured at the bottom
phase because the samplers rubber stopper prevents sampling of the last
2.54 cm(l in.) to the bottom of the container. This error decreases as
the bottom phase increases in volume as compared to the upper phases.
With less viscous and completely immiscible test liquids, the representa-
tiveness of the sample collected approaches unity.
Field Tests
The field tests consisted of sampling liquid wastes in drums and in
vacuum trucks. Drums of unknown liquid wastes are sampled at a hazardous
waste (Class I) disposal site in California. The sampler, which has a
4.8-cm (1 7/8-in.) O.D., easily cleared through the drum's bung holes.
Sampling was relatively fast. From the time a drum was opened, a
sample was collected and transferred into a container in less than 5
minutes. While a sample was in the sampler, no leakage was detected,
indicating a positive seal by the sampler's closing mechanism. On the
transfer of sample to a container, no splashings were observed, showing
that the sampler's content can be discharged at a regulated rate.
A drum containing a two-phase liquid waste mixture was also sampled.
Replicate samples were obtained, and each was placed in separate contain-
ers. The ratios of the liquid phases in each of the samples were deter-
mined and found to be approximately the same, indicating that reproducible
samples can be collected with the sampler.
i
Incoming vacuum trucks carrying liquid wastes to the disposal site
were sampled next with the Coliwasa. Again, the sampler was found to be
functional and very easy to use. Collection of samples was very fast,
minimizing the exposure of the sample collector to hazardous fumes and
other emissions from the wastes. Only one vacuum truck with a narrow hatch
opening and a total depth of about 163 cm(5.3 ft) was not successfully
sampled. The sampling tube of the Coliwasa is only 152 cm(5 ft) long.
A longer sampling tube (i.e.,183 cm(6 ft) long) could have remedied
the problem.
61
-------
APPENDIX B. PARTS FOR CONSTRUCTING THE COLIWASA
Item
Supplier
Approximate
Cost3
Sample tube, PVC plastic, trans-
lucent, 4.13 cm(l 5/8 in.) I.D. X
1.52 m(5 ft) long X 0.4 cm(5/32 in.)
Sample tube, glass borosilicate,
4.13 cm(l 5/8 in.) I.D. X 1.52 m
(5 ft) long, Code 72-1602.
Stopper, rubber, neoprene, #9,
modified as described in footnote.*3
Stopper rod, PVC, 0.95 cm(3/8 in.)
O.D. X 1.67 m(5h ft) long.
Stopper rod, Teflon, 0.95 cm(3/8 in.)
O.D. X 1.67 m(5% ft) long.
Locking block without sleeve, PVC,
3.8 ccadh in.) O.D. X 10.2 cm(4 in.)
long with l.ll-cm(7/16 in.) hole
drilled through center.
Sleeve, PVC, 4.13 cm(l 5/8 in.) I.D.
X 6.35 (2% in.) long.
T-handle, aluminum, 18 cm(7 in.)long
X 2.86 cm(l 1/8 in.) wide with 1.27
cm(% in.) wide channel.
Swivel, aluminum bar, 1.27 cm(^ in.)
square X 3.8 cm(l% in.)long with
3/8 National Coarse (NC) inside
thread to attach stopper rod.
Nut, PVC, 3/8 in. NC thread
Washer, PVC, 3/8 in.
Nut, SS, 3/8 in., NC
Washer, SS, 3/8 in.
Bolt, 3.12 cm(l k in.)long X 3/16 in.
Nut, 3/16 in., NC
Washer, lock 3/16 in.
Plastic supply houses $ 4.00 each
Corning Glass Works,
Corning, N.Y.
$18.00 each
Laboratory supply $ 6.00/0.45
kg(lb)
Plastic supply houses $ 5.00/6.1 m
(20 ft)
Plastic supply houses $30.00/3.05 m
(10 ft)
Fabricate. Rods avail-
able at plastic supply
houses. Can be bought
in 30.48 cm(l ft)length
Fabricate from stock of $ .80/30.48
4.13 cm(l 5/8 in.) I.D. cm(ft)
PVC pipe. Available at
plastic supply houses
Fabricate. Aluminum bar $3.00/1.83
stock available at hard- m(6 ft)
ware stores.
Fabricate. Aluminum bar $ 3.00/1.83
stock available at hard- m(6
ware stores.
Plastic supply houses
Plastic supply houses
Hardware stores
Hardware stores
Hardware stores
Hardware stores
Hardware stores
$ .03 each
$ .03 each
$ .10 each
$ .10 each
$ .10 each
$ .03 each
$ .03 each
1977 prices
62
-------
'Shape the stopper into a cone as follows: Bore a 0.95-cm(3/8-in.)
diameter center hole through the stopper. Insert a short piece of
0.95-cm(3/S in.) O.D. handle through the hole until the end of the
handle is flush against the bottom (smaller diameter) surface of
the stopper. Carefully and uniformly turn the stopper into a cone
against a grinding wheel. This is done by turning the stopper with
the handle and grinding it down conically from about 0.5 cm(3/16 in.)
of the top (larger diameter) surface to the edge of the 0.95-cm(3/8-in.)
hole on the bottom surface.
APPENDIX C. CHECKLIST OF ITEMS REQUIRED IN THE FIELD SAMPLING OF HAZARDOUS
WASTES.
Quantity
Item
Use
Supplier
Approximate
Cost
Coliwasa,
plastic
type
(Section 4)
Coliwasa,
glass type
(Section 4)
Soil samp-
ler > auger
(Section 4)
Grain
sampler
(Section 4)
Scoop,
stainless
steel blade
(Section 4)
To sample liquid
wastes, except
ketones, nitro-
benzene, dimethyl-
foraraide, tetra-
hydrofuran and
pesticides
Fabricate; Parts
can be purchased
from hardware
stores (see
Section 4)
$ 16.00
To sample liquid Fabricate; Glass $ 25.00
waste with pesticides, tube available from
and other wastes that Corning Glass Co.
cannot be sampled with Corning, N.Y. 14830
plastic Coliwasa ex- (see Section 4)
cept strong alkali and
hydrofluoric acid
solution
To sample contaminated Weyco Distributor $ 70.00
soil, dried ponds, etc. 1417 Heskett Way
Sacramento,Calif.
95825
To sample powdered
or granular wastes
Laboratory supply $ 50.00
houses
To sample top soil or Cole-Parmer
shallow layers of solid Instruments
wastes Chicago, 111.
$ 25.00
63
-------
APPENDIX C (continued).
Quantity Item
Use
Supplier
Apprgxima
te
1 Veihmeyer
soil samp-
ler
(Section 4)
1 Pond samp-
ler
(Section 4)
To collect soil core Hansen Machine, $
samples 334 N. 125h St.,
Sacramento , Calif .
95815
To sample ponds, pits, Fabricate (see $
etc. Section 4). Clamps
available at
Cole-Parmer
Instrument
3060 Gibraltar Ave.
Costa Mesa, Calif .
92626
200.00
9.00
1
pair
Trier,
single slot
(Section 4)
Waste pile
sampler
(Section 4)
To sample granular
and powdered material
in piles, sacks,
fiberdrums, etc.
To sample waste piles
1000-,2000- To contain solid and
ml(l-qt,2-qt) liquid samples except
linear pesticides and ch]or-
polyethylene inated hydrocarbons
Coverall, Protective garment
long-
sleeved,
cotton
Suit,
neoprene
rubber, long-
sleeved
Protective garment
Gloves, neo- Protective garment
prene rubber
Telescoping handle $ 16.24
available at
swimming pool
supply houses
Curtin-Matheson $ 25.00
Scientific
470 Valley Drive
P.O.Box 386
Brisbane,Calif.
94005
Fabricate. PVC $ 3.00
pipe available at
hardware stores
(see Section 4)
Laboratory supply $ 11.OO/
houses pkg.6
Clothing stores $ 14.00
MSA,Catalog #33496 $ 210.00
Laboratory supply $ 4.20
houses
64
-------
APPENDIX C (continued).
Quantity
Item
Use
Supplier
Approximate
Cost
1
pair
1
1
1
12
each
Self-contained For use in atmospheres MSA,Catalog
breathing deficient in oxygen #461704,
apparatus or otherwise immedi- Model 401 or
ately dangerous to equivalent
life.
Respirator, For use in atmospheres Comfo 11 Respir-
chemical not immediately ator, MSA,Catalog
cartridge dangerous to life #460968
type or equivalent
Cartridges for For use in atmospheres CMC Cartridge,MSA
respirator not immediately Cat.#459317 & GMD
dangerous to life Cartridge, MSA
Cat.#459318 or
equivalent
Goggles
Portable
eyewash
Fire extin-
quisher
Hard hat
Gas mask
18.9-liter
(5-gal) water
in cubitainer
or equivalent
with spigot
Teflon liners
for Bakelite
caps
Eye protection
For emergency
eyewash
Fire suppression
Head protection
For use in contami-
nated atmospheres
immediately dangerous
to life
For miscellaneous
washing purposes
To provide inert cap
liners
Sample labels, To document sample
seals, sample
analysis re-
quest sheets,
chain of
custody
records
65
MSA,Cat.#79179 or
equivalent
Laboratory supply
houses
Scientific Products
S1365-1 or equiva-
lent
MSA, Cat.#454740 or
equivalent
MSA, Cat.#448983 or
equivalent
Laboratory supply
houses
Scientific Special-
ties, P.O.Box 352
Randallstown, Md.
21133 or other
suppliers
Design using infor-
mation from Section
6
$580.00
$ 9.00
$ 5.00
$ 5.00
$ 4.00
$ 60.00
$ 5.00
$ 70.00
$ 5.50
$ 9.00
-------
APPENDIX C (continued).
Quantity Item
1
1
1
6
12
4
1
1
1
1
Field log book
(Section 6)
Weighted bottom
sampler
(Section 4)
Disposable
towels or
rags
Large poly-
ethylene bags
Polyethylene
bags
Waterproof
pens
Technical
grade
trichloro-
ethylene
Apron, oil
and acid
proof
Face mask
18.9 liter
(5-gal)
can
Use
To keep sample records
To sample storage
tanks or similar
containers
To clean sampling
equipment
To store waste papers,
rags, etc.
To store sample
containers
To complete records
and labels
To clean samplers
Protective garment
Protective garment
To store used
cleaning solvent
Supplier Approximate
Cost
Office supply
stores
Fabricate (see
Section 4 and
Figure 9)
Terry towels or
$ 2.00
$ 25.00
$ 4.00/
equivalent. Avail- pkg.
able at chemical
supply houses
Plastic supply
houses
Plastic supply
houses
Stationery stores
Chemical supply
stores
McMaster-Carr Co.
P.O.Box 4355
Chicago, 111.
MSA
400 Penn Center
Blvd.
Pittsburg, Pa.
15235
Hardware stores
$ 11. OO/
pkg/100
$ 4.00/
pkg/100
$ 3.00
$ 22. 00 /
gal.
$ 9.00
$ 4.00
$ 5.00
66
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APPENDIX D. RANDOM SAMPLING
Random Numbers
03
97
16
12
55
16
84
63
33
57
18
26
23
52
37
70
56
99
16
31
47
74
76
56
59
22
42
01
21
60
18
62
42
36
85
29
62
49
08
16
43
24
62
85
56
77
17
63
12
86
07
38
40
28
94
17
18
57
15
93
73
67
27
99
35
94
53
78
34
32
92
97
64
19
35
12
37
22
04
32
86
62
66
26
64
39
31
59
29
44
46
75
74
95
12
13
35
77
72
43
36
42
56
96
38
49
57
16
78
09
44
84
82
50
83
40
96
88
33
50
96
81
50
96
54
54
24
95
64
47
17
16
97
92
39
33
83
42
27
27
47
14
26
68
82
43
55
55
56
27
16
07
77
26
50
20
50
95
14
89
36
57
71
27
46
54
06
67
07
96
58
44
77
11
08
38
87
45
34
87
61
20
07
31
22
82
88
19
82
54
09
99
81
97
30
26
75
72
09
19
46
42
32
05
31
17
77
98
52
49
79
83
07
00
42
13
97
16
45
20
98
53
90
03
62
37
04
10
42
17
83
11
45
56
34
89
12
64
59
15
63
32
79
72
43
93
74
50
07
46
86
46
32
76
07
51
25
36
34
37
71
37
78
93
09
23
47
71
44
09
19
32
14
31
96
03
93
16
68
00
i
62
32
53
15
90
78
67
75
38
62
62
24
08
38
88
74
47
00
49
49
HOW TO USE THE TABLE OF RANDOM NUMBERS:
1. Based on available information, segregate the containers (i.e., drums,
sacks, etc.) according to waste types.
2. Number the containers containing the same waste types consecutively,
starting from 01.
3. Decide on how many samples you wish to take. This number is usually
determined by the objective of the sampling. For regular surveil-
lance sampling, the collection of one or two samples is usually
adequate. In this case, random sampling is not necessary. But for
regulatory or research purposes, more samples (such as one sample for
every group of five containers) taken at random will generate more
statistically valid data. Hence if there were 20 drums containing
the same type of waste, 5 drums have to be sampled.
4. Using the set of random numbers above, choose any number as a starting
point.
5. From this number, go down the column, then to the next column to the
right, or go in any predetermined direction until you have selected
five numbers between 01 and 20, with no repetitions. Larger numbers
are ineligible.
Example: If you were to choose 19 as the starting point on column
four, the next eligible numbers as you go down this
oolumn are 12 and 04. So far you have chosen only three
67
-------
eligible numbers. Proceed to the next column to the right.
Going down and starting from the top of this column, the
next eligible numbers are 12 and 13. But 12 is already
chosen. Proceeding to the sixth column, the next eligible
number is 16. Your five random numbers, therefore, are
19, 12, 04, 13 and 16. Thus the drums with corresponding
numbers have to be sampled.
APPENDIX E. SYSTEMATIC ERRORS USING THE COLIWASA
Certain systematic errors may occur in the determination of relative
phase composition of waste when using the Coliwasa. This error, in which
certain phases are disproportionately represented, results from the use
of a straight-sided sample tube to sample a container (tank truck) with
a circular cross section. On the basis of a two-phase system, error is
at a minimum when the phase interface is at the tank center and at a
maximum when the interface is near the bottom or top of the tank. These
errors do not occur when sampling a drum or other container when sampling
is done down the axis of the container (cylinder).
Errors in relative phase composition encountered in sampling the
typical cylindrical vacuum truck may be estimated using Table E-l. Num-
bers given in the table are representative values calculated from the
equations given below, which relate the geometry of the sample tube to
the geometry of the tank truck.
% A(tank) =
% A(sample) = 1-Cos % 9
(9-Sin 9)
2TT
Vacuum truck
Sample tube
68
-------
TABLE E-l. SAMPLE VOLUME CORRECTION FACTORS WHEN
SAMPLING CYLINDRICAL TANKS WITH COLIWASA
% A in sample % A in tank Correction (%)
10
20
30
40
50
60
70
80
90
100
5.20
14.2
25.2
37.4
50
62.6
74.8
85.8
94.8
100
+ 4.80
+ 5.8
+ 4.8
+ 2.6
0
- 2.6
- 4.8
- 5.8
- 4.8
0
69
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO. 2.
EPA-600/2-80-018
4. TITLE ANDSUBTITLE
Samplers and Sampling Procedures for Hazardous
Waste Streams
7' AUTHORifmil R. deVera, Bart P. Simmons, Robert D.
Stephens and David L. Storm,
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Hazardous Materials Laboratory
California Department of Health Services
2151 Berkeley Way
Berkeley, CA 94704
12. SPONSORING AGENCY NAME AND ADDRESS Cln . , UH
Municipal Environmental Research Laboratory —
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
3. RECIPIENT'S ACCESSIOWNO.
5. REPORT DATE
January 1980 (Issuing Date
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT
10. PROGRAM ELEMENT NO.
C73D1C, SOS #1, Task 32
11. CONTRACT/GRANT NO.
R 804692010
13. TYPE OF REPORT AND PERIOD COVEI
Final
14. SPONSORING AGENCY CODE
EPA/ 600/14
15. SUPPLEMENTARY NOTES
Richard A. Games, Project Officer (513/684-7871)
16. ABSTRACT
The goal of this project was to develop simple but effective sampling equipment
and procedures for collecting, handling, storing, and recording samples of hazardous
wastes. The report describes a variety of sampling devices designed to meet the nee
of those who regulate and manage hazardous wastes. Particular emphasis is given to
the development of a composite liquid waste sampler, the Coliwasa. This simple devi
is designed for use on liquid and semi-liquid wastes in a variety of containers, tan
and ponds. Devices for sampling solids and soils are also described.
In addition to the sampling devices, the report describes procedures for develc
ment of a sampling plan, sample handling, safety precautions, proper recordkeeping
and chain of custody, and sample containment, preservation, and transport. Also
discussed are certain limitations and potential sources of error that exist in the
sampling equipment and the procedures. The statistics of sampling are covered
briefly, and additional references in this area are given.
17. KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS
Samplers
Lagoons (ponds) — waste disposal
Hazardous materials
13. DISTRIBUTION STATEMENT
Release unlimited
b. IDENTIFIERS/OPEN ENDED TERMS
Representative sampling
Composite sampling
Sampling plans
Sampling procedures
Hazardous waste
Composite liquid waste
sampler
19. SECURITY CLASS (This Report)
unclassified
20. SECURITY CLASS (This page)
unclassified
c. COSATI Field/Grou
68C
21. NO. OF PAGES
78
22. PRICE
EPA Form 2220-1 (9-73)
70
- U S €0VCffl¥M£iV7 WW7WG OfFICf 1960-6 57-1
-------
APPENDIX II
FOR CHEMICAL ANALYSIS
OF WATER AND WASTES
-------
EI'A-600 4-79-020
METHODS FOR CHEMICAL ANALYSIS
OF WATER AND WASTES
March 1979
ENVIRONMENTAL MONITORING AND SUPPORT
LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
-------
DISCLAIMER
The mention of trade names or commercial products in this manual is for illustration purposes, and
does not constitute endorsement or recommendation for use by the U.S. Environmental Protection
Agency.
-------
FOREWORD
The accomplishment of our objective in protecting the environment requires a reliable assessment of
the present condition and a determination of the effectiveness of corrective measures. Decisions
which must be made on the need for pollution abatement and the most efficient means of achieving
environmental quality depend upon the availability of sound data. Test procedures for measurement
of the presence and concentration of substances hazardous to human health as well as an evaluation
of the quality of the environment are essential to satisfactory decision-making.
This manual of chemical methods was prepared by the staff of the Environmental Monitoring and
Support Laboratory of the Environmental Research Laboratory, Cincinnati to provide procedures
for monitoring water supplies, waste discharges, and the quality of ambient waters. These, test
methods have been carefully selected to meet the needs of Federal Legislation and to provide
guidance to laboratories engaged in protecting human health and the aquatic environment. The
contributions and counsel of scientists in other EPA laboratories are gratefully acknowledged.
Test procedures contained herein, that are approved for water and waste monitoring under the Safe
Drinking Water Act (SOWA) and the National Pollutant Discharge Elimination System (NPDES),
of PL 92-500 are so indicated at the bottom of each title page. These approved methods are also
recommended for ambient monitoring needs of Section 106 and 208 of PL 92-500. Methods without
this stated approval are presented for information only. Correspondence on these methods is invited.
Dwight G. Ballinger
Director, Environmental Monitoring and
Support Laboratory, Cincinnati, Ohio 45268
111
-------
ABSTRACT
This manual provides test procedures approved for the monitoring of water supplies, waste
discharges, and ambient waters, under the Safe Drinking Water Act, the National Pollutant
Discharge Elimination System, and Ambient Monitoring Requirements of Section 106 and 208
of Public Law 92-500. The test methods have been selected to meet the needs of federal legislation
and to provide guidance to laboratories engaged in the protection of human health and the
aquatic environment.
IV
-------
CONTENTS
Foreword iii
Abstract iv
Introduction xiii
Sample Preservation xv
EPA Quality Assurance Coordinators xx
100 Physical Properties
Color
Colorimetric, ADMI Method 110.1
Colorimetric, Platinum-Cobalt Method 110,2
Spectrophotometric Method. 110.3
Conductance
Specific Conductance Method 120.1
Hardness, Total (mg/1 as CaCO3)
Colorimetric, Automated EDTA Method 130.1
Titrimetric, EDTA Method 130.2
Odor
Threshold Odor (Consistent Series) Method 140,1
PH
Electrometric Method 150.1
Residue
Filterable
Gravimetric, Dried at 180°C Method 160.1
Non-Filterable
Gravimetric, Dried at 103-105°C Method 160.2
Total
Gravimetric, Dried at 103-105°C Method 160.3
Volatile
Gravimetric, Ignition at 550°C Method 160.4
Settleable Matter
Volumetric, Imhoff Cone Method 160.5
-------
Temperature
Thermometric Method 170.1
Turbidity
Nephelometric Method 180.1
200 Metals
Atomic Absorption Methods Section 200.0
Aluminum
AA, Direct Aspiration Method 202.1
AA, Furnace Method 202.2
Antimony
AA, Direct Aspiration Method 204.1
AA, Furnace Method 204.2
Arsenic
AA, Furnace Method 206.2
AA, Hydride Method 206.3
Spectrophotometric, SDDC Method 206.4
Digestion Method for Hydride and SDDC Method 206.5
Barium
AA, Direct Aspiration Method 208.1
AA, Furnace Method 208.2
Beryllium
AA, Direct Aspiration Method 210.1
AA, Furnace Method 210.2
Boron
Colorimetric, Curcumin Method 212.3
Cadmium
AA, Direct Aspiration Method 213.1
AA, Furnace Method 213.2
Calcium
AA, Direct Aspiration Method 215.1
Titrimetric, EDTA Method 215.2
VI
-------
Chromium
AA, Direct Aspiration Method 218.1
AA, Furnace Method 218.2
Chelation-Extraction Method 218.3
Hexavalent, Chelation-Extraction Method 218.4
Cobalt
AA, Direct Aspiration Method 219.1
AA, Furnace Method 219.2
Copper
AA, Direct Aspiration Method 220.1
AA, Furnace Method 220.2
Gold
AA, Direct Aspiration Method 231.1
AA, Furnace Method 231.2
Iridium
AA, Direct Aspiration Method 235.1
AA, Furnace Method 235.2
Iron
AA, Direct Aspiration Method 236.1
AA, Furnace Method 236.2
Lead
AA, Direct Aspiration Method 239.1
AA, Furnace Method 239.2
Magnesium
AA, Direct Aspiration Method 242.1
Manganese
AA, Direct Aspiration Method 243.1
AA, Furnace Method 243.2
Mercury
Cold Vapor, Manual Method 245.1
Cold Vapor, Automated Method 245.2
Cold Vapor, Sediments Method 245.5
vn
-------
Molybdenum
AA, Direct Aspiration Method 246.1
AA, Furnace Method 246.2
Nickel
AA, Direct Aspiration Method 249.1
AA, Furnace Method 249.2
Osmium
AA, Direct Aspiration Method 252.1
AA, Furnace Method 252.2
Palladium
AA, Direct Aspiration Method 253.1
AA, Furnace Method 253.2
Platinum
AA, Direct Aspiration Method 255.1
AA, Furnace Method 255.2
Potassium
AA, Direct Aspiration Method 258.1
Rhenium
AA, Direct Aspiration Method 264.1
AA, Furnace Method 264.2
Rhodium
AA, Direct Aspiration Method 265.1
AA, Furnace Method 265.2
Ruthenium
AA, Direct Aspiration Method 267.1
AA, Furnace Method 267.2
Selenium
AA, Furnace Method 270.2
AA, Hydride Method 270.3
Silver
AA, Direct Aspiration Method 272.1
AA, Furnace Method 272.2
VIM
-------
Sodium
AA, Direct Aspiration Method 273.1
Thallium
AA, Direct Aspiration Method 279.1
AA, Furnace Method 279.2
Tin
AA, Direct Aspiration Method 282.1
AA, Furnace Method 282.2
Titanium
AA, Direct Aspiration Method 283.1
AA, Furnace Method 283.2
Vanadium
AA, Direct Aspiration Method 286.1
AA, Furnace Method 286.2
Zinc
AA, Direct Aspiration Method 289.1
AA, Furnace Method 289.2
300 Inorganic, Non-metallics
Acidity
Titrimetric Method 305.1
Alkalinity
Titrimetric (pH 4.5) Method 310.1
Colorimetric, Automated Methyl Orange Method 310.2
Bromide
Titrimetric Method 320.1
Chloride
Colorimetric, Automated Ferricyanide, AA I Method 325.1
Colorimetric, Automated Ferricyanide, AA II Method 325.2
Titrimetric, Mercuric Nitrate Method 325.3
Chlorine, Total Residual
Titrimetric, Amperometric Method 330.1
Titrimetric, Back-Iodometric Method 330.2
IX
-------
Titrimetric, lodometric Method 330.3
Titrimetric, DPD-FAS Method 330.4
Spectrophotometric, DPD Method 330.5
Cyanide
Amenable to Chlorination
Titrimetric, Spectrophotometric Method 335.1
Total
Titrimetric, Spectrophotometric Method 335.2
Colorimetric, Automated UV Method 335.3
Fluoride
Colorimetric, SPADNS with Bellack
Distillation Method 340.1
Potentiometric, Ion Selective Electrode Method 340.2
Colorimetric, Automated Complexone Method 340.3
Iodide
Titrimetric Method 345.1
Nitrogen
Ammonia
Colorimetric, Automated Phenate Method 350.1
Colorimetric; Titrimetric; Potentiometric -
Distillation Procedure Method 350.2
Potentiometric, Ion Selective Electrode Method 350.3
Kjeldahl, Total
Colorimetric, Automated Phenate Method 351.1
Colorimetric, Semi-Automated
Block Digester AAII Method 351.2
Colorimetric; Titrimetric; Potentiometric Method 351.3
Potentiometric, Ion Selective Electrode Method 351.4
Nitrate
Colorimetric, Brucine Method 352.1
Nitrate-Nitrite
Colorimetric, Automated Hydrazine
Reduction Method 353.1
Colorimetric, Automated Cadmium Reduction Method 353.2
Colorimetric, Manual Cadmium Reduction Method 353.3
-------
Nitrite
Spectrophotometric Method 354.1/
Oxygen, Dissolved
Membrane Electrode Method 360.1
Modified Winkler (Full Bottle Technique) Method 360.2
Phosphorus
All Forms
Colorimetric, Automated, Ascorbic Acid Method 365.1
Colorimetric, Ascorbic Acid,
Single Reagent Method 365.2
Colorimetric, Ascorbic Acid,
Two Reagent Method 365.3
Total
Colorimetric, Automated, Block Digestor, AA II Method 365.4
Silica, Dissolved
Colorimetric Method 370.1
Sulfate
Colorimetric, Automated Chloranilate Method 375.1
Colorimetric, Automated Methyl Thymol Blue, AA II Method 375.2
Gravimetric Method 375.3
Turbidimetric Method 375.4
Sulfide
Titrimetric, Iodine Method 376.1
Colorimetric, Metnylene Blue Method 376.2
Sulfite
Titrimetric Method 377.1
400 Organics
Biochemical Oxygen Demand
BOD (5 day, 20°C) Method 405.1
Chemical Oxygen Demand
Titrimetric, Mid-Level Method 410.1
Titrimetric, Low Level Method 410.2
Titrimetric, High Level for Saline Waters Method 410.3
Colorimetric, Automated; Manual Method 410.4
XI
-------
Oil and Grease, Total Recoverable
Gravimetric, Separatory Funnel Extraction .-. Method 413.1
Spectrophotometric, Infrared Method 413.2
Organic Carbon, Total
Combustion or Oxidation Method 415.1
Petroleum Hydrocarbons, Total, Recoverable
Spectrophotometric, Infrared Method 418.1
Phenolics, Total Recoverable
Spectrophotometric, Manual 4-AAP with Distillation Method 420.1
Colorimetric, Automated 4-AAP with Distillation Method 420.2
Spectrophotometric, MBTH with Distillation Method 420.3
Methylene Blue Active Substances (MbAS)
Colorimetric Method 425.1
NTA
Colorimetric, Manual, Zinc-Zincon Method 430.1
Colorimetric, Automated, Zinc-Zincon, Method 430.2
xn
-------
INTRODUCTION
This third edition of "Methods for Chemical Analysis of Water and Wastes" contains the chemical
analytical procedures used in U.S. Environmental Protection Agency (EPA) laboratories for the
examination of ground and surface waters, domestic and industrial waste effluents, and treatment
process samples. Except where noted under "Scope and Application", the methods are applicable to
both water and wastewaters, and both fresh and saline water samples. The manual provides test
procedures for the measurement of physical, inorganic, and selected organic constituents and
parameters. Methods for pesticides, industrial organic waste materials, and sludges are given in other
publications of the Agency. The methods were chosen through the combined efforts of the EPA
Regional Quality Assurance Coordinators, the staff of the Physical and Chemical Methods Branch,
Environmental Monitoring and Support Laboratory, and other senior chemists in both federal and
state laboratories. Method selection was based on the following criteria:
(1) The method should measure the desired property or constituent with precision,
accuracy, and specificity sufficient to meet the data needs of EPA, in the presence of the
interfering materials encountered in water and waste samples.
(2) The procedure should utilize the equipment and skills available in modern water
pollution control laboratories.
(3) The selected method is in use in many laboratories or has been sufficiently tested to
establish its validity.
(4) The method should be rapid enough to permit routine use for the examination of a large
number of samples.
•
Instrumental methods have been selected in preference to manual procedures because of the
improved speed, accuracy, and precision. In keeping with this policy, procedures for the Technicon
AutoAnalyzer have been included for laboratories having this equipment available. Other
continuous flow automated systems using these identical procedures are acceptable.
Intralaboratory and interlaboratory precision and accuracy statements are provided where such data
are available. These interlaboratory statements are derived from interlaboratory studies conducted
by the Quality Assurance Branch, Environmental Monitoring and Support Laboratory; the
American Society for Testing Materials; or the Analytical Reference Service of the US Public Health
Service, DHEW. These methods may be used for measuring both total and dissolved constituents of
the sample. When the dissolved concentration is to be determined, the sample is filtered through a
0.45-micron membrane filter and the filtrate analyzed by the procedure specified. The sample should
be filtered as soon as possible after it is collected, preferably in the field. Where field filtration is not
practical, the sample should be filtered as soon as it is received in the laboratory.
Many water and waste samples are unstable. In situations where the interval between sample
collection and analysis is long enough to produce changes in either the concentration or the physical
state of the constituent to be measured, the preservation practices in Table I are recommended.
xiii
-------
This manual is a basic reference for monitoring water and wastes in compliance with the
requirements of the Federal Water Pollution Control Act Amendments of 1972. Although other test
procedures may be used, as provided in the Federal Register issue of October 16,1973 (38FR 28758)
and in subsequent amendments, the methods described in this manual will be used by the
Environmental Protection Agency in determining compliance with applicable water and effluent
standards established by the Agency.
Although a sincere effort has been made to select methods that are applicable to the widest range of
sample types, significant interferences may be encountered in certain isolated samples. In these
situations, the analyst will be providing a valuable service to EPA by defining the nature of the
interference with the method and bringing this information to the attention of the Director,
Environmental Monitoring and Support Laboratory, through the appropriate Quality Assurance
Coordinator.
xiv
-------
SAMPLE PRESERVATION
Complete and unequivocal preservation of samples, either domestic sewage, industrial wastes, or
natural waters, is a practical impossibility. Regardless of the nature of the sample, complete stability
for every constituent can never be achieved. At best, preservation techniques can only retard the
chemical and biological changes that inevitably continue after the sample is removed from the parent
source. The changes that take place in a sample are either chemical or biological. In the former case,
certain changes occur in the chemical structure of the constituents that are a function of physical
conditions. Metal cations may precipitate as hydroxides or form complexes with other constituents;
cations or anions may change valence states under certain reducing or oxidizing conditions; other
constituents may dissolve or volatilize with the passage of time. Metal cations may also adsorb onto
surfaces (glass, plastic, quartz, etc.), such as, iron and lead. Biological changes taking place in a
sample may change the valence of an element or a radical to a different valence. Soluble constituents
may be converted to organically bound materials in cell structures, or cell lysis may result in release
of cellular material into solution. The well known nitrogen and phosphorus cycles are examples of
biological influence on sample composition. Therefore, as a general rule, it is best to analyze the
samples as soon as possible after collection. This is especially true when the analyte concentration is
expected to be in the low ug/1 range.
Methods of preservation are relatively limited and are intended generally to (1) retard biological
action, (2) retard hydrolysis of chemical compounds and complexes, (3) reduce volatility of
constituents, and (4) reduce absorption effects. Preservation methods are generally limited to pH
control, chemical addition, refrigeration, and freezing.
The recommended preservative for various constituents is given in Table 1. These choices are based
on the accompanying references and on information supplied by various Quality Assurance
Coordinators. As more data become available, these recommended holding times will be adjusted to
reflect new information. Other information provided in the table is an estimation of the volume of
sample required for the analysis, the suggested type of container, and the maximum recommended
holding times for samples properly preserved.
xv
-------
TABLE 1
RECOMMENDATION FOR SAMPLING AND PRESERVATION
OF SAMPLES ACCORDING TO MEASUREMENT'15
Measurement
100 Physical Properties
Color
Conductance
Hardness
Odor
pH
Residue
Filterable
Non-
Filterable
Total
Volatile
Settleable Matter
Temperature
Turbidity
200 Metals
Dissolved
Suspended
Total
Vol.
Req.
(ml)
50
100
100
200
25
100
100
100
100
1000
1000
100
200
200
100
Container(2)
P,G
P,G
P,G
G only
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
Preservative
Cool, 4°C
Cool, 48C
Cool, 4'C
HNO3 to pH<2
Cool, 4°C
Det. on site
Cool, 4°C
Cool, 4°C
Cool, 4°C
Cool, 4°C
None Req.
Det. on site
Cool, 4°C
Filter on site
HNO3 to pH<2
Filter on site
HNO3 to pH<2
Holding
Time(3)
24 Hrs.
24 Hrs.(4)
6 Mos.<5)
24 Hrs.
6 Hrs.
7 Days
7 Days
7 Days
7 Days
24 Hrs.
No Holding
7 Days
6 Mos.(!)
6 Mos.
6 Mos.("
XVI
-------
TABLE 1 (CONT)
Vol.
Req.
Measurement (ml)
Mercury
Dissolved
Total
300 Inorganics, Non-Metallics
Acidity
Alkalinity
Bromide
Chloride
Chlorine
Cyanides
Fluoride
Iodide
Nitrogen
Ammonia
Kjeldahl, Total
Nitrate plus Nitrite
Nitrate
Nitrite
100
100
100
100
100
50
200
500
300
100
400
500
100
100
50
Container"' Preservative
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
P,G
Filter on site
HNO3 to pH<2
HNO3 to pH<2
None Req
Cool, 4°C
Cool, 48C
None Req.
Det. on site
Cool, 4°C
NaOH to pH 12
None Req.
Cool, 4°C
Cool,4°C
H2SO4 to pH<2
Cool, 4°C
H2SO4 to pH<2
Cool, 4°C
H2SO4 to pH<2
Cool, 4°C
Cool, 4°C
Holding
Time(3)
38 Days
(Glass)
13 Days
(Hard
Plastic)
38 Days
(Glass)
13 Days
(Hard
Plastic)
24 Hrs.
24 Hrs.
24 Hrs.
7 Days
No Holding
24 Hrs.
7 Days
24 Hrs.
24 Hrs.
24 Hrs.(6)
24 Hrs.(6)
24 Hrs.
48 Hrs.
XVII
-------
TABLE 1 (CONT)
Measurement
Dissolved Oxygen
Probe
Winkler
Phosphorus
Ortho-
phosphate,
Dissolved
Hydrolyzable
Total
Total,
Dissolved
Silica
Sulfate
Sulfide
Sulfite
400 Organics
BOD
COD
Oil & Grease
Organic carbon
Phenolics
Vol.
Req.
(ml)
300
300
50
50
50
50
50
50
500
50
1000
50
1000
25
500
Container"'
G only
G only
P,G
P,G
P,G
P,G
P only
P,G
P,G
P,G
P,G
P,G
G only
P,G
G only
Preservative
Det. on site
Fix on site
Filter on site
Cool, 4°C
Cool, 4'C
H2SO4 to pH<2
Cool, 4'C
H2S04 to pH<2
Filter on site
Cool, 4'C
H2SO4 to pH<2
Cool, 4°C
Cool, 4°C
2 ml zinc
acetate
Det. on site
Cool, 4°C
H2SO4 to pH<2
Cool, 4"C
H2SO4 or HC1 to pH<2
Cool, 4°C
H2SO4 or HC1 to pH<2
Cool, 4°C
Holding
Time(3)
No Holding
4-8 Hours
24 Hrs.
24 Hrs.(<)
24 Hrs.("
24 Hrs.'"
7 Days
7 Days
24 Hrs.
No Holding
24 Hrs.
7 Days'"
24 Hrs.
24 Hrs.
24 Hrs.
MBAS
250
P,G
H3PO4 to pH<4
1.0 g CuSO4/l
Cool, 4°C
24 Hrs.
XVlll
-------
TABLE 1 (CONT)
Vol.
Req. Holding
Measurement (ml) Container*2* Preservative Time(3)
NTA 50 P,G Cool, 4°C 24 Hrs.
1. More specific instructions for preservation and sampling are found with each procedure as
detailed in this manual. A general discussion on sampling water and industrial wastewater may
be found in ASTM, Part 31, p. 72-82 (1976) Method D-3370.
2. Plastic (P) or Glass (G). For metals, polyethylene with a polypropylene cap (no liner) is
preferred.
3. It should be pointed out that holding times listed above are recommended for properly
preserved samples based on currently available data. It is recognized that for some sample
types, extension of these times may be possible while for other types, these times may be too
long. Where shipping regulations prevent the use of the proper preservation technique or the
holding time is exceeded, such as the case of a 24-hour composite, the final reported data for
these samples should indicate the specific variance.
4. If the sample is stabilized by cooling, it should be warmed to 25°C for reading, or temperature
correction made and results reported at 25°C.
5. Where HNO3 cannot be used because of shipping restrictions, the sample may be initially
preserved by icing and immediately shipped to the laboratory. Upon receipt in the laboratory,
the sample must be acidified to a pH <2 with HNO3 (normally 3 ml 1:1 HNO3/liter is
sufficient). At the time of analysis, the sample container should be thoroughly rinsed with 1:1
HNO3 and the washings added to the sample (volume correction may be required).
6. Data obtained from National Enforcement Investigations Center-Denver, Colorado, support a
four-week holding time for this parameter in Sewerage Systems. (SIC 4952).
xix
-------
ENVIRONMENTAL PROTECTION AGENCY
REGIONAL QUALITY ASSURANCE COORDINATORS
REGION I
Warren H. Oldaker
New England Region Laboratory
60 Westview Street
Lexington, MA 02173
(617-861-6700)
FTS 861-6700
REGION II
Gerard F. McKenna
Edison Environmental Lab.
Edison, NJ 08817
(201-321-6645)
FTS 340-6645
REGION V
David Payne
Central Regional Lab.
536 South Clark Street
Chicago, IL 60605
(312-353-9351)
FTS 353-9351
REGION VI
Myron Knudson
1201 Elm St., First Int'l Bldg.
Dallas, TX 75270 •
(214-767-2697)
FTS 729-2697
REGION IX
Dr. Ho Young
215 Fremont Street
San Francisco, CA 94102
(415-556-2270)
FTS 555-2270
REGION X
Arnold R. Gahler
1555 Alaskan Way, South
Bldg 3
Seattle, WA 98134
(206-442-5840)
FTS 399-5840
REGION III
Charles Jones, Jr.
(3SA60)
6th & Walnut Streets
Philadelphia, PA 19106
(215-597-9162)
FTS 597-9162
REGION IV
Bobby J. Carroll
Southeast Envr. Res. Lab.
College Station Road
Athens, GA 30601
(404-546-3111)
FTS 250-3111
REGION VII
Dr. Harold G. Brown
25 Funston Road
Kansas City, KS 66115
(816-374-4285)
FTS 758-4285
REGION VIII
Douglas M. Skie
1860 Lincoln St.
Denver, CO 80295
(303-837-4935)
FTS 327-4935
xx
-------
pH
Method 150.1 (Electrometric)
STORET NO.
Determined on site 00400
Laboratory 00403
1. Scope and Application
1.1 This method is applicable to drinking, surface, and saline waters, domestic and industrial
wastes.
2. Summary of Method
2.1 The pH of a sample is determined electrometrically using either a glass electrode in
combination with a reference potential or a combination electrode.
3. Sample Handling and Preservation
3.1 Samples should be analyzed as soon as possible preferably in the field at the time of
sampling.
3.2 High-purity waters and waters not at equilibrium with the atmosphere are subject to
changes when exposed to the atmosphere, therefore the sample containers should be
filled completely and kept sealed prior to analysis.
4. Interferences
4.1 The glass electrode, in general, is not subject to solution interferences from color,
turbidity, colloidal matter, oxidants, reductants or high salinity.
4.2 Sodium error at pH levels greater than 10 can be reduced or eliminated by using a "low
sodium error" electrode.
4.3 Coatings of oily material or paniculate matter can impair electrode response. These
coatings can usually be removed by gentle wiping or detergent washing, followed by
distilled water rinsing. An additional treatment with hydrochloric acid (1+9) may be
necessary to remove any remaining film.
4.4 Temperature effects on the electrometric measurement 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 is the change of pH inherent in the sample at various temperatures. 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.
5. Apparatus
5.1 pH Meter-laboratory or field model. A wide variety of instruments are commercially
available with various specifications and optional equipment.
Approved for NPDES
Issued 1971
Editorial revision 1978
150.1-1
-------
5.2 Glass electrode.
5.3 Reference electrode-a calomel, silver-silver chloride or other reference electrode of
constant potential may be used.
NOTE 1: Combination electrodes incorporating both measuring and reference
functions are convenient to use and are available with solid, gel type filling materials that
require minimal maintenance.
5.4 Magnetic stirrer and Teflon-coated stirring bar.
5.5 Thermometer or temperature sensor for automatic compensation.
6. Reagents
6.1 Primary standard buffer salts are available from the National Bureau of Standards and
should be used in situations where extreme accuracy is necessary.
6.1.1 Preparation of reference solutions from these salts require 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.
6.2 Secondary standard buffers may be prepared from NBS salts or purchased as a solution
from commercial vendors. Use of these commercially available solutions, that have been
validated by comparison to NBS standards, are recommended for routine use.
7. Calibration
7.1 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.
7.2 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.
7.2.1 Various instrument designs may involve use of a "balance" or "standardize" dial
and/or a slope adjustment as outlined in the manufacturer's instructions. Repeat
adjustments on successive portions of the two buffer solutions as outlined in
procedure 8.2 until readings are within 0.05 pH units of the buffer solution value.
8. Procedure
8.1 Standardize the meter and electrode system as outlined in Section 7.
8.2 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.
8.2.1 If field measurements are being made the electrodes may be immersed directly in
the sample stream to an adequate depth and moved in a manner to insure sufficient
sample movement across the electrode sensing element as indicated by drift free
( < 0.1 pH) readings.
8.3 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
"'National Bureau of Standards Special Publication 260.
150.1-2
-------
compensators that electronically adjust for temperature differences. Refer to
manufacturer's instructions.
8.4 After rinsing and gently wiping the electrodes, if necessary, immerse them into the
sample beaker or sample stream and stir at a constant rate to provide homogeneity and
suspension of solids. Rate of stirring should minimize the air transfer rate at the air water
interface of the sample. Note and record sample pH and temperature. Repeat
measurement on successive volumes of sample until values differ by less than 0.1 pH
units. Two or three volume changes are usually sufficient.
9. Calculation
9.1 pH meters read directly in pH units. Report pH to the nearest 0.1 unit and temperature
to the nearest °C.
10. Precision and Accuracy
10.1 Forty-four analysts in twenty laboratories analyzed six synthetic water samples
containing exact increments of hydrogen-hydroxyl ions, with the following results:
pH Units
3.5
3.5
7.1
7.2
8.0
8.0
Standard Deviation
pH Units
0.10
0.11
0.20
0.18
0.13
0.12
Bias,
%
Accuracy as
-0.29
-0.00
+ 1.01
-0.03
-0.12
+0.16
Bias,
pH Units
-0.01
+0.07
-0.002
-0.01
+0.01
(FWPCA Method Study 1, Mineral and Physical Analyses)
10.2 In a single laboratory (EMSL), using surface water samples at an average pH of 7.7, the
standard deviation was ±0.1.
Bibliography
1. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 460, (1975).
2. Annual Book of ASTM Standards, Part 31, "Water", Standard D1293-65, p 178 (1976).
150.1-3
-------
METALS
(Atomic Absorption Methods)
1. Scope and Application
1.1 Metals in solution may be readily determined by atomic absorption spectroscopy. The
method is simple, rapid, and applicable to a large number of metals in drinking, surface,
and saline waters, and domestic and industrial wastes. While drinking waters free of
particulate matter may be analyzed directly, domestic and industrial wastes require
processing to solubilize suspended material. Sludges, sediments and other solid type
samples may also be analyzed after proper pretreatment.
1.2 Detection limits, sensitivity and optimum ranges of the metals will vary with the various
makes and models of satisfactory atomic absorption spectrophotometers. The data
shown in Table 1, however, provide some indication of the actual concentration ranges
measurable by direct aspiration and using furnace techniques. In the majority of
instances the concentration range shown in the table by direct aspiration may be
extended much lower with scale expansion and conversely extended upwards by using a
less sensitive wavelength or by rotating the burner head. Detection limits by direct
aspiration may also be extended through concentration of the sample and/or through
solvent extraction techniques. Lower concentrations may also be determined using the
furnace techniques. The concentration ranges given in Table 1 are somewhat dependent
on equipment such as the type of spectrophotometer and furnace accessory, the energy
source and the degree of electrical expansion of the output signal. When using furnace
techniques, however, the analyst should be cautioned as to possible chemical'reactions
occurring at elevated temperatures which may result in either suppression or
enhancement of the analysis element. To insure valid data with furnace techniques, the
analyst must examine each matrix for interference effects (see 5.2.1) and if detected, treat
accordingly using either successive dilution, matrix modification or method of standard
additions (see 8.5).
1.3 Where direct aspiration atomic absorption techniques do not provide adequate
sensitivity, in addition to the furnace procedure, reference is made to specialized
procedures such as the gaseous hydride method for arsenic and selenium, the cold vapor
technique for mercury, and the chelation-extraction procedure for selected metals.
Reference to approved colorimetric methods is also made.
1.4 Atomic absorption procedures are provided as the methods of choice; however, other
instrumental methods have also been shown to be capable of producing precise and
accurate analytical data. These instrumental techniques include emission spectroscopy,
X-ray fluorescence, spark source mass spectroscopy, and anodic stripping to name but a
few. The analyst should be cautioned that these methods are highly specialized
techniques requiring a high degree of skill to interpret results and obtain valid data.
Approved for NPDES and SDWA
Issued 1969
Editorial revision 1974 and 1978
METALS-1
-------
These above mentioned techniques are presently considered as alternate test procedures
and approval must be obtained prior to their use.
2. Summary of Method
2.1 In direct aspiration atomic absorption spectroscopy a sample is aspirated and atomized
in a flame. A light beam from a hollow cathode lamp whose cathode is made of the
element to be determined is directed through the flame into a monochromator, and onto
a detector that measures the amount of light absorbed. Absorption depends upon the
presence of free unexcited ground state atoms in the flame. Since the wavelength of the
light beam is characteristic of only the metal being determined, the light energy absorbed
by the flame is a measure of the concentration of that metal in the sample. This principle
is the basis of atomic absorption spectroscopy.
2.2 Although methods have been reported for the analysis of solids by atomic absorption
spectroscopy (Spectrochim Acta, 24B 53, 1969) the technique generally is limited to
metals in solution or solubilized through some form of sample processing.
2.2.1 Preliminary treatment of wastewater and/or industrial effluents is usually
necessary because of the complexity and variability of the sample matrix.
Suspended material must be subjected to a solubilization process before analysis.
This process may vary because of the metals to be determined and the nature of the
sample being analyzed. When the breakdown of organic material is necessitated,
the process should include a wet digestion with nitric acid.
2.2.2 In those instances where complete characterization of a sample is desired, the
suspended material must be analyzed separately. This may be accomplished by
filtration and acid digestion of the suspended material. Metallic constituents in this
acid digest are subsequently determined and the sum of the dissolved plus
suspended concentrations will then provide the total concentrations present. The
sample should be filtered as soon as possible after collection and the filtrate
acidified immediately.
2.2.3 The total sample may also be treated with acid without prior filtration to measure
what may be termed "total recoverable" concentrations.
2.3 When using the furnace technique in conjunction with an atomic absorption
spectrophotometer, a representative aliquot of a sample is placed in the graphite tube in
the furnace, evaporated to dryness, charred, and atomized. As a greater percentage of
available analyte atoms are vaporized and dissociated for absorption in the tube than the
flame, the use of small sample volumes or detection of low concentrations of elements is
possible. The principle is essentially the. same as with direct aspiration atomic absorption
except a furnace, rather than a flame, is used to atomize the sample. Radiation from a
given excited element is passed through the vapor containing ground state atoms of that
element. The intensity of the transmitted radiation decreases in proportion to the amount
of the ground state element in the vapor.
METALS-2
-------
TABLE 1
Atomic Absorption Concentration Ranges'"
Direct Aspiration Furnace Procedure'415)
Metal
Aluminum
Antimony
Arsenic'21
Barium(p)
Beryllium
Cadmium
Calcium
Chromium
Cobalt
Copper
Gold
Iridium(p)
Iron
Lead
Magnesium
Manganese
Mercury"1
Molybdenum(p)
Nickel(p)
Osmium
Palladium(p)
Platinum(p)
Potassium
Rhenium(p)
Rhodium(p)
Ruthenium
Selenium"1
Silver
Sodium
Thallium
Tin
Titanium (p)
Vanadium (p)
Zinc
Detection
Limit
mg/1
0.1
0.2
0.002
0.1
0.005
0.005
0.01
0.05
0.05
0.02
0.1
3
0.03
0.1
0.001
0.01
0.0002
0.1
0.04
0.3
0.1
0.2
0.01
5
0.05
0.2
0.002
0.01
0.002
0.1
0.8
0.4
0.2
0.005
Optimum
Concentration
Sensitivity
mg/1
1
0.5
_
0.4
0.025
0.025
0.08
0.25
0.2
0.1
0.25
8
0.12
0.5
0.007
0.05
-
0.4
0.15
1
0.25
2
0.04
15
0.3
0.5
_
0.06
0.015
0.5
4
2
0.8
0.02
5
1
0.002
1
0.05
0.05
0.2
0.5
0.5
0.2
0.5
20
0.3
1
0.02
0.1
0.0002
1
0.3
2
0.5
5
0.1
50
1
1
0.002
0.1
0.03
1
10
5
2
0.05
Range
mg/1
50
40
0.02
20
2
2
7
10
5
5
20
- 500
_ 5
20
0.5
3
0.01
40
5
- 100
15
75
^
- 1000
30
50
0.02
4
1
20
- 300
- 100
- 100
1
Detection
Limit
ug/1
3
3
1
2
0.2
0.1
-
1
1
1
1
30
1
1
-
0.2
-
1
1
20
5
20
-
200
5
20
2
0.2
-
1
5
10
4
0.05
Optimum
Concentration
Range
ug/1
20
20
5
10
1 —
0.5 -
-
5
5
5
5
100
5
5
-
1
-
3
'5
50
20
100
-
500
20
100
5
1
-
5
20
50
10
0.2 -
200
300
too
200
30
10
-
100
100
100
100
1500
100
too
-
30
-
60
100
500
400
2000
5000
400
2000
100
25
-
100
300
500
200
4
(1) The concentrations shown are not contrived values and should be obtainable with any satisfactory atomic absorption
spectrophotometer.
(2) Gaseous hydride method.
(3) Cold vapor technique.
(4) For furnace sensitivity values consult instrument operating manual.
(5) The listed furnace values are those expected when using a 20 ul injection and normal gas flow except in the case of arsenic and
selenium where gas interrupt is used. The symbol (p) indicates the use of pyrolytic graphite with the furnace procedure.
METALS-3
-------
The metal atoms to be measured are placed in the beam of radiation by increasing the temperature of
the furnace thereby causing the injected specimen to be volatilized. A monochromator isolates the
characteristic radiation from the hollow cathode lamp and a photosensitive device measures the
attenuated transmitted radiation.
3. Definition of Terms
3.1 Optimum Concentration Range: A range, defined by limits expressed in concentration,
below which scale expansion must be used and above which curve correction should be
considered. This range will vary with the sensitivity of the instrument and the operating
condition employed.
3.2 Sensitivity: The concentration in milligrams of metal per liter that produces an
absorption of 1%.
3.3 Detection Limit: Detection limits can be expressed as either an instrumental or method
parameter. The limiting factor of the former using acid water standards would be the
signal to noise ratio and degree of scale expansion used; while the latter would be more
affected by the sample matrix and preparation procedure used. The Scientific Apparatus
Makers Association (SAMA) has approved the following definition for detection limit:
that concentration of an element which would yield an absorbance equal to twice the
standard deviation of a series of measurements of a solution, the concentraton of which is
distinctly detectable above, but close to blank absorbance measurement. The detection
limit values listed in Table I and on the individual analysis sheets are to be considered
minimum working limits achievable with the procedures given in this manual. These
values may differ from the optimum detection limit reported by the various instrument
manufacturers.
3.4 Dissolved Metals: Those constituents (metals) which will pass through a 0.45 u
membrane filter.
3.5 Suspended Metals: Those constituents (metals) which are retained by a 0.45 u membrane
filter.
3.6 Total Metals: The concentration of metals determined on an unfiltered sample following
vigorous digestion (Section 4.1.3), or the sum of the concentrations of metals in both the
dissolved and suspended fractions.
3.7 Total Recoverable Metals: The concentration of metals in an unfiltered sample following
treatment with hot dilute mineral acid (Section 4.1.4).
4. Sample Handling and Preservation
4.1 For the determination of trace metals, contamination and loss are of prime concern. Dust
in the laboratory environment, impurities in reagents and impurities on laboratory
apparatus which the sample contacts are all sources of potential contamination. For
liquid samples, containers can introduce either positive or negative errors in the
measurement of trace metals by (a) contributing contaminants through leaching or
surface desorption and (b) by depleting concentrations through adsorption. Thus the
collection and treatment of the sample prior to analysis requires particular attention. The
sample bottle whether borosilicate glass, linear polyethylene, polyproplyene or Teflon
should be thoroughly washed with detergent and tap water; rinsed with 1:1 nitric acid,
METALS-4
-------
tap water, 1:1 hydrochloric acid, tap water and finally deionized distilled water in that
order.
NOTE 1: Chromic acid may be useful to remove organic deposits from glassware;
however, the analyst should be cautioned that the glassware must be thoroughly rinsed
with water to remove the last traces of chromium. This is especially important if
chromium is to be included in the analytical scheme. A commercial product—
NOCHROMIX—available from Godax Laboratories, 6 Varick St. New York, N.Y.
10013, may be used in place of chromic acid. [Chromic acid should not be used with
plastic bottles.]
NOTE 2: If it can be documented through an active analytical quality control program
using spiked samples, reagent and sample blanks, that certain steps in the cleaning
procedure are not required for routine samples, those steps may be eliminated from the
procedure.
Before collection of the sample a decision must be made as to the type of data desired, i.e.,
dissolved, suspended, total or total recoverable. For container preference, maximum
holding time and sample preservation at time of collection see Table 1 in the front part of
this manual. Drinking water samples containing suspended and setteable material should
be prepared using the total recoverable metal procedure (section 4.1.4).
4.1.1 For the determination of dissolved constituents the sample must be filtered
through a 0.45 u membrane filter as soon as practical after collection. (Glass or
plastic filtering apparatus using plain, non-grid marked, membrane filters are
recommended to avoid possible contamination.) Use the first 50-100 ml to rinse
the filter flask. Discard this portion and collect the required volume of filtrate.
Acidify the filtrate with 1:1 redistilled HNO3 to a pH of <2. Normally, 3 ml of
(1:1) acid per liter should be sufficient to preserve the sample (See Note 3). If
hexavalent chromium is to be included in the analytical scheme, a portion of the
filtrate should be transferred before acidification to a separate container and
analyzed as soon as possible using Method 218.3. Analyses performed on a sample
so treated shall be reported as "dissolved" concentrations.
NOTE 3: If a precipitate is formed upon acidification, the filtrate should be digested
using 4.1.3. Also, it has been suggested (International Biological Program, Symposium
on Analytical Methods, Amsterdam, Oct. 1966) that additional acid, as much as 25 ml of
cone. HCl/liter, may be required to stabilize certain types of highly buffered samples if
they are to be stored for any length of time. Therefore, special precautions should be
observed for preservation and storage of unusual samples intended for metal analysis.
4.1.2 For the determination of suspended metals a representative volume of unpreserved
sample must be filtered through a 0.45 u membrane filter. When considerable
suspended material is present, as little as 100 ml of a well mixed sample is filtered.
Record the volume filtered and transfer the membrane filter containing the
insoluble material to a 250 ml Griffin beaker and add 3 ml cone, redistilled HNO3.
Cover the beaker with a watch glass and heat gently. The warm acid will soon
dissolve the membrane. Increase the temperature of the hot plate and digest the
material. When the acid has nearly evaporated, cool the beaker and watch glass
and add another 3 ml of cone, redistilled HNO3. Cover and continue heating until
METALS-5
-------
the digestion is complete, generally indicated by a light colored digestate.
Evaporate to near dryness (DO NOT BAKE), add 5 ml distilled HC1 (1:1) and
warm the beaker gently to dissolve any soluble material. (If the sample is to be
analyzed by the furnace procedure, 1 ml of 1:1 distilled HNO3 per 100 ml dilution
should be substituted for the distilled 1:1 HC1.) Wash down the watch glass and
beaker walls with deionized distilled water and filter the sample to remove silicates
and other insoluble material that could clog the atomizer. Adjust the volume to
some predetermined value based on the expected concentrations of metals present.
This volume will vary depending on the metal to be determined. The sample is now
ready for analysis. Concentrations so determined shall be reported as "suspended"
(See Note 4.)
NOTE 4: Certain metals such as antimony arsenic, gold, iridium, mercury,
osmium, palladium, platinium, rhenium, rhodium, ruthenium, selenium, silver,
thallium, tin and titanium require modification of the digestion procedure and the
individual sheets for these metals should be consulted.
4.1.3 For the determination of total metals the sample is acidified with 1:1 redistilled
HNO3 to a pH of less than 2 at the time of collection. The sample is not filtered
before processing. Choose a volume of sample appropriate for the expected level of
metals. If much suspended material is present, as little as 50-100 ml of well mixed
sample will most probably be sufficient. (The sample volume required may also
vary proportionally with the number of metals to be determined.)
Transfer a representative aliquot of the well mixed sample to a Griffin beaker and
add 3 ml of cone, redistilled HNO3. Place the beaker on a hot plate and evaporate
to near dryness cautiously, making certain that the sample does not boil. (DO NOT
BAKE.) Cool the beaker and add another 3 ml portion of cone, redistilled HNO3.
Cover the beaker with a watch glass and return to the hot plate. Increase the
temperature of the hot plate so that a gentle reflux action occurs. Continue heating,
adding additional acid as necessary, until the digestion is complete (generally
indicated when the digestate is light in color or does not change in appearance with
continued refluxing). Again, evaporate to near dryness and cool the beaker. Add a
small quantity of redistilled 1:1 HC1 (5 ml/100 ml of final solution) and warm the
beaker to dissolve any precipitate or residue resulting from evaporation. (If the
sample is to be analyzed by the furnace procedure, substitute distilled HNO3 for 1:1
HC1 so that the final dilution contains 0.5% (v/v) HNO3.) Wash down the beaker
walls and watch glass with distilled water and filter the sample to remove silicates
and other insoluble material that could clog the atomizer. Adjust the volume to
some predetermined value based on the expected metal concentrations. The sample
is now ready for analysis. Concentrations so determined shall be reported as
"total" (see Note 4).
4.1.4 To determine total recoverable metals, acidify the entire sample at the time of
collection with cone, redistilled HNO3, 5 ml/1. At the time of analysis a 100 ml
aliquot of well mixed sample is transferred to a beaker or flask. Five ml of distilled
HC1 (1:1) is added and the sample heated on a steam bath or hot plate until the
METALS-6
-------
volume has been reduced to 15-20 ml making certain the samples do not boil. (If
the sample is being prepared for furnace analysis, the same process should be
followed except HC1 should be omitted.) After this treatment the sample is filtered
to remove silicates and other insoluble material that could clog the atomizer and
the volume adjusted to 100 ml. The sample is then ready for analysis.
Concentrations so determined shall be reported as "total". (See Notes 4, 5, and 6.)
NOTE 5: The analyst should be cautioned that this digestion procedure may not be
sufficiently vigorous to destroy certain metal complexes if a colorimetric procedure
is to be employed for the final determination. When this is suspect, the more
vigorous digestion given in 4.1.3 should be followed.
NOTE 6: For drinking water analyses by direct aspiration, the final volume may be
reduced to effect up to a 10X concentration of the sample, provided the total
dissolved solids in the original sample do not exceed 500 mg/1, the determination
is corrected for any non-specific absorbance and there is no loss by precipitation.
5. Interferences
5.1 Direct Aspiration
5.1.1 The most troublesome type of interference in atomic absorption
spectrophotometry is usually termed "chemical" and is caused by lack of
absorption of atoms bound in molecular combination in the flame. This
phenomenon can occur when the flame is not sufficiently hot to dissociate the
molecule, as in the case of phosphate interference with magnesium, or because the
dissociated atom is immediately oxidized to a compound that will not dissociate
further at the temperature of the flame. The addition of lanthanum will overcome
the phosphate interference in the magnesium, calcium and barium determinations.
Similarly, silica interference in the determination of manganese can be eliminated
by the addition of calcium.
5.1.2 Chemical interferences may also be eliminated by separating the metal from the
interfering material. While complexing agents are primarily employed to increase
the sensitivity of the analysis, they may also be used to eliminate or reduce
interferences.
5.1.3 The presence of high dissolved solids in the sample may result in an interference
from non-atomic absorbance such as light scattering. If background correction is
not available, a non-absorbing wavelength should be checked. Preferably, high
solids type samples should be extracted (see 5.1.1 and 9.2).
5.1.4 lonization interferences occur where the flame temperature is sufficiently high to
generate the removal of an electron from a neutral atom, giving a positive charged
ion. This type of interference can generally be controlled by the addition, to both
standard and sample solutions, of a large excess of an easily ionized element.
5.1.5 Although quite rare, spectral interference can occur when an absorbing
wavelength of an element present in the sample but not being determined falls
within the width of the absorption line of the element of interest. The results of the
determination will then be erroneously high, due to the contribution of the
interfering element to the atomic absorption signal. Also, interference can occur
METALS-7
-------
when resonant energy from another element in a multi-element lamp or a metal
impurity in the lamp cathode falls within the bandpass of the slit setting and that
metal is present in the sample. This type of interference may sometimes be reduced
by narrowing the slit width.
5.2 Flameless Atomization
5.2.1 Although the problem of oxide formation is greatly reduced with furnace
procedures because atomization occurs in an inert atmosphere, the technique is
still subject to chemical and matrix interferences. The composition of the sample
matrix can have a major effect on the analysis. It is those effects which must be
determined and taken into consideration in the analysis of each different matrix
encountered. To help verify the absence of matrix or chemical interference use the
following procedure. Withdraw from the sample two equal aliquots. To one of the
aliquots add a known amount of analyte and dilute both aliquots to the same
predetermined volume. [The dilution volume should be based on the analysis of the
undiluted sample. Preferably, the dilution should be 1:4 while keeping in mind the
optimum concentration range of the analysis. Under no circumstances should the
dilution be less than 1:1]. The diluted aliquots should then be analyzed and the
unspiked results multiplied by the dilution factor should be compared to the
original determination. Agreement of the results (within ±10%) indicates the
absence of interference. Comparison of the actual signal from the spike to the
expected response from the analyte in an aqueous standard should help confirm the
finding from the dilution analysis. Those samples which indicate the presence of
interference, should be treated in one or more of the following ways.
a. The samples should be successively diluted and reanalyzed to
determine if the interference can be eliminated.
b. The matrix of the sample should be modified in the furnace. Examples
are the addition of ammonium nitrate to remove alkali chlorides,
ammonium phosphate to retain cadmium, and nickel nitrate for
arsenic and selenium analyses [ATOMIC ABSORPTION
NEWSLETTER Vol. 14, No. 5, p 127, Sept-Oct 1975]. The mixing of
hydrogen with the inert purge gas has also been used to suppress
chemical interference. The hydrogen acts as a reducing agent and aids
in molecular dissociation.
c. Analyze the sample by method of standard additions while noting the
precautions and limitations of its use (See 8.5).
5.2.2 Gases generated in the furnace during atomization may have molecular absorption
bands encompassing the analytical wavelength. When this occurs, either the use of
background correction or choosing an alternate wavelength outside the absorption
band should eliminate this interference. Non-specific broad band absorption
interference can also be compensated for with background correction.
5.2.3 Interference from a smoke-producing sample matrix can sometimes be reduced by
extending the charring time at a higher temperature or utilizing an ashing cycle in
METALS-8
-------
the presence of air. Care must be taken, however, to prevent loss of the analysis
element.
5.2.4 Samples containing large amounts of organic materials should be oxidized by
conventional acid digestion prior to being placed in the furnace. In this way broad
band absorption will be minimized.
5.2.5 From anion interference studies in the graphite furnace it is generally accepted that
nitrate is the preferred anion. Therefore nitric acid is preferable for any digestion or
solubilization step. If another acid in addition to HN03 is required a minimum
amount should be used. This applies particularly to hydrochloric and to a lesser
extent to sulfuric and phosphoric acids.
5.2.6 Carbide formation resulting from the chemical environment of the furnace has
been observed with certain elements that form carbides at high temperatures.
Molybdenum may be cited as an example. When this takes place, the metal will be
released very slowly from the carbide as atomization continues. For molybdenum,
one may be required to atomize for 30 seconds or more before the signal returns to
baseline levels. This problem is greatly reduced and the sensitivity increased with
the use of pyrolytically-coated graphite.
5.2.7 lonization interferences have to date not been reported with furnace techniques.
5.2.8 For comments on spectral interference see section 5.1.5.
5.2.9 Contamination of the sample can be a major source of error because of the extreme
sensitivities achieved with the furnace. The sample preparation work area should
be kept scrupulously clean. All glassware should be cleaned as directed in part 6.9
of the Atomic Absorption Methods section of this manual. Pipet tips have been
known to be a source of contamination. If suspected, they should be acid soaked
with 1:5 HNO3 and rinsed thoroughly with tap and deionized water. The use of a
better grade pipet tip can greatly reduce this problem. It is very important that
special attention be given to reagent blanks in both analysis and the correction of
analytical results. Lastly, pyrolytic graphite because of the production process and
handling can become contaminated. As many as five to possibly ten high
temperature burns may be required to clean the tube before use.
Apparatus
6.1 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.
6.2 Burner: The burner recommended by the particular instrument manufacturer should be
used. For certain elements the nitrous oxide burner is required.
6.3 Hollow cathode lamps: Single element lamps are to be preferred but multi-element lamps
may be used. Electrodeless discharge lamps may also be used when available.
6.4 Graphite furnace: Any furnace device capable of reaching the specified temperatures is
satisfactory.
METALS-9
-------
6.5 Strip chart recorder: A recorder is strongly recommended for furnace work so that there
will be a permanent record and any problems with the analysis such as drift, incomplete
atomization, losses during charring, changes in sensitivity, etc., can be easily recognized.
6.6 Pipets: Microliter with disposable tips. Sizes can range from 5 to 100 microliters as
required. NOTE 7: Pipet tips which are white in color, do not contain CdS, and have
been found suitable for research work are available from Ulster Scientific, Inc. 53 Main
St. Highland, NY 12528 (914) 691-7500.
6.7 Pressure-reducing valves: The supplies of fuel and oxidant shall be maintained at
pressures somewhat higher than the controlled operating pressure of the instrument by
suitable valves.
6.8 Separatory flasks: 250 ml, or larger, for extraction with organic solvents.
6.9 Glassware: All glassware, linear polyethylene, polyproplyene or Teflon containers,
including sample bottles, should be washed with detergent, rinsed with tap water, 1:1
nitric acid, tap water, 1:1 hydrochloric acid, tap water and deionized distilled water in
that order. [See Notes 1 and 2 under (4.1) concerning the use of chromic acid and the
cleaning procedure.]
6.10 Borosilicate glass distillation apparatus.
7. Reagents
7.1 Deionized distilled water: Prepare by passing distilled water through a mixed bed of
cation and anion exchange resins. Use deionized distilled water for the preparation of all
reagents, calibration standards, and as dilution water.
7.2 Nitric acid (cone.): If metal impurities are found to be present, distill reagent grade
nitric acid in a borosilicate glass distillation apparatus or use a spectrograde acid.
Caution: Distillation should be performed in hood with protective sash in place.
7.2.1 Nitric Acid (1:1): Prepare a 1:1 dilution with deionized, distilled water by
adding the cone, acid to an equal volume of water.
7.3 Hydrochloric acid (1:1): Prepare a 1:1 solution of reagent grade hydrochloric acid and
deionized distilled water. If metal impurities are found to be present, distill this mixture
from a borosilicate glass distillation apparatus or use a spectrograde acid.
7.4 Stock standard metal solutions: Prepare as directed in (8.1) and under the individual
metal procedures. Commercially available stock standard solutions may also be used.
7.5 Calibration standards: Prepare a series of standards of the metal by dilution of the
appropriate stock metal solution to cover the concentration range desired.
7.6 Fuel and oxidant: Commercial grade acetylene is generally acceptable. Air may be
supplied from a compressed air line, a laboratory compressor, or from a cylinder of
compressed air. Reagent grade nitrous oxide is also required for certain determinations.
Standard, commercially available argon and nitrogen are required for furnace work.
7.7 Special reagents for the extraction procedure.
7.7.1 Pyrrolidine dithiocarbamic acid (PDCA) "see footnote": Prepare by adding 18
ml of analytical reagent grade pyrrolidine to 500 ml of chloroform in a liter flask.
The name pyrrolidine dithiocarbamic acid (PDCA), although commonly referenced in the scientific
literature is ambiguous. From the chemical reaction of pyrrolidine and carbon disulfide a more
proper name would be 1-pyrrolidine carbodithioic acid, PCD A (CAS Registry No. 25769-03-3).
METALS-10
-------
(See Note 8) Cool and add 15 ml of carbon disulfide in small portions and with
swirling. Dilute to 1 liter with chloroform. The solution can be used for several
months if stored in a brown bottle in a refrigerator.
NOTE 8: An acceptable grade of pyrrolidine may be obtained from the Aldrich
Chemical Co., 940 West St. Paul Ave., Milwaukee, WI. 53233 (414,273-3850).
7.7.2 Ammonium hydroxide, 2N: Dilute 3 ml cone. NH4OH to 100 ml with deionized
distilled water.
7.7.3 Bromph'enol blue indicator (Ig/liter): Dissolve O.lg bromphenol blue in 100 ml
of 50 percent ethanol or isopropanol.
7.7.4 HC1, 2.5% v/v: Dilute 2 ml redistilled HC1 to 40 ml with deionized distilled water.
8. Preparation of Standards and Calibration
8.1 Stock standard solutions are prepared from high purity metals, oxides or nonhygroscopic
reagent grade salts using deionized distilled water and redistilled nitric or hydrochloric
acids. (See individual analysis sheets for specific instruction.) Sulfuric or phosphoric
acids should be avoided as they produce an adverse effect on many elements. The stock
solutions are prepared at concentrations of 1000 mg of the metal per liter. Commercially
available standard solutions may also be used.
8.2 Calibration standards are prepared by diluting the stock metal solutions at the time of
analysis. For best results, calibration standards should be prepared fresh each time an
analysis is to be made and discarded after use. Prepare a blank, and at least four
calibration standards in graduated amounts in the appropriate range. The calibration
standards should be prepared using the same type of acid or combination of acids and at
the same concentration as will result in the samples following processing. As filtered
water samples are preserved with 1:1 redistilled HNO3 (3 ml per liter), calibration
standards for these analyses should be similarly prepared with HNO3. Beginning with
the blank and working toward the highest standard, aspirate the solutions and record the
readings. Repeat the operation with both the calibration standards and the samples a
sufficient number of times to secure a reliable average reading for each solution.
Calibration standards for furnace procedures should be prepared as described on the
individual sheets for that metal.
8.3 Where the sample matrix is so complex that viscosity, surface tension and components
cannot be accurately matched with standards, the method of standard addition must be
used. This technique relies on the addition of small, known amounts of the analysis
element to portions of the sample—the absorbance difference between those and the
original solution giving the slope of the calibration curve. The method of standard
addition is described in greater detail in (8.5).
METALS-11
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8.4 For those instruments which do not read out directly in concentration, a calibration
curve is prepared to cover the appropriate concentration range. Usually, this means the
preparation of standards which produce an absorption of 0 to 80 percent. The correct
method is to convert the percent absorption readings to absorbance and plot that value
against concentration. The following relationship is used to convert absorption values to
absorbance:
absorbance = log (100/%T) = 2-log % T
where % T = 100-% absorption
As the curves are frequently nonlinear, especially at high absorption values, the number
of standards should be increased in that portion of the curve.
8.5 Method of Standard Additions: In this method, equal volumes of sample are added to a
deionized distilled water blank and to three standards containing different known
amounts of the test element. The volume of the blank and the standards must be the
same. The absorbance of each solution is determined and then plotted on the vertical axis
of a graph, with the concentrations of the known standards plotted on the horizontal
axis. When the resulting line is extrapolated back to zero absorbance, the point of
interception of the abscissa is the concentration of the unknown. The abscissa on the left
of the ordinate is scaled the same as on the right side, but in the opposite direction from
the ordinate. An example of a plot so obtained is shown in Fig. 1.
Concentration
I Cone, of
Sample
Addn 0
No Addn
Addn I
Addn of 50%
of Expected
Amount
Addn 2
Addn of 100%
of Expected
Amount
Addn 3
Addn of 150%
of Expected
Amount
FIGURE 1. STANDARD ADDITION PLOT
METALS-12
-------
The method of standard additions can be very useful, however, for the results to be valid
the following limitations must be taken into consideration:
a) the absorbance plot of sample and standards must be linear over the
concentration range of concern. For best results the slope of the plot should
be nearly the same as the slope of the aqueous standard curve. If the slope is
significantly different (more than 20%) caution should be exercised.
b) the effect of the interference should not vary as the ratio of analyte
concentration to sample matrix changes and the standard addition should
respond in a similar manner as the analyte.
c) the determination must be free of spectral interference and corrected for non-
specific background interference.
9. General Procedure for Analysis by Atomic Absorption
9.1 Direct Aspiration: Differences between the various makes and models of satisfactory
atomic absorption spectrophotometers prevent the formulation of detailed instructions
applicable to every instrument. The analyst should follow the manufacturer's operating
instructions for his particular instrument. In general, after choosing the proper hollow
cathode lamp for the analysis, the lamp should be allowed to warm up for a minimum of
15 minutes unless operated in a double beam mode. During this period, align the
instrument, position the monochromator at the correct wavelength, select the proper
monochromator slit width, and adjust the hollow cathode current according to the
manufactuerer's recommendation. Subsequently, light the flame and regulate the flow of
fuel and oxidant, adjust the burner and nebulizer flow rate for maximum percent
absorption and stability, and balance the photometer. Run a series of standards of the
element under analysis and construct a calibration curve by plotting the concentrations
of the standards against the absorbance. For those instruments which read directly in
concentration set the curve corrector to read out the proper concentration. Aspirate the
samples and determine the concentrations either directly or from the calibration curve.
Standards must be run each time a sample or series of samples are run.
9.1.1 Calculation - Direct determination of liquid samples: Read the metal value in
mg/1 from the calibration curve or directly from the readout system of the
instrument.
9.1.1.1 If dilution of sample was required:
mg/1 metal in sample = A
where:
A = mg/1 of metal indiluted aliquot from calibration curve
B = ml of deionized distilled water used for dilution
C = ml of sample aliquot
METALS-13
-------
9.1.2 For samples containing particulates:
mg/1 metal in sample = A I -X-
where:
A = mg/1 of metal in processed sample from calibration curve
V = final volume of the processed sample in ml
C= ml of sample aliquot processed
9.1.3 For solid samples: report all concentrations as mg/kg dry weight
9.1.3.1 Dry sample:
mg metal/kg sample = — —
where:
A — mg/1 of metal in processed sample from calibration curve
V = final volume of the processed sample in ml
D = weight of dry sample in grams
9.1.3.2 Wet sample:
A x V
mg metal/kg sample = ^ -
where:
A = mg/1 of metal in processed sample from calibration curve
V = final volume of the processed sample in ml
W = weight of wet sample in grams
P = % solids
9.2 Special Extraction Procedure: When the concentration of the metal is not sufficiently
high to determine directly, or when considerable dissolved solids are present in the
sample, certain metals may be chelated and extracted with organic solvents. Ammonium
pyrrolidine dithiocarbamate (APDC) (see footnote) in methyl isobutyl ketone (MIBK) is
widely used for this purpose and is particularly useful for zinc, cadmium, iron,
manganese, copper, silver, lead and chromium+6. Tri-valent chromium does not react
with APDC unless it has first been converted to the hexavalent form [Atomic Absorption
Newsletter 6, p 128 (1967)]. This procedure is described under method 218.3.
The name ammonium pyrrolidine dithiocarbamate (APDC) is somewhat ambiguous and should more
properly be called ammonium, 1-pyrollidine carbodithioate (APCD), CAS Registry No. 5108-96-3.
METALS-14
-------
Aluminum, beryllium, barium and strontium also do not react with APDC. While the
APDC-MIBK chelating-solvent system can be used satisfactorily, it is possible to
experience difficulties. (See Note 9.)
NOTE 9: Certain metal chelates, manganese-APDC in particular, are not stable in
MIBK and will redissolve into the aqueous phase on standing. The extraction of other
metals is sensitive to both shaking rate and time. As with cadmium, prolonged extraction
beyond 1 minute, will reduce the extraction efficiency, whereas 3 minutes of vigorous
shaking is required for chromium.
Also, when multiple metals are to be determined either larger sample volumes must be
extracted or individual extractions made for each metal being determined. The acid form
of APDC-pyrrolidine dithiocarbamic acid prepared directly in chloroform as described
by Lakanen, [Atomic Absorption Newsletter 5, p 17 (1966)], (see 7.7.1) has been found
to be most advantageous. In this procedure the more dense chloroform layer allows for
easy combination of multiple extractions which are carried out over a broader pH range
favorable to multielement extractions. Pyrrolidine dithiocarbamic acid in chloroform is
very stable and may be stored in a brown bottle in the refrigerator for months. Because
chloroform is used as the solvent, it may not be aspirated into the flame. The following
procedure is suggested.
9.2.1 Extraction procedure with pyrrolidine dithiocarbamic acid (PDCA) in
chloroform.
9.2.1.1 Transfer 200 ml of sample into a 250 ml separatory funnel, add 2 drops
bromphenol blue indicator solution (7.7.3) and mix.
9.2.1.2 Prepare a blank and sufficient standards in the same manner and adjust
the volume of each to approximately 200 ml with deionized distilled
water. All of the metals to be determined may be combined into single
solutions at the appropriate concentration levels.
9.2.1.3 Adjust the pH by addition of 2N NH4OH solution (7:7.2) until a blue
color persists. Add HC1 (7.7.4) dropwise until the blue color just
disappears; then add 2.0 ml HC1 (7.7.4) in excess. The pH at this point
should be 2.3. (The pH adjustment may be made with a pH meter
instead of using indicator.)
9.2.1.4 Add 5 ml of PDCA-chloroform reagent (7.7.1) and shake vigorously
for 2 minutes. Allow the phases to separate and drain the chloroform
layer into a 100 ml beaker. (See NOTE 10.)
NOTE 10: If hexavalent chromium is to be extracted, the aqueous
phase must be readjusted back to a pH of 2.3 after the addition of
PDCA-chloroform and maintained at that pH throughout the
extraction. For multielement extraction, the pH may adjusted upward
after the chromium has been extracted.
METALS-15
-------
9.2.1.5 Add a second portion of 5 ml PDCA-chloroform reagent (7.7.1) and
shake vigorously for 2 minutes. Allow the phases to separate and
combine the chloroform phase with that obtained in step (9.2.1.4).
9.2.1.6 Determine the pH of the aqueous phase and adjust to 4.5.
9.2.1.7 Repeat step (9.2.1.4) again combining the solvent extracts.
9.2.1.8 Readjust the pH to 5.5, and extract a fourth time. Combine all extracts
and evaporate to dryness on a steam bath.
9.2.1.9 Hold the beaker at a 45 degree angle, and slowly add 2 ml of cone.
distilled nitric acid, rotating the beaker to effect thorough contact of
the acid with the residue.
9.2.1.10 Place the beaker on a low temperature hotplate or steam bath and
evaporate just to dryness.
9.2.1.11 Add 2 ml of nitric acid (1:1) to the beaker and heat for 1 minute. Cool,
quantitatively transfer the solution to a 10 ml volumetric flask and
bring to volume with distilled water. The sample is now ready for
analysis.
9.2.2 Prepare a calibration curve by plotting absorbance versus the concentration of the
metal standard 0/g/l) in the 200 ml extracted standard solution. To calculate
sample concentration read the metal value in ug/1 from the calibration curve or
directly from the readout system of the instrument. If dilution of the sample was
required use the following equation:
mg/1 metal in sample =
where:
Z = ug/1 of metal in diluted aliquot from calibration curve
B = ml of deionized distilled water used for dilution
C= ml of sample aliquot
9.3 Furnace Procedure: Furnace devices (flameless atomization) are a most useful means of
extending detection limits. Because of differences between various makes and models of
satisfactory instruments, no detailed operating instuctions can be given for each
instrument. Instead, the analyst should follow the instructions provided by the
manufacturer of his particular instrument and use as a guide the temperature settings
and other instrument conditions listed on the individual analysis sheets which are
recommended for the Perkin-Elmer HGA-2100. In addition, the following points may be
helpful.
9.3.1 With flameless atomization, background correction becomes of high importance
especially below 350 nm. This is because certain samples, when atomized, may
absorb or scatter light from the hollow cathode lamp. It can be caused by the
presence of gaseous molecular species, salt particules, or smoke in the sample
METALS-16
-------
beam. If no correction is made, sample absorbance will be greater than it should be,
and the analytical result will be erroneously high.
9.3.2 If during atomization all the analyte is not volatilized and removed from the
furnace, memory effects will occur. This condition is dependent on several factors
such as the volatility of the element and its chemical form, whether pyrolytic
graphite is used, the rate of atomization and furnace design. If this situation is
detected through blank burns, the tube should be cleaned by operating the furnace
at full power for the required time period as needed at regular intervals in the
analytical scheme.
9.3.3 Some of the smaller size furnace devices, or newer furnaces equipped with feedback
temperature control (Instrumentation Laboratories MODEL 555, Perkin-Elmer
MODELS HGA 2200 and HGA 76B, and Varian MODEL CRA-90) employing
faster rates of atomization, can be operated using lower atomization temperatures
for shorter time periods than those listed in this manual.
9.3.4 Although prior digestion of the sample in many cases is not required providing a
representative aliquot of sample can be pipeted into the furnace, it will provide for a
more uniform matrix and possibly lessen matrix effects.
9.3.5 Inject a measured microliter aliquot of sample into the furnace and atomize. If the
concentration found is greater than the highest standard, the sample should be
diluted in the same acid matrix and reanalyzed. The use of multiple injections can
improve accuracy and help detect furnace pipetting errors.
9.3.6 To verify the absence of interference, follow the procedure as given in part 5.2.1.
9.3.7 A check standard should be run approximately after every 10 sample injections.
Standards are run in part to monitor the life and performance of the graphite tube.
Lack of reproducibility or significant change in the signal for the standard
indicates that the tube should be replaced. Even though tube life depends on
sample matrix and atomization temperature, a conservative estimate would be that
a tube will last at least 50 firings. A pyrolytic-coating would extend that estimate
by a factor of 3.
9.3.8 Calculation-For determination of metal concentration by the furnace: Read the
metal value in ug/1 from the calibration curve or directly from the readout system
of the instrument.
9.3.8.1 If different size furnace injection volumes are used for samples than for
standards:
ug/1 of metal in sample = Z I -TT- j
where:
Z = ug/1 of metal read from calibration curve or readout system
S = ul volume standard injected into furnace for calibration curve
U = ul volume of sample injected for analysis
METALS-17
-------
9.3.8.2 If dilution of sample was required but sample injection volume same as
for standard:
/ C^ 4- R
ug/1 of metal in sample = Z
where:
Z = ug/1 metal in diluted aliquot from calibration curve
B = ml of deionized distilled water used for dilution
C= ml of sample aliquot
9.3.9 For sample containing particulates:
ug/1 of metal in sample = Z
where:
(*)
Z = ug/1 of metal in processed sample from calibration curve (See 9.3.8.1)
V = final volume of processed sample in ml
C = ml of sample aliquot processed
9.3.10 For solid samples: Report all concentrations as mg Ag dry weight
9.3.10.1 Dry sample:
V 1,000 )V
mg metal/kg sample = -
D
where:
Z = ug/1 of metal in processed sample from calibration curve (See 9.3.8.1)
V = final volume of processed sample in ml
D = weight of dry sample in grams
9.3.10.2 Wet sample:
mg metal/ kg sample =
W x P
where:
Z = ug/1 of metal in processed sample from calibration curve (See 9.3.8. 1)
V = final volume of processed sample in ml
W = weight of wet sample in grams
P= % solids
METALS-18
-------
10 Quality Control For Drinking Water Analysis
10.1 Minimum requirements
10.1.1 All quality control data should be maintained and available for easy
reference or inspection.
10.1.2 An unknown performance sample (when available) must be analyzed once
per year for the metals measured. Results must be within the control limit
established by EPA. If problems arise, they should be corrected, and a
follow-up performance sample should be analyzed.
10.2 Minimum Daily control
10.2.1 After a calibration curve composed of a minimum of a reagent blank and
three standards has been prepared, subsequent calibration curves must be
verified by use of at least a reagent blank and one standard at or near the
MCL. Daily checks must be within ± 10 percent of original curve.
10.2.2 If 20 or more samples per day are analyzed, the working standard curve must
be verified by running an additional standard at or near the MCL every 20
samples. Checks must be within ± 10 percent of original curve.
10.3 Optional Requirements
10.3.1 A current service contract should be in effect on balances and the atomic
absorption spectrophotometer.
10.3.2 Class S weights should be available to make periodic checks on balances.
10.3.3 Chemicals should be dated upon receipt of shipment and replaced as needed
or before shelf life has been exceeded.
10.3.4 A known reference sample (when available) should be analyzed once per
quarter for the metals measured. The measured value should be within the
control limits established by EPA.
10.3.5 At least one duplicate sample should be run every 10 samples, or with each
set of samples to verify precision of the method. Checks should be within the
control limit established by EPA.
10.3.6 Standard deviation should be obtained and documented for all
measurements being conducted.
10.3.7 Quality Control charts or a tabulation of mean and standard deviation
should be used to document validity of data on a daily basis.
METALS-19
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ANTIMONY
Method 204.1 (Atomic absorption, direct aspiration)
STORET NO. Total 01097
Dissolved 01095
Suspended 01096
Optimum Concentration Range: 1-40 mg/1 using a wavelength of 217.6 nm
Sensitivity: 0.5 mg/1
Detection Limit: 0.2 mg/1
Preparation of Standard Solution
1. Stock Solution: Carefully weigh 2.7426 g of antimony potassium tartrate (analytical
reagent grade) and dissolve in deionized distilled water. Dilute to 1 liter with deionized
distilled water. 1 ml = 1 mgSb( 1000 mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using the same type of acid and at
the same concentration as will result in the sample to be analyzed either directly or after
processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 through 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Antimony hollow cathode lamp
2. Wavelength: 217.6 nm
3. Fuel: Acetylene
4. Oxidant: Air
5. Type of flame: Fuel lean
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES
Issued 1974
Editorial revision 1978
204.1-1
-------
Interferences
1. In the presence of lead (1000 mg/1), a spectral interference may occur at the 217.6 nm
resonance line. In this case the 231.1 nm antimony line should be used.
2. Increasing acid concentrations decrease antimony absorption. To avoid this effect, the
acid concentration in the samples and in the standards should be matched.
Notes
1. Data to be entered into STORET must be reported as ug/1.
2. For concentrations of antimony below 0.35 mg/1, the furnace procedure (Method 204.2)
is recommended.
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent at
concentrations of 5.0 and 15 mg Sb/1, the standard deviations were ±0.08 and ±0.1,
respectively. Recoveries at these levels were 96% and 97%, respectively.
204.1-2
-------
ANTIMONY
Method 204.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01097
Dissolved 01095
Suspended 01096
Optimum Concentration Range: 20-300 ug/1
Detection Limit: 3 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to be used for "standard additions".
3. The calibration standard should be diluted to contain 0.2% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.3 of the
Atomic Absorption Methods section of this manual should be followed including the
addition of sufficient 1:1 HC1 to dissolve the digested residue for the analysis of
suspended or total antimony. The sample solutions used for analysis should contain 2%
(v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30sec-125°C.
2. Ashing Time and Temp: 30 sec-800°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 217.6nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES
Issued 1978
204.2-1
-------
Notes
1. l he above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Nitrogen may also be used as the purge gas.
4. If chloride concentration presents a matrix problem or causes a loss previous to
atomization, add an excess of 5 mg of ammonium nitrate to the furnace and ash using a
ramp accessory or with incremental steps until the recommended ashing temperature is
reached.
5. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
6. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
7. Data to be entered in to STORET must be reported as ug/1.
Precision and Accuracy
1. Precision and accuracy data are not available at this time.
204.2-2
-------
ARSENIC
Method 206.2 (Atomic Absorption, furnace technique)
STORE! NO. Total 01002
Dissolved 01000
Suspended 01001
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Dissolve 1.320 g of arsenic trioxide, As203 (analytical reagent grade) in
100 ml of deionized distilled water containing 4 g NaOH. Acidify the solution with 20 ml
cone. HNO3 and dilute to 1 liter. 1 ml = 1 mg As (1000 mg/1).
2. Nickel Nitrate Solution, 5 %: Dissolve 24.780 g of ACS reagent grade Ni(NO3)2»6H2O in
deionized distilled water and make up to 100ml.
3. Nickel Nitrate Solution, 1%: Dilute 20 ml of the 5% nickel nitrate to 100 ml with
deionized distilled water.
4. Working Arsenic Solution: Prepare dilutions of the stock solution to be used as
calibration standards at the time of analysis. Withdraw appropriate aliquots of the stock
solution, add 1 ml of cone. HNO3, 2ml of 30% H2O2 and 2ml of the 5% nickel nitrate
solution. Dilute to 100 ml with deionized distilled water.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Transfer 100 ml of well-mixed sample to a 250 ml Griffin beaker, add 2 ml of 30% H2O2
and sufficient cone. HNO3 to result in an acid concentration of 1 %(v/v). Heat for 1 hour
at 95°C or until the volume is slightly less than 50 ml.
2. Cool and bring back to 50 ml with deionized distilled water.
3. Pipet 5 ml of this digested solution into a 10-ml volumetric flask, add 1 ml of the 1%
nickel nitrate solution and dilute to 10 ml with deionized distilled water. The sample is
now ready for injection into the furnace.
Approved for NPDES and SDWA
Issued 1978
206.2-1
-------
NOTE: If solubilization or digestion is not required, adjust the HNO3 concentration of
the sample to 1% (v/v) and add 2 ml of 30%H2O2 and 2 ml of 5% nickel nitrate to each
100 ml of sample. The volume of the calibration standard should be adjusted with
deionized distilled water to match the volume change of the sample.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30 sec-1100°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 193.7 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
4. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
5. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent
containing 15 ug/1 and spiked with concentrations of 2, 10 and 25 ug/1, recoveries of
85%, 90% and 88% were obtained respectively. The relative standard deviation at these
concentrations levels were ±8.8%, ±8.2%, ±5.4% and ±8.7%, respectively.
2. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 20, 50 and 100 ug As/1, the standard deviations were ±0.7, ±1.1 and ±1.6
respectively. Recoveries at these levels were 105%, 106% and 101%, respectively.
206.2-2
-------
ARSENIC
Method 206.3 (Atomic Absorption—gaseous hydride)
STORET NO. Total 01002
Dissolved 01000
Suspended 01001
1. Scope and Application
1.1 The gaseous hydride method determines inorganic arsenic when present in
concentrations at or above 2 ug/1. The method is applicable to drinking water and most
fresh and saline waters in the absence of high concentrations of chromium, cobalt,
copper, mercury, molybdenum, nickel and silver.
2. Summary of Method
2.1 Arsenic in the sample is first reduced to the trivalent form using SnCl2 and converted to
arsine, AsH3, using zinc metal. The gaseous hydride is swept into an argon-hydrogen
flame of an atomic absorption spectrophotometer. The working range of the mehtod is
2-20 ug/1. The 193.7 nm wavelength line is used.
3. Comments
3.1 In analyzing drinking water and most surface and ground waters, interferences are rarely
encountered. Industrial waste samples should be spiked with a known amount of arsenic
to establish adequate recovery.
3.2 Organic forms of arsenic must be converted to inorganic compounds and organic matter
must be oxidized before beginning the analysis. The oxidation procedure given in
Method 206.5 (Standard Methods, 14th Edition, Method 404B, p. 285, Procedure 4.a)
has been found suitable.
3.3 For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
3.4 For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
3.5 Data to be entered into STORET must be reported as ug/1.
4. Precision and Accuracy
4.1 Ten replicate solutions of o-arsenilic acid at the 5,10 and 20 ug/1 level were analyzed by
a single laboratory. Standard deviations were ±0.3, ±0.9 and ±1.1 with recoveries of 94,
93 and 85%, respectively. (Caldwell, J. S., Lishka, R. J., and McFarren, E. F.,
"Evaluation of a Low Cost Arsenic and Selenium Determination at Microgram per Liter
Levels", JAWWA., vol 65, p 731, Nov., 1973.)
Approved for NPDES and SDWA
Issued 1974
206.3-1
-------
5. References
5.1, Except for the perchloric acid step, the procedure to be used for this determination is
found in: Standard Methods for the Examination of Water and Wastewater, 14th
Edition, p!59, Method 301A(VII),(1975)
206.3-2
-------
BARIUM
Method 208.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01007
Dissolved 01005
Suspended 01006
Optimum Concentration Range; 1-20 mg/1 using a wavelength of 553.6 nm
Sensitivity: 0.4 mg/1
Detection Limit: 0.1 mg/1
Preparation of Standard Solution
1. Stock Solution: Dissolve 1.7787 g barium chloride (BaQ2»2H2O, analytical reagent
grade) in deionized distilled water and dilute to 1 liter. 1 ml = 1 mg Ba (1000 mg/1).
2. Potassium chloride solution: Dissolve 95 g potassium chloride, KC1, in deionized
distilled water and make up to 1 liter.
3. Prepare dilutions of the stock barium solution to be used as calibration standards at the
time of analysis. To each 100 ml of standard and sample alike add 2.0 ml potassium
chloride solution. The calibration standards should be prepared using the same type of
acid and the same concentration as will result in the sample to be analyzed either directly
or after processing.
i
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 through 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Barium hollow cathode lamp
2. Wavelength: 553.6 nm
3. Fuel: Acetylene
4. Oxidant: Nitrous oxide
5. Type of flame: Fuel rich
Approved for NPDES and SDWA
Issued 1974
208.1-1
-------
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Interferences
1. The use of a nitrous oxide-acetylene flame virtually eliminates chemical interference;
however, barium is easily ionized in this flame and potassium must be added (1000 mg/1)
to standards and samples alike to control this effect.
2. If the nitrous oxide flame is not available and acetylene-air is used, phosphate, silicon and
aluminum will severely depress the barium absorbance. This may be overcome by the
addition of 2000 mg/1 lanthanum.
Notes
1. Data to be entered into STORET must be reported as ug/1.
2. For concentrations of barium below 0.2 mg/1, the furnace procedure (Method 208.2) is
recommended.
3. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent at
concentrations of 0.40 and 2.0 mg Ba/1, the standard deviations were ±0.043
and ±0.13, respectively. Recovereis at these levels were 94% and 113%, respectively.
2. In a round-robin study reported by Standard Methods (13th Edition, p215, method
129A, 1971), three synthetic samples containing barium were analyzed by 13
laboratories. At concentrations of 500, 1000 and 5000 ug Ba/1, the reported standard
deviations were ±50, ±89 and ±185 ug, respectively. The relative error at these
concentrations was 8.6%, 2.7% and 1.4%, respectively.
208.1-2
-------
BARIUM
Method 208.2 (Atomic Absorption, furnace technique)
STORE! NO. 01007
Dissolved 01005
Suspended 01006
Optimum Concentration Range: 10-200 ug/1
Detection Limit: 2 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions, of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to used for "standard additions".
3. The calibration standard should be diluted to contain 0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30sec-1200°C.
3. Atomizing Time and Temp: 10 sec-2800°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 553.6 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and pyrolytic
graphite.
Approved for NPDES and SDWA
Issued 1978
208.2-1
-------
2. The use of halide acid should be avoided.
3. Because of possible chemical interaction, nitrogen should not be used as a purge gas.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
6. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
7. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laborator y (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 500 and 1000 ug Ba/1, the standard deviations were +2.5 and ±2.2 ug, respectively.
Recoveries at these levels were 96% and 102%, respectively A dilution of 1:10 was
required to bring the spikes within the analytical range of the method.
208.2-2
-------
BERYLLIUM
Methods 210.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01012
Dissolved 01010
Suspended 01011
Optimum Concentration Range: 0.05-2 mg/1 using a wavelength of 234.9 nm
Sensitivity: 0.025 mg/1
Detection Limit: 0.005 mg/1
Preparation of Standard Solution
1. Stock solution: Dissolve 11.6586 g beryllium sulfate, BeSO4, in deionized distilled water
containing 2 ml cone, nitric acid and dilute to 1 liter. 1 ml = 1 mg Be (1000 mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using the same type of acid and at
the same concentration as will result in the sample to be analyzed either directly or after
processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 through 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Beryllium hollow cathode lamp
2. Wavelength: 234.9 nm
3. Fuel: Acetylene
4. Oxidant: Nitrous oxide
5. Type of flame: Fuel rich
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES
Issued 1974
210.1-1
-------
Interferences
1. Sodium and silicon at concentrations in excess of 1000 mg/1 have been found to severely
depress the beryllium absorbance.
2. Bicarbonate ion is reported to interfere; however, its effect is eliminated when samples
are acidified to a pH of 1.5.
3. Aluminum at concentrations of 500 ug/1 is reported to depress the sensitivity of
beryllium [Spectrochim Acta 22,1325 (1966)].
Notes
1. Data to be entered into STORET must be reported as ug/1.
2, The "aluminon color imetric method" may also be used (Standard Methods, 14th
Edition, p 177). The minimum detectable concentration by this method is 5 ug/1.
3. For concentrations of beryllium below 0.02 mg/1, the furnace procedure (Method 210.2)
is recommended.
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent at
concentrations of 0.01, 0.05 and 0.25 mg Be/1, the standard deviations were ±0.001,
±0.001 and ±0.002, respectively. Recoveries at these levels were 100%, 98% and 97%,
respectively.
210.1-2
-------
BERYLLIUM
Method 210.2 (Atomic Absorption, furnace technique)
STORET NO. 01012
Dissolved 01010
Suspended 01011
Optimum Concentration Range: 1-30 ug/1
Detection Limit: 0.2 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to be used for "standard additions".
3. The calibration standard should be diluted to contain 0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30 sec-1000°C.
3. Atomizing Time and Temp: 10 sec-2800°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 234.9 nm
6. The operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation see "Furnace Procedure" part 9.3 of the-
Atomic Absorption methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
Approved for NPDES
Issued 1978
210.2-1
-------
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Because of possible chemical interaction and reported lower sensitivity, nitrogen should
not be used as the purge gas.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Precision and Accuracy data are not available at this time.
210.2-2
-------
CADMIUM
Method 213.1 (Atomic Absorption, direct aspiration)
STORE! NO. Total 01027
Dissolved 01025
Suspended 01026
Optimum Concentration Range: 0.05-2 mg/1 using a wavelength of 228.8 nm
Sensitivity: 0.025 mg/1
Detection Limit: 0.005 mg/1
Preparation of Standard Solution
1. Stock Solution: Carefully weigh 2.282 g of cadmium sulfate (3CdSO4»8H2O, analytical
reagent grade) and dissolve in deionized distilled water. 1 ml = 1 mg Cd (1000 mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using the same type of acid and at
the same concentration as will result in the sample to be analyzed either directly or after
processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 through 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Cadmium hollow cathode lamp
2. Wavelength: 228.8 nm
3. Fuel: Acetylene
4. Oxidant: Air
5. Type of flame: Oxidizing
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974
213.1-1
-------
Notes
1. For levels of cadmium below 20 og/1, either the Special Extraction Procedure given in
Part 9.2 of the Atomic Absorption methods section as the furnace technique, Method
213.2 is recommended.
2. Data to be entered into STORET must be reported as(ug/l.
3. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
Precision and Accuracy
1. An interlaboratory study on trace metal analyses by atomic absorption was conducted by
the Quality Assurance and Laboratory Evaluation Branch of EMSL. Six synthetic
concentrates containing varying levels of aluminum, cadmium, chromium, copper, iron,
manganese, lead and zinc were added to natural water samples. The statistical results for
cadmium were as follows:
Number
of Labs
74
73
63
68
55
51
True Values
«g/liter
71
78
14
18
1.4
2.8
Mean Value
ug/liter
70
74
16.8
18.3
3.3
2.9
Standard
Deviation
ug/liter
21
18
11.0
10.3
5.0
2.8
Accuracy as
% Bias
-2.2
-5.7
19.8
1.9
135
4.7
213.1-2
-------
CADMIUM
Method 213.2 (Atomic Absorption, furnace technique)
STORE! NO. 01027
Dissolved 01025
Suspended 01026
Optimum Concentration Range: 0.5-10 og/1
Detection Limit: 0.1 ug/\
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Ammonium Phosphate solution (40%); Dissolve 40 grams of ammonium phosphate,
(NH4)2HPO4 (analytical reagent grade) in deionized distilled water and dilute to 100 ml.
3. Prepare dilutions of the stock cadmium solution to be used as calibration standards at the
time of analysis. To each 100 ml of standard and sample alike add 2.0 ml of the
ammonium phosphate solution. The calibration standards should be prepared to contain
0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30sec-125°C.
2. Ashing Time and Temp: 30 sec-500°C.
3. Atomizing Time and Temp: 10sec-1900°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 228.8 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1978
213.2-1
-------
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Contamination from the work area is critical in cadmium analysis. Use of pi pet tips
which are free of cadmium is of particular importance. (See part 5.5.7 of the Atomic
Absorption Methods section of this manual.)
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
6. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
7. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 2.5, 5.0 and 10.0 ug Cd/1, the standard deviations were ±0.10, ±0.16 and ±0.33,
respectively. Recoveries at these levels were 96%, 99% and 98%, respectively.
213.2-2
-------
CHROMIUM
Method 218.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01034
Dissolved 01030
Suspended 01031
Optimum Concentration Range: 0.5-10 mg/1 using a wavelength of 357.9 nm
Sensitivity: 0.25 mg/1
Detection Limit: 0.05 mg/1
Preparation of Standard Solution
1. Stock Solution: Dissolve 1.923 g of chromium trioxide (CrO3, reagent grade) in deionized
distilled water. When solution is complete, acidify with redistilled HNO3 and dilute to 1
liter with deionized distilled water. 1 ml = 1 mg Cr (1000 mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using the same type of acid and at
the same concentration as will result in the sample to be analyzed either directly or after
processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Chromium hollow cathode lamp
2. Wavelength: 357.9 nm
3. Fuel: Acetylene
4. Oxidant: Nitrous oxide
5. Type of flame: Fuel rich
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
218.1-1
-------
Notes
1. The following wavelengths may also be used:
359.3 nm Relative Sensitivity 1.4
425.4 nm Relative Sensitivity 2
427.5 nm Relative Sensitivity 3
428.9 nm Relative Sensitivity 4
2. The fuel rich air-acetylene flame provides greater sensitivity but is subject to chemical
and matrix interference from iron, nickel, and other metals. If the analysis is performed
in a lean flame the interference can be lessened but the sensitivity will also be reduced.
3. The suppression of both Cr (III) and Cr (VI) absorption by most interfering ions in fuel
rich air-acetylene flames is reportedly controlled by the addition of 1% ammonium
bifluoride in 0.2% sodium sulfate [Talanta 20, 631 (1973)]. A 1% oxine solution is also
reported to be useful.
4. For levels of chromium between 50 and 200 ug/1 where the air-acetylene flame can not
be used or for levels below 50 ug/1, either the furnace procedure or the extraction
procedure is recommended. See Method 218.2 for the furnace procedure and Method
218.3 for the chelation-extraction procedure.
5. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORE! must be reported as ug/1.
Precision and Accuracy
1. An interlaboratory study on trace metal analyses by atomic absorption was conducted by
the Quality Assurance and Laboratory Evaluation Branch of EMSL. Six synthetic
concentrates containing varying levels of aluminum, cadmium, chromium, copper, iron,
manganese, lead and zinc were added to natural water samples. The statistical results for
chromium were as follows:
Standard
Number True Values Mean Value Deviation Accuracy as
of Labs tig/liter ug/liter ug/liter % Bias
74 370 353 105 -4.5
76 407 380 128 -6.5
72 74 72 29 -3.1
70 93 84 35 -10.2
47 7.4 10.2 7.8 37.7
47 15.0 16.0 9.0 6.8
218.1-2
-------
CHROMIUM
Method 218.2 (Atomic Absorption, furnace technique)
STORET NO. 01034
Dissolved 01030
Suspended 01031
Optimum Concentration Range: 5-100ug/l
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Calcium Nitrate Solution: Dissolve 11.8 grams of calcium nitrate, Ca(NO3)2 • 4H2O
(analytical reagent grade) in deionized distilled water and dilute to 100 ml. 1 ml = 20 mg
Ca.
3. Prepare dilutions of the stock chromium solution to be used as calibration standards at
the time of analysis. The calibration standards should be prepared to contain 0.5% (v/v)
HNO3. To each 100 ml of standard and sample alike, add 1 ml of 30% H2O2 and 1 ml of
the calcium nitrate solution.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
secton of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% v/v HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30 sec-1000°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 357.9 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1978
218.2-1
-------
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injecton, continuous flow purge gas and non-pyrolytic
graphite.
2. Hydrogen peroxide is added to the acidified solution to convert all chromium to the
trivalent state. Calcium is added to a level above 200 mg/1 where its suppressive effect
becomes constant up to 1000 mg/1.
3. Background correction may be required if the sample contains high dissolved solids.
4. Nitrogen should not be used as a purge gas because of possible CN band interference.
5. Pipet tips have been reported to be a possible source of contamination. (See part 5.5.7 of
the Atomic Absorption Methods section of this manual.)
6. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
7. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manua-1.
8. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
9. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 19, 48, and 77 ug Cr/1, the standard deviations were ±0.1, ±0.2, and ±0.8,
respectively. Recoveries at these levels were 97%, 101%, and 102%, respectively.
218.2-2
-------
LEAD
Method 239.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01051
Dissolved 01049
Suspended 01050
Optimum Concentration Range: 1-20 mg/1 using a wavelength of 283.3 nm
Sensitivity: 0.5 mg/1
Detection Limit: 0.1 mg/1
Preparation of Standard Solution
1. Stock Solution: Carefully weigh 1.599 g of lead nitrate, Pb(NO3)2 (analytical reagent
grade), and dissolve in deionized distilled water. When solution is complete acidify with
10 ml redistilled HNO3 and dilute to 1 liter with deionized distilled water. 1 ml = 1 mg
Pb (1000 mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using the same type of acid and at
the same concentration as will result in the sample to be analyzed either directly or after
processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Lead hollow cathode lamp
2. Wavelength: 283.3 nm
3. Fuel: Acetylene
4. Oxidant: Air
5. Type of flame: Oxidizing
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
239.1-1
-------
Notes
1. The analysis of this metal is exceptionally sensitive to turbulence and absorption bands in
the flame. Therefore, some care should be taken to position the light beam in the most
stable, center portion of the flame. To do this, first adjust the burner to maximize the
absorbance reading with a lead standard. Then, aspirate a water blank and make minute
adjustments in the burner alignment to minimize the signal.
2. For levels of lead below 200 ug/1, either the Special Extraction Procedure given in part
9.2 of the Atomic Absorption Methods section or the furnace technique, Method 239.2,
is recommended.
3. The following lines may also be used:
217.0 nm Relative Sensitivity 0.4
261.4 nm Relative Sensitivity 10
4. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
5. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. An interlaboratory study on trace metal analyses by atomic absorption was conducted by
the Quality Assurance and Laboratory Evaluation Branch of EMSL. Six synthetic
concentrates containing varying levels of aluminum, cadmium, chromium, copper, iron,
manganese, lead and zinc were added to natural water samples. The statistical results for
lead were as follows:
Standard
Number True Values Mean Value Deviation Accuracy as
of Labs ug/liter ug/liter ug/liter % Bias
74 367 377 128 2.9
74 334 340 111 1.8
64 101 101 46 -0.2
64 84 85 40 1.1
61 37 41 25 9.6
60 25 31 22 25.7
239.1-2
-------
LEAD
Method 239.2 (Atomic Absorption, furnace technique)
STORE! NO. Total 01051
Dissolved 01049
Suspended 01050
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Lanthanum Nitrate Solution: Dissolve 58.64 g of ACS reagent grade La2O3 in 100 ml
cone. HNO3 and dilute to 1000 ml with deionized distilled water. 1 ml = 50 mg La.
3. Working Lead Solution: Prepare dilutions of the stock lead solution to be used as
calibration standards at the time of analysis. Each calibration standard should contain
0.5% (v/v) HNO3. To each 100 ml of diluted standard add 10 ml of the lanthanum
nitrate solution.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
2. To each 100 ml of prepared sample solution add 10 ml of the lanthanum nitrate solution.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30 sec-500°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 283.3 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure in the calculation see "Furnace Procedure", part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1978
239.2-1
-------
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Greater sensitivity can be achieved using the 217.0 nm line, but the optimum
concentration range is reduced. The use of a lead electrodeless discharge lamp at this
lower wavelength has been found to be advantageous. Also a lower atomization
temperature (2400°C) may be preferred.
4. To suppress sulfate interference (up to 1500 ppm) lanthanum is added as the nitrate to
both samples and calibration standards. (Atomic Absorption Newsletter Vol. 15, No. 3,
p 71, May-June 1976.)
5. Since glassware contamination is a severe problem in lead analysis, all glassware should
be cleaned immediately prior to use, and once cleaned, should not be open to the
atmosphere except when necessary.
6. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
7. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
8. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
9. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 25, 50, and 100 ug Pb/1, the standard deviations were ±1.3, ±1.6, and ±3.7,
respectively. Recoveries at these levels were 88%, 92%, and 95% respectively.
239.2-2
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MERCURY
Method 245.1 (Manual Cold Vapor Technique)
STORE! NO. Total 7190r
Dissolved 71890
Suspended 71895
1. Scope and Application
1.1 This method is applicable to drinking, surface, and saline waters, domestic and industrial
wastes.
1.2 In addition to inorganic forms of mercury, organic mercurials may also be present. These
organo-mercury compounds will not respond to the cold vapor atomic absorption
technique unless they are first broken down and converted to mercuric ions. Potassium
permanganate oxidizes many of these compounds, but recent studies have shown that a
number of organic mercurials, including phenyl mercuric acetate and methyl mercuric
chloride, are only partially oxidized by this reagent. Potasssium persulfate has been
found to give approximately 100% recovery when used as the oxidant with these
compounds. Therefore, a persulfate oxidation step following the addition of the
permanganate has been included to insure that organo-mercury compounds, if present,
will be oxidized to the mercuric ion before measurement. A heat step is required for
methyl mercuric chloride when present in or spiked to a natural system. For distilled
water the heat step is not necessary.
1.3 The range of the method may be varied through instrument and/or recorder expansion.
Using a 100 ml sample, a detection limit of 0.2 ug Hg/1 can be achieved; concentrations
below this level should be reported as < 0.2 (see Appendix 11.2).
2. Summary of Method
2.1 The flameless A A procedure is a physical method based on the absorption of radiation at
253.7 nm by mercury vapor. The mercury is reduced to the elemental state and aerated
from solution in a closed system. The mercury vapor passes through a cell positioned in
the light path of an atomic absorption spectrophotometer. Absorbance (peak height) is
measured as a function of mercury concentration and recorded in the usual manner.
3. Sample Handling and Preservation
3.1 Until more conclusive data are obtained, samples should be preserved by acidification
with nitric acid to a pH of 2 or lower immediately at the time of collection. If only
dissolved mercury is to be determined, the sample should be filtered through an all glass
apparatus before the acid is added. For total mercury the filtration is omitted.
4. Interference
4.1 Possible interference from sulfide is eliminated by the addition of potassium
permanganate. Concentrations as high as 20 mg/1 of sulfide as sodium sulfide do not
interfere with the recovery of added inorganic mercury from distilled water.
Approved for NPDES and SDWA
Issued 1974
245.1-1
-------
4.2 Copper has also been reported to interfere; however, copper concentrations as high as 10
mg/1 had no effect on recovery of mercury from spiked samples.
4.3 Sea waters, brines and industrial effluents high in chlorides require additional
permanganate (as much as 25 ml). During the oxidation step, chlorides are converted to
free chlorine which will also absorb radiation of 253 nm. Care must be taken to assure
that free chlorine is absent before the mercury is reduced and swept into the cell. This
may be accomplished by using an excess of hydroxylamine sulfate reagent (25 ml). In
addition, the dead air space in the BOD bottle must be purged before the addition of
stannous sulfate. Both inorganic and organic mercury spikes have been quantitatively
recovered from sea water using this technique.
4.4 Interference from certain volatile organic materials which will absorb at this wavelength
is also possible. A preliminary run without reagents should determine if this type of
interference is present (see Appendix 11.1).
5. Apparatus
5.1 Atomic Absorption Spectrophotometer: (See Note 1) Any atomic absorption unit having
an open sample presentation area in which to mount the absorption cell is suitable.
Instrument settings recommended by the particular manufacturer should be followed.
Note 1: Instruments designed specifically for the measurement of mercury using the cold
vapor technique are commercially available and may be substituted for the atomic
absorption spectfophotometer.
5.2 Mercury Hollow Cathode Lamp: Westinghouse WL-22847, argon filled, or equivalent.
5.3 Recorder: Any multi-range variable speed recorder that is compatible with the UV
detection system is suitable.
5.4 Absorption Cell: Standard spectrophotometer cells 10 cm long, having quartz end
windows may be used. Suitable cells may be constructed from plexiglass tubing, 1" O.D.
X 4-1/2". The ends are ground perpendicular to the longitudinal axis and quartz
windows (1" diameter X 1/16" thickness), are cemented in place. The cell is strapped to a
burner for support and aligned in the light beam by use of two 2" by 2" cards. One inch
diameter holes are cut in the middle of each card; the cards are then placed over each end
of the cell. The cell is then positioned and adjusted vertically and horizontally to give the
maximum transmittance.
5.5 Air Pump: Any peristaltic pump capable of delivering 1 liter of air per minute may be
used. A Masterflex pump with electronic speed control has been found to be satisfactory.
5.6 Flowmeter: Capable of measuring an air flow of 1 liter per minute.
5.7 Aeration Tubing: A straight glass frit having a coarse porosity. Tygon tubing is used for
passage of the mercury vapor from the sample bottle to the absorption cell and return.
5.8 Drying Tube: 6" X 3/4" diameter tube containing 20 g of magnesium perchlorate (see
Note 2). The apparatus is assembled as shown in Figure 1.
NOTE 2: In place of the magnesium perchlorate drying tube, a small reading lamp with
60W bulb may be used to prevent condensation of moisture inside the cell. The lamp is
positioned to shine on the absorption cell maintaining the air temperature in the cell
about 10°C above ambient.
245.1-2
-------
6. Reagents
6.1 Sulfuric Acid, Cone.: Reagent grade.
6.1.1 Sulfuric acid, 0.5 N: Dilute 14.0 ml of cone, sulfuric acid to 1.0 liter.
6.2 Nitric Acid, Cone: Reagent grade of low mercury content (See Note 3).
NOTE 3: If a high reagent blank is obtained, it may be necessary to distill the nitric acid.
6.3 Stannous Sulfate: Add 25 g stannous sulfate to 250 ml of 0.5 N sulfuric acid. This
mixture is a suspension and should be stirred continuously during use. (Stannous
chloride may be used in place of stannous sulfate.)
6.4 Sodium Chloride-Hydroxylamine Sulfate Solution: Dissolve 12 g of sodium chloride and
12 g of hydroxylamine sulfate in distilled water and dilute to 100 ml. (Hydroxylamine
hydrochloride may be used in place of hydroxylamine sulfate.)
6.5 Potassium Permanganate: 5% solution, w/v. Dissolve 5 g of potassium permanganate in
100 ml of distilled water.
6.6 Potassium Persulfate: 5% solution, w/v. Dissolve 5 g of potassium persulfate in 100 ml
of distilled water.
6.7 Stock Mercury Solution: Dissolve 0.1354 g of mercuric chloride in 75 ml of distilled
water. Add 10 ml of cone, nitric acid and adjust the volume to 100.0 ml. 1 ml = 1 mg
Hg-
O * BUBBLER
ABSORPTION
CELL
SAMPLE
IN BOO
SOLUTION
BOTTLE
SCRUBBER
CONTAINING
A MERCURY
ABSORBING
MEDIA
FIGURE 1. APPARATUS FOR FLAMELESS
MERCURY DETERMINATION
245.1-3
-------
6.8 Working Mercury Solution: Make successive dilutions of the stock mercury solution to
obtain a working standard containing 0.1 ug per ml. This working standard and the
dilutions of the stock mercury solution should be prepared fresh daily. Acidity of the
working standard should be maintained at 0.15% nitric acid. This acid should be added
to the flask as needed before the addition of the aliquot.
7. Calibration
7.1 Transfer 0, 0.5, 1.0, 2.0, 5.0 and 10.0 ml aliquots of the working mercury solution
containing 0 to 1.0 ug of mercury to a series of 300 ml BOD bottles. Add enough distilled
water to each bottle to make a total volume of 100 ml. Mix thoroughly and add 5 ml of
cone, sulfuric acid (6.1) and 2.5 ml of cone, nitric acid (6.2) to each bottle. Add 15 ml of
KMnO4 (6.5) solution to each bottle and allow to stand at least 15 minutes. Add 8 ml of
potassium persulfate (6.6) to each bottle and heat for 2 hours in a water bath mainlined at
95°C. Cool and add 6 ml of sodium chloride-hydroxylamine sulfate solution (6.4) to
reduce the excess permanganate. When the solution has been decolorized wait 30
seconds, add 5 ml of the stannous sulfate solution (6.3) and immediately attach the bottle
to the aeration apparatus forming a closed system. At this point the sample is allowed to
stand quietly without manual agitation. The circulating pump, which has previously
been adjusted to a rate of 1 liter per minute, is allowed to run continuously (See Note 4).
The absorbance will increase and reach maximum within 30 seconds. As soon as the
recorder pen levels off, approximately 1 minute, open the bypass valve and continue the
aeration until the absorbance returns to its minimum value (see Note 5). Close the bypass
valve, remove the stopper and frit from the BOD bottle and continue the aeration.
Proceed with the standards and construct a standard curve by plotting peak height
versus micrograms of mercury.
NOTE 4: An open system where the mercury vapor is passed through the absorption cell
only once may be used instead of the closed system.
NOTE 5: Because of the toxic nature of mercury vapor precaution must be taken to avoid
its inhalation. Therefore, a bypass has been included in the system to either vent the
mercury vapor into an exhaust hood or pass the vapor through some absorbing media,
such as:
a) equal volumes of 0.1 M KMnO4 and 10% H2SO4
b) 0.25 % iodine in a 3 % KI solution
A specially treated charcoal that will adsorb mercury vapor is also available from
Barnebey and Cheney, E. 8th Ave. and N. Cassidy St., Columbus, Ohio 43219,
Cat. #580-13 or #580-22.
8. Procedure
8.1 Transfer 100 ml, or an aliquot diluted to 100 ml, containing not more than 1.0 ug of
mercury, to a 300 ml BOD bottle. Add 5 ml of sulfuric acid (6.1) and 2.5 ml of cone.
nitric acid (6.2) mixing after each addition. Add 15 ml of potassium permanganate
solution (6.5) to each sample bottle. For sewage samples additional permanganate may
be required. Shake and add additional portions of potassium permanganate solution, if
necessary, until the purple color persists for at least 15 minutes. Add 8 ml of potassium
persulfate (6.6) to each bottle and heat for 2 hours in a water bath at 95°C. Cool and add 6
245.1-4
-------
MERCURY
Method 245.2 (Automated Cold Vapor Technique)
STORET NO. Total 71900
Dissolved 71890
Suspended 71895
1. Scope and Application
1.1 This method is applicable to surface waters. It may be applicable to saline waters,
wastewaters, effluents, and domestic sewages providing potential interferences are not
present (See Interference 4).
1.2 The working range is 0.2 to 20.0 ug Hg/1.
2. Summary of Method
2.1 The flameless AA procedure is a physical method based on the absorption of radiation at
253.7 nm by mercury vapor. The mercury is reduced to the elemental state and aerated
from solution. The mercury vapor passes through a cell positioned in the light path of an
atomic absorption spectrophotometer. Absorbance (peak height) is measured as a
function of mercury concentration and recorded in the usual manner.
2.2 In addition to inorganic forms of mercury, organic mercurials may also be present. These
organo-mercury compounds will not respond to the flameless atomic absorption
technique unless they are first broken down and converted to mercuric ions. Potassium
permanganate oxidizes many of these compounds but recent studies have shown that a
number of organic mercurials, including phenyl mercuric acetate and methyl mercuric
chloride, are only partially oxidized by this reagent. Potassium persulfate has been found
to give approximately 100% recovery when used as the oxidant with these compounds.
Therefore, an automated persulfate oxidation step following the automated addition of
the permanganate has been included to insure that organo-mercury compounds, if
present, will be oxidized to the mercuric ion before measurement.
3. Sample Handling and Preservation
3.1 Until more conclusive data are obtained, samples should be preserved by acidification
with nitric acid to a pH of 2 or lower immediately at the time of collection."' If only
dissolved mercury is to be determined, the sample should be filtered before the acid is
added. For total mercury the filtration is omitted.
4. Interference (See NOTE 1)
4.1 Some sea waters and waste-waters high in chlorides have shown a positive interference,
probably due to the formation of free chlorine.
4.2 Interference from certain volatile organic materials which will absorb at this wavelength
is also possible. A preliminary run under oxidizing conditions, without stannous sulfate,
would determine if this type of interference is present.
Approved for NPDES and SDWA
Issued 1974
245.2-1
-------
4.3 Formation of a heavy precipitate, in some wastewaters and effluents, has been reported
upon addition of concentrated sulfuric acid. If this is encountered, the problem sample
cannot be analyzed by this method.
4.4 Samples containing solids must be blended and then mixed while being sampled if total
mercury vlaues are to be reported.
NOTE 1: All the above interferences can be overcome by use of the Manual Mercury
method in this manual.
5. Apparatus
5.1 Technicon Auto Analyzer consisting of:
5.1.1 Sampler II with provision for sample mixing.
5.1.2 Manifold.
5.1.3 Proportioning Pump II or III.
5.1.4 High temperature heating bath with two distillation coils (Technicon Part
#116-0163) in series.
5.2 Vapor-liquid separator (Figure 1).
5.3 Absorption cell, 100 mm long, 10 mm diameter with quartz windows.
5.4 Atomic Absorption Spectrophotometer (See Note 2): Any atomic absorption unit having
an open sample presentation area in which to mount the absorption cell is suitable.
Instrument settings recommended by the particular manufacturer should be followed.
NOTE 2: Instruments designed specifically for the measurement of mercury using the
cold vapor technique are commercially available and may be substituted for the atomic
absorption Spectrophotometer.
5.5 Mercury Hollow Cathode Lamp: Westinghouse WL-22847, argon filled, or equivalent.
5.6 Recorder: Any multi-range variable speed recorder that is compatible with the UV
detection system is suitable.
5.7 Source of cooling water for jacketed mixing coil and connector A-7.
5.8 Heat lamp: A small reading lamp with 60W bulb may be used to prevent condensation of
moisture inside the cell: The lamp is positioned to shine on the absorption cell
maintaining the air temperature in the cell about 10°C above ambient.
6. Reagents
6.1 Sulfuric Acid, Cone: Reagent grade
6.1.1 Sulfuric acid, 2 N: Dilute 56 ml of cone, sulfuric acid to 1 liter with distilled water.
6.1.2 Sulfuric acid, 10%: Dilute 100 ml cone, sulfuric acid to 1 liter with distilled water.
6.2 Nitric acid, Cone: Reagent grade of low mercury content.
6.2.1 Nitric Acid, 0.5% Wash Solution: Dilute 5 ml of cone, nitric acid to 1 liter with
distilled water.
6.3 Stannous Sulfate: Add 50 g stannous sulfate to 500 ml of 2 N sulfuric acid (6.1.1). This
mixture is a suspension and should be stirred continuously during use.
NOTE 3: Stannous chloride may be used in place of stannous sulfate.
6.4 Sodium Chloride-Hydroxylamine Sulfate Solution: Dissolve 30 g of sodium chloride
and 30 g of hydroxylamine sulfate in distilled water to 1 liter.
245.2-2
-------
ml of sodium chloride-hydroxylamine sulfate (6.4) to reduce the excess permanganate.
After a delay of at least 30 seconds add 5 ml of stannous sulfate (6.3) and immediately
attach the bottle to the aeration apparatus. Continue as described under Calibration.
9. Calculation
9.1 Determine the peak height of the unknown from the chart and read the mercury value
from the standard curve.
9.2 Calculate the mercury concentration in the sample by the formula:
ugHg/l =
66
V aliquot I \ volume of aliquot in ml
)
9.3 Report mercury concentrations as follows: Below 0.2 ug/1, <0.2; between 1 and 10
ug/1, one decimal; above 10 ug/1, whole numbers.
10. Precision and Accuracy
10.1 In a single laboratory (EMSL), using an Ohio River composite sample with a
background mercury concentration of 0.35 ug/1, spiked with concentrations of 1.0, 3.0
and 4.0 ug/1, the standard deviations were ±0.14, ±0.10 and ±0.08, respectively.
Standard deviation at the 0.35 level was ±0.16. Percent recoveries at the three levels
were 89, 87, and 87%, respectively.
10.2 In a joint EPA/ASTM interlaboratory study of the cold vapor technique for total
mercury in water, increments of organic and inorganic mercury were added to natural
waters. Recoveries were determined by difference. A statistical summary of this study
follows:
Standard
Number True Values Mean Value Deviation Accuracy as
of Labs ug/liter ug/liter ug/liter % Bias
76 0.21 0.349 0.276 66
80 0.27 0.414 0.279 53
82 0.51 0.674 0.541 32
77 0.60 0.709 0.390 18
82 3.4 3.41 1.49 0.34
79 4.1 3.81 1.12 -7.1
79 8.8 8.77 3.69 -0.4
78 9.6 9.10 3.57 -5.2
11. Appendix
11.1 While the possibility of absorption from certain organic substances actually being present
in the sample does exist, EMSL has not encountered such samples. This is mentioned
only to caution the analyst of the possibility. A simple correction that may be used is as
follows: If an interference has been found to be present (4.4), the sample should be
analyzed both by using the regular procedure and again under oxidizing conditions only,
245.1-5
-------
that is without the reducing reagents. The true mercury value can then be obtained by
subtracting the two values.
11.2 If additional sensitivity is required, a 200 ml sample with recorder expansion may be used
provided the instrument does not produce undue noise. Using a Coleman MAS-50 with a
drying tube of magnesium perchlorate and a variable recorder, 2 mv was set to read full
scale. With these conditions, and distilled water solutions of mercuric chloride at
concentrations of 0.15, 0.10, 0.05 and 0.025 ug/1 the standard deviations
were +0.027, ±0.006, ±0.01 and ±0.004. Percent recoveries at these levels were 107,
83, 84 and 96%, respectively.
11.3 Directions for the disposal of mercury-containing wastes are given in ASTM Standards,
Part 31, "Water", p 349, Method D3223 (1976).
Bibliography
1. Kopp, J. F., Longbottom, M. C. and Lobring, L. B., "Cold Vapor Method for Determining
Mercury", AWWA, vol 64, p. 20, Jan., 1972.
2. Annual Book of ASTM Standards, Part 31, "Water", Standard D3223-73, p 343 (1976).
3. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 156 (1975).
245.1-6
-------
NOTE 4: Hydroxylamine hydrochloride may be used in place of hydroxylamine sulfate.
6.5 Potassium Permanganate: 0.5% solution, w/v. Dissolve 5 g of potassium permanganate
in 1 liter of distilled water.
6.6 Potassium Permanganate, 0.1 N: Dissolve 3.16 g of potassium permanganate in distilled
water and dilute to 1 liter.
6.7 Potassium Persulfate: 0.5% solution, w/v. Dissolve 5 g potassium persulfate in 1 liter of
distilled water.
6.8 Stock Mercury Solution: Dissolve 0.1354 g of mercuric chloride in 75 ml of distilled
water. Add 10 ml of cone, nitric acid and adjust the volume to 100.0 ml. 1.0 ml = 1.0
mgHg.
6.9 Working Mercury Solution: Make successive dilutions of the stock mercury solution
(6.8) to obtain a working standard containing 0.1 ug per ml. This working standard and
the dilutions of the stock mercury solution should be prepared fresh daily. Acidity of the
working standard should be maintained at 0.15% nitric acid. This acid should be added
to the flask as needed before the addition of the aliquot. From this solution prepare
standards containing 0.2,0.5, 1.0,2.0, 5.0, 10.0,15.0 and 20.0 ug Hg/1.
6.10 Air Scrubber Solution: Mix equal volumes of 0.1 N potassium permanganate (6.6) and
10% sulfuric acid (6.1.2).
7, Procedure
7.1 Set up manifold as shown in Figure 2.
7.2 Feeding all the reagents through the system with acid wash solution (6.2.1) through the
sample line, adjust heating bath to 105°C.
7.3 Turn on atomic absorption spectrophotometer, adjust instrument settings as
recommended by the manufacturer, align absorption cell in light path for maximum
transmittance and place heat lamp directly over absorption cell.
7.4 Arrange working mercury standards from 0.2 to 20.0 ug Hg/1 in sampler and start
sampling. Complete loading of sample tray with unknown samples.
7.5 Prepare standard curve by plotting peak height of processed standards against
concentration values. Determine concentration of samples by comparing sample peak
height with standard curve.
NOTE 5: Because of the toxic nature of mercury vapor, precaution must be taken to
avoid its inhalation. Venting the mercury vapor into an exhaust hood or passing the
vapor through some absorbing media such as:
a) equal volumes of 0.1 N KMnO4 (6.6) and 10% H2SO4 (6.1.2).
b) 0.25% iodine in a 3% KI solution, is recommended.
A specially treated charcoal that will adsorb mercury vapor is also available from
Barnebey and Cheney, E. 8th Ave. and North Cassidy St., Columbus, Ohio 43219,
Cat. #580-13 or #580-22.
7.6 After the analysis is complete put all lines except the H2SO4 line in distilled water to wash
out system. After flushing, wash out the H2SO4 line. Also flush the coils in the high
temperature heating bath by pumping stannous sulfate (6.3) through the sample lines
followed by distilled water. This will prevent build-up of oxides of manganese.
245.2-3
-------
8. Precision and Accuracy
8.1 In a single laboratory (SEWL), using distilled water standards at concentrations of 0.5,
1.0, 2.0, 5.0, 10.0 and 20.0 ug Hg/1, the standard deviations
were ±0.04, ±0.07, ±0.09, ±0.20, ±0.40 and ±0.84 ug/1, respectively.
8.2 In a single laboratory (SEWL), using surface water samples spiked with ten organic
mercurials at the 10 ug/1 level, recoveries ranged from 87 to 117%. Recoveries of the
same ten organic mercurials in distilled water at the 10 ug/1 level, ranged from 92% to
125%.
Bibliography
1. Wallace, R. A., Fulkerson, W., Shults, W. D., and Lyon, W. S., "Mercury in the Environment-
The Human Element", Oak Ridge National Laboratory, ORNL-NSF-EP-1, p 31, (January,
1971).
2. Hatch, W. R. and Ott, W. L., "Determination of Sub-Microgram Quantities of Mercury by
Atomic Absorption Spectrophotometry", Anal. Chem. 40, 2085 (1968).
3. Brandenberger, H. and Bader, H., "The Determination of Nanogram Levels of Mercury in
Solution by a Flameless Atomic Absorption Technique", Atomic Absorption Newsletter 6^
101 (1967).
4. Brandenberger, H. and Bader, H., "The Determination of Mercury by Flameless Atomic
Absorption II, A Static Vapor Method", Atomic Absorption Newsletter 7^53 (1968).
5. Goulden, P. D. and Afghan, B. K., "An Automated Method for Determining Mercury in
Water", Technicon, Adv. in Auto. Anal. 2, p 317 (1970).
245.2-4
-------
MERCURY IN SEDIMENT
Method 245.5 (Manual Cold Vapor Technique)
1. Scope and Application
1.1 This procedure'0 measures total mercury (organic f inorganic) in soils, sediments,
bottom deposits and sludge type materials.
1.2 The range of the method is 0.2 to 5 ug/g. The range may be extended above or below the
normal range by increasing or decreasing sample size or through instrument and
recorder control.
2. Summary of Method
2.1 A weighed portion of the sample is digested in aqua regia for 2 minutes at 95°C, followed
by oxidation with potassium permanganate. Mercury in the digested sample is then
measured by the conventional cold vapor technique.
2.2 An alternate digestion'2' involving the use of an autoclave is described in (8.2).
3. Sample Handling and Preservation
3.1 Because of the extreme sensitivity of the analytical procedure and the omnipresence of
mercury, care must be taken to avoid extraneous contamination. Sampling devices and
sample containers should be ascertained to be free of mercury; the sample should not be
exposed to any condition in the laboratory that may result in contact or air-borne
mercury contamination.
3.2 While the sample may be analyzed without drying, it has been found to be more
convenient to analyze a dry sample. Moisture may be driven off in a drying oven at a
temperature of 60°C. No mercury losses have been observed by using this drying step.
The dry sample should be pulverized and thoroughly mixed before the aliquot is
weighed.
4. Interferences
4.1 The same types of interferences that may occur in water samples are also possible with
sediments, i.e., sulfides, high copper, high chlorides, etc.
4.2 Volatile materials which absorb at 253.7 nm will cause a positive interference. In order to
remove any interfering volatile materials, the dead air space in the BOD bottle should be
purged before the addition of stannous sulfate.
5. Apparatus
5.1 Atomic Absorption Spectrophotometer (See Note 1): Any atomic absorption unit
having an open sample presentation area in which to irfount the absorption cell is
suitable. Instrument settings recommended by the particular manufacturer should be
followed.
NOTE 1: Instruments designed specifically for the measurement of mercury using the
cold vapor technique are commercially available and may be substituted for the atomic
absorption Spectrophotometer.
Issued 1974
245.5-1
-------
5.2 Mercury Hollow Cathode Lamp: Westinghouse WL-22847, argon filled, or equivalent.
5.3 Recorder: Any multi-range variable speed recorder that is compatible with the UV
detection system is suitable.
5.4 Absorption Cell: Standard spectrophotometer cells 10 cm long, having quartz end
windows may be used. Suitable cells may be constructed from plexiglass tubing, 1" O.D.
X 4-1/2". The ends are ground perpendicular to the longitudinal axis and quartz
windows (1" diameter X 1/16" thickness) are cemented in place. Gas inlet and outlet
ports (also of plexiglass but 1/4" O.D.) are attached approximately 1/2" from each end.
The cell is strapped to a burner for support and aligned in the light beam to give the
maximum transmittance.
NOTE 2: Two 2" X 2" cards with one inch diameter holes may be placed over each end
of the cell to assist in positioning the cell for maximum transmittance.
5.5 Air Pump: Any peristaltic pump capable of delivering 1 liter of air per minute may be
used. A Masterflex pump with electronic speed control has been found to be satisfactory.
(Regulated compressed air can be used in an open one-pass system.)
5.6 Flowmeter: Capable of measuring an air flow of 1 liter per minute.
5.7 Aeration Tubing: Tygon tubing is used for passage of the mercury vapor from the sample
bottle to the absorption cell and return. Straight glass tubing terminating in a coarse
porous frit is used for sparging air into the sample.
5.8 Drying Tube: 6" X 3/4" diameter tube containing 20 g of magnesium perchlorate (See
Note 3). The apparatus is assembled as shown in the accompanying diagram.
NOTE 3: In place of the magnesium perchlorate drying tube, a small reading lamp with
60W bulb may be used to prevent condensation of moisture inside the cell. The lamp is
positioned to shine on the absorption cell maintaining the air temperature in the cell
about 10°C above ambient.
6. Reagents
6.1 Aqua Regia: Prepare immediately before use by carefully adding three volumes of cone.
HC1 to one volume of cone. HNO3.
6.2 Sulfuric Acid, 0.5 N: Dilute 14.0 ml of cone, sulfuric acid to 1 liter.
6.3 Stannous Sulfate: Add 25 g stannous sulfate to 250 ml of 0.5 N sulfuric acid (6.2). This
mixture is a suspension and should be stirred continuously during use.
6.4 Sodium Chloride-Hydroxylamine Sulfate Solution: Dissolve 12 g of sodium chloride
and 12 g of hydroxylamine sulfate in distilled water and dilute to 100 ml.
NOTE 4: A 10% solution of stannous chloride may be substituted for (6.3) and
hydroxylamine hydrochloride may be used in place of hydroxylamine sulfate in (6.4).
6.5 Potassium Permanganate: 5% solution, w/v. Dissolve 5 g of potassium permanganate in
100 ml of distilled water.
6.6 Stock Mercury Solution: Dissolve 0.1354 g of mercuric chloride in 75 ml of distilled
water. Add 10 ml of cone, nitric acid and adjust the volume to 100.0 ml. 1.0 ml = 1.0
mgHg.
6.7 Working Mercury Solution: Make successive dilutions of the stock mercury solution
(6.6) to obtain a working standard containing 0.1 ug/ml. This working standard and the
dilution of the stock mercury solutions should be prepared fresh daily. Acidity of the
245.5-2
-------
AIR
OUT
AIR AND
SOLUTION
IN
W 7/25 T
J 0.7 cm ID
0.4cm ID
l4cm
SOLUTION
OUT
FIGURE 1. VAPOR LIQUID SEPARATOR
245.2-5
-------
245.2-6
-------
working standard should be maintained at 0.15% nitric acid. This acid should be added
to the flask as needed before the addition of the aliquot.
7. Calibration
7.1 Transfer 0, 0.5, 1.0, 2.0, 5.0 and 10 ml aliquots of the working mercury solution (6.7)
containing 0 to 1.0 ug of mercury to a series of 300 ml BOD bottles. Add enough distilled
water to each bottle to make a total volume of 10 ml. Add 5 ml of aqua regia (6.1) and
heat 2 minutes in a water bath at 95°C. Allow the sample to cool and add 50 ml distilled
water and 15 ml of KMnO4 solution (6.5) to each bottle and return to the water bath for
30 minutes. Cool and add 6 ml of sodium chloride-hydroxylamine sulfate solution (6.4)
to reduce the excess permanganate. Add 50 ml of distilled water. Treating each bottle
individually, add 5 ml of stannous sulfate solution (6.3) and immediately attach the
bottle to the aeration apparatus. At this point, the sample is allowed to stand quietly
without manual agitation. The circulating pump, which has previously been adjusted to
rate of 1 liter per minute, is allowed to run continuously. The absorbance, as exhibited
either on the spectrophotometer or the recorder, will increase and reach maximum
within 30 seconds. As soon as the recorder pen levels off, approximately 1 minute, open
the bypass value and continue the aeration until the absorbance returns to its minimum
value (See Note 5). Close the bypass value, remove the fritted tubing from the BOD
bottle and continue the aeration. Proceed with the standards and construct a standard
curve by plotting peak height versus micrograms of mercury.
NOTE 5: Because of the toxic nature of mercury vapor precaution must be taken to avoid
its inhalation. Therefore, a bypass has been included in the system to either vent the
mercury vapor into an exhaust hood or pass the vapor through some absorbing media,
such as:
a) equal volumes of 0.1 N KMnO4 and 10% H2SO4
b) 0.25% iodine in a 3% KI solution.
A specially treated charcoal that will absorb mercury vapor is also available from
Barnebey and Cheney, E. 8th Ave., and North Cassidy St., Columbus, Ohio 43219,
Cat. #580-13 or #580-22.
8. Procedure
8.1 Weigh triplicate 0.2 g portions of dry sample and place in bottom of a BOD bottle. Add 5
ml of distilled water and 5 ml of aqua regia (6.1). Heat 2 minutes in a water bath at 95°C.
Cool, add 50 ml distilled water and 15 ml potassium permanganate solution (6.5) to each
sample bottle. Mix thoroughly and place in the water bath for 30 minutes at 95°C. Cool
and add 6 ml of sodium chloride-hydroxylamine sulfate (6.4) to reduce the excess
permanganate. Add 55 ml of distilled water. Treating each bottle individually, add 5 ml
of stannous sulfate (6.3) and immediately attach the bottle to the aeration apparatus.
Continue as described under (7.1).
8.2 An alternate digestion procedure employing an autoclave may also be used. In this
method 5 ml of cone. H2SO4 and 2 ml of cone. HNO3 are added to the 0.2 g of sample. 5
ml of saturated KMnO4 solution is added and the bottle covered with a piece of
aluminum foil. The samples are autoclaved at 121°C and 15 Ibs. for 15 minutes. Cool,
make up to a volume of 100 ml with distilled water and add 6 ml of sodium chloride-
245.5-3
-------
hydroxylamine sulfate solution (6.4) to reduce the excess permanganate. Purge the dead
air space and continue as described under (7.1).
9. Calculation
9.1 Measure the peak height of the unknown from the chart and read the mercury value from
the standard curve.
9.2 Calculate the mercury concentration in the sample by the formula:
. _ ug Hg in the aliquot
tfgHg/g - wt Qf t^e ajjquot [n gms
9.3 Report mercury concentrations as follows: Below 0.1 ug/gm, <0.1; between 0.1 and 1
ug/gm, to the nearest 0.01 ug; between 1 and 10 ug/gm, to nearest 0.1 ug; above 10
ug/gm, to nearest ug.
10. Precision and Accuracy
10.1 The following standard deviations on replicate sediment samples were recorded at the
indicated levels; 0.29 ug/g ±0.02 and 0.82 ug/g ±0.03. Recovery of mercury at these
levels, added as methyl mercuric chloride, was 97% and 94%, respectively.
Bibliography
1. Bishop, J. N., "Mercury in Sediments", Ontario Water Resources Comm., Toronto, Ontario,
Canada, 1971.
2. Salma, M., private communication, EPA Cal/Nev Basin Office, Almeda, California.
245.5-4
-------
NICKEL
Method 249.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01067
Dissolved 01065
Suspended 01066
Optimum Concentration Range: 0.3-5 mg/1 using a wavelength of 232.0 nm
Sensitivity: 0.15 mg/1
Detection Limit: 0.04 mg/1
Preparation of Standard Solution
1. Stock Solution: Dissolve 4.953 g of nickel nitrate, Ni(NO3)2»6H2O (analytical reagent
grade) in deionized distilled water. Add 10 ml of cone, nitric acid and dilute to 1 liter
with deionized distilled water. 1 ml = 1 mg Ni (1000 mg/1).
2. Prepare dilutions of the stock nickel solution to be used as calibration standards at the
time of analysis. The calibration standards should be prepared using the same type of
acid and at the same concentration as will result in the sample to be analyzed either
directly or after processing.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.4 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory.
Instrumental Parameters (General)
1. Nickel hollow cathode lamp
2. Wavelength: 232.0 nm
3, Fuel: Acetylene
4. Oxidant: Air
5. Type of Flame: Oxidizing
Analysis Procedure
1. For analysis procedure and calculation, see "Direct Aspiration", part 9.1 of the Atomic
Absorption Methods section of this manual.
Approved for NPDES
Issued 1974
Editorial revision 1978
249.1-1
-------
Interferences
1. The 352.4 nm wavelength is less susceptible to spectral interference and may be used.
The calibration curve is more linear at this wavelength; however, there is some loss of
sensitivity.
Notes
1. For levels of nickel below 100 ug/1, either the Special Extraction Procedure, given in
part 9.2 of the Atomic Absorption Methods section or the furnace technique, Method
249.2, is recommended.
2. Data to be entered into STORET must be reported as ug/1.
3. The heptoxime method may also be used (Standard Methods, 14th Edition, p 232).
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent at
concentrations of 0.20,1.0and5.0mgNi/l, the standard deviations were ±0.011, ±0.02
and ±0.04, respectively. Recoveries at these levels were 100%, 97% and 93%,
respectively.
249.1-2
-------
NICKEL
Method 249.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01067
Dissolved 01065
Suspended 01066
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to used for "standard additions".
3. The calibration standard should be diluted to contain 0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Temp: 30 sec-900°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 232.0 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and pyrolytic
Approved for NPDES
Issued 1978
249.2-1
-------
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Nitrogen may also be used as the purge gas.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Precision and accuracy data are not available at this time.
249.2-2
-------
SELENIUM
Method 270.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01147
Dissolved 01145
Suspended 01146
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 2 ug/1
Preparation of Standard Solution
1. Stock Selenium Solution: Dissolve 0.3453 g of selenous acid (actual assay 94.6% H2SeO3)
in deionized distilled water and make up to 200 ml. 1 ml = 1 mg Se (1000 mg/1).
2. Nickel Nitrate Solution, 5%: Dissolve 24.780 g of ACS reagent grade Ni(NO3)2»6H2O in
deionized distilled water and make up to 100 ml.
3. Nickel Nitrate Solution, 1%: Dilute 20 ml of the 5% nickel nitrate to 100 ml with
deionized distilled water.
4. Working Selenium Solution: Prepare dilutions of the stock solution to be used as
calibration standards at the time of analysis. Withdraw appropriate aliquots of the stock
solution, add 1 ml of cone. HNO3, 2 ml of 30% H2O2 and 2 ml of the 5% nickel nitrate
solution. Dilute to 100 ml with deionized distilled water.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Transfer 100 ml of well-mixed sample to a 250 ml Griffin beaker, add 2 ml of 30% H2O2
and sufficient cone. HNO3 to result in an acid concentration of l%(v/v). Heat for 1 hour
at 95°C or until the volume is slightly less than 50 ml.
2. Cool and bring back to 50 ml with deionized distilled water.
3. Pipet 5 ml of this digested solution into a 10-ml volumetric flask, add 1 ml of the 1%
nickel nitrate solution and dilute to 10 ml with deionized distilled water. The sample is
now ready for injection into the furnace. NOTE: If solubilization or digestion is not
required adjust the HNO3 concentration of the sample to 1 % (v/v) and add 2 ml of 30%
H2O2 and 2 ml of 5% nickel nitrate to each 100 ml of sample. The volume of the
calibration standard should be adjusted with deionized distilled water to match the
volume change of the sample.
Approved for NPDES and SDWA
Issued 1978
270.2-1
-------
Instrument Parameters
1. Drying time and temperature: 30 sec @ 125°C
2. Charring time and temperature: 30 sec © 1200°C
3. Atomizing time and temperature: 10 sec @ 2700°C
4. Purge Gas Atmosphere: Argon
5. Wavelength: 196.0 nm.
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HG A-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Selenium analysis suffers interference from chlorides (> 800 mg/1) and sulfate (> 200
mg/1). For the analysis of industrial effluents and samples with concentrations of sulfate
from 200 to 2000 mg/1, both samples and standards should be prepared to contain 1%
nickel.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
6. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
7. Data to entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Using a sewage treatment plant effluent containing <2 ug/1 and spiked with a
concentration of 20 ug/1, a recovery of 99% was obtained.
2. Using a series of industrial waste effluents spiked at a 50 ug/1 level, recoveries ranged
from 94 to 112%.
3. Using a 0.1% nickel nitrate solution as a synthetic matrix with selenium concentrations
of 5, 10, 20, 40, 50, and 100 ug/1, relative standard deviations of 14.2, 11.6, 9.3, 7.2, 6.4
and 4.1 %, respectively, were obtained at the 95% confidence level.
270.2-2
-------
4. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 5, 10, and 20 ug Se/1, the standard deviations were ±0.6, ±0.4, and ±0.5,
respectively. Recoveries at these levels were 92%, 98%, and 100%, respectively.
Reference:
"Determining Selenium in Water, Wastewater, Sediment and Sludge By Flameless Atomic
Absorption Spectroscopy", Martin, T. D., Kopp, J. F. and Ediger, R. D. Atomic Absorption
Newsletter 14,109 (1975).
270.2-3
-------
SELENIUM
Method 270.3 (Atomic Absorption, gaseous hydride)
STORE! NO. Total 01147
Dissolved 01145
Suspended 01146
1. Scope and Application
1.1 The gaseous hydride method determines inorganic selenium when present in
concentrations at or above 2 ug/1. The method is applicable to drinking water and most
fresh and saline waters, in the absence of high concentrations of chromium, cobalt,
copper, mercury, molybdenum, nickel and silver.
2. Summary of Method
2.1 Selenium in the sample is reduced from the + 6 oxidation state to the + 4 oxidation state
by the addition of SnCl2. Zinc is added to the acidified sample, producing hydrogen and
converting the selenium to the hydride, SeH2. The gaseous selenium hydride is swept into
an argon-hydrogen flame of an atomic absorption spectrophotometer. The working
range of the method is 2-20 ug/1 using the 196.0 nm wavelength.
3. Comments
3.1 In analyzing drinking water and most surface and ground waters, interferences are rarely
encountered. Industrial waste samples should be spiked with a known amount of
selenium to establish adequate recovery.
3.2 Organic forms of selenium must be converted to an inorganic form and organic matter
must be oxidized before beginning the analysis. The oxidation procedure given in method
206.5 (Standard Methods, Uth Ed. 404B, p 285, Procedure 4.1) should be used.
3.3 For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
3.4 For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
3.5 Data to be entered into STORET must be reported as ug/1.
4. Precision and Accuracy
4.1 Ten replicate solutions of selenium oxide at the 5, 10 and 15 ug/1 level were analyzed by
a single laboratory. Standard deviations at these levels were ±0.6, ±1.1 and ±2.9 with
recoveries of 100, 100 and 101%. (Caldwell, J. S., Lishka, R. J., and McFarren, E. F.,
"Evaluation of a Low-Cost Arsenic and Selenium Determination at Microgram per Liter
Levels", JAWWA, vol 65, p. 731, Nov. 1973.)
Approved for NPDES and SDWA
Issued 1974
270.3-1
-------
5. References
5.1 Except for the perchloric acid step, the procedure to be used for this determination is
found in:
Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 159,
Method 301A(VII), (1975)
270.3-2
-------
SILVER
Method 272.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01077
Dissolved 01075
Suspended 01076
Optimum Concentration Range: 1-25 ug/1
Detection Limit: 0.2 ug/1
Preparation of Standard Solution
1. Stock Solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to be used for "standard additions".
3. The calibration standard should be diluted to contain 0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125°C.
2. Ashing Time and Tem p: 30 sec-400°C.
3. Atomizing Time and Temp: 10 sec-2700°C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 328.1 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES and SDWA
Issued 1978
272.2-1
-------
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. Background correction may be required if the sample contains high dissolved solids.
3. The use of halide acids should be avoided.
4. If adsorption to container walls or formation of AgCl is suspected, see NOTE 3 under the
Direct Aspiration Method 272.1.
5. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
6. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
7. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
8. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy:
1. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 25, 50, and 75 ug Ag/1, the standard deviations were tO.4, iO.7, and +0.9,
respectively. Recoveries at these levels were 94%, 100% and 104%, respectively.
272.2-2
-------
SILVER
Method 272.1 (Atomic Absorption, direct aspiration)
STORET NO. Total 01077
Dissolved 01075
Suspended 01076
Optimum Concentration Range: 0.1-4 mg/1 using a wavelength of 328.1 nm
Sensitivity: 0.06 mg/1
Detection Limit: 0.01 nig/1
Preparation of Standard Solution
1. Stock Solution: Dissolve 1.575 g of AgNO3 (analytical reagent grade) in deionized
distilled water, add 10 ml cone. HNO3 and make up to 1 liter. 1 ml = 1 mg Ag (1000
mg/1).
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. The calibration standards should be prepared using nitric acid and at the same
concentration as will result in the sample to be analyzed either directly or after
processing.
3. Iodine Solution, 1 N: Dissolve 20 grams of potassium iodide, KI (analytical reagent
grade) in 50 ml of deionized distilled water, add 12.7 grams of iodine, I2 (analytical
reagent grade) and dilute to 100 ml. Store in a brown bottle.
4. Cyanogen Iodide (CNI) Solution: To 50 ml of deionized distilled water add 4.0 ml cone.
NH4OH, 6.5 grams KCN, and 5.0 ml of 1.0 NI2 solution. Mix and dilute to 100 ml with
deionized distilled water. Fresh solution should be prepared every two weeks.'0
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.3 of the
Atomic Absorption Methods section of this manual have been found to be satisfactory;
however, the residue must be taken up in dilute nitric acid rather than hydrochloric to
prevent precipitation of AgCl.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974
Technical revision 1978
272.1-1
-------
Instrumental Parameters (General)
1. Silver hollow cathode lamp
2. Wavelength: 328.1 nm
3. Fuel: Acetylene
4. Oxidant: Air
5. Type of flame: Oxidizing
Analysis Procedure
1. For the analysis-procedure and the calculation, see "Direct Aspiration", part 9.1 of the
Atomic Absorption Methods section of this manual.
Notes
1. For levels of silver below 30 ug/1, either the Special Extraction Procedure, given in part
9.2 of the Atomic Absorption Methods section or the furnace procedure, Method 272.2,
is recommended.
2. Silver nitrate standards are light sensitive. Dilutions of the stock should be discarded
after use as concentrations below 10 mg/1 are not stable over long periods of time.
3. If absorption to container walls or the formation of AgCl is suspected, make the sample
basic using cone. NH4OH and add 1 ml of (CNI) solution per 100 ml of sample. Mix the
sample and allow to stand for 1 hour before proceeding with the analysis.'"
4. The 338.2 nm wavelength may also be used. This has a relative sensitivity of 2.
5. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a round-robin study reported by Standard Methods, a synthetic sample containing 50
ug Ag/1 was analyzed by 50 laboratories with a reported standard deviation of ±8.8 and
a relative error 10.6%.
References *
1. "The Use of Cyanogen Iodide (CNI) as a Stabilizing Agent for Silver in Photographic
Processing Effluent Sample", Owerbach, Daniel, Photographic Technology Division,
Eastman Kodak Company, Rochester, N.Y. 14650.
2. Standard Methods for Examination of Water and Wastewater, 14th Edition, p. 148,
Method 301A.
272.1-2
-------
APPENDIX III
METHODS FOR BENZIDINE, CHLORINATED ORGANIC COMPOUNDS,
PENTACHLOROPHENOL AND PESTICIDES
IN WATER AND WASTEWATER
-------
METHODS FOR BENZIDINE, CHLORINATED ORGANIC COMPOUNDS,
PENTACHLOROPHENOL AND PESTICIDES
IN WATER AND WASTEWATER
INTERIM
Pending Issuance of
Methods for Organic Analysis
of Water and Wastes
U.S. ENVIRONMENTAL PROTECTION AGENCY
ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
CINCINNATI, OHIO 42568
September 1978
-------
FOREWORD
This collection of methods for the determination of benzidine,
chlorinated organic compounds, pentachlorophenol and pesticides has been
assembled by the staff of the Environmental Monitoring and Support
Laboratory - Cincinnati (EMSL-Cinti.) for use by the NPDES Permits
Program.
These methods are as referenced in the Federal Register of
December 1, 1976 and are being provided only for the interim period until
the manual "Methods for Organic Analysis of Water and Wastes" becomes
available.
Dwight G. Ballinger, Director
Environmental Monitoring and Support Laboratory - Cincinnati
-------
DISCLAIMER
The mention of trade names or commercial products in this manual is for
illustration purposes, and does not constitute endorsement or recommendatior
by the U. S. Environmental Protection Agency.
111
-------
TABLE OF CONTENTS
Page
Method for Benzidine and Its Salts in Water and Wastewater 1
Method for Chlorinated Hydrocarbons in Water and Wastewater 7
Method for Organophosphorus Pesticides in Water and Waste-
water 25
Method for Polychlorinated Biphenyls (PCBs) in Water and Waste-
water 43
Method for Triazine Pesticides in Water and Wastewater 83
Method for 0-aryl Carbamate Pesticides in Water and Wastewater 94
Method for N-aryl Carbamate and Urea Pesticides in Water and
Wastewater 104
Method for Chlorophenoxy Acid Pesticides in Water and Wastewater 115
Method for Volatile Chlorinated Organic Compounds in Water and
Wastewater 130
Method for Pentachlorphenol in Water and Wastewater 140
Appendix I 141
Appendix II 146
Appendix III 149
Appendix IV 151
Bibliography 154
IV
-------
PAGE REFERENCES
F.R.# Parameter
14
94
95
Benzidine
EPA
This Manual
1
14th ed.
Std. Methods
ASTM
(1975)
USGS*
Chlorinated organic
compounds:
Benzylchloride 130
Carbon tetrachloride 130
Chlorobenzene 130
Chloroform 130
Epichlorohydrin 130
Heptachloro epoxide
Methylene chloride 130
PCB-1016 43
PCB-1221 43
PCB-1232 43
PCB-1242 43
PCB-1248 43
PCB-1254 43
PCB-1260 43
1,1,2,2-Tetrachloroethane 130
Tetrachloroethylene 130
1,2,4-Trichlorobenzene 130
1,1,2-Trichloroethane 130
Pentachlorophenol
Pesticides
Aldrin
Ametryn
Aminocarb
Atraton
Atrazine
Azinphos methyl
Bar ban
BHC
Captan
Carbaryl
Carbophenothion
Chlordane
Chlorpropham
2,4-D
140
7
83
94
83
83
25
104
7
7
94
7
104
115
555
529
30
555
529 • 30
555
555
555
529
529
30
35
STORE!
NUMBER
39120
32102
34301
32160
39420
34423
34671
39488
39492
39496
39500
39504
39508
34475
39032
39330
39033
39640
39750
39350
-------
F.R.# Parameter
EPA
This Manual
14th ed.
Std. Methods
ASTM
(1975)
USGS*
STORET
NUMBER
ODD
DDE
DDT
Demeton-0
Diazinon
Dicamba
Dichlorofenthion
Dichloran
Dicofol
Dieldrin
Dioxathion
Disulfoton
Diuron
Endosulfan
Edrin
Ethion
Fenuron
Fenuron - TCA
Heptachlor
Isodrin
Lindane
Linuron
Malath ion
Methiocarb
Methoxychlor
Mexacarbate
Mi rex
Monuron
Monuron-TCA
Neburon
Parathion methyl
Parathion ethyl
PCNB
Perthane
Prometon
Prometryn
Propazine
Propham
Proporur
Secbumeton
Siduron
Si 1 vex
Simazine
Strobane
Swep
2,4,5-T
Terbuthylazine
7
7
7
25
25
115
555
555
555
529
529
529
25
104
7
7
104
104
7
7
104
25
94
7
94
7
104
104
104
25
25
7
83
83
83
104
94
83
104
115
83
7
104
115
83
555
555
555
555
555
555
555
555
555
555
555
555
529
529
529
529
529
529
529
529
30
30
30
30"
30
30
30
30
30
30
30
30
30
30
30
35
35
39360
39365
39370
39560
39570
39780
39010
39650
39388
39390
39398
39410
39430
39782
39530
39489
39755
39600
39540
39029
39034
39056
39057
39024
39052
39760
39055
vi
-------
F.R.I Parameter EPA 14th ed. ASTM USGS* STORET
This Manual Std. Methods (1975) NUMBER
Toxaphene 7 555 529 30 39400
Trifluraline 7 — — — 39030
*Goerlitz, D. & Brown, E. "Methods for Analysis of Organic Substances In
Water," U.S. Geological Survey Techniques of Water-Resources Inv. Book 5, Ch.
A3 (1972).
vii
-------
METHOD FOR BENZIDINE AND ITS SALTS IN WASTEWATERS
1. Scope and App1i cat i on
1.1 This method covers the determination for benzidine and its salts
in water and wastewaters. The method can be modified to apply
also to the determination of closely related materials as des-
cribed under Interferences (4.2).
1.2 The salts of benzidine, such as benzidine sulfate, are measured
and reported as benzidine, STORET NO. 39120.
1.3 The method detection limit is 0.2 ;jg/l when analyzing 1 liter of
sample.
2. Summary
2.1 The water sample is made basic and the benzidine is extracted
with ethyl acetate. Cleanup is accomplished by extracting the
benzidine from the ethyl acetate with hydrochloric acid.
Chloramine-T is added to the acid solution to oxidize the benzi-
dine. The yellow oxidation product is extracted with ethyl
acetate and measured with a scanning spectrophotometer. The
spectrum from 510 nm to 370 nm is used for qualitative identi-
fication.
3. Hazards
3.1 Benzidine is a known carcinogen. All manipulations of this
method should be carried out in a hood with protection
1
-------
provided for the hands and arms of the analyst. Consult OSHA
regulations (1) before working with benzidine.
4. Interferences
4.1 The multiple extractions effectively limit the interferences to
organic bases. The oxidation with Chloramine-T to form a yellow
product is very selective and has been described in detail
(2,3). The use of the absorption spectrum for the identi-
fication of benzidine results in a highly specific procedure.
4.2 Some compounds having a structure very similar to benzidine will
interfere with the quantification, if present. Examples of
these interfering compounds are dichlorobenzidine, o-tolidine,
and dianisidine.
4.3 A general yellow background color in the extract will limit the
cell pathlength that can be employed and thus limit the sensi-
tivity of the method.
5. Apparatus and Materials
5.1 Spectrophotometer-visible, scanning (510-370 nm).
5.2 Separatory Funnels - 125 ml, 250 ml, 2000 ml.
5.3 Cells - 1 to 5 cm pathlength, 20 ml volume maximum.
6. Reagents, Solvents and Standards
6.1 Ethyl acetate
6.2 Hydrochloric acid (IN)- Add 83 ml cone, hydrochloric acid to
water and dilute to one liter.
6.3 Chloramine-T - 10% solution. Prepare fresh daily by dissolving
l.Og Chloramine-T in 10 ml distilled water.
-------
6.4 Stock standard (0.2 jjg/jjl) - Dissolve 100.0 mg purified benzi-
dine in about 30 ml 1 N HC1. Dilute to 500 ml with water.
7. Preparation of Calibration Curve
7.1 To a series of 125-ml separatory funnels, add 45 ml of hydro-
chloric acid and 10 ml of ethyl acetate. Shake for one minute
to saturate the acid layers. Discard the solvent layers. Dose
the series with volumes from 1.0 to 20.0 /jl of stock standard,
using syringes.
7.2 Treat standards according to the Procedure beginning with 8.5.
8. Quality Control
8.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts should be developed
and used as a check on the analytical system. Quality control
check samples and performance evaluation samples should be
analyzed on a regular basis.
8.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
9. Procedure
9.1 Adjust the sample pH to 8.5 to 9.0 with dilute NaOH or HC1.
9.2 Transfer 1 liter of sample to a 2000-ml separatory funnel. Add
150 ml ethyl acetate and shake for two minutes. Allow the
layers to separate, then drain the water layer into a
-------
second 2-liter separatory funnel. Drain the solvent layer into
a 250-ml separatory funnel.
9.3 Repeat the extraction of the water layer twice more with
50-ml portions of ethyl acetate. Combine all solvent layers,
then discard the water layer.
9.4 Extract the solvent layer three times with 15-ml portions of
hydrochloric acid by shaking 2 minutes and allowing the phases
to separate. Combine the acid layers in a glass stoppered
container for cold storage until time is available for analysis,
or transfer the layers directly into a 125-ml separatory funnel.
9.5 Prepare the spectrophotometer so it is warmed and ready to use.
The remaining steps of the procedure must be performed rapidly
on one sample at a time.
9.6 To the hydrochloric acid solution in a 125 ml separatory funnel,
add 1.0 ml chloramine-T solution and mix. Add
25.0-ml ethyl acetate with a pipet and shake for two minutes.
Allow the layers to separate, then discard the aqueous phase.
9.7 Filter the solvent layer through coarse filter paper and fill a
5-cm cell with the filtrate.
9.8 Scan the solvent from 510 nm to 370 nm. Ethyl acetate is used
for a blank with double beam instruments. Shorter pathlength
cells should be used in cases where absorbance exceeds 0.8.
10. Calculation of Results
10.1 Benzidine is identified by its absorbance maximum at
436 nm. Dichlorobenzidine gives similar response but has its
absorbance maximum at 445 nm.
-------
10.2 Construct a baseline from the absorbance minimum at about 470 nm
to the minimum at 390 nm (or 420 nm minimum for samples with a
high background). Record the absorbance ofthe peak maximum and
the absorbance of the constructed baseline at the 436 nm. Treat
samples and standards in the same fashion.
10.3 Using the net absorbance values, prepare a calibration plot from
the standards. Determine the total micrograms in each sample
from this plot.
10.4 Divide the total micrograms by the sample volume, in liters, to
determine /jg/1. Correct results for cell pathlength if
necessary.
11. Reporting Results
11.1 Report results in micrograms per liter as benzidine without
correction for recovery data. When duplicate and spike samples
are analyzed all data obtained should be reported.
12. Accuracy and Precision
12.1 When 1 liter samples of river water were dosed with 1.80 pg of
benzidine, an average of 1.24;jg was recovered. The standard
deviation was 0.092 ^g/1 (n«8).
-------
REFERENCES:
1. Federal Register. Volume 39, Page 3779, Paragraph 1910.93; (January 29,
T974Y:
2. Glassman, J. M., and Meigs, 0. W., "Benzidine (4,4'-Diaminobiphenyl) and
Substituted Benzidines", Arch. Industr. Hyg.. i, 519, (1951).
3. Butt, L. T. and Strafford, N., "Papilloma of the Bladder in the Chemical
Industry. Anlaytical Methods for the Determination of Benzidine and
B-Naphtylamine, Recommended by A.B.C.M. Sub-Committee", J. Appl. Chem.,
6, 525 (1956).
-------
METHOD FOR CHLORINATED HYDROCARBONS IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of various organo-
chlorine pesticides and heptachlor epoxide in water and
wastewater.
1.2 The following pesticides may be determined individually by this
method:
Parameter Storet No.
Aldrin 39330
BHC
Captan 39640
Chlordane 39350
ODD 39360
DDE 39365
DDT 39370
Dichloran
Dieldrin 39380
Endosulfan 39388
Endrin 39390
Heptachlor 39410
Lindane 39782
Methoxychlor 39480
Mi rex 39755
PCNB 39029
Strobane
Toxaphene 39400
Trifluralin 39030
1.3 The following chlorinated organic compound may be determined
individually by this method:
Compound Storet No.
Heptachlor epoxide
-------
2. Summary
2.1 The method offers several analytical alternatives, dependent on
the analyst's assessment of the nature and extent of interfer-
ences and/or the complexity of the pesticide mixtures found.
Specifically, the procedure describes the use of an effective
co-solvent for efficient sample extraction; provides, through
use of column chromatography and liquid-liquid partition,
methods for elimination of non-pesticide interferences and the
pre-separation of pesticide mixtures. Identification is made
by selective gas chromatographic separations and may be corro-
borated through the use of two or more unlike columns.
Detection and measurement is accomplished by electron capture,
microcoulometric or electrolytic conductivity gas chromato-
graphy. Results are reported in micrograms per liter.
2.2 Confirmation of the identity of the compounds should be made by
6C-MS when a new or undefined sample type is being analyzed and
the concentration is adequate for such determination.
2.3 This method is recommended for use only by experienced pesti-
cide analysts or under the close supervision of such qualified
persons.
3. Interferences
3.1 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. Specific
selection of reagents and purification of solvents by distill-
ation in all-glass systems may be required. Refer to Appendix
I.
3.2 The interferences in industrial effluents are high and varied
and often pose great difficulty in obtaining accurate and
precise measurement of organochlorine pesticides. Sample
clean-up procedures are generally required and may result in
the loss of certain organochlorine pesticides. Therefore,
great care should be exercised in the selection and use of
methods for eliminating or minimizing interferences. It is not
possible to describe procedures for overcoming all of the
interferences that; may be encountered in industrial effluents.
3.3 Polychlorinated Biphenyls (PCBs) - Special attention is called
to industrial plasticizers and hydraulic fluids such as the
PCBs, which are a potential source of interference in pesticide
analysis. The presence of PCBs is indicated by a large number
of partially resolved or unresolved peaks which may occur
throughout the entire chromatogram. Particularly severe PCB
interference will require special separation procedures (1, 2).
3.4 Phthalate Esters - These compounds, widely used as
plasticizers, respond to the electron capture detector and are
a source of interference in the determination of organochlorine
pesticides using this detector. Water leaches these materials
from plastics, such as polyethylene bottles and tygon tubing.
-------
The presence of phthalate esters is implicated in samples that
respond to electron capture but not to the microcoulometric or
electrolytic conductivity halogen detectors or to the flame
photometric detector.
3.5 Organophosphorus Pesticides - A number of organophosphorus
pesticides, such as those containing a nitro group, e.g., para-
thion, also respond to the electron capture detector and may
interfere with the determination of the organochlorine pesti-
cides. Such compounds can be identified by their response to
the flame photometric detector (3).
4. Apparatus and Materials
4.1 Gas Chromatograph - Equipped with glass lined injection port.
4.2 Detector Options:
4.2.1 Electron Capture - Radioactive (tritium or nickel-63)
4.2.2 Microcoulometric Titration
4.2.3 Electrolytic Conductivity
4.3 Recorder - Potentiometric strip chart (10 in.) compatible with
the detector.
4.4 Gas Chromatographic Column Materials:
4.4.1 Tubing - Pyrex (180 cm long X 4 mm ID)
4.4.2 Glass Wool - Silanized
4.4.3 Solid Support - Gas-Chrom-Q (100-120 mesh)
4.4.4 Liquid Phases - Expressed as weight percent coated on
solid support.
4.4.4.1 OV-1, 3%
4.4.4.2 OV-210, 5%
10
-------
4.4.4.3 OV-17, 1.5* plus QF-1 or OV-210, 1.95%
4.4.4.4 QF-1, 6% plus SE-30, 4%
4.5 Kuderna-Danish (K-D) Glassware
4.5.1 Snyder Column - three-ball (macro) and two-ball
(micro)
4.5.2 Evaporative Flasks - 500 ml
4.5.3 Receiver Ampuls - 10 ml, graduated
4.5.4 Ampul Stoppers
4.6 Chromatographic Column - Chromaflex (400 mm long x 19 mm ID)
with coarse fritted plate on bottom and Teflon stopcock; 250-ml
reservoir bulb at top of column with flared out funnel shape at
top of bulb - a special order (Kontes K-420540- 9011).
4.7 Chromatographic Column - pyrex (approximately 400 mm long x 20
mm ID) with coarse fritted plate on bottom.
4.8 Micro Syringes - 10, 25, 50 and 100 ;jl.
4.9 Separatory funnels - 125 ml, 1000 ml and 2000 ml with Teflon
stopcock.
4.10 Blender - High speed, glass or stainless steel cup.
4.11 Graduated cylinders - 100 and 250 ml.
4.12 Florisil - PR Grade (60-100 mesh); purchase activated at
1250°F and store in the dark in glass containers with glass
stoppers or foil-lined screw caps. Before use, activate each
batch overnight at 130°C in foil-covered glass container.
Determine lauric-acid value (See Appendix II).
5. Reagents, Solvents, and Standards
5.1 Sodium Chloride - (ACS) Saturated solution in distilled water
11
-------
(pre-rinse NaCl with hexane).
5.2 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.3 Sodium Sulfate - (ACS) Granular, anhydrous (conditioned at 400
C for 4 hrs.).
5.4 Sulfuric Acid - (ACS) Mix equal volumes of cone. f^SO^ with
distilled water.
5.5 Diethyl Ether - Nanograde, redistilled in glass, if necessary.
5.5.1 Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM Laboratories,
Inc., 500 Executive Blvd., Elmsford, N.Y. 10523.)
5.5.2 Procedures recommended for removal of peroxides are
provided with the test strips.
5.6 Acetonitrile, Hexane, Methanol, Methylene Chloride, Petroleum
Ether (boiling range 30-60aC) - nanograde, redistill in glass
if necessary.
5.7 Pesticide Standards - Reference grade.
6. Calibration
6.1 Gas chromatographic operating conditions are considered accept-
able if the response to dicapthon is at least 50% of full scale
when < 0.06 ng is injected for electron capture detection and
^100 ng is injected for microcoulometric or electrolytic
conductivity detection. For all quantitative measurements, the
detector must be operated within its linear response range and
the detector noise level should be less than 2% of full scaJe.
6.2 Standards are injected frequently as a check on the stability
of operating conditions. Gas chromatograms of several standard
12
-------
pesticides are shown in Figures 1, 2, 3 and 4 and provide
reference operating conditions for the four recommended columns.
6.3 The elution order and retention ratios of various organo-
chlorine pesticides are provided in Table 1, as a guide.
7. Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts (4) should be developed
and used as a check on the analytical system. Quality control
check samples and performance evaluation samples should be
analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
8. Sample Preparation
8.1 The sample size taken for analysis is dependent on the type of
sample and the sensitivity required for the purpose at hand.
Background information on the pesticide levels previously
detected at a given sampling site will assist in determining
the sample size required, as well as the final volume to which
the extract needs to be concentrated. A 1-liter sample is
usually taken for drinking water and ambient water analysis to
provide a detection limit of O.OBOto 0.100/jg/l. One-hundred
milliliters is usually adequate to provide a detection limit of
1 ;jg/l for industrial effluents.
13
-------
1N3A10S
wiviimiiu
3H8* —
8N3d
NIH01V
]QIXOd3 id3H
NIH013IO
M01H3AXON13N
14
-------
IS 10 5 0
RETENTION TIME IN MINUTES
Figure 2. Column Packing: 5% OV-210, Carrier Gas: Argon/Methane
at 70 ml/ffiin, Column Temperature: 180 C, Detector:
Electron Capture.
15
-------
1N3A10S
3H9»
9N3d
NIHQ1V
Niivanuiai
NVbOlHOIQ
NIUQ13IQ
I NVJinSOQN]
1QQ .(I'd,
H01H3AXOHJL3W
X]8IW
CO
a>
as ..
a> •»—
"r- o
v_ a»
ae co
oo cs
CM
ra
a>
a>
3
CxO
16
-------
1N3A10S
Nnvinuiu
jHa>»
NVH01H3IO + 8N3d
NIHQ13IO
« „;
a> co
NNQH1
o - CJ
X3HIN
17
-------
Table 1
RETENTION RATIOS OF VARIOUS ORGANOCHLORINE PESTICIDES RELATIVE TO ALDRIN
Liquid
Phase1 1
Column Temp.
Argon/Methane
Carrier Flow
Pesticide
Trifluralin
«-BHC
PCNB
Lindane
Dichloran
Heptachlor
Aldrin
Heptachlor Epoxide
Endosulfan I
p,p'-DDE
Dieldrin
Captan
Endrin
o,p'-DDT
p,p'-DDD
Endosulfan II
p,p'-DDT
Mi rex
Methoxychlor
Aldrin
(Win. absolute)
1.5% OV-17
+
.95% QF-12
200 C
60 ml/min
RR
0.39
0.54
0.68
0.69
0.77
0.82
1.00
1.54
1.95
2.23
2.40
2.59
2.93
3.16
3.48
3.59
4.18
6.1
7.6
3.5
5%
OV-210
180 C
70 ml/min
RR
1.11
0.64
0.85
0.81
1.29
0.87
1.00
1.93
2.48
2.10
3.00
4.09
3.56
2.70
3.75
4.59
4.07
3.78
6.5
2.6
3%
OV-1
180 C
70 ml/min
RR
0.33
0.35
0.49
0.44
0.49
0.78
1.00
1.28
1.62
2.00
1.93
1.22
2.18
2.69
2.61
2.25
3.50
6.6
5.7
4.0
6% QF-1
+
4% SE-30
200 C
60 ml/min
RR
0.57
0.49
0.63
0.60
0.70
0.83
1.00
1.43
1.79
1.82
2.12
1.94
2.42
2.39
2.55
2.72
3.12
4.79
4.60
5.6
columns glass, 180 cm x 4 mm ID, solid support Gas-Chrom Q (100/120
mesh)
ZOV-210 also may be used
18
-------
8.2 Quantitatively transfer the proper aliquot of sample from the
sample container into a two-liter separatory funnel. If less
than 800 ml is analyzed, dilute to one liter with interference
free distilled water.
9. Extraction
9.1 Add 60 ml of 15% methylene chloride in hexane (v:v) to the
sample in the separatory funnel and shake vigorously for two
minutes.
9.2 Allow the mixed solvent to separate from the sample, then draw
the water into a one-liter Erlenmeyer flask. Pour the organic
layer into a 100 ml beaker and then pass it through a column
containing 3-4 inches of anhydrous sodium sulfate, and collect
it in a 500 ml K-D flask equipped with a 10 ml ampul. Return
the water phase to the separatory funnel. Rinse the Erlenmeyer
flask with a second 60-ml volume of solvent; add the solvent to
the separatory funnel and complete the extraction procedure a
second time. Perform a third extraction in the same manner.
9.3 Concentrate the extract in the K-D evaporator on a hot water
bath.
9.4 Analyze by gas chromatography unless a need for cleanup is
indicated (See Section 10).
10. Clean-up and Separation Procedures
10.1 Interferences in the form of distinct peaks and/or high back-
ground in the initial gas chromatographic analysis, as well as
the physical characteristics of the extract (color, cloudiness,
viscosity) and background knowledge of the sample will indicate
19
-------
whether clean-up is required. When these interfere with
measurement of the pesticides, or affect column life or
detector sensitivity, proceed as directed below.
10.2 Acetonitrile Partition - This procedure is used to isolate fats
and oils from the sample extracts. It should be noted that not
all pesticides are quantitatively recovered by this procedure.
The analyst must be aware of this and demonstrate the effi-
ciency of the partitioning for specific pesticides. All of the
pesticides listed in Scope (1.2) with the exception of mirex
,are efficiently recovered.
10.2.1 Quantitatively transfer the previously concentrated
extract to a 125-ml separatory funnel with enough
hexane to bring the final volume to 15 ml. Extract the
sample four times by shaking vigorously for one minute
with 30-ml portions of hexane-saturated acetonitrile.
10.2.2 Combine and transfer the acetonitrile phases to a
one-liter separatory funnel and add 650 ml of distilled
water and 40 ml of saturated sodium chloride solution.
Mix thoroughly for 30-45 seconds. Extract with two
100-ml portions of hexane by vigorously shaking about
15 seconds.
10.2.3 Combine the hexane extracts in a one-liter separatory
funnel and wash with two 100-ml portions of distilled
water. Discard the water layer and pour the hexane
layer through a 3-4 inch anhydrous sodium sulfate
column into a 500-ml K-D flask equipped with a 10-ml
20
-------
ampul. Rinse the separatory funnel and column with
three 10-ml portions of hexane.
10.2.4 Concentrate the extracts to 6-10 ml in the K-D evapor-
ator in a hot water bath.
10.2.5 Analyze by gas chromatography unless a need for further
cleanup is indicated.
10.3 Florisil Column Adsorption Chromatography
10.3.1 Adjust the sample extract volume to 10 ml.
10.3.2 Place a charge of activated Florisil (weight determined
by lauric-acid value, see Appendix II) in a Chromaflex
column. After settling the Florisil by tapping the
column, add about one-half inch layer of anhydrous
granular sodium sulfate to the top.
10.3.3 Pre-elute the column, after cooling, with 50-60 ml of
petroleum ether. Discard the eluate and just prior to
exposure of the sulfate layer to air, quantitatively
transfer the sample extract into the column by
decantation and subsequent petroleum ether wash- ings.
Adjust the elution rate to about 5 ml per minute and,
separately, collect up to three eluates in 500-ml K-D
flasks equipped with 10-ml ampuls (see Eluate
Composition 10.4.). Perform the first elution with
200 ml of 6% ethyl ether in petroleum ether, and the
second elution with 200 ml of 15% ethyl ether in
21
-------
petroleum ether. Perform the third elution with 200 ml
of 50% ethyl ether - petroleum ether and the fourth
elution with 200 ml of 100* ethyl ether.
10.3.4 Concentrate the eluates to 6-10 ml in the K-D eva-
porator in a hot water bath.
10.3.5 Analyze by gas chromatography.
10.4 Eluate Composition - By using an equivalent quantity of any
batch of Florisil, as determined by its lauric acid value, the
pesticides will be separated into the eluates indicated below:
6% Eluate
Aldrin DDT Mi rex
BHC Heptachlor PCNB
Chlordane Heptachlor Epoxide Strobane
ODD Lindane Toxaphene
DDE Methoxychlor Trifluralin
15% Eluate 50% Eluate
Endosulfan I Endosulfan II
Endrin Captan
Dieldrin
Dichloran
Certain thiophosphate pesticides will occur in each of the
above fractions as well as the 100% fraction. For additional
information regarding eluate composition, refer to the FDA
Pesticide Analytical Manual (5).
11. Calculation of Results
11.1 Determine the pesticide concentration by using the absolute
calibration procedure described below or the relative cali-
bration procedure described in Appendix III.
22
-------
(1) Micrograms/liter = (Aj (Bj (vt)
(V-i) (Vs)
A - ng standard
Standard area
B = Sample aliquot area
V-j = Volume of extract injected (pi)
Vt= Volume of total extract (jul)
Vs= Volume of water extracted (ml)
12. Reporting Results
12.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are ana-
lyzed., all data obtained should be reported.
23
-------
REFERENCES:
1. Monsanto Methodology for Aroclors - Analysis of Environmental Materials
for Biphenyls, Analytical Chemistry Method 71-35, Monsanto Company,
St. Louis, Missouri, 63166, 1970.
2. "Method for Polychlorinated Biphenyls in Water and Wastewater", this
manual, p. 43.
3. "Method for Organophosphorus Pesticides in Water and Wastewater", this
manual, p. 25.
4. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories", Chapter 6, Section 6.4, U. S. Environmental Protection
Agency, National Environmental Research Center, Analytical Quality Con*
trol Laboratory, Cincinnati, Ohio, 45268, 1973.
5. "Pesticide Analytical Manual", U. S. Dept. of Health, Education and
Welfare, Food and Drug Administration, Washington, D. C.
24
-------
METHOD FOR ORGANOPHOSPHORUS PESTICIDES IN WATER AND WASTEWATER
Scope and Application
1.1 This method covers the determination of various organophosphorus
pesticides in water and wastewater.
J.2 The following pesticides may be determined individually by this
method:
Parameter Storet No.
Azinphos methyl —
Demeton-0 39560
Demeton-S —
Diazinon 39570
Disulfoton 39010
Malath ion 39530
Parathion methyl 39600
Parathion ethyl 39540
Summary
2.1 The method offers several analytical alternatives, dependent on
the analyst's assessment of the nature and extent of interferences
and the complexity of the pesticide mixtures found. Specifically,
the procedure describes the use of an effective co-solvent for
efficient sample extraction; provides, through use of the column
chromatography and liquid-liquid partition, methods for the
elimination of non-pesticide interferences and the preseparation
of pesticide mixtures. Identification is made by selective gas
chromatographic separation and may be corroborated through the use
of two or more unlike columns. Detection and measurement are best
accomplished by flame photometric gas chromatography using a
25
-------
phosphorus specific filter. The electron capture detector, though
non-specific, may also be used for those compounds to which it
responds. Results are reported in micrograms per liter.
2.2 Confirmation of the identity of the compounds should be made by
GC-MS when a new or undefined sample type is being analyzed and
the concentration is adequate for such determination.
2.3 This method is recommended for use only by experienced pesticide
analysts or under the close supervision of such qualified persons.
3. Interferences
3.1 Solvents, reagents, glassware, and other sample processing hard-
ware may yield discrete artifacts and/or elevated baselines, caus-
ing misinterpretation of gas chromatograms. All of these
materials must be demonstrated to be free from interferences under
the conditions of the analysis. Specific selection of reagents
and purification of solvents by distillation in all-glass systems
may be required. Refer to Appendix I.
3.2 The interferences in industrial effluents are high and varied and
often pose great difficulty in obtaining accurate and precise
measurement of organophosphorus pesticides. Sample clean-up
procedures are generally required and may result in the loss of
certain organophosphorus pesticides. Therefore, great care should
be exercised in the selection and use of methods for eliminating
or minimizing interferences. It is not possible to describe
procedures for overcoming all of the interferences that may be
encountered in industrial effluents.
26
-------
3.3 Compounds such as organochlorine pesticides, polychlorinated
biphenyls and phthalate esters interfere with the analysis of
organophosphorus pesticides by electron capture gas chro-
matography. When encountered, these interferences are overcome by
the use of the phosphorus specific flame photometric detector. If
such a detector is not available, these interferences may be
removed from the sample by using the clean-up procedures described
in the EPA methods for those compounds (1, 2).
3.4 Elemental sulfur will interfere with the determination of organo-
phosphorus pesticides by flame photometric and electron capture
gas chromatography. The elimination of elemental sulfur as an
interference is described in Section 10.5, Clean-up and Separation
Procedures.
4. Apparatus and Materials
4.1 Gas Crhomatograph - Equipped with glass lined injection port.
4.2 Detector options:
4.2.1 Flame Photometric - 526 mu phosphorus filter.
4.2.2 Electron Capture - Radioactive (tritium or nickel-63).
4.3 Recorder - Potentiometric strip chart (10 in.) compatible with the
detector.
4.4 Gas Chromatographic Column Materials:
4.4.1 Tubing - Pyrex (180 cm long x 4 mm ID)
4.4.2 Glass Wool - Silanized
4.4.3 Solid Support - Gas Chrom Q (100-120 mesh)
4.4.4 Liquid Phases - Expressed as weight percent coated on
solid support.
27
-------
4.4.4.1 OV-1, 3%
4.4.4.2 OV-210, 5%
4.4.4.3 OV-17, 1.5% plus QF-1 or OV-210, 1.95%
4.4.4.4 QF-1 or OV-210, 6% plus SE-30, 4%
4.5 Kuderna-Dam'sh (K-D) Glassware
4.5.1 Snyder Column - three ball (macro) and two ball (micro)
4.5.2 Evaporative Flasks - 500 ml
4.5.3 Receiver Ampuls - 10 ml, graduated
4.5.4 Ampul Stoppers
4.6 Chromatographic Column - Chromaflex (400 mm x 19 mm ID) with
coarse fritted plate and Teflon stopcock on bottom; 250 ml
reservoir bulb at top of column with flared out funnel shape at
top of bulb - a special order (Kontes K-420540-9011).
4.7 Chromatographic Column - pyrex (approximately 400 mm long x 20 mm
ID) with coarse fritted plate on bottom.
4.8 Micro Syringes - 10, 25, 50 and 100 >jl.
4.9 Separatory funnels - 125 ml, 1000 ml and 2000 ml with Teflon
stopcock.
4.10 Micro-pipets - disposable (140 mm long x 5 mm ID).
4.11 Blender - High speed, glass or stainless steel cup.
4.12 Graduated cylinders - 100 and 250 ml.
4.13 Florisil - PR Grade (50-100 mesh); purchase activated at 1250°F
and store in the dark in glass containers with glass stoppers, or
foil-lined screw caps. Before use, activate each batch overnight
at 130°C in foil-covered glass container. Determine lauric-acid
value (See Appendix II).
28
-------
4.14 Alumina - Woelm, neutral; deactivate by pipeting 1 ml of distilled
water into 125 ml ground glass-stoppered Erlenmeyer flask. Rotate
flask to distribute water over surface of glass. Immediately add
19.0 g fresh alumina through small powder funnel. Shake flask
containing mixture for two hours on a mechanical shaker (3).
5. Reagents, Solvents, and Standards
5.1 Sodium Chloride - (ACS) Saturated solution in distilled water
(pre-rinse NaC'l with hexane).
5.2 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.3 Sodium Sulfate - (ACS) Granular, anhydrous (conditioned at 400°C
for 4 hrs.).
5.4 Sulfuric Acid - (ACS) Mix equal volumes of cone. H2S04 with
distilled water.
5.5 Diethyl Ether - Nanograde, redistilled in glass, if necessary.
5.5.1 Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM
*
Laboratories, Inc., 500 Executive Blvd., Emslford, N.Y.
10523.)
5.5.2 Procedures recommended for removal of peroxides are
provided with the test strips.
5.6 Acetonitrile, Hexane, Methanol, Methylene Chloride, Petroleum
Ether (boiling range 30-60°C) - nanograde, redistill in glass if
necessary.
5.7 Pesticide Standards - Reference grade.
29
-------
6. Calibration
6.1 Gas chromatographic operating conditions are considered acceptable
if the response to dicapthon is at least 50% of full scale when <1.5
ng is injected for flame photometric detection and < 0.06 ng is
injected for electron capture detection. For all quantitative
measurements, the detector must be operated within its linear
response range and the detector noise level should be less than 2%
of full scale.
6.2 Standards are injected frequently as a check on the stability of
operating conditions. Gas chromatograms of several standard
pesticides are shown in Figures 1, 2, 3 and 4 and provide reference
operating conditions for the four recommended columns.
6.3 The elution order and retention ratios of various organophosphorus
pesticides are provided in Table 1, as a guide.
7. Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts (4) should be developed and
used as a check on the analytical system. Quality control check
samples and performance evaluation samples should be analyzed on a
regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that used
to dilute the sample.
8. Sample Preparation
8.1 The sample size taken for analysis is dependent on the type of
sample and the sensitivity required for the purpose at hand.
30
-------
10
12
2 4 & 8
RETENTION TIME IN MINUTES
Figure 1. Column Packing: 1.5% OY-17 + 1.95 % QF-1,
Carrier Gas: Nitrogen at 70 ml/min, Column Temperature: 215 C,
Detector: Flame Photometric (Phosphorus).
31
-------
rxi
0
10
2468
RETENTION TIME IN MINUTES
Figure 2. Column Packing: 5% OY-210, Carrier Gas: Nitrogen
at 60 ml/min, Column Temperature: 200 C, Detector:
Flame Photometric (Phosphorus).
32
-------
0
2
10
12
4 6 8
RETENTION TIME IN MINUTES
Figure 3. Column Packing: 6% QF-1 +4% SE-30, Carrier Gas: Nitrogen
at 70 ml/min, Column Temperature: 215 C, Detector: Flame
Photometric (Phosphorus).
33
-------
I I I I I I I
2
10
4 6 8
RETENTION TIME IN MINUTES
Figure 4. Column Packing: 3% OV-1, Carrier Gas: Nitrogen at
60 ml/min, Column Temperature: 200 C, Detector: Flame
Photometric (Phosphorus).
34
-------
TABLE 1
RETENTION TIMES OF SOME ORGANOPHOSPHOROUS PESTICIDES
RELATIVE TO PARATHION
Liquid Phase1
Column Temp.
Nitrogen
Carrier Flow
Pesticide
Demeton3
Diazinon
Disulfoton
Ma lath ion
Parathion methyl
Parathion ethyl
Azinphos methyl
1.5% OV-17
+
1.95% QF-12
215 C
70 ml/min
RR
0.46
0.40
0.46
0.86
0.82
1.00
6.65
6% QF-1*
+
4% SE-30
215 C
70 ml/min
RR
0.26
0.43
0.38
0.45
0.78
0.80
1.00
4.15
5%
OV-210
200 C
60 ml/min
RR
0.20
0.38
0.25
0.31
0.73
0.81
1.00
4.44
7%
OV-1
200 C
60 ml/min
RR
0.74
0.59
0.62
0.92
0.79
1.00
4.68
Parathion
(min absolute)
I AT 1 rnlnmnc n
4,. 5
6.6
5.7
3.1
'All columns glass, 180 xm x 4 mm ID, solid support Gas-Chrom Q, 100/120
mesh.
*May substitute OV-210 for QF-1.
^Anomalous, multipeak response often encountered.
35
-------
Background information on the pesticide levels previously detected
at a given sampling site will assist in determining the sample
size required, as well as the final volume to which the extract
needs to be concen- trated. A 1-liter sample is usually taken for
drinking water and ambient water analysis to provide a detection
limit of 0.050 to 0.100^g/l. One-hundred milliliters is usually
adequate to provide a detection limit of 1 ^ig/1 for industrial
effluents.
8.2 Quantitatively transfer the proper aliquot of sample from the
sample container into a two-liter separatory funnel. If less than
a 800 ml is analyzed, dilute to one liter with interference free
distilled water.
9. Extraction
9.1 Add 60 ml of 15% methylene chloride in hexane (v:v) to the sample
in the separatory funnel and shake vigorously for two minutes.
9.2 Allow the mixed solvent to separate from the sample, then draw the
water into a one-liter Erlenmeyer flask. Pour the organic layer
into a 100 ml beaker and then pass it through a column containing
3-4 inches of anhydrous sodium sulfate, and collect it in a 500 ml
K-D flask equipped with a 10 ml ampul. Return the water phase to
the separatory funnel. Rinse the Erlenmeyer flask with a second
60 ml volume of solvent; add the solvent to the separatory funnel
and complete the extraction procedure a second time. Perform a
third extraction in the same manner.
9.3 Concentrate the extract in the K-D evaporator on a hot water bath.
36
-------
9.4 Analyze by gas chromatography unless a need for cleanup is indi-
cated. (See Section 10).
10. Clean-up and Separation Procedures
10.1 Interferences in the form of distinct peaks and/or high background
in the initial gas chromatographic analysis, as well as the
physical characteristics of the extract (color, cloudiness,
viscosity) and background knowledge of the sample source will
indicate whether clean-up is required. When these interfere with
measurement of the pesticides, or affect column life or detector
sensitivity, proceed as directed below. The use of these
procedures is not required for samples free of interferences.
They are provided as options to the analyst to be used when needed.
10.2 Acetonitrile Partition - This procedure is used to separate fats
and oils from the sample extracts. It should be noted that not
all pesticides are quantitatively recovered by this procedure.
The analyst must be aware of this and demonstrate the efficiency
of the partitioning for specific pesticides.
10.2.1 Quantitatively transfer the previously concentrated
extract to a 125-ml separatory funnel with enough hexane
to bring the final volume to 15 ml. Extract the sample
four times by shaking vigorously for one minute with 30
ml portions of hexane-saturated acetonitrile.
10.2.2 Combine and transfer the acetonitrile phases to a
one-liter separatory funnel and add 650 ml of distilled
water and 40 ml of saturated sodium chloride solution.
Mix thoroughly for 30-45 seconds. Extract with two
37
-------
TOO ml portions of hexane by vigorously shaking about 15
seconds.
10.2.3 Combine the hexane extracts in a one-liter separatory
funnel and wash with two 100 ml portions of distilled
water. Discard the water layer and pour the hexane
layer through a 3-4 inch anhydrous sodium sulfate column
into a 500-ml K-D flask equipped with a 10-ml ampul.
Rinse the separatory funnel and column with three 10 ml
portions of hexane.
10.2.4 Concentrate the extracts to 6-10 ml in the K-D
evaporator in a hot water bath.
10.2.5 Analyze by gas chromatography unless a need for further
clean-up is indicated.
10.3 Florisil Column Adsorption Chromatography
10.3.1 Adjust the sample extract volume to 10 ml.
10.3.2 Place a charge of activated Florisil (weight determined
by lauric-acid value, see Appendix II) in a Chromaflex
column. After settling the Florisil by tapping the
column, add about one-half inch layer of anhydrous
granular sodium sulfate to the top.
10.3.3 Pre-elute the column, after cooling, with 50-60 ml of
petroleum ether. Discard the eluate and just prior to
exposure of the sulfate layer to air, quantitatively
transfer the sample extract into the column by
decantation and subsequent petro- leum ether washings.
Adjust the elution rate to about 5 ml per minute and,
38
-------
separately, collect up to four eluates in 500-ml K-D
flasks equipped with l(Hnl ampuls. (See Eluate Compos-
ition, 10.4.) Perform the first elution with 200 ml of
6% ethyl ether in petroleum ether, and the second
elution with 200 ml of 15% ethyl ether in petroleum
ether. Perform the third elution with 200 ml of 50%
ethyl ether - petroleum ether and the fourth elution
with 200 ml of 100% ethyl ether.
10.3.4 Concentrate the eluates to 6-10 ml in the K-D evaporator
in a hot water bath.
10.3.5 Analyze by gas chromatography.
10.4 Eluate Composition - By using an equivalent quantity of any batch
of Florisil as determined by its 1auric-acid value, the pesticides
will be separated into the eluates indicated below:
6% Eluate 15% Eluate
Demeton Diazinon
Disulfoton Malathion (trace)
Parathion Methyl
50% Eluate 100% Eluate
Malathion Azinphos methyl (80%)
Azinphos methyl (20%)
For additional information regarding eluate composition, refer
to the FDA Pesticide Analytical Manual (5).
10.5 Removal of Sulfur - If elemental sulfur interferes with the gas
chromatographic analysis, it can be removed by the use of an
alumina microcolumn.
10.5.1 Adjust the sample extract volume to 0.5 ml in a K-D
39
-------
apparatus, using a two-ball Snyder mlcrocolumn.
10.5.2 Plug a disposable pipet with a small quantity of glass
wool. Add enough alumina to produce a 3-cm column after
settling. Top the alumina with a 0.5-cm layer of
anhydrous sodium sulfate.
10.5.3 Quantitatively transfer the concentrated extract to the
alumina microcolumn using a 100 yl syringe. Rinse the
ampul with 200 ^1 of hexane and add to the microcolumn.
10.5.4 Elute the microcolumn with 3 ml of hexane and discard the
first eluate which contains the elemental sulfur.
10.5.5 Next elute the column with 5 ml of 10% hexane in
methylene chloride. Collect the eluate in a 10 ml
graduated ampul.
10.5.6 Analyze by gas chromatography.
NOTE: If the electron capture detector is to be used methylene
chloride must be removed. To do this, attach the ampul
to a K-D apparatus (500-ml flask and 3-ball Snyder
column) and concentrate to about 0.5 ml. Adjust volume
as required prior to analysis.
11. Calculation of Results
11.1 Determine the pesticide concentration by using the absolute
calibration procedure described below or the relative cali-
bration procedure described in Appendix III.
40
-------
(1) Micrograms/liter = (A) (B) (Vt)
W (VS)
A - ng standard
Standard area
B * Sample aliquot area
V^ » Volume of extract injected (nl)
Vt = Volume of total extract (jjl)
Vs = Volume of water extracted (ml)
12. Reporting Results
12.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are analyzed
all data obtained should be reported.
41
-------
REFERENCES:
1. "Method for Chlorinated Hydrocarbons in Water and Wastewater", this
manual, p. 7.
2. "Method for Polychlorinated Biphenyls (PCBs) in Water and Wastewater",
this manual, p. 43.
3. Law, L. M. and Georlitz, D. F., "Microcolumn Chromatographic Clean-up
for the Analysis of Pesticides in Water", Journal of the Association
for Analytical Chemists. 53_, 1276 (1970).
4. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories", Chapter 6, Section 6.4, U. S. Environmental Protection
Agency, National Environmental Research Center, Analytical Quality Con-
trol Laboratory, Cincinnati, Ohio, 45268, 1973.
5. "Pesticide Analytical Manual", U. S. Dept. of Health, Education and
Welfare, Food and Drug Administration, Washington, D. C.
42
-------
METHOD FOR POLYCHLORINATED BIPHENYLS (PCBs) IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of various polychlorinated
bipheny! (PCB) mixtures in water and wastewater.
1.2 The following mixtures of chlorinated biphenyls (Aroclors) may be
determined by this method:
Parameter Storet No.
PCB-1016 34671
PCB-1221 39488
PCB-1232 39492
PCB-1242 39496
PCB-1248 39500
PCB-1254 39504
PCB-1260 39508
1.3 The method is an extension of the Method for Chlorinated
Hydrocarbons in Water and Wastewater (1). It is designed so
that determination of both the PCBs and the organochlorine
pesticides may be made on the same sample.
2. Summary
2.1 The PCBs and the organochlorine pesticides are co-extracted by
liquid-liquid extraction and, insofar as possible, the two
classes of compounds separated from one another prior to gas
chromatographic determination. A combination of the standard
Florisil column cleanup procedure and a silica gel microcolumn
separation procedure (2)(3) are employed. Identification is
43
-------
made from gas chromatographic patterns obtained through the use
of two or more unlike columns. Detection and measurement is
accomplished using an electron capture, microcoulometric, or
electrolytic conductivity detector. Techniques for confirming
qualitative identification are suggested.
3. Interferences
3.1 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. Specific selection of
reagents and the purification of solvents by distillation in
all-glass systems may be required. Refer to Appendix I.
3.2 The interferences in industrial effluents are high and varied
and pose great difficulty in obtaining accurate and precise
measurement of PCBs and organochlorine pesticides. Separation
and clean-up procedures are generally required and may result
in the loss of certain organochlorine compounds. Therefore,
great care should be exercised in the selection and use of
methods for eliminating or minimizing interferences. It is not
possible to describe procedures for overcoming all of the
interferences that may be encountered in industrial effluents.
3.3 Phthalate esters, certain organophosphorus pesticides, and
elemental sulfur will interfere when using electron capture for
detection. These materials do not interfere when the
44
-------
microcoulometric or electrolytic conductivity detectors are
used in the halogen mode.
3.4 Organochlorine pesticides and other halogenated compounds
constitute interferences in the determination of PCBs. Most of
these are separated by the method described below. However,
certain compounds, if present in the sample, will occur with
the PCBs. Included are: Sulfur, Heptachlor, aldrin, DDE,
technical chlordane, mirex, and to some extent, o,p'-DDT and
p,p'-DDT.
4. Apparatus and Materials
4.1 Gas Chromatograph - Equipped with glass lined injection port.
4.2 Detector Options:
4.2.1 Electron Capture - Radioactive (tritium or nickel-63)
4.2.2 Microcoulometric Titration
4.2.3 Electrolytic Conductivity
4.3 Recorder - Potentiometric strip chart (10 in.) compatible with
the detector.
4.4 Gas Chromatographic Column Materials:
4.4.1 Tubing - Pyrex (180 cm long X 4 mm ID)
4.4.2 Glass Wool - Silanized
4.4.3 Solid Support - Gas-Chrom Q (100-120 mesh)
4.4.4 Liquid Phases - Expressed as weight percent coated on
solid support.
4.4.4.1 SE-30 or OV-1, 3%
4.4.4.2 OV-17, 1.5* + QF-1 or OV-210, 1.95%
-------
4.5 Kuderna-Danish (K-D) Glassware
4.5.1 Snyder Column - three-ball (macro) and two-ball (micro)
4.5.2 Evaporative Flasks - 500 ml
4.5.3 Receiver Ampuls - 10 ml, graduated
4.5.4 Ampul Stoppers
4.6 Chromatographic Column - Chromaflex (400 mm long x 19 mm ID)
with coarse fritted plate on bottom and Teflon stopcock; 250-ml
reservoir bulb at top of column with flared out funnel shape at
top of bulb - a special order (Kontes K-420540-9011).
4.7 Chromatographic Column - pyrex (approximately 400 mm long x 20
mm ID) with coarse fritted plate on bottom.
4.8 Micro Column Pyrex - constructed according to Figure 1.
4.9 Capillary pipets disposable (5-3/4 in.) with rubber bulb
(Scientific Products P5205-1).
4.10 Low pressure regulator - 0 to 5 PSIG - with low-flow needle
valve (see Figure 1, Matheson Model 70).
4.H Beaker - 100 ml
4.12 Micro Syringes - 10, 25, 50 and 100 ul.
4.13 Separatory funnels - 125 ml, 1000 ml and 2000 ml with Teflon
stopcock.
4.14 Blender - High speed, glass or stainless steel cup.
4.15 Graduated cylinders - 100 and 250 ml.
4.16 Florisil - PR Grade (60-100 mesh); purchase activated at
1250°F and store in the dark in glass containers with glass
stoppers or foil-lined screw caps. Before use, activate each
46
-------
COMPRESSED
AIR
SUPPLY
. ^PRESSURE
L^GAUGE
3X1
SHUT-OFF
VALVE
0-5
PSIG
REGULATOR
NEEDLE
VALVE
I cm
FLEXIBLE
TUBING
SILICA GEL
5 cm
I cm
§ 10/30
_ 15ml
RESERVOIR
§ 10/30
r
23cm x 4.2 mm I.D.
2 cm x 2 mm I.D.
FIGURE I. MICROCOLUMN SYSTEM
-------
batch overnight at 130°C in foil-covered glass container.
Determine 1 auric-acid value (See Appendix II).
4.17 Silica gel - Davison code 950-08008-226 (60/200 mesh).
4.18 Glass Wool - Hexane extracted.
4.19 Centrifuge Tubes - Pvrex calibrated (15 ml).
5. Reagents, Solvents, and Standards
5.1 Sodium Chloride - (ACS) Saturated solution in distilled water
(pre-rinse NaCl with hexane).
5.2 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.3 Sodium Sulfate - (ACS) Granular, anhydrous (conditioned at 400
C for 4 hrs.).
5.4 SuIfuric Acid - (ACS) Mix equal volumes of cone. H^SO^ with
distilled water.
5.5 Diethyl Ether - Nanograde, redistilled in glass, if necessary.
5.5.1 Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM Labora-
tories, Inc., 500 Executive Blvd., Elmsford, N.Y.
10523).
5.5.2 Procedures recommended for removal of peroxides are
provided with the test strips.
5,6 n-Hexane - Pesticide quality (NOT MIXED HEXANES).
5.7 Acetonitrile, Hexane, Methanol, Methylene Chloride, Petroleum
Ether (boiling range 30-60°C) - pesticide quality, redistill in
glass if necessary.
5.8 Standards - Aroclors 1221, 1232, 1242, 1248, 1254, 1260, and
1016.
48
-------
5.9 Anti-static Solution - STATNUL, Daystrom, Inc., Weston Instru-
ment Division, Newark, N.J., 95212.
6. Calibration
6.1 Gas chromatographic operating conditions are considered accept-
able if the response to dicapthon is at least 50% of full scale
when < 0.06 ng is injected for electron capture detection and <
100 ng is injected for microcoulometric or electrolytic con-
ductivity detection. For all quantitative measurements, the
detector must be operated within its linear response range and
the detector noise level should be less than 2% of full scale.
6.2 Standards are injected frequently as a check on the stability
of operating conditions, detector and column. Example chro-
matograms are shown in Figures 3 through 8 and provide
reference operating conditions.
7. Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts (4) should be developed
and used as a check on the analytical system. Quality control
check samples and performance evaluation samples should be
analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
8. Samp 1e Preparat i on
8.1 Blend the sample if suspended matter is present and adjust pH
49
-------
to near neutral (pH 6.5-7.5) with 50# sulfuric acid or 10 N
sodium hydroxide.
8.2 For sensitivity requirement of 1 ug/1, when using micro-
coulometric or electrolytic conductivity methods for detection
take 1000 ml of sample for analysis. If interferences pose no
problem, the sensitivity of the electron capture detector
should permit as little as 100 ml of sample to be used. Back-
ground information on the extent and nature of interferences
will assist the analyst in choosing the required sample size
and preferred detector.
8.3 Quantitatively transfer the proper aliquot into a two-liter
separatory funnel and dilute to one liter.
9. Extraction
9.1 Add 60 ml of 15% methylene chloride in hexane (v:v) to the
sample in the separatory funnel and shake vigorously for two
minutes.
9.2 Allow the mixed solvent to separate from the sample, then draw
the water into a one-liter Erlenmeyer flask. Pour the organic
layer into a 100-ml beaker and then pass it through a column
containing 3-4 inches of anhydrous sodium sulfate, and collect
it in a 500-ml K-D flask equipped with a 10 mi-ampul. Return
the water phase to the separatory funnel. Rinse the Erlenmeyer
flask with a second 60-ml volume of solvent; add the solvent to
the separatory funnel and complete the extraction procedure a
second time. Perform a third extraction in the same manner.
50
-------
9.3 Concentrate the extract in the K-D evaporator on a hot water
bath.
9.4 Qualitatively analyze the sample by gas chromatography with an
electron capture detector. From the response obtained decide:
a. If there are any organochlorine pesticides present.
b. If there are any PCBs present.
c. If there is a combination of a and b.
d. If elemental sulfur is present.
e. If the response is too complex to determine a, b or c.
f. If no response, concentrate to 1.0 ml or less, as required,
and repeat the analysis looking for a, b, c, d, and e.
Samples containing Aroclors with a low percentage of
chlorine, e.g., 1221 and 1232, may require this concentra-
tion in order to achieve the detection limit of 1 yg/1.
Trace quantities of PCBs are often masked by background
which usually occur in samples.
9.5 If condition
-------
9.8 If condition d^ exists, separate the sulfur from the sample
using the method outlined in 10.3 followed by the method in
10.5.
9.9 If condition e exists, the following macro cleanup and separa-
tion procedures (10.2 and 10.3) should be employed and, if
necessary, followed by the micro separation procedures (10.4
and 10.5).
10. Cleanup and Separation Procedures
10.1 Interferences in the form of distinct peaks and/or high back-
ground in the initial gas chromatographic analysis, as well as
the physical characteristics of the extract (color, cloudiness,
viscosity) and background knowledge of the sample will indicate
whether clean-up is required. When these interfere with
measurement of the PCBs, or affect column life or detector
sensitivity, proceed as directed below.
10.2 Acetonitrile Partition - This procedure is used to remove fats
and oils from the sample extracts. It should be noted that not
all pesticides are quantitatively recovered by this procedure.
The analyst must be aware of this and demonstrate the effi-
ciency of the partitioning for the compounds of interest.
10.2.1 Quantitatively transfer the previously concentrated
extract to a 125-ml separatory funnel with enough hexane
to bring the final volume to 15 ml. Extract the sample
four times by shaking vigorously for one minute with
30-ml portions of hexane-saturated acetonitrile.
52
-------
10.2.2 Combine and transfer the acetonitrile phases to a
one-liter separatory funnel and add 650 ml of distilled
water and 40 ml of saturated sodium chloride solution.
Mix thoroughly for 30-45 seconds. Extract with two
100-ml portions of hexane by vigorously shaking about 15
seconds.
10.2.3 Combine the hexane extracts in a one-liter separatory
funnel and wash with two 100-ml portions of distilled
water. Discard the water layer and pour the hexane
layer through a 3-4 inch anhydrous sodium sulfate column
into a 500-ml K-D flask equipped with a 10-ml ampul.
Rinse the separatory funnel and column with three 10-ml
portions of hexane.
10.2.4 Concentrate the extracts to 6-10 ml in the K-D eva-
porator in a hot water bath.
10.2.5 Analyze by gas chromatography unless a need for further
cleanup is indicated.
10.3 Florisil Column Adsorption Chromatography
10.3.1 Adjust the sample extract volume to 10 ml.
10.3.2 Place a charge of activated Florisil (weight determined
by lauric-acid value, see Appendix II) in a Chromaflex
column. After settling the Florisil by tapping the
column, add about one-half inch layer of anhydrous
granular sodium sulfate to the top.
53
-------
10.3.3 Pre-elute the column, after cooling, with 50-60 ml of
petroleum ether. Discard the eluate and just prior to
exposure of the sulfate layer to air, quantitatively
transfer the sample extract into the column by
decantation and subsequent petroleum ether washings.
Adjust the elution rate to about 5 ml per minute and,
separately, collect up to three eluates in 500-ml K-D
flasks equipped with 10-ml ampuls (see Eluate Composi-
tion 10.4.). Perform the first elution with 200 ml of
6% ethyl ether in petroleum ether, and the second
elution with 200 ml of 15% ethyl ether in petroleum
ether. Perform the third elution with 200 ml of 50%
ethyl ether - petroleum ether and the fourth elution
with 200 ml of 100% ethyl ether.
10.3.3.1 Eluate Composition - By using an equivalent
quantity of any batch of Florisil as deter-
mined by its lauric acid value, the pesti-
cides will be separated into the eluates
indicated as follows.
6% Eluate
Aldrin DDT Pentachloro-
BHC Heptachlor nitrobenzene
Chlordane Heptachlor Epoxide Strobane
ODD Lindane Toxaphene
DDE Methoxychlor Trifluralin
Mirex PCBs
15% Eluate 50% Eluate
Endosulfan I Endosulfan II.
Endrin Captan
Dieldrin
Dichloran
Phthalate esters
54
-------
Certain thiophosphate pesticides will occur in
each of the above fractions as well as the 100%
fraction. For additional information regarding
eluate composition, refer to the FDA Pesticide
Analytical Manual (5).
10.3.4 Concentrate the eluates to 6-10 ml in the K-D evaporator
in a hot water bath.
10.3.5 Analyze by gas chromatography.
10.4 Silica Gel Micro-Column Separation Procedure (6)
10.4.1 Activation for Silica Gel
10.4.1.1 Place about 20 gm of silica gel in a 100-ml
beaker. Activate at 180°C for approximately
16 hours. Transfer the silica gel to a 100-ml
glass-stoppered bottle. When cool, cover with
about 35 ml of 0.50% diethyl ether in benzene
(volume:volume). Keep bottle well sealed. If
silica gel collects on the ground glass
surfaces, wash off with the above solvent
before resealing. Always maintain an excess of
the mixed solvent in bottle (aproximately 1/2
in. above silica gel). Silica gel can be
effectively stored in this manner for several
days.
10.4.2 Preparation of the Chromatographic Column
10.4.2.1 Pack the lower 2 mm ID section of the micro-
column with glass wool. Permanently mark
55
-------
the column 120 mm above the glass wool. Using
a clean rubber bulb from a disposable pipet
seal the lower end of the microcolumn. Fill
the microcolumn with 0.50% ether in benzene
(v:v) to the bottom of the 10/30 joint (Figure
1). Using a disposable capillary pipet,
transfer several aliquots of the silica gel
slurry into the microcolumn. After approxi-
mately 1 cm of silica gel collects in the
bottom of the microcolumn, remove the rubber
bulb seal, tap the column to insure that the
silica gel reaches the 120 + 2 mm mark. Be
sure that there are no air bubbles in the
column. Add about 10 mm of sodium sulfate to
the top of the silica gel. Under low humidity
conditions, the silica gel may coat the sides
of the column and not settle properly. This
can be minimized by wiping the outside of the
column with an anti-static solution.
10.4.2.2 Deactivation of the Silica Gel
a. Fill the microcolumn to the base of the
10/30 joint with the 0.50% ether-benzene
mixture, assemble reservoir (using spring
clamps) and fill with approximately 15 ml
of the 0.50% ether-benzene mixture. Attach
the air pressure device (using spring
56
-------
clamps) and adjust the elation rate to
approximately 1 ml/min. with the air
pressure control. Release the air pressure
and detach reservoir just as the last of
the solvent enters the sodium sulfate.
Fill the column with n-hexane (not mixed
hexanes) to the base of the 10/30 fitting.
Evaporate all residual benzene from the
reservoir,, assemble the reservoir section
and fill with 5 ml of n-hexane. Apply air
pressure and remove the reservoir just as
the n-hexane enters the sodium sulfate.
The column is now ready for use.
b. Pipet a 1.0 ml aliquot of the concentrated
sample extract (previously reduced to a
total volume of 2.0 ml) on to the column.
As the last of the sample passes into the
sodium sulfate layer, rinse down the
internal wall of the column twice with 0.25
ml of n-hexane. Then assemble the upper
section of the column. As the last of the
n-hexane rinse reaches the surface of the
sodium sulfate, add enough n-hexane (volume
predetermined, see 10.4.3) to just elute
all of the PCBs present in the sample.
Apply air pressure and adjust until the
57
-------
flow is 1 ml/min. Collect the desired
volume of eluate (predetermined, see
10.4.3) in an accurately calibrated ampul.
As the last of the n-hexane reaches the
surface of the sodium sulfate, release the
air pressure and change the collection
ampul.
c. Fill the column with 0.50% diethyl ether in
benzene, again apply air pressure and
adjust flow to 1 ml/min. Collect the
eluate until all of the organochlorine
pesticides of interest have been eluted
(volume predetermined, see 10.4.3).
d. Analyze the eluates by gas chromatography.
10.4.3 Determination of Elution Volumes
10.4.3.1 The elution volumes for the PCBs and the
pesticides depend upon a number of factors
which are difficult to control. These include
variation in:
a. Mesh size of the silica gel
b. Adsorption properties of the silica gel
c. Polar contaminants present in the eluting
solvent
d. Polar materials present in the sample and
sample solvent
58
-------
e. The dimensions of the microcolumns
Therefore, the optimum elution volume must
be experimentally determined each time a
factor is changed. To determine the
elution volumes, add standard mixtures of
Aroclors and pesticides to the column and
serially collect 1-ml elution volumes.
Analyze the individual eluates by gas
chromatography and determine the cut-off
volume for n-hexane and for ether-benzene.
Figure 2 shows the retention order of the
various PCB components and of the pesti-
cides. Using this information, prepare the
mixtures required for calibraton of the
microcolumn.
10.4.3.2 In determining the volume of hexane required to
elute the PCBs the sample volume (1 ml) and the
volume of n-hexane used to rinse the column
wall must be considered. Thus, if it is
determined that a 10.0-ml elution volume is
required to elute the PCBs, the volume of
hexane to be added in addition to the sample
volume but including the rinse volume should be
9.5 ml.
59
-------
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60
-------
10.4.3.3 Figure 2 shows that as the average chlorine
content of a PCB mixture decreases the solvent
volume for complete elution increases. Quali-
tative determination (9.4) indicates which
Aroclors are present and provides the basis for
selection of the ideal elution volume. This
helps to minimize the quantity of organo-
chlorine pesticides which will elute along with
the low percent chlorine PCBs and insures the
most efficient separations possible for
accurate analysis.
10.4.3.4 For critical analysis where the PCBs and pesti-
cides are not separated completely, the column
should be accurately calibrated according to
(10.4.3.1) to determine the percent of material
of interest that elutes in each fraction. Then
flush the column with an additional 15 ml of
0.50% ether in benzene followed by 5 ml of
n-hexane and use this reconditioned column for
the sample separation. Using this technique
one can accurately predict the amount (%) of
materials in each micro column fraction.
10.5 Micro Column Separation of Sulfur, PCBs, and Pesticides
10.5.1 See procedure for preparation and packing micro column
in PCB analysis section (10.4.1 and 10,4.2).
61
-------
10.5.2 Microcolumn Calibration
10.5.2.1 Calibrate the microcolumn for sulfur and PCB
separation by collecting 1.0-ml fractions and
analyzing them by gas chromatography to
determine the following:
1) The fraction with the first eluting PCBs
(those present in 1260),
2) The fraction with the last eluting PCBs
(those present in 1221),
3) The elution volume for sulfur,
4) The elution volume for the pesticides of
interest in the 0.50% ether-benzene
fraction.
From these data determine the following:
1) The eluting volume containing only sulfur
(Fraction I),
2) The eluting volume containing the last of
the sulfur and the early eluting PCBs
(Fraction II),
3) The eluting volume containing the remaining
PCBs (Fraction III),
4) The ether-benzene eluting volume containing
the pesticides of interest (Fraction IV).
10.5.3 Separation Procedure
10.5.3.1 Carefully concentrate the 6% eluate from the
62
-------
florisil column to 2.0 ml in the graduated
ampul on a warm water bath.
10.5.3.2 Place 1.0 ml (50%) of the concentrate into the
microcolumn with a 1-ml pipet. Be careful not
to get any sulfur crystals into the pipet.
10.5.3.3 Collect Fractions I and II in calibrated
centrifuge tubes. Collect Fractions III and IV
in calibrated ground glass stoppered ampuls.
10.5.3.4 Sulfur Removal (7) - Add 1 to 2 drops of
mercury to Fraction II stopper and place on a
wrist-action shaker. A black precipitate
indicates the presence of sulfur. After
approximately 20 minutes the mercury may become
entirely reacted or deactivated by the
precipitate. The sample should be quanti-
tatively transferred to a clean centrifuge tube
and additional mercury added. When crystals
are present in the sample, three treatments may
be necessary to remove all the sulfur. After
all the sulfur has been removedfrom Fraction II
(check using gas chromatography) combine
Fractions II and III. Adjust the volume to 10
ml and analyze by gas chromatography. Be sure
no mercury is transferred to the combined
Fractions II and III, since it can react with
certain pesticides.
63
-------
By combining Fractions II and III, if PCBs are
present, it is possible to identify the
Aroclor(s) present and a quantitative analysis
can be performed accordingly. Fraction I can
be discarded since it only contains the bulk of
the sulfur. Analyze Fractions III and IV for
the PCBs and pesticides. If DDT and its
homologs, aldrin, heptachlor, or technical
chlordane are present along with the PCBs, an
additional microcolumn separation can be
performed which may help to further separate
the PCBs from the pesticides (See 10.4).
11. Quantitative Determination
11.1 Measure the volume of n-hexane eluate containing the PCBs and
inject 1 to 5/
-------
Microgram/liter = K| \P|/{v)W x [N ]
A _ ng of Standard Injected _ ng2
mm
2
B = of Sample Peak Areas - (mm )
V^ = Volume of sample injected (yl)
V. = Volume of Extract (vl) from which sample
is injected into gas chromatograph
V = Volume of water sample extracted (ml)
N = 2 when micro column used
1 when micro column not used
Peak Area = Peak height (mm x Peak Width at 1/2
height
11.2.2 For complex situatons, use the calibration method
described below (8). Small variations in components
between different Aroclor batches make it necessary to
obtain samples of several specific Aroclors. These
reference Aroclors can be obtained from the Southeast
Environmental Research Laboratory, EPA, Athens, Georgia,
30601. The procedure is as follows:
11.2.2.1 Using the OV-1 column, chromatograph a known
quantity of each Aroclor reference standard.
Also chromatograph a sample of p,p'-DDE.
Suggested concentration of each standard is 0.1
ng/yl for the Aroclors and 0.02 ng/ul for the
p,p'-DDE.
65
-------
11.2.2.2 Determine the relative retention time (RRT) of
each PCB peak in the resulting chromatograms
using p,p'-DDE as 100.
RT x 100
RRT =
RTDDE
RRT = Relative Retention Time
RT = Retention time of peak of interest
RTDDE = Retention tinie °f P»P'-DDE
Retention time is measured as that distance in
mm between the first appearance of the solvent
peak and the maximum for the compound.
11.2.2.3 To calibrate the instrument for each PCB
measure the area of each peak.
Area = Peak height (mm) x Peak width at 1/2
height. Using Tables 1 through 6 obtain the
proper mean weight factor, then determine the
2
response factor ng/mm .
ng/mm =
(ng.) (mean weight percent)
1 100
(Area)
rig,- = ng of Aroclor Standard Injected
Mean weight percent - obtained from Tables 1
through 6.
11.2.2.4 Calculate the RRT value and the area for each
PCB peak in the sample chromatogram. Compare
the sample chromatogram to those obtained for
each reference Aroclor standard. If it is
66
-------
Table 1
Composition of Aroclor 1221 (8)
RRTa
11
14
16
19
21
28
32
37
40
Mean
Weight
Percent
31.8
19.3
10.1
2.8
20.8
5.4
1.4
1.7
Relative
Std. Dev.b
15.8
9.1
9.7
9.7
9.3
13.9
30.1
48.8
Number of
Chlorines0
1
1
2
2
2
2 85%
3 15%
2 10%
3 90%
3
Detention time relative to p,p'-DDE=100. Measured from first appearance
of solvent. Overlapping peaks that are quantitated as one peak are
bracketed.
''Standard deviation of seventeen results as a percentage of the mean of
the results.
cFrom GC-MS data. Peaks containing mixtures of isomers of different
chlorine numbers are bracketed.
67
-------
Table 2
Composition of Aroclor 1232 (8)
RRTa
11
14
16
20
21
28
32
37
40
47
54
58
70
78
Mean
Weight
Percent
16.2
9.9
7.1
17.8
9.6
3.9
6.8
6.4
4.2
3.4
2.6
4.6
1.7
Relative .
Std. Dev.
3.4
2.5
6.8
2.4
3.4
4.7
2.5
2.7
4.1
3.4
3.7
3.1
7.5
Number of
Chlorines
1
1
2
2
2
3
3
3
3
4
3
4
4
4
5
4
40%
60*
33%
67%
90%
10%
Total 94.2
Detention time relative to p.p'-DDE^lOO. Measured from first appearance
of solvent. Overlapping peaks that are quantitated as one peak are
bracketed.
^Standard deviation of four results as a mean of the results.
cFrom GC-MS data. Peaks containing mixtures of isomers of different
chlorine numbers are bracketed.
68
-------
Table 3
Composition of Aroclor 1242 (8)
RRTa
11
16
21
28
32
37
40
47
54*
.
58
70
78
84
98
104
125
146
Mean
Weight
Percent
1.1
2.9
11.3
11.0
6.1
11.5
11.1
8.8
6.8
5.6
10.3
3.6
2.7
1.5
2.3
1,6
1.0
Relative .
Std. Dev.
35.7
4.2
3.0
5.0
4.7
5.7
6.2
4.3
2.9
3.3
2.8
4.2
9.7
9.4
16.4
20.4
19.9
Number of
Chlorines
1
2
2
2
3
3
3
3
4
3
4
4
4
5
4
5
5
5
5
6
5
6
25%
75%
33%
67%
90%
10%
85%
15%
75%
25%
Detention time relative to p,p'-DDE=100. Measured from first appearance
of solvent.
^Standard deviation of six results as a percentage of the mean of the
results.
cFrom GC-MS data. Peaks containing mixtures of isomers of different
chlorine numbers are bracketed.
69
-------
Table 4
Composition of Aroclor 1248 (8)
RRTa
21
28
32
47
40
47
54
58
70
78
84
98
104
112
125
146
Mean
Weight
Percent
1.2
5.2
3.2
8.3
8.3
15.6
9.7
9.3
19.0
6.6
4.9
3.2
3.3
1.2
2.6
1.5
Relative .
Std. Dev.
23.9
3.3
3.8
3.6
3.9
1.1
6.0
5.8
1.4
2.7
2.6
3.2
3.6
6.6
5.9
10.0
Number of
Chlorines0
2
3
3
3
3
4
4
3
4
4
4
5
4
5
5
4
5
5
5
6
5
6
85*
15%
10%
90%
80tf
20%
10%
90%
90%
10%
85%
15%
Total 103.1
Detention time relative to p,p'-DDE=100. Measured from first appearance
of solvent.
''Standard deviation of six results as a percentage of the mean of the.
results.
cFrom GC-MS data. Peaks containing mixtures of isomers of different
chlorine numbers are bracketed.
70
-------
Table 5
Composition of Aroclor 1254 (8)
RRTa
47
54
58
70
84
98
104
125
(
146
160
174
203
232
Mean
Weight
Percent
6.2
2.9
1.4
13.2
17.3
7.5
13.6
15.0
10.4
1.3
8.4
1.8
1.0
Relative .
Std. Dev.
3.7
2.6
2.8
2.7
1.9
5.3
3.8
2.4
2.7
8.4
5.5
18.6
26.1
Number of
Chlorines
4
4
4
4 25%
5 75*
5
5
5
5 70%
6 80*
5 30%
6 70515
6
6
6
7
Total 100.0
Detention time relative to p,p'-DDE=100. Measured from first appearance
of solvent.
bStandard deviation of six results as a percentage of the mean of the
results.
cFrom 6C-MS data. Peaks containing mixtures of isomers are bracketed.
71
-------
Table 6
Composition of Aroclor 1260 (8)
RRTa
70
84
98
104
117
125
146
160
174
203
232
244
280
332
372
448
528
Mean
Weight
Percent
2.7
4.7
3.8
3.3
12.3
14.1
4.9
12.4
9.3
9.8
11.0
4.2
4.0
.6
1.5
Relative ,
Std. Dev.
6.3
1.6
3.5
6.7
3.3
3.6
2.2
2.7
4.0
3.4
2.4
5.0
8.6
25.3
10.2
Number of
Chlorines
5
5
d
5
6
6
5
6
6
6
7
6
6
7
e
6
7
7
7
8
8
8
60%
40%
15%
85%
50%
50%
i
10%
90%
10%
90%
Total 98.6
Detention time relative to p,p'-DDE=100. Measured from first appearance
of solvent. Overlapping peaks that are quantitated as one peak are
bracketed.
^Standard deviation of six results as a mean of the results.
cFrom SC-MS data. Peaks containing mixtures of isomers of different
chlorine numbers are bracketed.
Composition determined at the center of peak 104.
Composition determined at the center of peak 232.
72
-------
apparent that the PCB peaks present are due to
only one Aroclor, then calculate the concen-
tration of each PCB using the following formula:
2
ng PCB = ng/mm x Area
2
Where Area = Area (mm ) of sample peak
2
ng/mm = Response factor for that peak
measured.
Then add the nanograms of PCBs present in the
injection to get the total number of nanograms
of PCBs present. Use the following formula to
calculate the concentration of PCBs in the
sample:
Micrograms/Liter =
V = volume of water extracted (ml)
V. = volume of extract (pi)
V.. = volume of sample injected (pi)
ng = sum of all the PCBs in nanograms for that
Aroclor identified
N = 2 when microcolumn used
N = 1 when microcolumn not used
The value can then be reported as micro-
grams/liter PCBs or as the Aroclor. For
samples containing more than one Aroclor, use
Figure 9 chromatogram divisional flow chart to
assign a proper response factor to each peak
and also identify the "most likely" Aroclors
73
-------
present. Calculate the ng of each PCB isomer
present and sum them according to the
divisional flow chart. Using the formula
above, calculate the concentration of the
various Aroclors present in the sample.
12. Reporting Results
12.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are analyzed,
all data obtained should be reported.
74
-------
37
AROCLOR 1242
146
Figure 3. Column: 3% OV-1, Carrier Gas: Nitrogen at 60 ml/min,
Column Temperature: 170 C, Detector: Electron Capture
75
-------
70
AROCLOR 1254
104
125
232
Figure 4. Column: 3% OV-1, Carrier Gas: Nitrogen at 60 ml/min,
Column Temperature: 170 C, Detector: Electron Capture.
76
-------
•\J
AROCLOR 1260
280
Figure 5. Column: 3% OV-1, Carrier Gas: Nitrogen at 60 ml/min,
Column Temperature: 170 C, Detector: Electron Capture,
77
-------
AROCLOR 1242
I
I
I
1
j
3
21
24
6 9 12 15 18
RETENTION TIME IN MINUTES
Figure 6. Column: 1.5% OV-17 + 1.95% QF-1, Carrier Gas: Nitrogen
at 60 ml/min, Column Temperature: 200 C, Detector: Electron Capture.
78
-------
AROCLOR 1254
(0
Figure 7. Column: 1.5% OV-17 + 1.95%
Detector: Electron Capture.
RETENTION TIME IN MINUTES
QF-1, Carrier Gas: Nitrogen at 60 ml/min, Column Temperature: 200 C,
-------
CO
AROCLOR 1260
_L
J
54
3
12 15
18
36
39
42
45
48
51
21 24 27 30 33
RETENTION TIME IN MINUTES
Figure ft. Column: 1.5% OV-17 + 1.95% QF-1, Carrier Gas: Nitrogen at 60 ml/min, Column Temperature: 200C, Detector: Electron Capture
-------
RRT of first peak < 47? |
YES/ \HO
Is there a distinct
peak with RRT 78?
RRT
YES
/ V
YES\
47-58? [
NO
Use 1242 for
peaks! RRT 84
Use 1242 for
peaks- RRT 70
Use 1254
for peaks
1 RRT 104
|RRT-i70?j
Is there a distinct
peak with RRT 117?
Use 1260 for
all peaks
YES
NO
Use 1254 for all
peaks! RRT 174
Use 1260 for
all other peaks
Figure 9. Chromatogram Division Flowchart (8).
81
-------
REFERENCES:
1. "Method for Chlorinated Hydrocarbons in Water and Wastewater", this
manual, p. 7.
2. Leoni, V., "The Separation of Fifty Pesticides and Related Compounds and
Polychlorinated Biphenyls into Four Groups by Silica Gel Microcolumn
Chromatography", Journal of Chromatography, 62, 63 (1971).
3. McClure, V. E., "Precisely Deactivated Adsorbents Applied to the Separa-
tion of Chlorinated Hydrocarbons", Journal of Chromatography. 70, 168
(1972).
4. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories", Chapter 6, Section 6.4, U. S. Environmental Protection
Agency, National Environmental Research Center, Analytical Quality
Control Laboratory, Cincinnati, Ohio, 45268, 1972.
5. "Pesticide Analytical Manual", U. S. Dept. of Health, Education and
Welfare, Food and Drug Administration, Washington, D. C.
6. Bellar, T. A. and Lichtenberg, J. J., "Method for the Determination of
Polychlorinated Biphenyls in Water and Sediment", U. S. Environmental
Protection Agency, National Environmental Research Center, Analytical
Quality Control Laboratory, Cincinnati, Ohio, 45268, 1973.
7. Goerlitz, D. F. and Law, L. M., "Note on Removal of Sulfur Interferences
from Sediment Extracts for Pesticide Analysis", Bulletin of Environmental
Contamination and Toxicology, £, 9 (1971).
8. Webb, R. G. and McCall, A. C., "Quantitative PCB Standards for Electron
Capture Gas Chromatography", Journal of Chromatographic Science, 11, 366
(1973).
82
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METHOD FOR TRIAZINE PESTICIDES IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of various symmetrical triazine
pesticides in water and wastewaters.
1.2 The following pesticides may be determined individually by this
method:
Parameter Storet No.
Ametryn —
Altraton —
Atrazine 39033
Prometon 39056
Prometryn 39057
Propazine 39024
Secbumeton —
Simazine 39055
Terbuthylazine —
2. Summary
2.1 The method describes an efficient sample extraction procedure
and provides, through use of column chromatography, a method
for the elimination of non-pesticide interferences and the
pre-separation of pesticide mixtures. Identification is made
by selective gas chromatographic separation, and measurement
is accomplished by the use of an electroytic conductivity
detector (CCD) in the nitrogen mode or a nitrogen specific
thermionic detector. Results are reported in micrograms per
liter.
83
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2.2 This method is recommended for use only by experienced
pesticide analysts or under the close supervision of such
qualified persons.
3. Interferences
3.1 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. Specific
selection of reagents and purification of solvents by
distillation in all-glass systems may be required. Refer to
Appendix I.
3.2 The interferences in industrial effluents are high and varied
and often pose great difficulty in obtaining accurate and
precise measurement of triazine pesticides. The use of a
specific detector supported by an optional column cleanup
procedure will eliminate many of these interferences.
3.3 Nitrogen containing compounds other than the triazines may
interfere.
4. Apparatus and Materials
4.1 Gas Chromatograph - Equipped with glass lined injection port.
4.2 Detector Options
4.2.1 Electrolytic Conductivity.
4.2.2 Nitrogen specific thermionic
4.3 Recorder - Potentiometric strip chart (10 in.) compatible
with the detector.
84
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4.4 Gas Chromatographic Column Materials:
4.4.1 Tubing - Pyrex (180 cm long x 4 mm ID)
4.4.2 Glass Wool - Silanized
4.4.3 Solid Support - Gas Chrom Q (100-120 mesh)
4.4.4 Liquid Phases - Expressed as weight percent coated on
solid support.
4.4.4.1 Carbowax 20M, IX
4.5 Kuderna-Danish (K-D) Glassware
4.5.1 Snyder Column - three ball (macro) and two ball (micro)
4.5.2 Evaporative Flasks - 500 ml
4.5.3 Receiver Ampuls - 10 ml, graduated
4.5.4 Ampul Stoppers
4.6 Chromatographic Column - Chromaflex (400 mm x 19 mm ID) with
coarse fritted plate and Teflon stopcock on bottom; 250 ml
reservoir bulb at top of column with flared out funnel shape
at top of bulb - a special order (Kontes K-420540-9011).
4.7 Chromatographic Column - Pyrex (approximately 400 mm long x
20 mm ID) with coarse fritted plate on bottom.
4.8 Micro Syringes - 10, 25, 50 and 100 ;u1.
4.9 Separatory funnels - 2000 ml with Teflon stopcock.
4.10 Blender - High speed, glass or stainless steel cup.
4.11 Graduated Cylinders - 1000 ml.
4.12 Florisil - PR Grade (60-100 mesh); purchase activated at
1250°F and store in the dark in glass containers with glass
stoppers or foil-lined screw caps. Before use, activate each
85
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batch overnight at 130°C in foil-covered glass container.
Determine lauric acid value (See Appendix II).
5. Reagents, Solvents, and Standards
5.1 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.2 Sodium Sulfate - (ACS) Granular, anhydrous (conditioned at
400 C for 4 hrs.).
5.3 Sulfuric Acid - (ACS) Mix equal volumes of cone. H2S04
with distilled water.
5.4 Diethyl Ether - Pesticide Quality, redistilled in glass, if
necessary
5.4.1 Must be free of peroxides as indicated by EM Quant
Test strips. (Test strips are available from EM
Laboratories, Inc., 500 Executive Blvd., Elmsford,
N.Y. 10523.)
5.4.2 Procedures recommended for removal of peroxides are
provided with the test strips.
5.5 Hexane, Methanol, Methylene Chloride, Petroleum Ether
(boiling range 30-60°C) - pesticide quality, redistill in
glass if necessary.
5.6 Pesticide Standards - Reference grade.
6. Calibration
6.1 Gas chromatographic operating conditions are considered
optimum when an injection of ^ 20 ng of each triazine will
yield a peak at least 50% of full scale deflection with the
modified Coulson detector (1). For all quantitative
86
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measurements, the detector must be operated within its linear
response range and the detector noise level should be less
than 2% of full scale.
»
6.2 Inject standards frequently as a check on the stability of
operating conditions. A chromatogram of a mixture of several
pesticides is shown in Figure 1 and provides reference
operating conditions for the recommended column.
6.3 The elution order and retention ratios of various
organophosphorus pesticides are provided in Table 1, as a
guide.
7. QualityControl
7.1 Duplicate and spiked sample analyses are recommended as
quality control checks. Quality control charts (2) should be
developed and used as a check on the analytical system.
Quality control check samples and performance evaluation
samples should be analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
8. Sample Preparation
8.1 Blend the sample if suspended matter is present and adjust pH
to near neutral (pH 6.5-7.5) with 50% sulfuric acid or ION
sodium hydroxide.
8.2 Quantitatively transfer a 1000 ml aliquot into a two-liter
separatory funnel.
87
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6 8 10
RETENTION TIME IN MINUTES
14
Figure 1. Column Packing: 1% Carbowax 20M on Gas-Chrom Q (100/120 mesh),
Column Temperature: 155 C, Carrier Gas: Helium at 80 ml/min,
Detector: Electrolytic Conductivity.
-------
TABLE 1
RETENTION RATIOS OF VARIOUS TRIAZINE
PESTICIDES RELATIVE TO ATRAZINE
Pesticide
Prometon
Atraton
Propazine
Terbuthylazirie
Secbumeton
Atrazine
Prometryne
Simazine
Ametryne
Retention Ratio
0.52
0.67
0.71
0.78
0.88
1.00
1.10
1.35
1.48
Absolute retention time of atrazine = 10.1 minutes
89
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9. Extraction
9.1 Add 60 ml methylene chloride to the sample in the separatory
funnel and shake vigorously for two minutes.
9.2 Allow the solvent to separate from the sample, draw the
organic layer into a 100-ml beaker, then pass the organic
layer through a chromatographic column containing 3-4 inches
anhydrous sodium sulfate, and collect it in a 500-ml K-D
flask equipped with a 10 ml ampul. Add a second 60-ml volume
of solvent to the separatory funnel and complete the
extraction procedure a second time. Perform a third
extraction in the same manner.
9.3 Concentrate the extract to 10 ml in a K-D evaporator on a hot
water bath. Disconnect the Snyder column just long enough to
add 10 ml hexane to the K-D flask and then continue the
concentration to about 5-6 ml. This operation is to displace
methylene chloride and give a final hexane solution. If the
need for cleanup is indicated, continue to Florisil Column
Cleanup (10 below).
9.4 If further cleanup is not required, replace the Snyder column
and flask with a micro-Snyder column and continue the
concentration to 0.5-1.0 ml. Analyze this final concentrate
by gas chromatography.
10. Florisil Column Adsorption Chromatography
10.1 Adjust the sample extract volume to 10 ml.
90
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10.2 Place a charge of activated Florisil (weight determined by
lauric acid value, see Appendix II) in a Chromaflex column.
After settling the Florisil by tapping the column, add about
one-half inch layer of anhydrous granular sodium sulfate to
the top.
10.3 Pre-elute the column, after cooling, with 50-60 ml of
petroleum ether. Discard the eluate and just prior to
exposure of the sulfate layer to air, quantitatively
transfer the sample extract into the column by decantation
and subsequent petroleum ether washings. Adjust the elution
rate to about 5 ml per minute and, separately, collect up to
four eluates in 500-ml K-D flasks equipped with 10-ml
ampuls. (See Eluate Composition, 10.4.) Perform the first
elution with 200 ml of 6% ethyl ether in petroleum ether,
and the second elution with 200 ml of 15% ethyl ether in
petroleum ether. Perform the third elution with 200 ml of
50% ethyl ether - petroleum ether and the fourth elution
with 200 ml of 100% ethyl ether.
10.4 Eluate Composition - By using an equivalent quantity of any
batch of Florisil as determined by its lauric acid value,
the pesticides will be separated into the eluates indicated
as follows:
91
-------
15% Eluate 50% Equate 100% Eluate
Propazine (90%) Propazine (10%) Atraton
Terbuthylazine (30%) Terbuthylazine(70%) Secbumeton
Atrazine (20%) Atrazine (80%) Prometon
Ametryne
Prometryne
Slmazine
10.5 Concentrate the eluates to 6-10 ml in the K-D evaporator in a
hot water bath. Change to the micro-Snyder column and continue
concentration to 0.5-1.0 ml.
10.6 Analyze by gas chromatography.
11. Calculation of Results
11.1 Determine the pesticide concentration by using the absolute
calibration procedure described below or the relative
calibration procedure described in Appendix III.
(1) Micrograms/liter = (A) (B) (V>1
)
(Vs
A = ng standard
Standard area
B = Sample aliquot area
Vj = Volume of extract injected
Vt = Volume of total extract
Vs = Volume of water extracted (ml)
12. Reporting Results
12.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are analyzed
all data obtained should be reported.
92
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REFERENCES:
1. Patchett, G. G., "Evaluation of the Electrolytic Conductivity Detector
for Residue Analyses of Nitrogen-Containing Pesticides", Journal of
Chroma tog raphic Science. 8., 155 (1970).
2. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories", Chapter 6, Section 6.4, U. S. Environmental Protection
Agency, National Environmental Research Center, Analytical Quality
Control Laboratory, Cincinnati, Ohio, 45268, 1972. (Revised edition
to be available soon.)
93
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METHOD FOR 0-ARYL CARBAMATE PESTICIDES IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of various 0-aryl carbamate
pesticides in water and wastewater.
1.2 The following, pesticides may be determined individually by this
method:
Parameter Storet No.
Aminocarb —
Carbaryl 39750
Methiocarb —
Mexacarbate —
Propoxur —
2. Summary
2.1 A measured volume of water is extracted with methylene
chloride. The concentrated extract is cleaned up with a
Florisil column. Appropriate fractions from the column are
concentrated and portions are separated by thin-layer
chromatography. The carbamates are hydrolyzed on the layer and
the hydrolysis products are reacted with 2,6-dibromoquinone
chlorimide to yield specific colored products. Quantitative
measurement is achieved by visually comparing the responses of
sample extracts to the responses of standards on the same
thin-layer. Identifications are confirmed by changing the pH
of the layer and observing color changes of the reaction
products. Results are reported in micrograms per liter.
94
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2.2 • This method is recommended for use only by experienced
pesticide analysts or under the close supervision of such
qualified persons.
3. Interferences
3.1 Direct interferences may be encountered from phenols that may
be present in the sample. These materials react with the
chromogenic reagent and yield reaction products similar to
those of the carbamates. In cases where phenols are suspected
of interfering with a determination, a different solvent system
should be used to attempt to isolate the carbamates.
3.2 Indirect interferences may be encountered from naturally
colored materials whose presence masks the chromogenic reaction.
4. Apparatus and Materials
4.1 Thin-layer plates - Glass plates (200 x 200 mm) coated with
0.25 mm layer of Silica Gel G (gypsum binder).
4.2 Spotting Template
4.3 Developing Chamber
4.4 Sprayer - 20 ml capacity
4.5 Kuderna-Danish (K-D) Glassware (Kontes)
4.5.1 Snyder Column - three ball (K-503000)
4.5.2 Micro-Snyder Column - two ball (K-569001)
4.5.3 Evaporative Flasks - 500 ml (K-570001)
4.5.4 Receiver Ampuls - 10 ml graduated (K-570050)
4.5.5 Ampul Stoppers
95
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4.6 Chromatographic Column - Chromaflex (400 mm long x 19 mm ID) with
coarse fritted plate on bottom and Teflon stopcock; 250 ml
reservoir bulb at top of column with flared out funnel shape at
top of bulb - a special order (Kontes K-420540-9011).
4.7 Chromatographic Column - Pyrex (approximately 400 mm long x 20 mm
ID) with coarse fritted plate on bottom.
4.8 Micro Syringes - 10, 25, 50 and 100 ;ul.
4.9 Separatory Funnel - 2000 ml, with Teflon stopcock.
4.10 Blender - High speed, glass or stainless steel cup.
4.11 Florisil - PR Grade (60-80 mesh); purchase activated at 1250°F
and store in the dark in glass containers with glass stoppers or
foil-lined screw caps. Before use activate each batch overnight
at 130°C in foil-covered glass container. Determine lauric
acid value (see Appendix II).
5. Reagents, Solvents, and Standards
5.1 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.2 Sodium Sulfate - (ACS) Granular, anhydrous.
5.3 Sulfuric Acid - (ACS) Mix equal volumes of cone. H2S04 with
distilled water.
5.4 Diethyl Ether - Nanograde, redistilled in glass, if necessary.
5.4.1 Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM Laboratories,
Inc., 500 Executive Blvd., Elmsford, N.Y. 10523.)
5.4.2 Procedures recommended for removal of peroxides are
provided with the test strips.
96
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5.5 Hexane, Methanol, Methylene Chloride, Petroleum Ether -
nanograde, redistill in glass if necessary.
5.6 Pesticide Standards - Reference grade.
5.8.1 TLC Standards - 0.100^g/ul in chloroform.
5.7 Chromogenic agent - Dissolve 0.2 g 2,6-dibromoquinone chlorimide
in 20 ml chloroform.
5.8 Buffer solution - 0.1 N sodium borate in water.
6. Calibration
6.1 To insure even solvent travel up the layer, the tank used for
layer development must be thoroughly saturated with developing
solvent before it is used. This may be achieved by lining the
inner walls of the tank with chromatography paper and introducing
the solvent 1-2 hours before use.
6.2 Samples and standards should be introduced to the layer using a
syringe, micropipet or other suitable device that permits all the
spots to be about the same size and as small as possible. An air
stream directed on the layer during spotting will speed solvent
evaporation and help to maintain small spots.
6.3 For qualitative and quantitative work, spot a series representing
0.1-1.0 pg of a pesticide. Tables 1 and 2 present color
responses and R.f values for several solvent systems.
7. Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts should be developed
and used as a check on the analytical system. Quality control
97
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Table 1
Values of 0-Aryl Carbamate Pesticides in Several Solvent Systems
Carbaryl
Aminocarb
Mexacarbate
Methiocarb
Propoxur
A
0.26
0.26
0.34
0.31
0.27
B
0.22
0.02
0.22
0.31
0.10
C
0.48
0.46
0.54
0.55
0.53
D
0.41
0.52
0.53
0.55
0.59
E
0.58
0.54
0.60
0.59
0.60
F
0.24
0.04
0.24
0.28
0.13
Solvent Systems:
A. Hexane/acetone (3:1)
B. Methylene chloride
C. Benzene/acetone (4:1)
D. Benzene/cyclohexane/diethylamine (5:2:2)
E. Ethyl acetate
F. Chloroform
98
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Table 2
Color Responses and Detection Limit for 0-Aryl Carbamates
Colors:Detection
Before After Limit
Buffer Buffer (ug)
Carbaryl Brown Red-Purple 0.1
Aminocarb Gray Green 0.1
Mexacarbate Gray Green 0.1
Methiocarb Brown Tan 0.2
Propoxur Blue Blue 0.1
99
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check samples and performance evaluation samples should be
analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that used
to dilute the sample.
8. Sample Preparation
8.1 Blend the sample if suspended matter is present and adjust pH to
near neutral (pH 6.5-7.5) with 5Q% sulfuric acid or 10 N sodium
hydroxide.
8.2 Quantitatively transfer a one-liter aliquot into a two-liter
separatory funnel.
9. Extraction
9.1 Add 60 ml of methylene chloride to the sample in the separatory
funnel and shake vigorously for two minutes.
9.2 Allow the solvent to separate from the sample, draw the organic
layer into a 100-ml beaker, then pass the organic layer through a
chromatographic column containing 3-4 inches anhydrous sodium
sulfate, and collect it in a 500-ml K-D flask equipped with a
10-ml ampul. Add a second 60-ml volume of solvent to the
separatory funnel and complete the extraction procedure a second
time. Perform a third extraction in the same manner.
9.3 Concentrate the extract to 10 ml in a K-D evaporator on a hot
water bath. Disconnect the Snyder column just long enough to add
10 ml of hexane to the K-D flask and then continue the
concentration to about 5-6 ml. If the need for cleanup is
indicated, continue to Florisil Column Cleanup (10 below).
100
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9.4 If further cleanup is not required, replace the Snyder column and
t
flask with a micro-Snyder column and continue the concentration
to 0.5-1.0 ml. Analyze this final concentrate by thin-layer
chromatography (Section 11).
10. Florisil Column Cleanup
10.1 Adjust the sample extract to 10 ml with hexane.
10.2 Place a charge of activated Florisil (weight determined by
lauric-acid value, see Appendix II) in a Chromaflex column.
After settling the Florisil by tapping the column, add about
one-half inch layer of anhydrous granular sodium sulfate to the
top.
10.3 Pre-elute the column, after cooling, with 50-60 ml of petroleum
ether. Discard the eluate and just prior to exposure of the
sulfate layer to air, quantitatively transfer the sample extract
into the column by decantation and subsequent petroleum ether
washings. Adjust the elution rate to about 5 ml per minute and,
separately collect the four eluates in 500-ml K-D flasks equipped
with 10-ml ampuls. Perform the first elution with 200 ml of 6%
ethyl ether in petroleum ether, and the second elution with 200
ml of 15% ethyl ether in petroleum ether. Perform the third
elution with 200 ml of 50% ethyl ether - petroleum ether and the
fourth elution with 200 ml of 100% ethyl ether.
10.3.1 Eluate Composition - By using an equivalent quantity of
any batch of Florisil as determined by its lauric acid
value, the pesticides will be separated into the eluates
indicated as follows:
101
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50% Eluate 100% Eluate
Carbaryl (70%) Carbaryl (30%)
Mexacarbate Aminocarb
Propoxur
10.4 Concentrate the eluates to 6 - 10 ml in the K-D evaporator in a
hot water bath. Change to the micro-Snyder column and continue
concentration to 0.5-1.0 ml.
10.5 Analyze according to 11. below.
11. Separation and Detection
11.1 Carefully spot 10% of the extract on a thin layer. On the same
plate spot several pesticides or mixtures for screening
purposes, or a series of 1, 2, 4, 6, 8 and 10 ^1 of specific
standards for quantitative analysis.
11.2 Develop the layers 10 cm in a tank saturated with solvent
vapors. Remove the plate and allow it to dry.
11.3 Spray the layer rapidly and evenly with about 10-15 ml
£
chromogenic reagent. Heat the layer in an oven at 110 C for 15
minutes. The pesticides will appear with colors as indicated in
Table 2. Make quantitative estimates by visually comparing the
intensity and size of the spots with those of the series of
standards.
11.4 Spray the layer with sodium borate reagent and observe the color
shift of the reaction products. The color shift must be the
same for sample and standard for identification to be confirmed.
102
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12. Calculation of Results
12.1 Determine the concentration of pesticide in a sample by
comparing the response in a sample to that of a quantity Of
f
standard treated on the same layer. Divide the result, in
micrograms, by the fraction of extract spotted to convert to
micrograms per liter.
13. Reporting Results
13.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are analyzed
all data obtained should be reported.
103
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METHOD FOR N-ARYL CARBAMATE AND UREA PESTICIDES IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of various N-aryl carbamate
and urea pesticides in water and wastewater.
1.2 The following pesticides may be determined individually by this
method:
Parameter Storet No.
Barban —
Chlorpropham —
Diuron 39650
Fenuron —
Fenuron-TCA —
Linuron —
Monuron —
Monuron-TCA —
Neburon —
Propham 39052
Siduron —
Swep —
2. Summary
2.1 A measured volume of water is extracted with methylene chloride
and the concentrated extract is cleaned up with A Florisil
column. Appropriate fractions from the column are concentrated
and portions are separated by thin-layer chromatography. The
pesticides are hydrolyzed to primary amines, which in turn are
chemically converted to diazonium salts. The layer is sprayed
with 1-naphthol and the products appear as colored spots.
Quantitative measurement is achieved by visually comparing the
104
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responses of sample extracts to the responses of standards on the
same thin layer. Results are reported in micrograms per liter.
2.2 This method is recommended for use only by experienced pesticide
analysts or under the close supervision of such qualified persons.
3. Interferences
3.1 Direct interferences may be encountered from aromatic amines that
may be present in the sample. These materials react with the
chromogenic reagent and yield reaction products similar to those
of the pesticides. In cases where amines are suspected of
interfering with a determination, a different solvent system
should be used to attempt to isolate the pesticides on the layer.
3.2 Indirect interferences may be encountered from naturally colored
materials whose presence masks the chromogenic reaction.
4. Apparatus and Materials
4.1 Thin-layer plates - Glass plates (200 x 200 mm) coated with 0.25
mm layer of Silica Gel G (gypsum binder).
4.2 Spotting Template
4.3 Developing Chamber
4.4 Sprayer - 20 ml capacity
4.5 Kuderna-Danish (K-D) Glassware (Kontes)
4.5.1 Snyder Column - three ball (K-503000)
4.5.2 Micro-Snyder Column - two ball (K-569001)
4.5.3 Evaporative Flasks - 500 ml (K-570001)
4.5.4 Receiver Ampuls - 10 ml graduated (K-570050)
4.5.5 Ampul Stoppers
105
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4.6 Chromatographic Column - Chromaflex (400 mm long x 19 mm ID) with
coarse fritted plate on bottom and Teflon stopcock; 250 ml
reservoir bulb at top of column with flared out funnel shape at
top of bulb - a special order (Kontes K-420540-9011).
4.7 Chromatographic Column - Pyrex (approximately 400 mm long x 20 mm
ID) with coarse fritted plate on bottom.
4.8 Micro Syringes - 10, 25, 50 and 100 jjl.
4.9 Separatory Funnel - 2000 ml, with Teflon stopcock.
4.10 Blender - High speed, glass or stainless steel cup.
4.11 Florisil - PR Grade (60-80 mesh); purchase activated at 1250°F
and store in the dark in glass containers with glass stoppers or
foil-lined screw caps. Before use activate each batch overnight
at 130°C in foil-covered glass container. Determine lauric acid
value (see Appendix II).
5. Reagents, Solvents, and Standards
5.1 Sodium Chloride - (ACS) Saturated solution in distilled water
(pre-rinse NaCl with hexane).
5.2 Sodium Hydroxide - (ACS) 10 N in distilled water.
5.3 Sodium Sulfate - (ACS) Granular, anhydrous (conditioned at 400 C
for 4 hrs.).
5.4 Sulfuric Acid - (ACS) Mix equal volumes of cone. H^SO* with
distilled water.
5.5 Diethyl Ether - Nanograde, redistilled in glass, if necessary.
5.5.1 Must be free of peroxides as indicated by EM Quant test
strips. (Test strips are available from EM Laboratories,
Inc., 500 Executive Blvd., Elmsford, N.Y. 10523.)
106
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5.5.'2 Procedures recommended for removal of peroxides are
provided with the test strips.
5.6 Hexane, Methanol, Methylene Chloride, Petroleum Ether - nanograde,
redistill in glass if necessary.
5.7 Pesticide Standards - Reference grade.
5.9.1 TLC Standards - O.lOOjug/jul in chloroform.
5.8 Nitrous acid - prepare just before use by mixing 1 g NaNC^ with
20 ml 0.2 N HC1.
5.9 Chromogenic agent - dissolve 1.0 g 1-Naphthol in 20 ml ethanol.
Prepare fresh daily.
6. Calibration
6.1 To insure even solvent travel up the layer, the tank used for
layer development must be thoroughly saturated with developing
solvent before it is used. This may be achieved by lining the
inner walls of the tank with chromatography paper and introducing
the solvent 1-2 hours before use.
6.2 Samples and standards should be introduced to the layer using a
syringe, micropipet or other suitable device that permits all the
spots to be about the same size and as small as possible. An air
stream directed on the layer during spotting will speed solvent
evaporation and help to maintain small spots.
6.3 For qualitative and quantitative work, spot a series representing
0.1-l.Ojjg of a pesticide. Tables 1 and 2 present color responses
and R~ values for. several solvent systems.
107
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TABLE 1
Rf VALUES OF N-ARYL CARBAMATE AND UREA PESTICIDES
IN SEVERAL SOLVENT SYSTEMS
Carbamates
Propham
Chloropropham
Barban
Swep
Urea
Fenuron
Fenuron-TCA
Monuron
Monuron-TCA
Diuron
Linuron
Neburon
Siduron
A
0.49
0.57
0.61
0.48
0.03
0.03
0.04
0.04
0.05
0.40
0.21
0.02
B
0.54
0.60
0.59
0.44
0.04
0.04
0.05
0.06
0.09
0.43
0.28
0.07
C
0.73
0.73
0.72
0.70
0.38
0.36
0.37
0.34
0.38
0.62
0.64
0.68
D
0.48
0.49
0.41
0.41
0.22
0.22
0.24
0.24
0.28
0.39
0.41
0.39
E
0.36
0.37
0.28
0.28
0.10
0.10
0.10
0.10
a. 13
0.24
0.26
0.25
F
0.68
0.70
0.70
0.67
0.41
0.41
0.47
0.46
0.54
0.66
0.68
0.62
G
0.69
0.73
0.74
0.66
0.30
0.30
0.34
0.34
0.44
0.64
0.65
0.55
Solvent Systems:
A. Methylene chloride
B. Chloroform
C. Ethyl Acetate
D. Hexane/acetone (2:1)
E. Hexane/acetone (4:1)
F. Chloroform/acetonitrile (2:1)
6. Chloroform/acetonitrile (5:1)
108
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TABLE 2
COLOR RESPONSES AND DETECTION LIMIT FOR THE N-ARYL CARBAMATES
AND UREAS
Carbamates
Propham
Chlorpropham
Bar ban
Swep
Ureas
Fenuron
Fenuron-TCA
Monuron
Monuron-TCA
Diuron
Linuron
Neburon
Siduron
Color
Red-purple
Purple
Purple
Blue-purple
Red-purple
Red-purple
Pink -orange
Pink-orange
Blue-purple
Blue-purple
Blue-purple
Red-purple
Detection Limit (ug)
0.2
0.1
0.05
0.2
0.05
0.1
0.05
0.1
0.1
0.1
0.1
0.05
109
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7- Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts (1) should be developed
and used as a check on the analytical system. Quality control
check samples and performance evaluation samples should be
analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that used
to dilute the sample.
8. Sample Preparation
8.1 Blend the sample if suspended matter is present and adjust pH to
near neutral (pH 6.5-7.5) with 50% sulfuric acid or 10 N sodium
hydroxide.
8.2 Quantitatively transfer a one-liter aliquot into a two-liter
separatory funnel.
9. Extraction
9.1 Add 60 ml of methylene chloride to the sample in the separatory
funnel and shake vigorously for two minutes.
9.2 Allow the solvent to separate from the sample, draw the organic
layer into a 100-ml beaker, then pass the organic layer through a
chromatographic column containing 3-4 inches anhydrous sodium
sulfate, and collect it in a 500-ml K-D flask equipped with a
10-ml ampul. Add a second 60-ml volume of solvent to the
separatory funnel and complete the extraction procedure a second
time. Perform a third extraction in the same manner.
110
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9.3 Concentrate the extract to 10 ml in a K-D evaporator on a hot
water bath. Disconnect the Snyder column just long enough to add
10-ml hexane to the K-D flask and then continue the concentration
to about 5-6 ml. If the need for cleanup is indicated, continue
to Florisil Column Cleanup (10 below).
9.4 If further cleanup is not required, replace the Snyder column and
flask with a micro-Snyder column and continue the concentration to
0.5-1.0 ml. Analyze this final concentrate by thin-layer
chromatography (Section 11).
10. Florisil Column Cleanup
10.1 Adjust the sample extract to 10 ml with hexane.
10.2 Place a charge of activated Florisil (weight determined by lauric
acid value, see Appendix II) in a Chromaflex column. After
settling the Florisil by tapping the column, add about one-half
inch layer of anhydrous granular sodium sulfate to the top.
10.3 Pre-elute the column, after cooling, with 50-60 ml of petroleum
ether. Discard the eluate and just prior to exposure of the
sulfate layer to air, quantitatively transfer the sample extract
into the column by decantation and subsequent petroleum ether
washings. Adjust the elution rate to about 5 ml per minute and,
separately, collect up to four eluates in 500-ml K-D flasks
equipped with 10-ml ampuls. (See Eluate Composition, 10.3.1.)
Perform the first elution with 200 ml of 6% ethyl ether in
petroleum ether, and the second elution with 200 ml of 15% ethyl
ether in petroleum ether. Perform the third elution with 200 ml
111
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of 50% ethyl ether - petroleum ether and the fourth elution with
200 ml of 100% ethyl ether.
10.3.1 Eluate Composition - By using an equivalent quantity of any
batch of Florisil as determined by its lauric acid value,
the pesticides will be separated into the eluates indicated
below:
15% Eluate 50% Eluate 100% Eluate
Chlorpropham Barban (5%) Neburon (92%)
Propham Linuron Diuron
Barban (95%) Neburon (8%) Monuron
Siduron
CAUTION: Fenuron and Fenuron-TCA are not recovered from
the Florisil column.
10.4 Concentrate the eluates to 6-10 ml in the K-D evaporator in a hot
water bath. Change to the micro-Snyder column and continue
concentration to 0.5-1.0 ml.
10.5 Analyze according to 11. below.
11. Separation and Detection
11.1 Carefully spot 10% of the extract on a thin layer. On the same
plate spot several pesticides or mixtures for screening purposes,
or a series of 1, 2, 4, 6, 8 and 10 ;jl of specific standards for
quantitative analysis.
11.2 Develop the layers 10 cm in a tank saturated with solvent vapors.
Remove the plate and allow it to dry.
11.3 Spray the layer rapidly and evenly with about 10-15 ml sulfuric
acid solution. Heat the layer in an oven at 110°C for 15
minutes.
112
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11.4 When the layer is cool, spray it with nitrous acid reagent and
allow it to dry. Spray the layer with 1-naphthol reagent and
allow it to dry again. The pesticides will appear as purple spots
(see Table 2). Identifications are made by comparison of colors
and Rf values. Quantitative estimates are made by visually
comparing the intensity and size of the spots with those of the
series of standard.
12. Calculation of Results
12.1 Determine the concentration of pesticide in a sample by comparing
the response in a sample to that of a quantity of standard treated
on the same layer. Divide the result, in micrograms, by the
fraction of extract spotted to convert to micrograms per liter.
13. Reporting Results
13.1 Report results in micrograms per liter without correction for
recovery data. When duplicate and spiked samples are analyzed all
data obtained should be reported.
113
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REFERENCES:
1. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories", Chapter 6, Section 6.4, U. S. Environmental Protection
Agency, National Environmental Research Center, Analytical Quality
Control Laboratory, Cincinnati, Ohio, 45268, 1972.
114
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METHOD FOR CHLOROPHENOXY ACID PESTICIDES IN WATER AND WASTEWATERS
1. Scope and Application
1.1 This method covers the determination of various chlorinated
phenoxy acid pesticides in water and wastewater.
1.2 The following pesticides may be determined individually by this
method:
Parameter Storet No.
2,4-D
Dicamba —
Silvex 39760
2,4,5-T
1.3 Since these compounds may occur in water in various forms
(i.e., acid, salt, ester, etc.) a hydrolysis step is included
to permit the determination of the active part of the herbicide.
2. Summary
2.1 Chlorinated phenoxy acids and their esters are extracted from
the acidified water sample with ethyl ether. The esters are
hydrolyzed to acids and extraneous organic material is removed
by a solvent wash. The acids are converted to methyl esters
which are extracted from the aqueous phase. The extract is
cleaned by passing it through a micro-adsorption column.
Identification of the esters is made by selective gas
chromatographic separations and may be corroborated through the
use of two or more unlike columns. Detection and measurement
is accomplished by electron capture, microcoulometric or
115
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electrolytic conductivity gas chromatography (1). Results are
reported in micrograms per liter.
2.2 This method is recommended for use only by experienced
pesticide analysts or under the close supervision of such
qualified persons.
3. Interferences
3.1 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 interference
under the conditions of the analysis. Specific selection of
reagents and purification of solvents by distillation in
all-glass systems may be required. Refer to Appendix I.
3.2 The interferences in industrial effluents are high and varied
and often pose great difficulty in obtaining accurate and
precise measurement of chlorinated phenoxy acid herbicides.
Sample clean-up procedures are generally required and may
result in loss of certain of these herbicides. It is not
possible to describe procedures for overcoming all of the
interferences that may be encountered in industrial effluents.
3.3 Organic acids, especially chlorinated acids, cause the most
direct interference with the determination. Phenols including
chlorophenols will also interfere with this procedure.
3.4 Alkaline hydrolysis and subsequent extraction eliminates many
of the predominant chlorinated insecticides which might
otherwise interfere with the test.
116
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3.5 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Glassware
and glass wool should be acid-rinsed and sodium sulfate should
be acidified with sulfuric acid to avoid this possibility.
4. Apparatus and Materials
4.1 Gas Chromatograph - Equipped with glass lined injection port.
4.2 Detector Options:
4.2.1 Electron Capture - Radioactive (tritium or nickel-63)
4.2.2 Microcoulometric Titration
4.2.3 Electrolytic Conductivity
4.3 Recorder - Potentiometric strip chart (10 in.) compatible with
the detector.
4.4 Gas Chromatographic Column Materials:
4.4.1 Tubing - Pyrex (180 cm long X 4 mm ID)
4.4.2 Glass Wool - Silanized
4.4.3 Solid Support - Gas-Chrom-Q (100-120 mesh)
4.4.4 Liquid Phases - Expressed as weight percent coated on
solid support.
4.4.4.1 OV-210, 5%
4.4.4 2 OV-17, 1.5% plus QF-1 or OV-210, 1.95%
4.5 Kuderna-Danish (K-D) Glassware
4.5.1 Snyder Column - three ball (macro) and two ball
(micro)
4.5.2 Evaporative Flasks - 250 ml
117
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4.5.3 Receiver Ampuls - 10 ml, graduated
4.5.4 Ampul Stoppers
4.6 Blender - High speed, glass or stainless steel cup.
4.7 Graduated cylinders - 100 and 250 ml.
4.8 Erlenmeyer flasks - 125 ml, 250 ml ground glass * 24/40
4.9 Microsyringes - 10, 25, 50 and 100 1.
4.10 Pipets - Pasteur, glass disposable (140 mm long X 5 mm ID).
4.11 Separatory Funnels - 60 ml and 2000 ml with Teflon stopcock.
4.12 Glass wool - Filtering grade, acid washed.
4.13 Diazald Kit - Recommended for the generation of diazomethane
(available from Aldrich Chemical Co., Cat. #210,025-2).
5. Reagents, Solvents and Standards
5.1 Boron Trifluoride-Methanol-esterification-reagent, 14 percent
boron trifluoride by weight.
5.2 N-methyl-N-nitroso-p-toluenesulfonamide (Diazald) - High
purity, melting point range 60-62°C. Precursor for the
generation of diazomethane (see Appendix IV).
5.3 Potassium Hydroxide Solution - A 37 percent aqueous solution
prepared from reagent grade potassium hydroxide pellets and
reagent water.
5.4 Sodium Chloride - (ACS) Saturated solution (pre-rinse NaCl with
hexane) in distilled water.
5.5 Sodium Hydroxide - (ACS) 10 N in distilled water.
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5.6 Sodium Sulfate, Acidified - (ACS) granular sodium sulfate,
treated as follows: Add 0.1 ml of cone, sulfuric acid to lOOg
of sodium sulfate slurrled with enough ethyl ether to just
cover the solid. Remove the ether with the vacuum. Mix 1 g of
the resulting solid with 5 ml of reagent water and ensure the
mixture to have a pH below 4. Store at 130CC.
5.7 Sulfuric add - (ACS) concentrated, Sp. Gr. 1.84.
5.8 Flor1s1l - PR grade (60-100 mesh) purchased activated at 1250°F
and stored at 130°C.
5.9 Carbltol (dlethylene glycol monoethyl ether).
5.10 Dlethyl Ether - Nanograde, redistilled 1n glass, 1f necessary.
5.1,0.1 Must be free of peroxides as Indicated by EM Quant test
strips. (Test strips are available from EM
Laboratories, Inc., 500 Executive Blvd., Elmsford, N.Y.
10523.)
5.10.2 Procedures recommended for removal of peroxides are
provided with the test strips.
5.11 Benzene Hexane - Nanograde, redistilled 1n glass, if necessary.
5.12 Pesticide Standards - Adds and Methyl Esters, reference grade.
5.12.1 Stock; standard solutions - Dissolve 100 mg of each
herbicide in 60 ml ethyl ether; then make to 100 ml with
redistilled hexane. Solution contains 1 mg/ml.
5.12.2 Working standard - Pipet 1.0 ml of each stock solution
Into a single 100 ml volumetric flask. Make to volume
with a mixture of ethyl ether and hexane (1:1).
Solution contains 10 /ig/ml of each standard.
119
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5.12.3 Standard for Chromatography (Diazomethane Procedure) -
Pipet 1.0 ml of the working standard into a glass
stoppered test tube and evaporate the solvent using a
steam bath. Add 2 ml diazomethane to the residue. Let
stand 10 minutes with occasional shaking, then allow the
solvent to evaporate spontaneously. Dissolve the
residue in 200 ^il of hexane for gas chromatography.
5.12.4 Standard for Chromatgraphy (Boron Trifluoride Proce-
dure) - Pipet 1.0 ml of the working standard into a
glass stoppered test tube. Add 0.5 ml of benzene and
evaporate to 0.4 ml using a two-ball Snyder microcolumn
and a steam bath. Proceed as in 11.3.1. Esters are
then ready for gas chromatography.
6. Calibration
6.1 Gas chromatographic operating conditions are considered
acceptable if the response to dicapthon is at least 50% of full
scale when < 0.06 ng is injected for electron capture detection
and < 100 ng is injected for microcoulometric or electrolytic
conductivity detection. For all quantitative measurements, the
detector must be operated within its linear response range and
the detector noise level should be less than 2% of full scale.
6.2 Standards, prepared from methyl esters of phenoxy acid
herbicides calculated as the acid equivalent, are injected
frequently as a check on the stability of operating
conditions. Gas chromatograms of several chlorophenoxys are
shown in Figure 1.
120
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6 4
RETENTION
TIME
2
IN
0
MINUTES
Fig. I Column: 1.5% 0V -17 + 1.95% QF- I,
Carrier Gas : Argon (5%) / Methane: 70ml/min.,
Column Temp. 185 C, Detector: Electron Capture .
121
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6.3 The elution order and retention ratios of methyl esters of
chlorinated phenoxy acid herbicides are provided in Table 1, as
a guide.
7. Quality Control
7.1 Duplicate and spiked sample analyses are recommended as quality
control checks. Quality control charts (2) should be developed
and used as a check on the analytical system. Quality control
check samples and performance evaluation samples should be
analyzed on a regular basis.
7.2 Each time a set of samples is extracted, a method blank is
determined on a volume of distilled water equivalent to that
used to dilute the sample.
8. Sample Preparation
8.1 The sample size taken for analysis is dependent on the type of
sample and the sensitivity required for the purpose at hand.
Background information on the pesticide levels previously
detected at a given sampling site will assist in determining
the sample size required, as well as the final volume to which
the extract needs to be concentrated, A 1-liter sample is
usually taken for drinking water and ambient water analysis to
provide a detection limit of 0.050 to O.lOOjug/1. One-hundred
milliliters is usually adequate to provide a detection limit of
1 >jg/l for industrial effluents.
122
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Table 1
RETENTION RATIOS FOR METHYL ESTERS OF SOME CHLORINATED
PHENOXY ACID HERBICIDES RELATIVE TO 2,4-D
Liquid Phase1
Column Temp.
Argon /Methane
Carrier Flow
Herbicide
dicamba
2,4-D
sllvex
2,4, 5-T
2,4-D
(minutes absolute)
1.5% OV-17
1.95% OF-12
185°C
70 ml/min
RR
0.60
1.00
1.34
1.72
2.00
5% OV-210
185°C
70 ml/min
RR
0.61
1.00
1.22
1.51
1.62
columns glass, 180 cm x 4 mm ID, solid support
Gas Chrom Q (100/120 mesh)
2OV-210 may be substituted
123
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8.2 Quantitatively transfer the proper aliquot of sample from the
sample container into a two-liter separatory funnel. If less
than 800 ml is analyzed, dilute to one liter with interference
free distilled water.
9. Extraction
9.1 Add 150 ml of ether to the sample in the separatory funnel and
shake vigorously for one minute.
9.2 Allow the contents to separate for at least ten minutes. After
the layers have separated, drain the water phase into a 1-liter
Erlenmeyer flask. Then collect the extract in a 250-ml
ground-glass Erlenmeyer flask containing 2 ml of 37 percent
aqueous potassium hydroxide.
9.3 Extract the sample two more times using 50 ml of ether each
time, and combine the extracts in the Erlenmeyer flask. (Rinse
the 1-liter flask with each additional aliquot of extracting
solvent.)
10. Hydrolysis
10.1 Add 15 ml of distilled water and a small boiling stone to the
flask containing the ether extract, and fit the flask with a
3-ball Snyder column. Evaporate the ether on a steam bath and
continue heating for a total of 60 minutes.
10.2 Transfer the concentrate to a 60-ml separatory funnel. Extract
the basic solution two times with 20 ml of ether and discard
the ether layers. The herbicides remain in the aqueous phase.
124
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10.3 Acidify the contents of the separatory funnel by adding 2 ml of
cold (4°C) 25 percent sulfuric acid (5.9). Extract the
herbicides once with 20 ml of ether and twice with 10 ml of
ether. Collect the extracts in a 125-ml Frlenmeyer flask
containing about 0.5 g of acidified anhydrous sodium sulfate
(5.8). Allow the extract to remain in contact the the sodium
sulfate for approximately two hours.
11. Esterification (3.4)
11.1 Transfer the ether extract, through a funnel plugged with glass
wool, into a Kuderna-Danish flask equipped with a 10-ml
graduated ampul. Use liberal washings of ether. Using a glass
rod, crush any caked sodium sulfate during the transfer.
11.1.1 If esterification is to be done with diazomethane,
evaporate to approximately 4 ml on a steam bath (do not
immerse the ampul in water) and proceed as directed in
Section 11.2. Prepare diazomethane as directed in
Appendix IV.
11.1.2 If esterification is to be done with boron trifluoride,
add 0.5 ml benzene and evaporate to about 5 ml on a
steam bath. Remove the ampul from the flask and further
concentrate the extract to 0.4 ml using a two-ball
Snyder microcolumn and proceed as in 11.3.
11.2 Diazomethane Esterification
11.2.1 Disconnect the ampul from the K-D flask and place in a
hood, away from steam bath. Adjust volume to 4 ml with
125
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ether, add 2 ml diazomethane, and let stand 10 minutes
with occasional swirling.
11.2.2 Rinse inside wall of ampul with several hundred
micro liters of ethyl ether. Take sample to
approximately 2 ml to remove excess diazomethane by
allowing solvent to evaporate spontaneously (room
temperature.
11.2.3 Dissolve residue in.5 ml of hexane. Analyze by gas
chromatography.
11.2.4 If further clean-up of the sample is required, proceed
as in 11.3.4 substituting hexane for benzene.
11.3 Boron Trifluoride Ester ification
11.3.1 After the benzene solution in the ampul has cooled, add
0.5 ml of borontrifluoride-methanol reagent. Use the
two-ball Snyder microcolumn as an air-cooled condenser
and hold the contents of the ampul at 50°C for 30
minutes on the steam bath.
11.3.2 Cool and add about 4.5 ml of a neutral 5 percent aqueous
sodium sulfate solution so that the benzene-water
interface is in +he neck of the Kuderna-Danish ampul.
Seal the flask with a ground glass stopper and shake
vigorously for about one minute. Allow to stand for
three minutes for phase separation.
11.3.4 Pipet the solvent layer from the ampul to the top of a
small column prepared by plugging a disposable Pasteur
126
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pipet with glass wool and packing with 2.0 cm of sodium
sulfate over 1.5 cm of Florisil adsorbent. Collect the
eluate in a graduated ampul. Complete the transfer by
repeatedly rinsing the ampul with small quantities of
benzene and passing the rinses through the column until
a final volume of 5.0 ml of eluate is obtained. Analyze
by gas chromatography.
12. Calculation of Results
12.1 Determine the methyl ester concentration by using the absolute
calibration procedure described below or the relative calibra-
tion procedure described in Appendix III.
(1) Micrograms/liter = lALJll
(V-j) (Vs)
A = ng standard
standard area
B = Sample aliquot area
V]= Volume of extract injected
Vt= Volume of total extract QJ!)
Vs= Volume of water extracted (ml)
12.2 Molecular weights for the calculation of methyl esters as the
acid equivalents.
2,4-D 222.0 Dicamba 221.0
2,4-D methyl ester 236.0 Dicamba methyl ester 236.1
Silvex 269.5 2,4,5-T 255.5
Silvex methyl ester 283.5 2,4,5-T methyl ester 269.5
127
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13. Reporting Results
13.1 Report results in micrograms per liter as the acid equivalent
without correction for recovery data. When duplicate and
spiked samples are analyzed all data obtained should be
reported.
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REFERENCES:
1. Goerlitz, D. 6., and Lamar, W. L., "Determination of Phenoxy and
Herbicides in Water by Electron-Capture and Microcoulometric Gas
Chromatography", U. S. Geol. Survey Water-Supply Paper 1817-C (1967).
2. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories" (1972), U. S. Environmental Protection Agency, National
Environmental Research Center, Analytical Quality Control Laboratory,
Cincinnati, Ohio, 45268.
3. Metcalf, L. D. and Schmitz, A. A., "The Rapid Preparation of Fatty Acid
Esters for Gas Chromatographic Analysis", Analytical Chemistry, 33,
363 (1961).
4. Schlenk, H. and Gellerman, J. L., "Esterification of Fatty Acids with
Diazomethane on a Small Scale", Analytical Chemistry. 32, 1412 (1960).
129
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METHOD FOR VOLATILE CHLORINATED ORGANIC COMPOUNDS IN WATER AND WASTEWATERS
1. Scope and Application
1.1 This method covers the determination of various chlorinated organic
compounds in water and wastewater.
1.2 The following chlorinated organic compounds may be determined
individually by this method:
Parameter Storet No.
Benzylchloride —
Carbon tetrachloride 32102
Chlorobenzene 34301
Chloroform 32106
Epichlorohydrin —
Methylene Chloride 34423
1,1,2,2-Tetrachloroethane —
Tetrachloroethylene 34475
1,2,4-Trichlorobenzene —
1,1,2-Trichloroethane —
2. Summary
2.1 If the sample is turbid, it is initially centrifuged or filtered
through a fiber glass filter in order to remove suspended matter.
A three to ten microliter aliquot of the sample is injected into
the gas chromatograph equipped with a halogen specific detector.
The resulting chromatogram is used to identify and quantitate
specific components in the sample. Results are reported in
micrograms per liter. Confirmation of qualitative identifications
are made using two or more dissimilar columns.
130
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3. Interferences
3,1 The use of a halogen specific detector minimizes the possibility of
interference from compounds not containing chlorine, bromine, or
iodine. Compounds containing bromine or iodine will interfere with
the determination of organochlorine compounds. The use of two
dissimilar chromatographic columns helps to eliminate this
interference and, in addition, this procedure helps to verify all
qualitative identifications. When concentrations are sufficiently
high, unequivocal identifications can be made using infrared or
mass spectroscopy. Though non-specific, the flame ionization
detector may be used for known systems where interferences are not
a problem.
3.2 Ghosting is usually attributed to the history of the
chromatographic system. Each time a sample is injected, small
amounts of various compounds are adsorbed on active sites in the
inlet and at the head of the column. Subsequent injections of
water tend to steam clean these sites resulting in
non-representative peaks or displacement of the baseline. This
phenomenon normally occurs when an analysis of a series of highly
concentrated samples is followed by a low level analysis. The
system should be checked for ghost peaks prior to each quantitative
analysis by injecting distilled water in a manner identical to the
sample analysis (1). If excessive ghosting occurs, the following
corrective measures should be applied, as required, in the order
listed:
1) Multiple flushes with distilled water
131
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2) Clean or replace the glass injector liner
3) Replace the chromatographic column.
4. Apparatus and Materials
4.1 Gas Chromatograph - Equipped with programmed oven temperature
controls and glass-lined injection port. The oven should be
equipped with a column exit port and heated transfer line for
convenient attachment to the halogen specific detector.
4.2 Detector Options:
4.2.1 Microcoulometric Titration
4.2.2 Electrolytic Conductivity
4.2.3 Flame lonization
4.3 Recorder - Potentiometric strip chart recorder (10 in) compatible
with the detector.
4.4 Syringes - 1 ^il, 10>jl, and 50 jul.
4.5 BOD type bottle or 40 ml screw cap vials sealed with Teflon faced
silicone septa.
4.6 Volumetric Flasks - 500 ml, 1000 ml.
4.7 Syringe - Hypodermic Lur-lock type (30 ml).
4.8 Filter glass fiber filter - Type A (13 mm).
4.9 Filter holder - Swinny-type hypodermic adapter (13 mm).
4.10 Glass stoppered ampuls - 10 ml
4.11 Chromatographic columns
4.11.1 Moderately-Polar Column - 23 ft x 0.1 in ID x 0.125 in 00
stainless steel column #304 packed with 5% Carbowax 20 M
on Chromosorb-W (60-80 mesh).
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4.11.2 Highly-Polar Column - 23 ft x 0.1 in ID x 0.125 in OD
stainless steel #304 packed with 5% l,2,3-Tris-(2-cyano-
ethoxy) propane on Chromosorb-W (60-80 mesh).
4.11.3 Porous Polymer Column - 6 ft x 0.1 in ID x 0.125 in OD
stainless steel #304 packed with Chromosorb-101 (60-80
mesh).
4.11.4 Carbopack Column - 8 ft x 0.1 in ID x 0.125 in OD
stainless steel #304 packed with Carbopack-C (80-100
mesh) + 0.2% Carbowax 1500.
5. Reagents
5.1 Chlorinated hydrocarbon reference standards
5.1.1 Prepare standard mixtures in volumetric flasks using
contaminant-free distilled water as solvent. Add a
known amount of the chlorinated compounds with a
microliter syringe. Calculate the concentration of
each component as follows:
mg/1 = (Density of Compound) (^il injected) K - -^- - '
(Dilution Volume (ml))
6. Quality Control
6.1 Duplicate quantitative analysis on dissimilar columns should be
performed. The duplicate quantitative data should agree within
experimental error (+6 percent). If not, analysis on a third
dissimilar column should be performed. Spiked sample analyses
should be routinely performed to insure the integrity of the method.
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7. Selection Gas Chromatographic Column
7.1 No single column can efficiently resolve all chlorinated
hydrocarbons. Therefore, a specific column must be selected to
perform a given analysis. Columns providing only partially or
non-resolved peaks are useful only for confirmatory
identifications. If the qualitative nature of the sample is known>
an efficient column selection can be made by reviewing the
literature (2). In doing this, one must remember that injection of
large volumes of water can cause two serious problems not normally
noted using common gas Chromatographic techniques:
1) Water can cause early column failure due to liquid phase
displacement.
2) Water passing through the column causes retention times and
orders to change when compared to common sample solvent media,
i.e., hexane or air.
For these reasons, column life and the separations obtained by
direct aqueous injection may not be identical to those suggested in
literature.
8. Sample Collection and Handling
8.1 The sample containers should have a total volume in excess of 25 to
40 ml , although larger narrow-mouth bottles may be used.
8.1.1 Narrow mouth screw cap bottles with the TFE fluorocarbon
t
face silicone septa cap liners are strongly recommended.
Crimp-seal serum vials with TFE fluorocarbon faced septa or
134
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ground glass stoppered bottles are acceptable if the seal is
properly made and maintained during shipment.
8.2 Sample Bottle Preparation
8.2.1 Wash all sample bottles and TFE seals in detergent. Rinse
with tap water and finally with distilled water.
8.2.2 Allow the bottles and seals to air dry at room temperature.
8.2.3 Place the bottle in a 200°C oven for one hour, then allow
to cool in an area known to be free of organics.
8.2.4 When cool, seal the bottles using the TFE seals that will be
used for sealing the samples.
8.3 The sample is best preserved by protecting it from phase
separation. Since the majority of the chlorinated solvents are
volatile and relatively insoluble in water, it is important that
the sample bottle be filled completely to minimize air space over
the sample. Acidification will minimize the formation of
nonvolatile salts formed from chloroorganic acids and certain
chlorophenols. However, it may interfere with the detection of
acid degradable compounds such as chloroesters. Therefore, the
sample history must be known before any chemical or physical
preservation steps can be applied. To insure sample integrity, it
1s best to analyze the sample within 1 hour of collection.
6.4 Collect all samples in duplicate.
8.5 Fill the sample bottles in such a manner that no air bubbles pass
through the sample as the bottle is filled.
8.6 Seal the bottles so that no air bubbles are entrapped in it.
8.7 Maintain the hermetic seal on the sample bottle until analysis.
135
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8.8 Sampling from a water tap.
8.8.1 Turn on water and allow the system to flush. When the
temperature of the water has stabilized, adjust the flow to
about 50Q-ml/minute and collect duplicate samples from the
flowing stream.
8.9 Sampling from an open body of water.
8.9.1 Fill a 1-quart wide-mouth bottle with sample from a repre-
sentative area. Carefully fill duplicate 25 to 40 mi-sample
bottles from the 1-quart bottle.
9. Sample Preparation
9.1 If the sample is turbid, it should be filtered or centrifuged to
prevent syringe plugging or excessive ghosting problems. Filtering
the sample is accomplished by filling a 30-ml hypodermic syringe
with sample and attaching the Swinny-type hypodermic filter adaptor
with a glass fiber filter "Type A" installed. Discard the first 5
ml of sample then collect the filtered sample in a glass stoppered
sample filled to the top. (One should occasionally analyze the
non-filtered sample to insure that the filtering technique does not
adversely affect the sample).
10. Method of Analysis
10.1 Daily, analyze a standard containing 10.0 mg/1 of each compound to
be analyzed as a quality check sample before any samples are
analyzed. Instrument status checks and lower limit of detection
estimations based upon response factor calculations at two times
the signal to noise ratio are obtained from these data. In
136
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addition, response factor data obtained from this standard can be
used to estimate the concentration of the unknowns.
10.2 Analyze the filtered sample of unknown composition by injecting 3 to
10 ^il into the gas chromatograph. Record the injection volume and
detector sensitivity.
10.3 Prepare a standard mixture consisting of the same compounds in
concentrations approximately equal to those detected in the sample.
Chromatograph the standard mixture under conditions identical to the
unknown.
11. Calculation or Results
11.1 Measure the area of each unknown peak and each reference standard
peak as follows:
Area = (Peak Height)(Width of Peak at 1/2 Height)
11.2 Calculate the concentration of each unknown as follows:
(Area of Sample peak)(jul of Standard Injected)(Conc'n of Standard)
mg/1 = (jul of Sample injected)(Area of Standard Peak)
12. Reporting Results
12.1 Report results in mg/1. If a result is negative, report the minimum
detectable limit (see 10.1). When duplicate and spiked samples are
analyzed, all data obtained should be reported.
137
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_l i 1 I I I I I I I I I I I I I
16
24
32
RETENTION TIME IN MINUTES
Figure 1. Column: Chromosorb-101, Temperature Program: 125C
for 4 min then 4C/min up to 280 C., Carrier Gas: Nitrogen at
36 ml/min, Detector: Microcoulometric.
138
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REFERENCES:
1. Dressman, R. C., "Elimination of Memory Peaks Encountered in Aqueous-
Injection Gas Chromatography", Journal of Chromatographic Science, 8_,
265 (1970).
2. "Gas Chromatography Abstracts", Knapman, C. E. H., Editor, Institute of
Petroleum, 61 New Cavendish Street, London W1M8AR, Annually 1958 to date,
since 1970, also includes Liquid Chromatography Abstracts.
139
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METHOD FOR PENTACHLOROPHENOL IN WATER AND WASTEWATER
1. Scope and Application
1.1 This method covers the determination of pentachlorophenol (PCP) in
water and wastewater.
2. Summary
2.1 Pentachlorophenol is extracted from the acidified water sample (pH
3) with toluene, methylated with diazomethane, and analyzed by
electron-capture gas chromatography, using the columns listed in
the organochlorine pesticide method. (Page 7, this manual)
2.2 Further identification of pentachlorophenol is made with a mass
spectrometer.
3. Interferences
3.1 Chlorinated pesticides and other high boiling chlorinated organic
compounds may interfere with the analysis of PCP.
3.2 Injections of samples not treated with diazonmethane indicate, to a
certain degree, whether interfering substances are present.
4. Precision and Accuracy
4.1 Single laboratory accuracy and precision reported for this method
when analyzing five replicates of tap water spiked with 0.05 to
0.07 .ug/1 of PCP is as follows:
Recovery - mean 95.9%, range 88.1 to 100.2%
Standard Deviation - 6.0%
140
-------
REFERENCE:
1. "Analysis of Pentachlorophenol Residues in Soil, Water and Fish," Stark,
A., Agricultural and Food Chemistry, J7» 871 (July/August 1969).
141
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APPENDIX I
CONSIDERATIONS FOR GLASSWARE AND REAGENTS USED IN ORGANIC ANALYSIS*
1. Glassware
1.1 Cleaning Procedure - It is particularly important that glassware
used in trace organic analyses be scrupulously cleaned before
initial use as well as after each analysis. The glassware should
be cleaned as soon as possible after use, first rinsing with water
or the solvent that was last used in it. This should be followed
by washing with hot soap water, rinsing with tap water, distilled
water, redistilled acetone and finally with pesticide quality
hexane. Heavily contaminated glassware ,may require muffling at
e
400C for 15-to 30-minutes. High boiling materials, such as some of
the polychlorinated biphenyls (PCBs) may not be eliminated by such
heat treatment. NOTE: Volumetric ware should not be muffled. The
glassware should be stored immediately after drying to prevent
accumulation of dust or other contaminants. Store inverted or
cover mouth with foil.
1.2 Calibration - Individual Kuderna-Danish concentrator tubes and/or
centrifuge tubes used for final concentration of extracts must be
*Methods for Organic Pesticides in Water and Wastewater," 1971,
Environmental Protection Agency, National Environmental Research Center,
Cincinnati, Ohio, 45268
142
-------
accurately calibrated at the working volume. This is especially
important at volumes below 1 ml. Calibration should be made using
a precision microsyringe, recording the volume required to bring
the liquid level to the individual graduation marks. Glass A
volumetric ware should be used for preparing all standard solutions.
2. Standards, Reagents and Solvents
2.1 Analytical Standards and Other Chemicals - Analytical reference
grade standards should be used whenever available. They should be
stored according to the manufacturer's instructions. Standards and
reagents sensitive to light should be stored in dark bottles and/or
in a cool dark place. Those requiring refrigeration should be
allowed to come to room temperature before opening. Storing of
such standards under nitrogen is advisable.
2.1.1 Stock Standards - Pesticide stock standards solutions
should be prepared in 1 jjg/jjl concentrations by
dissolving 0.100-grams of the standard in pesticide-
quality hexane or other appropriate solvent (Acetone
should not be used since some pesticides degrade on
standing in this solvent) and diluting to volume in a 100
ml ground glass stoppered volumetric flask. The stock
solution is transferred to ground glass stoppered reagent
bottles. These standards should be checked frequently
for signs of degradation and concentration, especially
just prior to preparing working standards from them.
143
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2.1.2 Working Standards - Pesticide working standards are
prepared from the stock solutions using a micro syringe
preferably equipped with a Chaney adapter. The
concentration of the working standards will vary
depending on the detection system employed and the level
of pesticide in the samples to be analyzed. A typical
concentration (0.1 ng/jul) may be prepared by diluting 1
jul of the 1 jug/ul stock to volume in a 10-ml ground glass
stoppered volumetric flask. The standard solutions
should be transferred to ground glass stoppered reagent
bottles. Preparation of a fresh working standard each
day will minimize concentration through evaporation of
solvent. These standards should be stored in the same
manner as the stock solutions.
2.1.3 Identification of Reagents - All stock and working
standards should be labeled as follows: name of
compound, concentration, date prepared, solvent used, and
name of person who prepared it.
2.1.4 Anhydrous sodium sulfate used as a drying agent for
solvent extracts should be prewashed with the solvent or
solvents that it comes in contact with in order to remove
any interferences that may be present.
2.1.5 Glass wool used at the top of the sodium sulfate column
must be pre-extracted for about 40-hours in soxhlet using
the appropriate solvent.
144
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2.2 Solvents - Organic solvents must be of pesticide quality and
demonstrated to be free of interferences in a manner compatible
with whatever analytical operation is to be performed. Solvents
can be checked by analyzing a volume equivalent to that used in the
analysis and concentrated to the minimum final volume.
Interferences are noted in terms of gas chromatographic response -
relative retention time, peak geometry, peak intensity and width of
solvent response. Interferences noted under these conditions can
be considered maximum. If necessary, a solvent must be redistilled
in glass using a high efficiency distillation system. A 60-cm
column packed with 1/8 inch glass helices is effective.
2.2.1 Ethyl Ether - Hexane - It is particularly important that
these two solvents, used for extraction of organochlorine
pesticides from water, be checked for interferences just
prior to use. Ethyl ether, in particular, can produce
troublesome interferences. (NOTE: The formation of
peroxides in ethyl ether creates a potential explosion
hazard. Therefore it must be checked for peroxides
before use..) It is recommended that the solvents be
mixed just prior to use and only in the amount required
for immediate use since build-up of interferences often
occurs on standing.
2.2.2 The great sensitivity of the electron capture detector
requires that all solvents used for the analysis be of
pesticide quality. Even these solvents sometimes require
145
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redistillation in an all glass system prior to use. The
quality of the solvents may vary from lot to lot and even
within the same lot, so that each bottle of solvent must
be checked before use.
146
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APPENDIX II
STANDARDIZATION OF FLORISIL COLUMN BY WEIGHT ADJUSTMENT BASED ON
ADSORPTION OF LAURIC ACID
1. Scope
1.1 A rapid method for determining adsorptive capacity of
Florisil is based on adsorption of lauric acid from hexane
solution. An excess of lauric acid is used an'd amount not
adsorbed is measured by alkali titration. Weight of lauric
acid adsorbed is used to calculate, by simple proportion,
equivalent quantities of Florisil for batches having
different adsorptive capacities.
2. Apparatus
2.1 Buret — 25 ml with 1/10 ml graduations.
2.2 Erlenmeyer flasks — 125 ml narrow mouth and 25 ml, glass
stoppered.
2.3 Pipet — 10 and 20 ml transfer.
2.4 Volumetric flasks — 500 ml.
3. Reagents and Solvents
3.1 Alcohol, ethyl. -- USP or absolute, neutralized to
phenolphthalein.
3.2 Hexane -- Distilled from all glass apparatus.
3.3 Lauric acid -- Purified, CP.
147
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3.4 Laurie acid solution - Transfer 10.000 g lauric acid to 500
ml volumetric flask, dissolve in hexane, and dilute to 500 ml
(1 ml =20 mg).
3.5 Phenolphthalein Indicator - Dissolve 1 g in alcohol and
dilute to 100 ml.
3.6 Sodium hydroxide - Dissolve 20 g NaOH (pellets, reagent
grade) in water and dilute to 500 ml'(IN.)- Dilute 25 ml IN^
NaOH to 500 ml with water (0.05J^). Standardize as follows:
Weigh 100-200 mg lauric acid into 1250 ml Erlenmeyer flask.
Add 50 ml neutralized ethyl alcohol and 3 drops
phenolphthalein indicator; titrate to permanent end point.
Calculate mg lauric acid/ml 0.05 N NaOH (about 10 mg/ml).
4. Procedure
4.1 Transfer 2.000 g Florisil to 25 ml glass stoppered Erlenmeyer
flasks. Cover loosely with aluminum foil and heat overnight
at 130°C. Stopper, cool to room temperature, add 20.0 ml
lauric acid solution (400 mg), stopper, and shake
occasionally for 15 min. Let adsorbent settle and pipet 10.0
ml of supernatant into 125 ml Erlenmeyer flask. Avoid
inclusion of any Florisil.
4.2 Add 50-ml neutral alcohol and 3 drops indicator solution;
titrate with 0.05N; to a permanent end point.
5. Calculation of Lauric Acid Value and Adjustment of Column Weight
5.1 Calculate amount of lauric acid adsorbed on Florisil as
follows:
148
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Laurie Add value * mg lauric ac1d/g Florlsil » 200 - (ml
required for titration X mg lauric acid/ml 0.05IN NaOH).
5.2 To obtain an equivalent quantity of any batch of Florlsil,
divide 110 by lauric acid value for that batch and multiply
by 20 g. Verify proper elution of pesticides by 6.
6. Test for Proper Elution Pattern and Recovery of Pesticides
6.1 Prepare a test mixture containing aldrin, heptachlor epoxide,
p,p'-DDE, dieldrin, Parathion and malathion. Dieldrin and
Parathion should elute 1n the 15% eluate; all but a trace of
malathion in the 50% eluate and others in the 6% eluate.
7. References
7.1 "Pesticide Analytical Manual," U.S. Department of Health,
Education and Welfare, Food and Drug Administration,
Washington, D.C.
7.2 Mills, P.A., "Variation of Florisil Activity: Simple Method
for Measuring Adsorbent Capacity and Its Use in Standardizing
Florisil Columns," Journal of the Association of Official
Analytical Chemists. 51, 29 (1968).
149
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APPENDIX III
CHROMATOGRAPHIC CALIBRATION TECHNIQUE
Relative Calibration (Internal Standardization);
A relative calibration curve is prepared by simultaneously chromatographing
mixtures of the previously identified sample constituent and a reference
standard in known weight ratios and plotting the weight ratios against area
ratios. An accurately known amount of the reference material is then added
to the sample and the mixture chromatographed. The area ratios are
calculated and the weight ratio is read from the curve. Since the amount of
reference material added is known, the amount of the sample consitituent can
be calculated as follows:
Rw x Us.
micrograms/liter = ys
Rw = Weight ratio of component to standard
obtained from calibration curve
Ws = Weight of internal standard added to
sample in nanograms
Vs = Volume of sample in mi Hi liters
Using this method, injection volumes need not be accurately measured the
detector response need not remain constant since changes in response will
not alter the ratio. This method is preferred when the internal standard
meets the following conditions:
a) we 11-resolved from other peaks
b) elutes close to peaks of interest
150
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c) approximates concentration of unknown
d) structurally similar to unknown.
"Methods for Organic Pesticides in Water and Wastewater," U.S.
Environmental Protection Agency, National Environmental Research
Center, Cincinnati, Ohio 45268
151
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APPENDIX IV
PREPARATION OF DIAZOMETHANE IN ETHER
1. Scope
1.1 Diazomethane is prepared by reaction of Carbitol and Diazald in the
presence of KOH. Solutions of diazomethane decompose rapidly in
the presence of solid material such as copper powder, calcium
chloride, boiling stones, etc. These solid materials cause solid
polymethylene and nitrogen gas to form.
2. Apparatus
2.1 Distilling flask with condenser, 125 ml, long neck with dropping
funnel.
2.2 Erlenmeyer flasks - 500 ml and 125 ml.
2.3 Water bath.
3. Reagents and Solvents
3.1 Ether
3.2 Potassium hydroxide pellets.
3.3 Carbitol (diethylene glycol monoethyl ether).
3.4 Diazald in ether. Dissolve 21.5 g of Diazald in 140 ml ether.
4. Procedure
4.1 Use a we 11-ventilated hood and cork stoppers for all connections.
Fit a 125-ml long-neck distilling flask with a dropping funnel and
an efficient condenser set downward for distillation. Connect the
condenser to two receiving flasks in a series - a 500-ml Erlenmeyer
152
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followed by a 125-ml Erlenmeyer containing oO ml ether. The inlet
to the 125-ml Erlenmeyer should dip below the ether. Cool both
receivers to 0°C. As a water bath for the distilling flask, set
up a 2-liter beaker on a stirplate (hot plate and stirrerl,
maintaining temperature at 70°C.
4.2 Dissolve 6-g KOH in 10 ml water in the distilling- flask (no heat).
Ad 35 ml Carbitol (diethylene glycol monoethyl ether), stirring
bar, and another 10 ml ether. Connect the distilling flask to the
condenser and immerse distilling flask in water bath. By means of
the dropping funnel, add a solution of 21.5 g Diazald in 140 ml
ether over a period of 20 minutes. After distillation is
apparently complete, add another 20 ml ether and continue
distilling until distillate is colorless. Combine the contents of
the two receivers in a glass bottle (WITHOUT ground glass neck),
stopper with cork, and freeze overnight. Decant the diazomethane
from the ice crystals into a glass bottle, stopper with cork, and
store in freezer until ready for use. The final solution may be
stored up to six months without marked deterioration. The 21.5 g
of Diazald reacted in this manner produce about 3 g of Diazomethane.
5. Cautions
5.1 Diazomethane is very toxic. It can explode under certain
conditions. The following precautions should be observed.
5.1.1 Use only in we 11-ventilated hood.
5.1.2 Use safety screen.
5.1.3 Do not pipette solution of diazomethane by mouth.
153
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5.1.4 For pouring solutions of diazomethane, use of gloves is
optional.
5.1.5 Do not heat solutions at 100°C (EXPLOSIONS).
5.1.6 Store solutions of gas at low temperatures (freezer
compartment of explosion-proof refrigerators).
5.1.7 Avoid ground glass apparatus, glass stirrers and sleeve
bearings where grinding may occur (EXPLOSIONS).
5.1.8 Keep solutions away from alkali metals (EXPLOSIONS).
6. Reference
6.1 "Pesticide Analytical Manual," U.S. Department of Health, Education
and Welfare, Food and Drug Administration, Washington, D.C.
154
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BIBLIOGRAPHY
1. "Analysis of Pesticide Residues in Human and Environmental
Samples," U.S. Environmental Protection Agency, Perrine Primate
Research Laboratories, Perrine, Florida, 33157, 1971.
2. Mills, P.A., "Variation of Florisil Activity: Simple Method for
Measuring Adsorbent Capacity and Its Use in Standardizing Florisil
Columns," Journal of the Association of Official Analytical
Chemists. 51., 29 (1968).
3. Goerlitz, D.F. and Brown, E., "Methods for Analysis of Organic
Substances in Water," Techniques of Water Resources Investigations
of the United States Geological Survey, Book 5, Chapter A3, U.S.
Department of the Interior, Geological Survey, Washington, D.C.
20242, 1972, pp. 24-40.
4. Steere, N.V., editor, "Handbook of Laboratory Safety," Chemical
Rubber Company, 18901 Cranwood Parkway, Cleveland, Ohio, 44128,
1971, pp. 250-254.
3. Cochrane, W. P. and Wilson, B. P., "Electrolytic conductivity
detection of some nitrogen-containing herbicides," Journal of
Chromatography. 63_, 364 (1971).
4. "Standard Practice for Measuring Volatile Organic Matter in Water
by Aqueous - Injection Gas Chromatography," D2908 Annual Book of
ASTM Standards, Part 31, Water; American Society for Testing and
Materials, 1916 Race Street, Philadelphia, PA, 19103.
5. Gas-liquid Chromatographic Techniques for Petro Chemical
Wastewater Analysis, Sugar, J.W. and Conway, R.A., Journal of
WPCF. 40, (Annual Conference Issue) 1622 (1968).
6. "Handbook of Chemistry and Physics," 48th Edition, the Chemical
Rubber Company, 18901 Cranwood Parkway, Cleveland, Ohio, 44128.
(1967-1968).
155
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APPENDIX IV
GUIDELINES ESTABLISHING TEST PROCEDURES
FOR THE ANALYSIS OF POLLUTANTS;
PROPOSED REGULATIONS
-------
Monday
December 3, 1979
Part 111
Environmental
Protection Agency
Guidelines Establishing Test Procedures
for the Analysis of Pollutants; Proposed
Regulations
-------
69468
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
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Appendix I—-Gas Chromatographic and
HPLC Methods—Methods 601 through
612
Purgeable Halocarbon$—Method601
1. Scope and Application.
1.1 This method covers the
determination of 29 purgeable
halocarbons. The following parameters
may be determined by this method:
PvanwMr STORETNo.
32104
32101
34413
32102
34301
34311
34578
32108
34418
34105
34538
34566
34571
34688
34498
34531
34501
34S48
34541
34S6t
34561
34423
34518
34473
34508
34511
39180
34488
39175
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest. If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rathe_r than instrumental
limitations. The limits of detection listed
in Table 1 represent sensitivities that
can be achieved in wastewaters under
optimum operating conditions.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of. such qualified persons.
2. Summary of Method.
2.1 An inert gas is bubbled through a
5 ml water sample contained in a
specially-designed purging chamber.
The halocarbons are efficiently
transferred from the aqueous phase to
the vapor phase. The vapor is swept
through a short sorbent tube where the
halocarbons are trapped. After the purge
is completed, the trap is heated and
backflushed with gas to desorb the
halocarbons into a gas chromatographic
system. A temperature program is used
in the GC system to separate the
halocarbons before detection with a
halide-specific detector.
2.2 If interferences are encountered,
the method provides an optional gas
chromatographic column that may be
helpful in resolving the compounds of
interest from the interferences.
3. Interferences.
' 3.1 Impurities in the purge gas and
organic compounds out-gasing 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 method blanks.
Method blanks are run by charging the
purging device with organic-free water
and analyzing it in a normal manner.
The use of non-TFE plastic tubing, non-
TFE thread sealants, or flow controllers
with rubber components in the purging
device should be avoided.
3.2 Samples can be contaminated by
diffusion of volatile organics
(particularly freons and methylene
chloride) through the septum seal into
the sample during shipment and storage.
A sample blank prepared from organic-
free 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 level and low level
samples are sequentially analyzed. To
reduce the likelihood of this, the purging
device and sample syringe should be
rinsed out twice between samples with
organic-free water. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of organic-free water to check
-for cross contamination. For samples
containing large amounts of water-
soluble materials, suspended solids,
high boiling compounds or high
organohalide levels, it may be necessary
to wash out the purging device with a
soap solution, rinse with distilled water.
and then dry in a 105° C oven between
analyses.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
sampling.
4.1. Vial, with cap—40 ml capacity
screw cap (Pierce #13075 or equivalent).
Detergent wash and dry at 105° C before
use.
4.1.2 Septum—Teflon—faced
silicone (Pierce #12722 or equivalent).
Detergent wash, rinse with tap and
distilled water, and dry at 105"C for one
hour before use.
4.2 Purge and trap device—The
purge and trap equipment consists of
three separate pieces-of apparatus: the
purging device, trap, and desorber.
Several complete devices are now
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
available commercially. The device
must meet the following specifications:
The unit mint be completely compatible
with the gas chromatographic system;
the purging chamber must be designed
for a 5 ml volume and be modeled after
Figure 1; the dimensions for the sorbant
portion of the trap must meet or exceed
those in Figure 2. Figures 3 and 4
illustrate the complete system in the
purge and the desorb mode.
4.3 Gas chromatograph—Analytical
system complete with programmable gas
chromatograph suitable for on-column
injection and all required accessories
including halide-specific detector,
column supplies, recorder, and gases. A
data system for measuring peak areas is
recommended.
4.4 Syringes—5-ml glass hypodermic
with lueriok tip (2 each}.
4.5 Micro syringes—10, 25,100 /A!.
4.6 Z-way syringe valve with Luer
ends (3 each).
4.7 ' Syringe—5-ml gas-tight with
shut-off valve.
4.3 Bottle—15-mi screw-cap, with
Teflon cap liner.
5. Regents.
5.1 Sodium thiosulfate—(ACS)
Granular.
5.2 Trap Materials
5.2.1 Porus polymer packing 60/80
mesh chromatographic grade Tenax GC
(2.6-diphenylene oxide).
5.2.2 Three percent OV-l on
Chromosorb-W 60/80 mesh.
5.2.3. Silica gel—(35/80 mesh}—
Davison, grade-15 or equivalent
5.2.4 Coconut charcoal 6/10 mesh
Barnaby Chaney, CA-580-28 lot #• M-
2649 or equivalent
5.3 Activated carbon—Filtrasorb-
200 (Calgon Corp.) or equivalent
5.4 Organic-free water
5.4.1 Organic-free water is defined
as water free of interference when
employed in the purge and trap
procedure described herein. It is
generated by passing tap water through
a carbon filter bed containing about 1 Ib.
of
5.4.2 A water purfication system
(Millipore Super-Q or equivalent) may
be used to generate organic-free
deionized water.
5.4.3 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 one hour. While
still hot, transfer the water to a narrow
mouth screw cap bottle and seal with a
Teflon line septum and cap.
5.5 Stock standards—Prepare stock
standard solutions in methyl alcohol
using assayed liquids or gas cylinders 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 toxic gas respirator should be
used when the analyst handles high
concentrations of such materials.
5.5.1 Place about 9.8 ml of methyl
alcohol into a 10 ml ground glass
stoppered volumetric flask. Allow die
flask to stand, unstoppered for about 10
minutes or until all alcohol wetted
surfaces have dried. Weigh the flask to
the nearest 0.1 mg.
5.5.2 Add the assayed reference
material:
jui.2.1 Liquids—Using a 100 pi
syringe, immediately add 2 drops of
assayed reference material to the flask,
then reweigh. Be sure that the 2 drops
fall directly into the alcohol without
contacting the neck of the flask.
5.5.2.2 Gases—To prepare standards
for any of the six haiocarbons that boil
below 30* C (bromomethane,
chloroethane, chloromethane.
dichJorodifluoromethane,
trichlorodifluoromethane, 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 methyl alcohol menicus.
Slowly inject the reference standard
above the surface of the liquid (the
heavy gas will rapidly dissolve into the
methyl alcohol).
5.5.3 Reweigh, dilute to volume,
stopper, then mix by inverting the flask
several times. Transfer die standard
solution to a 15 ml screw-cap bottle with
a Teflon cap liner.
5.5.4 Calculate the concentration in
micrograms per microliter from the net
gain in weight
5.5.5 Store stock standards at 4* C
Prepare fresh standards weekly for the
six gases and 2-chloroethylvinyl ether.
All other standards must be replaced
with fresh standard each month.
8. Calibration.
8.1 Using stock standards, prepare
secondary dilution standards in methyl
alcohol that contain the compounds of
interest either singly or mixed together.
The standards shoud be prepared at
concentrations such that the aqueous
standards prepared in 6.2 will
completely bracket the working range of
the analytical system.
6.2 Using secondary dilution
standards, prepare calibration
standards by carefully adding 20.0 p.1 of
standard in methyl alcohol to 100, 500,
or 1000 ml of organic-free water. A 25 ul
syringe (Hamilton 702N or equivalent)
should be used for this operation. These
aqueous standards must be prepared
fresh daily.
6.3 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table 1.
By injecting secondary dilution
standards, establish, the sensitivity limit
and the linear range of the analytical
system for each compound..
8.4 Assemble die necessary purge
and trap device. The trap must meet the
minimum specifications as shown in
Figure 2 to achieve satisfactory results.
Condition the trap overnight at 180* C
by backflushing with an inert gas flow
of at least 20 ml/min. Prior to use, daily
condition, traps 10 minutes while
backflushing at 180* C Analyze aqueous
calibration standards (6,2) according to
the purge and trap procedure in Section
8. Compare the responses to those
obtained by injection of standards (8.3).
to determine purging efficiency and also
calculate analytical precision. The
purging efficiencies and analytical
precision of die analysis of aqueous
standards must be comparable to data
presented by Bellar and Lichtenberg
(1978) before reliable sample analysis
may begin.
6.5 By analyzing calibration
standards, establish the sensitivity limit
and linear range of the entire analytical
system for each compound.
7. Quality Control.
7.1 Before processing any samples,
the analyst should daily demonstrate
through the analysis of an organic-free
water method blank that the entire-
analytical system is interference-free.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the gas
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
7.3 The analyst should maintain
constant surveillance of 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
blank with surrogate haiocarbons. A
combination of bromochloromethane, 2-
bromo-1-chloropropane, and 1,4-
dichlorobutane is recommended to
encompass the boiling range covered by
this method. From stock standard
solutions prepared as above, add a
volume to give 1000 jxg of each surrogate
to 45 ml of organic-free water contained
in a 50-ml volumetric flask, mix and
dilute to volume (20 ng/ul). Dose 5.0 pi
of this surrogate spiking solution
-------
69470
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
directly Into the 5 ml syringe with every
sample and reference standard
analyzed. Prepare a fresh surrogate
spiking solution on a weekly basis.
8. Sampb Collection, Preservation,
and Handling.
8.1 Grab samples must be collected
in glass containers having a total
volume in excess of 40 ml. Fill the
sample bottles in such a manner that no
air bubbles pass through the sample as
the bottle is being filled. Seal the bottle
so that no air bubbles are entrapped in
it. Maintain the hermetic seal on the
sample bottle until time of analysis.
8.2. The samples must be iced or
refrigerated from the time of collection
until extraction. If the sample contains
free or combined chlorine, add sodium
thiosulfate preservative (10 mg/40 mi
will suffice for up to 5 ppm CU) to the
empty sample bottles just prior to
shipping to the sampling site, fill with
sample just to overflowing, seal the
bottle, and shake vigorously for 1
minute.
8.3 All samples must be analyzed
within 14 days of collection.
9. Sampb Extraction and Gca
Chromatograph,
9.1 Ad)ust the purge gas (nitrogen or
helium) flow rate to 40 ml/min. Attach
the trap Inlet to the purging device, and
set the device to purge. Open the syringe
valve located on the purging device
sample introduction needle.
9.2 Remove the plunger from a 5 ml
syringe and attach a closed syringe
valve. Open the sample bottle (or
standard) and carefully pour the water
into the syringe barrel until it overflows.
Replace-the syringe plunger and
compress the sample. Open the syringe
valve and vent any residual air while
adjusting the samples volume to 5.0 ml
Since this process of taking an aliquot
destroys the validity of the sample for
future analysis, the analyst should fill a
second syringe at this time to protect
against possible loss of data. Add 5.0 ul
of the surrogate spiking solution (7.3)
through the valve bore, then close the
valve.
9.3 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.
9.4 Close both valves and purge the
sample for 11.0 ± .05 minutes.
9.5 After the 11 minute purge time,
attach the trap to the chromatograph,
and adjust the device to the desorb
mode. Introduce the trapped materials to
the GC column by rapidly heating the
trap to 180'C while back-flushing the
trap with an inert gas between 20 and 60
ml/min for 4 minutes. If rapid heating
cannot be achieved, the gas
chromatographic column must be used
as a secondary trap by cooling it to 30* C
(or sub/ambient if problems persist)
instead of the initial program
temperature of 45*C
9.6 While the trap is being desorbed
into the gas chromatograph, empty the
purging chamber using the sample
introduction syringe. Wash the chamber
with two 5 ml flushes of organic-free
water.
9.7 After desorbing the sample for
approximately four minutes recondition
the trap by returning the purge and trap
device to the purge mode. Wait 15
seconds then dose the syringe valve on
the purging device to begin gas flow
through the trap. Maintain the trap
temperature at 180'C. After
approximately seven 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.
9.8 Table 1 summarizes some
recommended gas chromatographic
column material and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should bt>
achieved by-this method. An example of
the separation achieved by column 1 is
shown in Figure 5. Calibrate the system
daily by analysis of a minimum of three
concentration levels of calibration
standards.
10. Calculations.
10.1 Determine the concentration of
individual compounds directly from
calibrations plots of concentration (ju.g/1)
vs. peak height or area units.
10.2 Reports results in micrograras
per liter. When duplicate and spiked
samples are sadples are analyzed, all
data obtained should be reported.
11. Accuracy and Precision. The U.S.
EPA Environmental Monitoring and
Support Laboratory in Cincinnati is in
the process of conducting an inter-
laboratory method study to determine
the accuracy and precision of this test
procedure.
Bibliography
1. Bellar, T. A., and J. ]. Uchtenberg, journal
American Water Works Astociatioa Vol. 96.
No. 12. Dec. 1974, pp. 730-744.
2. Bellar, T. A., and J. J. Uchtenberg, "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds," Proceeding
from ASTM Symposium on Measurement of
Organic Pollutants in Water and Wastewater.
June 1978 (In Press).
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).
Tabto l—Orytnotmtidt* TMMrf Using Purgt tnd
(mln.)
0»t«rton
COI.1 « Col 2 »
CntOroflWlneWsj". ,„ _. «_. »n.» ™
OtehlorodMuoromMniM-
Vinyl cntond*
CnloroHnmi... ..»
Mothyton* oMorld*
Trientoranuoremitfwrai..
1,1 -Oiehleraetnino .._
1.M
tran»1;i
CMorotomfs.
1,1,1-TrlchloroolfMno.
Cirbontm
BromodlehloromMMM
TneMonMihi
Obron
1,1,2-TrtonlorocttMn*.....
Cl*' 1,34lohloraprop«M.
1,1,2^-T«t«ohtoro«N
TcmohloroMnini.._
l.S-Otei
1,44tonlerebonxirM
1.50
2.17
2.82
2.87
3.33
US
7.18
7.93
9.30
10.1
10.7
11.4
12.8
13.0
13.7
14.g
1J.2
ISJ
18.S
16.S
184
18.0
2l!s
21.7
24.2
34.0
34.9
3S.4
US
7.0S
(1
US
S.SS
10.1
(1
7.72
12.8
9.38
12.1
1S.4
13.1
14.4
14.8
18.8
18.8
13.1
16.8
18,1
18.0
(1
19.2
(1
18.0
18.9
22.4
23J
22.3
0.0009
0.03
0.03
0.01
0.01
0.01
0.008
0.004
0.008
0.008
0.008
O.OOS
0.007
0.008
0.004
0.008
0.008
0.01
0.008
0.008
0.08
0.02
0.008
0.007
0.03
0.04
044
0.04
limit ii eueuand (ram ttw minimum
QC ruponM Mng *qu* to flv« am* tt» QC HwKgreuM
n«M. wing • Hi* Mod* 700-A OoMOler.
'CtfbopMk. S-80/80 m*h OMMd wttfl 1* SP-tOOO
pMlud In in 8 tt x 0.1 in ID iMnMw MM or glMt column
with nHhm earn* gw tl 40 ml/mm flow nm. Column two-
ptritur* h*« *! tt'C tor 3 irtn. thin programmed II S'C/
mm. to 220* thm nMd tor 18 irtn.
•PorW-C 100/120 mud ooeMd wnh nooun* peekod m i
8 (I x 0.1 In 10 MMM8 Meet or glaw ootumn wttti heUom
oirrlor QH M 40 ml/mtn now tut. Ootumn t«nnp«nUuri n*W
II WC (or 3 mln ttwn pnjgrimm»a it e'C/mto to 170* lhw>
hold (or 4 mtn.
•Not doMrmlnid
SltUrM COM SSCB-ei-M
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3,1979 / Proposed Rules
68471
OPTIONAL
FOAM
TRAP
•EXIT '/• IN.
0. 0.
—14MM 0. 0.
INLET '/4 IN.
0. 0.
'/« IN.
0. 0. EXIT
INLET
3—2-WAY SYRINGE VALVE
j[——17CM. 20 GAUGE SYRINGE NEEDLE
. 0. D. RUBBER SEPTUM
1/16 IN. O.D.
\SSTAINLESS STEEL
13X MOLECULAR
SIEVE PURGE
RLTER
PURGE GAS
FLOW
CONTROL
10MM GLASS FRIT
MEDIUM POROSITY
Figure 1. Purging device
PACKING PROCEDURE
CONSTRUCTION
„ 7 A/FOOT
2 RESISTANCE
ACTIVATED I ^ W1RE wRAppE3
CHARCOAL7.7ai^i SOLJD<
CHARCOAL p (DOUBLE LAYER)
COMPRESSION
GRADE 15
SILICA
t
7.7CT'
TEN AX 7.7 Oft \
3?'- OV-1 j p-j
GLASS WOOL100 * -
15CM
7-^/FOOT.,
RESISTANCE
WIRE WRAPPED
SOUD
(SINGLE LAYER)
8 CM-
^
/-
^^.
—.
c
c;
. <.
_J
•—
^*
r^-
— •
•M>
--
»«
^__<
•)
4
7
k
:>
•••ii
;j
Mill
AND
niu IMU i
FERRULES
THERMOCOUPLE/
CONTROLLER
SENSOR
ELECTRONIC
JTEMPERATURE
< ^AND
^^
PYROMETER
TUBING 25CM
0.105 IN. I.D.
0.125 IN. O.D.
STAINLESS STEEL
'i
•»'
Figure 2. Trap packings and construction to include
desorb capability
-------
69472
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
REGULATOR
PURGE GAS ,
FLOW CONTROL ,L
13X MOLECULAR
SIEVE FILTER &
COLUMN OVEN
1 CONFIRMATORY COLUMN
ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN.
SRECTION VALVE
e-PORT TRAP INI ET
VALVE J RESISTANCE WIRE
PURGING
DEVICE
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 80°C.
Figure 3. Schematic of purge and trap device - purge mode
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
REGULATOR
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
COLUMN OVEN
CONFIRMATORY COLUMN
DETECTOR
ANALYTICAL COLUMN
^ OPTIONAL 4-PORT COLUMN
SELECTION VALVE
S-PORT TRAP INLET
VALVE j RESISTANCE WIRE HEATgH
"jHApji" """" ' CONTROL
PURGING
DEVICE
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD SE HEATED
TO 80° C.
Figure 4. Schematic of purge and trap device • desorb mode
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3, 1979 / Proposed Rules
89473
3N3ZN3aoa01HOIQ-fr 'I
< «o
«£j
i|<
o So
12°
MS • <
3§S
O gtu
^ &M O
3^ez^aaoao^HO
3NVH130a01H3Vai31-2 7 'I 'L
•^••M
3NVdoadoao"iHOiai-t: 7 'L
^NVHI30a01HOVai31-2'l' L' L
a
*~
2
^
"a
a
3
3)
o
(Q
o
at
in
a
-------
69474 Federal Register / Vol. 44, No. 233 / Monday. December 3, 1979 / Proposed Rules
Purgeable Afomatics—Method 602
1. Scope and Application.
1.1 This method covers the
determination of various purgeable
aromatics. The following parameters
may be determined by this method:
Straw,.
obi
u-oia
1,3-Ofcl
Town*
34030
34301
34S36
34S86
3*571
34371
34010
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table 1 represent sensitivities that
can be achieved in wastewaters under
optimum operating conditions.
1.4 This method is recommended for
use only by experienced residue
analysts or under the dose supervision
of such qualified persons.
2. Summary of Method.
2.1 An inert gas is bubbled through a
5 ml water sample contained in a
specially-designed purging chamber.
The aromatics are efficiently transferred
from the aqueous phase to the vapor
phase. The vapor is swept through a
short sorbent tube where the aromatics
are trapped. After the purge is
completed, the trap is heated and
backflushed with gas to desorb the
aromatic compounds into a gas
chromatographic system. A temperature
program is used in the GC system to
separate the aromatics before'detection
with a photoionization detector.
3. Interferences.
3.1 Impurities in the purge gas and
organic compounds out-gasing from the
plumbing ahead of the trap account for
the majority of contamination problems.
The analytical system must be
demonstrated to be free from
interferences under the conditions of the
analysis by running method blanks.
Method blanks are run by charging the
purging device with organic-free water
and analyzing it in a normal manner.
The use of non-TFE plastic tubing, non-
TFE thread sealants or How controllers
with rubber components in the purging
device should be avoided.
3.2 Samples can be contaminated by
diffusion of volatile organics through the
septum seal into the sample during
shipment and storage. A sample blank
prepared from organic free 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 level and low level
samples are sequentially analyzed. To
reduce the Likelihood of this, the purging
device and sample syringe should be
rinsed out twice between samples with
organic-free water. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of organic-free water to check
for cross contamination. For samples
containing large amounts of water
soluble materials, suspended solids,
high boiling compounds, or high levels of
aromatics, it may be necessary to wash
out the purging device with a soap
solution, rinse with distilled water, and
then dry in a 105* C oven between
analyses.
* 4. Apparatus and Materials.
4.1 Sampling equipment for discrete
sampling.
4.1.1 Vial, with cap—40 ml capacity
screw cap (Pierce #13075 or equivalent).
Detergent wash and dry at 105* C before
use,
4.1.2 Septum-Teflon-faced silicons
(Pierce §12722 or equivalent). Detergent
wash, rinse, with tap and distilled
water, and dry at 105* C for one hour
before use.
4.2 Purge and trap device—The
purge and trap equipment consists of-
three separate pieces of apparatus: the
purging device, trap, and desorber.
Several complete devices are available
commercially. The device must meet the
following specifications: The unit must
be completely compatible with the gas
chromatograhpic system: the purging
chamber must be designed for a 5 ml
volume and be modeled after Figure 1;
the dimensions for the sorbant portion
of the trap most meet or exceed those in
Figure 2. Figures 3 and 4 illustrate the
complete system in the purge and the
desorb mode.
4.3 Gas chromatograph—Analytical
system complete with programmable gas
chromatograph suitable for on-column
injection and all required accessories
including Model PI-51-02
photoionization detector fh-nu Systems,
Inc.), column supplies, recorder, and
gases. A data system for measuring
peak areas is recommended
4.4 Syringes—5-ml glass hyodennic
with luerlok tip (2 each).
4.5 Micro syringes—10. 25,100 jil
4.8 2-way syringe value with Luer
ends (3 each).
4.7 Bottle—15-ml screw-cap, with
Teflon cap liner.
5. Reagents.
5.1 Solium thiosulfate—(ACS)
Granular.
5.2 Trap Materials
5.2.1 Porous polymer packing 60/80
mesh chromatographic grade Tenax GC
(2,8-diphenylene oxide).
5.2.2 Three percent OV-1 on
Chromosorb-W 80/80 mesh.
5.3 Activated carbon—Filtrasorb-200
(Calgon Corp.) or, equivalent
5.4 Organic-free water
5.4.1 Organic-free water is defined
as water free of interference when
employed in die purge and trap
procedure described herein. It is
generated by passing tap water through
a carbon filter bed containing about 1 Ib.
of activated carbon.
5.4.2 A water purification system
(millipore Super-Q or equivalent) may
be used to generate organic-free
deioniaed water.
5.4.3 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 one hour. While
still hot, transfer the water to a narrow
mouth screw cap bottle and seal with a
Teflon lined septum and cap.
5.5 Stock standards—Prepare stock
standard solutions in methyl alcohol
using assayed liquids. Because benzene
an 1,4-dichlorobenzene are suspected
carcinogens, primary dilutions of these
compounds should be prepared in a
hood.
5.5.1 Place about 9.8 ml of methyl
alcoftol into a 10 ml 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 0.1 mg.
5.5.2 Using a 100 fii syringe,
immediately add 2^drops of assayed
reference material to the flask, then
reweigh. Be sure that the 2 drops fall
directly into the alcohol without
contacting the neck of the flask.
5.5.3 Dilute to volume, stopper, then
mix by inverting the flask several times.
Transfer the standard solution to a 15 ml
screw-cap bottle with a Teflon cap liner.
5.5.4 Calculate the concentration in
mircograms per microliter from the net
gain in weight
5.5.5 Store stock standards at 4*C.
All standards must be replaced with
fresh standard each month.
8. Calibration.
6.1 Using stock standards, prepare
secondary dilution standards in methyl
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules 69475
alcohol that contain the compounds of
interest either singly or mixed together.
The standards should be prepared at
concentrations such that the aqueous
standards prepared in 0.2 will
completely bracket the working range of
the analytical system.
6.2 Using secondary dilution
standards, prepare calibration
standards by carefully adding 20.0 (d of
standard in methyl alcohol to 100, 500,
or 1000 ml of organic-free water. A 25 jil
syringe (Hamilton 702N or equivalent)
should be used for this operation. These
aqueous standards must be prepared
fresh daily.
8.3 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table 1.
By injecting secondary dilution
standards, establish the sensitivity limit
and the linear range of the analytical
system for each compound.
6.4 Assemble th necessary purge and
trap device. The Trap must meet the
minimum specifications shown hi Figure
2 to achieve satisfactory results.
Condition the trap overnight at ISO'C by
backflushing with an inert gas flow of at
least 20 ml/min. Prior to use, daily
condition traps 10 minutes while
backflushing at 180'C. Analyze aqueous
calibration standards (8.2) according to
the purge and trap procedure in Section
8. Compare the responses to those
obtained by injection of standards (6.3),
to determine purging efficiency and also
to calculate analytical precision. The
purging efficiencies and analytical
precision of the analysis of aqueous
standards must be comparable to data
presented by Bellar and Uchtenberg
(1978) before reliable sample analysis
may begin.
6.S By analyzing calibration
standards, establish the sensitivity limit
and linear range of the entire analytical
system for each compound.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate daily
through the analysis of an organic-free
water method blank that the entire
analytical system is interference-free.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the gas
chromatogram, confirmatory techniques
such a» mass spectroscopy should be
used.
74 The analyst should maintain
constant surveillance of 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
blank with surrogate compounds (e.g.
aact-trifluorotoluene).
8. Sample Collection, Preservation,
and Handling.
8.1 Collect about 500 ml sample in a
clean container. Adjust the pH of the
sample to about 2 by adding 1:1 diluted
HC1 while stirring vigorously. If the
sample contains free or combined
chlorine, add 35 mg of sodium
thiosulfate per part per million of free
chlorine per liter of sample. Fill a 40 ml
sample bottle in such a manner that no
air bubbles pass through the sample as
the bottle is being filled. Seal the bottle
so that no air bubbles are entrapped in
it Maintain the hermetic seal on the
sample bottle until time of analysis.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction.
8.3 All samples must be analyzed
within 7 days of collection.
9. Sample Extraction and Gas »
Chromatography.
9.1 Adjust the purge gas (nitrogen or
helium) flow rate to 40 ml/min. Attach
the trap inlet to the purging device, and
set the device to purge. Open the syringe
valve located on the purging device
sample introduction needle.
9.2 Remove the plunger from a 5 ml
syringe and attach a dosed syringe
valve. Open the sample bottle (or
standard) and carefully pour the water
into the syringe barrel until it overflows.
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.
Since this process of taking an aliquot
destroys the validity of the sample for
future analysis, the analyst should fill a
second syringe at this time to protect
against possible loss of data. Add the
surrogate spiking solution (7.3) through
the valve bore, then close the valve.
9.3 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.
9.4 Close both valves and purge the
sample for 12.0 ± .05 minutes.
9.5 After the 12 minute purge time.
disconnect the purge chamber from the
trap. Dry the trap by maintaining a flow
rate of 40 cc/min dry purge gas for 6
min. Attach the trap to the
chromatograph, and adjust the device to
the desorb mode. Introduce the trapped
materials to the GC column by rapidly
heating the trap to 180'C while
backflushing die trap with an inert gas
between 20 and 60 ml/min for 4 minutes.
If rapid heating cannot be achieved, 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 initial program
temperature of 50'C.
9.6 While the trap is being desorbed
into the gas chromatograph, empty the
purging chamber using the sample
introduction syringe. Wash the chamber
with two 5 ml flushes of organic-fine
water.
9.7 After desorbing the sample for
approximately four 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. Maintain the trap
temperature at 180'C. After
approximately seven 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.
9.8 Table 1 summarized the
recommended gas chromatographic
column material and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method. An example of
the separation achieved by this column
is shown in Figure 5. Calibrate the
system daily by analysis of a minimi^
of three concentration levels of
calibration standards.
10. Calculations.
10.1 Determine the concentration of
individual compounds directly from
calibrations plots of concentration (/xg/1)
vs. peak height or area units.
10.2 Report results in micrograms per
liter. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
11. Accuracy and Precision. The U.S.
EPA Environmental Monitoring and
Support Laboratory in Cincinnati is in
the process of conducting an
interlaboratory method study to
determine the accuracy and precision of
this test procedure.
Bibliography
1. Bellar, T. A., and J. I. Uchtenberg. Journal
American Water Works Association, VoL 86,
No. 12, Dec. 1974, pp. 739-744.
2. Bellar, T. A., and J. J. Uchtenberg, "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds," Proceeding
from ASTM Symposium on Measurement of
Organic Pollutants in Water and Wastewater.
June 1978 (In Press).
3. Bellar. T. A., and J. J. Uchtenberg, "The
Determination of Purgeable Aromatic
-------
69478 Federal Register / VoL 44. No. 233 / Monday, December 3, 1979 / Proposed Rules
Compounds in Drinking Waters and
Industrial Wattes," (In preparation).
4. "Development and Application of Test
Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 11—
Purgeabies and Category 12—Acrolein,
Acrylonitnle", and Dicnlorodifluoromethane."
Report for EPA Contract 68-03-2635 (In
preparation).
Table l.—Cttromatognpfiy ofAromsUcs Using
Purg* arxi Trap Mfthod
GofnpouAQ
3f«tnt .»«... ,.,.—..
ToHitw* « —
Ethyl tMnzen*
Retention
«m»(min.)
Cot i '
..._. ... 3 33
5.75
.................. 8.2S
9 17
OMcttan
limit
rt/'1
<^
O
(1
(t
«
(I
«
'•Suptteoport 100/120 nwin eo*Md wild 5% SP-Z100 and
1.75% B*nton*-34 pvlwd in a < ft x 0.085 m 10 stanta*
SIM) eekmn wtti nMum can** g»* at 38 ee/mn (tow rat*.
Column Mmpcmura n*U at WC (or 2 mm. -
grafflnwd at 6'C/fnin. 10 90*C for * Hn« hoM.
:OMwt»n IMt is eatauMMd from ttw mnimwn iMMctaM*
OC iMponw P*mg tqutl to DM Mm tM QC backgraund
noM. using a h-nu MocM W-51-02 photonnxatton oanetor
wftna 10.2 av lamp;
'NolotMnnnad.
8IUINO CODE ISSO-01-M
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3, 1979 / Propoaed Rules
69477
[OPTIONAL
1FOAM
TRAP
'/4 IN.
0. 0. EXIT
-EXIT 1/« IN.
0. D.
-—14MM 0. D.
INLET % IN.
0.0.
< /
I I
I I
-SAMPLE INLET
•2-WAlf SYRINGE VALVE
-17QL 20 GAUGE SYRINGE NEEDLE
.0.0. RUBBER SEPTUM
1/16 IN. O.D.-
\ySTAINLESSSTEEL
13* MOLECULAR
SIEVE PURGE
GAS FILTER
PURGE GAS
FLOW
CONTROL
10MM GLASS FRIT
MEDIUM POROSm
Figure 1. Purging device
PACKING PROCEDURE
CONSTRUCTION
WOOL
TENAX 23CM
3T. OV-1
GLASS WOOL
101)
5UU
TRAP INLET
COMPRESSION FITTING
NUT AND FERRULES
14FT.7A/FOOT RESISTANCE
WIRE WRAPPED SOUO
THERMOCOUPLE/
CONTROLLER
SENSOR
! TUBING 25QI.
-0.105 IN. I.D.
0.125 IN. 0.0.
STAINLESS STEEL
Rgure 2. Trap packings and construction to include
desorb capability
-------
69478
Federal Register / Vol. 44. No. 233 / Monday, December 3,1979 / Proposed Rules
CARRIER GAS
ROW CONTROL LIQUID INJECTION PORTS
PRESSURE
REGULATOR
PURGE GAS
.
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER"
COLUMN OVEN
CONFIRMATORY COLUMN
^ T° DETECTOR
—ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SELECTION VAIVE
6-PORT TRAP INLET
VALVE J RESISTANCE WIRE
PURGING
DEVICE
HEATER
CONTROL
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 80° C.
Figure 3. Schematic of purge and trap device - purge mode
CARRIER GAS
FLOW CONTROL
PRESSORE
REGULATOR.
PURGE GAS
FLOW
13X MOLECULAR^
SIEVE FILTER &
UQU10 INJECTION PORTS
COLUMN OVEN
CONFIRMATORT COLUMN
-ANALYTICAL COLUMN
^OPTIONAL 4^>OHT COLUMN
SELECTION VALVE
6-PORT TRAP INLET
VALVE J RESISTANCE WIRE
TRAP ( QN >
180«C ^°N'
HEATER
CONTROL
PURGING
DEVICE
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 80° C.
Figure 4. Schematic of purge and trap device - desorb mode
WLUNO COM «MO-«1-C
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules 69479
UJ UJ
2 2
Ud UJ
2 2
SUJ
uj a
uj 2 £ O
9 uj 2 es
§3 ~3
^3 s» ty ••
^2 S3
I
o
1
s
T
Q
S
e'
1
1
UJ
;
^^Jw^w^ ->.
UJ
UJ
fsi
S
a
0
s
o
3
5
.
i
J
UJ
2
UJ
UJ
a
g
O
s.
U
5
r>
UJ
UJ
2
UJ
a
i
o
_J
u
o
A . .
2 4 6 8 10 12 14 16 18 20 22 24 25 28
RETENTION TIME-MINUTES
Figure 5. Gas chromatogram of purgeable aromatics
Acrolein and Acrylonitrile—Method 803
1. Scope and Application.
1.1 This method covers the
determination of acrolein and
acrylonitrile. The following parameters
may be determined by this method:
Parameter Slant No.
Acrolan - 3*210
AcrykxWnl. - - 32415
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest. If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table 1 represent sensitivities that
can be achieved in wastewaters under
optimum operating conditions.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons.
2. Summary of Method.
2.1 An inert gas is bubbled through a
5 ml water sample contained in a
specially-designed heated purging
chamber. Acrolein and acrylonitrile are
transferred from the aqueous phase to
the vapor phase. The vapor is passed
through a short sorbent rube where the
compounds are trapped. After the
extraction is completed, the trap is
heated and backflushed with gas to
desorb the compounds into a gas
chromatographic system. A temperature
program is used in the GC system to
separate the compounds before
detection with a flame ionization
detector.
3. Interferences.
3.1 Impurities in the purge gas and
organic compounds out-gasing from the
plumbing ahead of the trap account for
the majority of contamination problems.
The analytical system must be
demonstrated to be free from
interferences under the conditions of the
analysis by running method blanks.
Method blanks are run by charging the
purging device with organic-free water
and analyzing it in a normal manner.
The use of non-TFE plastic tubing, non-
TFE thread sealants, or flow controllers
with rubber components in the purging
device should be avoided.
-------
69480 Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
3.2 Samples can be contaminated by
diffusion of volatile organics
(particularly methylene chloride)
through the septum seal into the sample
during shipment and storage. A sample
blank prepared from organic-free 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 level and low level
samples are sequentially analyzed. To
reduce the likelihood of this, the purging
device and sample syringe should be
rinsed out twice between samples with
organic-free water. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of organic-free water to check
for cross-contamination. For samples
containing large amounts of water
soluble materials, suspended solids,
high boiling compounds or high
organohalide levels it may be necessary
to wash out the purging device with a
soap solution, rinse with distilled water,
and then dry in a 105* C oven between
analyses.
3.4 Interferences are sometimes
reduced or eliminated by first purging
the water samples for 5 minutes at room
temperature in 9.4. Then the purge
device is rapidly heated to 85* C and
purged as in 9.4. With such a
modification, approximately 5 to 10% of
the acrylonitrile and a trace of the
acrolein in the sample will be lost
Therefore, calibration must be
established for the compounds under the
conditions of this modified procedure.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
sampling.
4.1.1 Vial, with cap—40 ml capacity
screw cap (Pierce #13075 or equivalent).
Detergent wash and dry at 105* C before -
use.
4.1.2 Septum-Teflon-faced silicons
(Pierce #12722 or equivalent). Detergent
wash, rinse with tap and distilled water,
and dry at 105* C for one hour before
use.
4.2 Purge and trap device—The
purge and trap equipment consists of
three separate pieces of apparatus: the
purging device, trap, and desorber. The
purging device should be equipped for
heating in the same manner as the trap
(electrically) or with a circulating water
jacket. If electrical heating is used the
electrical parts must be protected so
that water will not drip on the
conductors, causing dangerous electrical
shock or shorts. All temperature
parameters must be carefully controlled.
Several complete devices are available
commercially although most are not
equipped to heat the purging chamber.
The device must meet the following
specifications: die unit must be
completely compatible with'the gas
chromatographic system the purging
chamber must be designed for a 5 ml
volume and be modeled after Figure 1;
the dimensions for the sorbant portion
of the trap must meet or exceed those in
figure 2. Figures 3 and 4 illustrate the
complete system in die purge and the
desorb mode.
4.3 Gas chromatograph—Analytical
system complete with programmable gas
chromatograph suitable for on-column
injection, equipped with matched
columns for dual column analysis and a
differential flame ionization detector. A
nitrogen specific detector (thermionic or
Hall) may be used if only acrylonitrile is
to be detected. Required accessories
include: column supplies, recorder, and
gases. A data system for measuring
peak areas is recommended.
4.4 Syringes—5-ml glass hypodermic
with luerlok tip (2 each).
4.5 Micro syringes—10, 25,100 uL
4.6 2-way syringe valve with Luer
ends (3 each).
4.7 Bottle—15-tnl screw-cap, with
Teflon cap liner.
5. Reagents.
5.1 Preservatives
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2. Sulfuric acid—(ACS). Mix
equal volumes of cone. H*SO4 with
distilled water.
. 5.1.3 Sodium thiosulfate—{ACS)
Granular.
5.2 Trap absorbent—Porous polymer
packing, 50/80 mesh chromatographic
grade Porapak N.
5.3 Activated carbon—Filtrasorb-200
(Calgon Corp.) or equivalent
5.4 Organic-free water.
5.4.1 Organic-free water is denned
as water free of interference when
employed in the purge and trap
procedure described herein. It is
generated by passing tap water through
a carbon filter bed containing about 1 Ib.
of activated carbon.
5.4.2 A water purification system
(Millipore Super-Q or equivalent) may
be used to generate organic-free
deionized water.
5.4.3 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 one hour. While
still hot transfer the water to a narrow
mouth screw cap bottle and seal with a
Teflon lined septum and cap.
5.5 Stock standards—Prepare stock
standard solutions daily in water using
assayed standards. Because of toxicity,
primary dilutions of these materials
should be prepared in a hood. A
NIOSH/MESA approved toxic gas
respirator should be used when the
analyst handles high concentrations of
the materials.
5.5.1 Place about 9.8 ml of water (pH
8.5 to 7.5) into a 10 ml ground glass
stoppered volumetric flask. Allow the
flask to stand, unstoppered, for about 10
minutes or until all water wetted
surfaces have dried. Weigh the flask to
the nearest 0.1 mg.
5.5.2 Using a 100 ul syringe,
immediately add 2 drops of assayed
reference material to the flask, then
reweigh. Be sure that the 2 drops fall
directly into the water without
contacting the neck of the flask
5.5.3 Dilute to volunle, stopper, then
mix by inverting the flask several times.
Transfer the standard solution to a 15 ml
screw-cap bottle with a Teflon cap liner.
5.5.4 Calculate the concentration in
micrograms per microliter from the net
gain in weight
6. Calibration.
6.1 Using stock standards, prepare
secondary dilution standards in water.
The standards should be prepared at
concentrations such that the aqueous
standards prepared in 6.2 will
completely bracket the working range of
the chromatographic system.
6.2 Using secondary dilution
standards, prepare calibration
standards by carefully adding 20 ul of
stock standard to 100, 500, or 1000 ml of
organic-free water.
6.3 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated hi Table 1.
By injecting secondary dilution
standards, establish the sensitivity limit
and the linear range of the analytical
system for each compound.
6.4 Assemble the necessary purge
and trap device. The trap must meet the
minimum specifications as shown in
Figure 2 to achieve satisfactory results.
Condition the trap overnight at 180* C
by backflushing with an inert gas flow
of at least 20 ml/min. Prior to use, daily
condition traps 10 minutes while
backflushing at 180* C. Analyze aqueous
calibration standards (6.2) according to
the purge and trap procedure in Section
9. Compare the responses to those
obtained by injection of standards (6.3),
to determine purging efficiency and also
to calculate analytical precision. The
purging efficiencies and analytical
precision of the analysis of aqueous
standards should be 85±5% for acrolein
and 98±5% for acrylonitrile.
6.5 By analyzing calibration
standards, establish the sensitivity limit
and linear range of the entire analytical
system for each compound.
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate daily
through the analysis of an organic-free
water method blank that the entire
analytical system is interference-free.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the gas
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
7.3 The analyst should maintain
-constant surveillance of 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
blank with surrogate compounds.
8. Sample Collection, Preservation,
and Handling.
8.1 Collect about 500 ml sample in a
clean container. Adjust the pH of the
sample to 6.5 to 7.5 by adding 1:1 diluted
HaSO« or NaOH while stirring
vigorously. If the sample contains
residual chlorine, add 35 mg of sodium
thiosulfate per part per million of free
chlorine per liter of sample. Fill a 40 ml
sample bottle and seal the bottle so that
no air bubbles are entrapped in it.
Maintain the hermetic seal on the
sample bottle until time of analysis.
8.2 The samples must be iced or
refrigerated at 4*C from the time of
collection until extraction.
8.3 All samples must be analyzed
within 3 days of collection.
9. Sample Extraction and Gas
Chromatography.
9.1 Adjust the helium purge gas flow
rate to 20±1 ml/min and the
temperature of the purge device to 85*C.
Attach, the trap inlet to the purging
device, and set the device to purge.
Open the syringe valve located on the
purging device sample introduction
needle.
9.2 Remove the plunger from a 5 ml
syringe and attach a closed syringe
valve. Open the sample bottle (or
standard] and carefully pour the water
into the syringe barrel until it oveflows.
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.
9.3 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.
9.4 Close both valves and purge the
sample for 30.0±0.1 minutes. Monitor
and control the temperature of the purge
device to obtain 85±1*C,
9.5 After the 30-minute purge time,
attach the trap to the chromatograph.
and adjust the device to the desorb
mode. Introduce the trapped materials to
the GC column by rapidly hearing the
trap to 170'C while backflushing the trap
with helium at 45 ml/min for 5 minutes.
The backflushing time and gas flow rate
must be carefully reproduced from
sample to sample. During blackflushing
the chromatographic column is held at
100'C. Record GC retention time from
the beginning of desorption.
9.6 While the trap is being desorbed
into the gas chromatograph, empty the
purging chamber using the sample
introduction syringe. Wash the chamber
with two 5 ml flushes of organic-free
water.
9.7 After desorbing the sample for 5
minutes recondition the trap by
returning the purge and trap device to
the purge mode and begin the GC
progrm. Wait 15 seconds then close the
syringe valve on the purging device to
begin gas flow through the trap.
Maintain the trap temperature at 170° C.
After approximately seven 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.
9.8 Table 1 summarizes some
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method. An example of
the separation achieved by this column
is shown hi Figure 5. Calibrate the
system daily by analysis of a minimum
of three concentrations levels of
calibration standards.
10. Calculations.
10.1 Determine the concentration of
individual compounds directly from
calibrations plots of concentration (ug/1)
vs. peak height or area units.
10.2 Report results in micrograms per
liter. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
11. Accuracy.and precision
The U.S. EPA Environmental
Monitoring and Support Laboratory in
Cincinnati is in the process of
conducting an interlaboratory method
study to determine the accuracy and
precision of this test procedure.
Bibliography
1. Bellar. TA., and}.}. Lichtenbeig. Journal
American Water Works Association. VoL 66,
No. 12, Dec. 1974. pp. 739-744.
2. Beilar. TA» and J.J. Uchtenberg, "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Pnrgeablv
Volatile Organic Compound*," Proceeding
from ASTM Symposium on Measurement of
Organic pollutants in Water and wastewater,
June 1978 (In Press).
3. "Development and application of Test
Procedures for Specific Organic Toxic
Substance* in Wastewaters. Category 11-
Purgeables and Category 12-Acrolein.
Acrylonitrile, and Dichiorodifluoromethane."
Report for EPA Contract 68-03-2633 (In
preparation).
4. Going, John, et al.. "Environmental
Monitoring Near Industrial Sites-
Acrylonitrile," EPA Report No. 560/6-79-003,
1979.
Table 1.—GasChromatogmpfiytjy Heated Puiy»
ana Trap
Retention Oatecabn
Compound ' Time (iMn.) Urn* uc/4 '
Acfotem
Acrytonitnle
7.8
8.9
'Column condition* • Chnxnoaorb 101 80/100
packed m a 8" x tt" 0.0. start*** Heel cotumn vyitfi heftum
earner 9a» at 46 ml/mm flow rate. Column temperature it
h«w at 100* C (or J minuMt during traD-deaorptton. then pro-
grammed at 10 • C/mn to 140 *C and held for S minute*.
'Detection limit ts «*kna«d. baaed upon «§ uee of r
(lam* wnizatton detector.
BILLING
-------
69482
Federal Register/Vol. 44, No. 233 / Monday. December 3,1979 / Proposed Rules
OPTIONAL
FOAM
TRAP
',4 IN.
0. D. EXIT
2-WAY SYRINGE VALVE
17CH. 20 GAUGE SYRINGE NEEDLE
. 0. D. RUBBER SEPTUM
ins IN. o.o.
\SSTAINLESS STEEL
13X MOLECULAR
SIEVE PURGE
GAS RLTEH
PURGE GAS
FLOW
CONTROL
10MM.GLASS FRIT
MEDIUM POROSITY
Figure 1. Purging device
PACKING PROCEDURE
WOOL
PORAPAK N
24CU
GLASS WOOL I
5MM
CONSTRUCTION
COMPRESSION
.FITTING NUT
AND FERRULES
14FT. 7A/PQOT
'RESISTANCE WIRE
WRAPPED SOLJD
TriESKOCOUPLE/
? CONTROLLER
C'T-T""" SENSOR
TRAP INLET
TUSING 25OB
0.105 IN. 1.0.
'0.125 IN. 0.0.
STAINLESS STEEL
Figure 2. Trap packings and construction to include
desorb capability
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3.1979 / Propoaed Rulea 69483
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
REGULATOR
PURGE GAS
FLOW CONTROL ,
13* MOLECULAR
SIEVE FILTER "
OPTIONAL 4 PORT COLUMN
SELECTION VALVE
6 PORT TRAP INLH
RESISTANCE WIRE
COLUMN OVEN
-1 1 CONFIRMATORY COLUMN
n n FfTO DETECTOR
"• I' " ANALYTICAL COLUMN
I RAP
22°C
I OFF
HEATER
CONTROL
Nole:
ALL LINES BETWEEN
PURGING DEVICE TRAP AND GC
HEATED 10 85°C SHOULD BE HEATED
TO 95°C.
Figure 3. Schematic of purge and trap device - purge mode
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
REGULATOR
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
COLUMN OVEN
r-innn I CONFTRIBATORY COLUMN
JU DETECTOR
- '^ - L
ANALY-nCAL COLUMN
\OPT10NAL 4-PORT COLUMN
SELECTION VALVE
TRAP INLET
VALVE J RESISTANCE WIRE ...,,—.»
_z HEATER
"CONTROL
PURGING
DEVICE
Note:
ALL LINES 3ETAEBI
TRAP AND GC
SHOULD 8E HEATED
TO 95° C.
Figure 4. Schematic of purge-and trap device • desorfa mode
SILU
cooe w«o-oi-c
-------
69484 Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules
COLUMN: CHROIMOSOR8 101
PROGRAM: 80*C-5 MINUTES.
8*C/ MINUTE TO 150SC
DETECTOR: FLAME IONIZAT10N
tu
«i
2
2463
RETENTION T11WE-.MINUTES
10
Figures. Gas chromatogram oi acrolein and acrylonitrile
Phenols—Method 604
\. Scope and Application.
1.1 This method covers the
determination of various phenolic
compounds. The following parameters
may be determined by this method:
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest. If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table I represent sensitivities that
can be achieved in wastewaters with a
flame ionization detector in the absence
of interferences. If the darivatization
cleanup is required, the sensitivity of the
method is 10 ju.g/1. This concentration
represents the minimum amount proven
to date to give reproducible and linear
response during derivatization.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons,
2. Summary of Method.
2.1 A 1-liter sample' of wastewater is
acidified and extracted with methylene
chloride using separatory funnel
techniques. The extract is dried and
concentrated to a volume of 10 ml or
less. Flame ionization gas
chroma tographic conditions are
described which allow for the
measurement of the compounds in the
extract.
2.2 The method also provides
for the preparation of
pentafluorobenzylbromide (PFB)
derivatives for electron capture gas
chromatography with additional cleanup
procedures to aid the analyst in the
elimination of interferences.
3. Interferences.
3.1 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 running method blanks.
Specific selection of reagents and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextracted from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled. While
general cleanup techniques are provided
as part of this method, unique samples
may require additional cleanup
approaches to achieve the sensitivities
stated in Table I.
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules 69485
4, Apparatus and Materials.
4.1 Sampling equipment for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or 1-quart volume. French
or Boston Round design is
recommended. .The container must be
washed.and solvent rinsed before use to
minimize interferences.
4.1.2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample -
containers for the collection of a
minumum of 250 ml. Sample containers
must be kept refrigerated during
sampling. No tygon or rubber tubing
may be used in this system.
4.2 Separatory funnel—2000 ml, with
Teflon stopcock.
4.3 Drying column—20 mm ID Pyrex
chromatographic column with coarse
frit.
4.4. Kudema-Danish (K-D)
Apparatus
4.4.1 Concentrator tube—10 ml
graduated (Kontes K-570050-1025 or
equivalent}. Calibration must be
checked. Ground glass stopper (size 19/
22 joint] is used to prevent evaporation
of extracts.
4.4.2 Evaporative flask—300 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-0012).
4.4.3 Snyder column—three-ball
macro (Kontes K-503000-0121 or
equivalent).
4.4.4 Snyder column—two-ball micro
(Kontes K-569001-0219 or equivalent).
4.4.5 Boiling chips—solvent
extracted, approximately 10/40 mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (±2°C). The bath
should be used in a hood.
4.6 Gas chromatograph—Analytical
system complete with gas
chromatograph suitable for on-column
injection and' all required accessories
Including flame ionization and electron
capture detector, column supplies,
recorder, gases, syringes. A data system
for measuring peak areas is
recommended.
4.7 Chromatographic column—10
mm 10 by 100 mm length, with Teflon
stopcock.
4.8 Reaction vial—20 ml, with
Teflon-lined cap.
S. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid—(1 + 1) Mix equal
volumes of cone. H»SO< (ACS) with
distilled water.
5.1.3 Sodium thiosulfate—(ACS)
Granular.
5.2 Methylene chloride, acetone, 2-
propanol, hexane, toluene—Pesticide
quality or equivalent.
5.3 Sodium sulfate—(ACS) Granular,
anhydrous (purified by heating at 400" C
for 4 hrs. in a shallow tray).
5.4 Stock standards—Prepare stock
standard solutions at a concentration of
1.00 jxg/^1 by dissolving 0.100 grams of
assayed reference material,in pesticide
quality 2-propanol and diluting to
volume in a 100 ml ground glass
stoppered volumetric flask. The stock
solution is transferred to ground glass
stoppered reagent bottles, stored in a
refrigerator, and checked frequently for
signs of degradation or evaporation,
especially just prior to preparing
working standards from them.
5.5 Sulfuric acid—(ACS) 1 N in
distilled water.
5.6 Potassium carbonate—(ACS)
powdered.
5.7 Pentafluorobenzyl bromide (a-
Bromopentafluoro toluene)—97%
minimum purity.
5.8 1,4.7,10,13,16—
Hexaoxacyclooctadecane (18-crown
6)—98% minimum purity.
5.9 Derivatization reagent—Add 1 ml
pentafluorobenzyi bromide and 1 gram
18 crown 6 to a 50 ml volumetric flask
and dilute to volume with 2-propanol.
Prepare fresh weekly.
5.10 Silica gel—(ACSJ100/200 mesh.
grade 923; activated at 130°C and stored,
in a sesiccator.
8. Calibration.
8.1 Prepare calibration standards for
the flame ionization detector that
contain the compounds of interest,
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 pig/1 in
the final extract, for example, prepare
standards at 10 ug/1, 50 ug/1,100 Mg/1,
500 fig/1, etc. so that injections of 1-5 ^il
of each calibration standard will define
the linearity of the detector in the
working range.
8.2 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table L
By injecting calibration standards,
establish the sensitivity limit of the
detector and the linear range of the
analytical system for each compound.
8.3 Before using the derivatization
clean up procedure, the analyst must
process a series of calibration standards
through the procedure to validate the
precision of the derivatization and the
absence of interferences from the
.reagents.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate through
the analysis of a distilled water method
blank, that all glassware and-reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
chronic laboratory contamination.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection. Preservation,
and Handling.
8.1 Grab samples must be collected
in glass containers. Conventional
sampling practices should be followed.
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. At the sampling
location fill the glass container with
sample. Add 35 mg of sodium thiosulfate
per part per million free chlorine per
liter. Adjust the sample pH to
approximately 2, as measured by pH
paper, using appropriate sulfuric acid
-------
69486 Federal Register / Vol. 44, No. 233 / Monday. December 3. 1979 / Proposed Rules
solution or ION sodium hydroxide.
Record the volume of acid used on the
sample identification tag so the sample
volume can be corrected later.
8.3 All samples, must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entice sample into a two-liter
separatory funnel Adjust the sample pH
to 12 with sodium hydroxide.
9-2 Add 60 ml methylene chloride to
the sample bottle, seal, and shake 30
seconds to rinse the inner walla.
Transfer the solvent into the. separatory
funnel, and extract the sample by
shaking the- funnel for one minute with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
of fon minutes. If the emulsion.
interface between layers is more than
one-third the size of the solvent layer,
the analyst must employ mechnical
techniques to complete the phase
separation. The optimum technique
depends upon the sample, but may
include stirring; filtration of the
emulsion through glass wool, or
centrifugation. Discard the methylene
chloride layer, and wash the sample
with an additional two 60ml portions of
methylene chloride in similar fashion.
9.3 Adjust the aqueous- layer to a pH
of 1-4 with sulfuric add.
9.4 Add CO ml of methylene chloride
to the sample and shake for two
minutes. Allow the solvent to separate
from the sample and collect the
methylene chloride in a 250 ml
Erlenmeyer flask.
9.5 Add a second 60 ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.6 Perform a third extraction in the
same manner. Pour the combined
extract the through a drying column
containing 3-4 inches of anhydrous-
sodium sulfate, and collect it in a 500-ml
Kuderna-Oanish (K-D) flask equipped
with a 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30 ml methylene chloride to complete
the quantitative transfer.
9.7 Add 1-2 clean boiling chips to
the 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 (60-65'C) so that the
concentrator tube is partially immersed
tn the hot water, and the entire lower
rounded surface of the flask is bathed in
vapor. Adjust the vertical position of the
appartus and the water temperature as.
required to complete the concentration
in 15-20 minutes. At the proper rate of
distillation the balls of tie column will
actively chatter but the chambers will
not flood. When the apparent volume of
liquid reaches 1 mi remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling.
9.8 Increase the temperature of the
hot water bath to 95-100'C. Remove the
Snyder column, and rinse the flask and
its lower joint into the concentrator tube
with 1-2 ml of 2-propanoL A 5-tnl
syringe is recommended for this
operation. Attach a micro-Snyder
column to the concentrator tube and
prewet the column by adding about 0.5
ml 2-propanol 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.
At the proper rate of distillation, the
balls of the column wilt actively chatter
but the chambers will not flood. When
the apparent volume of the liquid
reaches 2J5 ml, remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling. Add an
additional i ml of 2-propanol through
the top of the micro-Snyder column and
resume concentrating as before. When
the apparent volume of liquid reaches
0.5 ml. remove the K-D apparatus and
allow it to drain for at least 10 minutes
while, cooling. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with, a minimum
amount of 2-propanol. Adjust the extract
volume to 1.0 ml. Stopper the
concentrator tube and store in
refrigerator, if further processing will not
be performed immediately. If the- sample
extract requires no-further cleanup,
proceed with flame ionization gas
chroma tographic analysis. If the sample
requires cleanup, proceed to section 11.
9.9 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. After
correction for sulfuric acid preservative,
record the sample volume to the nearest
5mL
10. Gas Chromatography-Flanie
lonizatitm Detector.
10.1 Table I summarizes some
recommended gas chroma tographic
column materials and operating
conditions for the instrument Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method An example of
the separation achieved by one of these
columns is shown in Figure 1. Calibrate
the gas chromatographic system daily
with a minimum of three injections of
calibration standards.
10.2 Inject 2-5 pj of the sample
extract using the solvent-flush
technique. Smaller (1.0 /il) volumes can
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 ul and the resulting
peak size, in area units.
10.3 If the peak area exceeds the
linear-range of the system, dilute the
extract and reanalyze.
10.4 If the peak area measurement is
prevented by the presence of
interferences, the phenols must be
derivatized and analyzed by electron
capture gas chromatography.
11. Derivatization and Electron
Capture Gas Chromdtography.
11.1 Pipet a 1.0 ml aliquot of the 2-
propanol solution of standard or sample
extract into a glass reaction vial. Add
1.0 ml derivatization reagent This is a
sufficient amount of reagent to
derivatize a solution whose total
phenolic content does not exceed 0.3
mg/ml.
11.2 Add about 3 mg of potassium
carbonate to the solution and shake
gently.
11.3 Cap the mixture and heat it for 4
hours at 80°C in a hot water bath.
11.4 Remove the solution from the
hot water bath and allow it to cool.
11.5 Add 10 ml hexane to the
reaction vial and snake vigorously for
one minute. Add 3.0 ml of distilled,
deionized water to the reaction vial and
shake for two minutes.
11.8 Decant organic layer into a
concentrator tube and cap with a glass
stopper.
11.7 Pack a 1O mm ID
chromatographic column with 4.0 grams
of activated silica gel. After settling the
silica gel by tapping the column, add
about two grams of anhydrous sodium
sulfate to the top.
11.8 Pre-elute the column with 6 ml
hexane. Discard the eluate and just prior
to exposure of the sulfate layer to air,
pipet onto the column 2.0 ml of the
hexane solution (11.6) that contains the
derivatized sample or standard. Elute
the column with 10.0 ml of hexane
(Fraction 1) and discard this fraction.
Elute the column, in order, with: 10.0 mi
15% toluene in hexane (Fraction 2); 10.0
ml 40% toluene in hexane (Fraction 3);
10.0 ml 75% toluene in hexane (Fraction
4]; and 10.0 ml 15% 2-propanol in toluene
(Fraction 5). Elution patterns for the
phenolic derivatives are shown in Table
II. Fractions may be combined as
desired depending upon the specific
phenols of interest or level of
interferences.
11.9 Analyze the fractions by
electron capture gas chromatography.
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3, 1979 / Proposed Rules
69487
Table II summarizes some
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times that should be achieved by this
method. Examples of the separation
achieved by this column is shown in
Figure 2. Calibrate the system daily with
a minimum of three aliquots of
calibration standards, containing each
of the phenols of interest that are
derivatized according to the procedure.
11.10 Inject 2-5 p.1 of the column
fractions using the solvent-flush
technique. Smaller (1.0 JA!) volumes can
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 pL and the resulting
peak size, in area units. If the peak area
exceeds the linear range of the system.
dilute the extract and reanalyze.
12. Calculations
12.1 Determine the concentration of
individual compounds measured by the
flame ionization procedure (without
derivatization) according to the formula:
'*'roW
(Vj(VJ
Where:
\ a> Calibration factor for chromatographic
system, in nanograms material per area
unit.
B * Peak size in injection of sample extract.
in area units
V, — Volume of extract injected Oil)
V, - Volume of total extract Oil}
V, =» Volume of water extracted (p.1)
12.2 Determine the concentration of
individual compounds measured by the
derivatization and electron capture
procedure according to the following
procedure:
12.2.1 From the concentration of the
calibration standards that were
derivatized with the samples, calculate
the amounts, ia nanograms, of
underivatized phenols that were added
as 2-propanoi solution (11.1). From the
size of the injection into the electron
capture gas chromatograph, determine
the nanograms of material (calculated as
the underivatized phenol) injected onto
the column. Compare the detector
responses obtained to develop a
calibration factor for the
chromatographic system, in nanograms
of material per area unit.
12.2.2 Determine the concentration
of individual compounds according to^
the formula:
(VJnttrophenol . _ .....
2.4-C4chk»opnenol
2.4.8-Trtenlorephenol „
Pxitauh torophenol
(2,4.Dtnrtfophenol)
(2-MetnyM.64Mtrophenol
3.3
9 1
1.8 _. .
SB
70 „
48 ._
28 S „...
14.0
»48.9 _
»36.8 _
90
90
- - 95
.. . . 95
50 50 ....
g4
75 20
- -
10
7
>1
14
> 1
90
—
"
90
•Column condMom: Cftremoeore W-AW-OMCS 80/100 mean coated with 5% OV-17 pecked in a 1.8 m long x 2,0 mm 10
glace column with 5% methane/93% argon earner gas at 30 ml/mm flow rate. Column temperature t» 200'C.
'From: "Development and Application ol Teat Procedure! for Specific Organic Toxic Substaneea in Waatewaten. Catego-
nea 3-Chtonnat*d Hydroottont and Category 8-Phenol»."
•Retention ttnea included for qualitative information only. The lack ol accuracy and precision of the oenvaftation reacdoA
preclude* the uae of thn approach for quantitative purposes.
S1LUNG CODE 8SSO-01-M
-------
1 s
I s
cs wu
2rf 5
SSQ /-\
COLUMN: 1% SP-1240DA ON SUPELCOPORT
PROGRAM: 80*C.-0 MINUTES 8* /MINUTE TO 15Q*C.
DETECTOR: FLAME IONIZATION
S
O
UJ
O
i
-Z 5 _j2
csa >: o5
111 $-.
*" f CT
" 1
2
^«* i 35 ^™
•r o ** *•
5 I eg
» O 2 m
U
O.
O
O
z
0 4 8 12 16 20 24
RETENTION TIME-MINUTES
Figure 1. Gas chromatogram of phenols
Bit-UNO CODE wM-OI-C
COLUMN: 5% OV-17 ON CHROMOSORB W-AW
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
O
8
§
cr
8 12 16 20 24 28
RETENTION TIME-MINUTES
32
Figure 2. Gas chromatogram of PFB derivatives of phenols
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules 69491
Phthalate Esters—Method 806
1. Scope and Application.
1.1 This method covers the
determination of certain phthalate
esters. The following parameters may be
determined by this method:
StCfttHo.
34292
39100
34110
34S98
34336
34341
Bmyl but* pftttuMM
OWvoetyi p«hai«t« ..._
OMhyl pMhtUM
OkD*""* phmnat*
0 « 3 12
RETENTION TUBE-MINUTES
Figure 1, Liquid chromatogram of banzidinM
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the. National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest. If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table I represent sensitivities that
can be achieved in waste-waters in the
absence of interferences.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons.
2. Summary of Method.
2.1 A 1-liter sample of wastewater is
extracted with methylene chloride using
separatory funnel techniques. The
extract is dried and concentrated to a
volume of 10 ml or less.
Chromatographic conditions are
described which allow for the accurate
measurement of the compounds in the
extract.
2.2 If interferences are encountered,
the method provides selected general
purpose cleanup procedures to aid the
analyst in their elimination.
3. Interferences.
3.1 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 running method blanks.
Specific selection of reagents and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextracted from
the sample will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled. While
general cleanup techniques are provided
as part of this method, unique samples
may require additional cleanup
approaches to achieve the sensitivities
stated in Table I.
3.3 Phthalate esters contaminate
many types of products commonly found
in the laboratory. The analyst must
demonstrate that no phthalate residues
contaminate the sample or solvent
extract under the conditions of the
analysis. Of particular importance is the
avoidance of plastics because
phthalates are commonly used as
piasticizers and are easily extracted
from plastic materials. Serious phthalate
contamination may result at any time if
consistent quality control is not
practiced.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or l-quart volume. French
or Boston Round design is
recommended. The container must be
washed and solvent rinsed before use to
minimize interferences.
4.1.2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon. Foil may be
substituted if sample is not corrosive.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum ef 250 ml. Sample containers
must be kept refrigerated during
sampling. No tygon or rubber tubing
may be used in the system.
4.2 Separatory funnel—2000 mi. with
Teflon stopcock.
4.3 Drying column—20 mm ID pyrex
chromatographic column with coarse
frit.
4.4 Kuderna-Danish (K-D)
Apparatus
-------
69492 Federal Register / Vol. 44. No. 233 / Monday. December 3, 1979 / Proposed Rules
4.4.1 Concentrator tube—10 ml,
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked. Ground glass stopper (size 19/
22 joint) is used to prevent evaporation
of extracts.
4.4.2 . Evaporative flask—500 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-0012).
4.4.3 Snyder column—three-ball
macro (Kontes K503000-0121 or
equivalent).
4.4.4 Snyder column—two-ball micro
(Kontes K-569001-0219 or equivalent).
4.4.5 Boiling chips—solvent
extracted^ approximately 10/40 mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (± 2°Q. The bath
should be used in a hood.
4.6 Gas chromatograph—Analytical
system complete with gas
chromatograph suitable for on-column
injection and all required accessories
including electron capture or flame
ionization detector, column supplies,
recorder, gases, syringes. A data system
for measuring peak areas is
recommended
4.7 Chromatography column—300
mm long x 10 mm ID with coarse fritted
disc at bottom and Teflon stopcock
(Kontes K-420540-0213 or equivalent).
5. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid—(ACS) Mix equal
volumes of cone. H,SO, with distilled
water.
5.2 Methylene chloride—Pesticide
quality or equivalent.
5.3 Sodium Sulfate—(ACS) Granular,
anhydrous (purified by heating at 400°C
for 4 hrs. in a shallow tray).
5.4 Stock standards—Prepare stock
standard solutions at a concentration of
1.00 fig/i*l by dissolving 0.100 grams of
assayed reference material in pesticide
quality isooctane op-other appropriate
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation. especiaOy
just prior to preparing working
standards from them.
5.5 Diethyl Ether—Nanograde,
redistilled in glass if necessary.
5.5.1 Must be free of peroxides as
indicated by EM Quant test strips. (Test
strips are available from EM
Laboratories, Inc., 500 Executive Blvd.,
Elmsford. N.Y. 10523.)
5.5.2 Procedures recommended for
removal of peroxides are provided with
the test strips. After cleanup, 20 ml ethyl
alcohol preservative must be added to
each liter of ether.
5.8 Florisil—PR grade (80/100 mesh);
purchase activated at 1250*F and store
in dark in glass container with ground
glass stoppers or foil-lined screw caps.
5.7 Alumina—Activity Super I,
Neutral. W200 series, (ICN Life Sciences
Group, No. 404583).
5.8 Hexane—Pesticide quality.
6. Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest,
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitudes that will
completely bracket the working range of
the chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 ng/1
in the final extract for example, prepare
standards at 10 j*g/l, 50 fig/1,100 pg/1.
500 fig/1, etc. so that injections of 1-5 >il
of each calibration standard will define
the linearity of the detector in the
working range.
&2 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table 1.
By injecting calibration standards,
establish-the sensitivity limit of the
detector and the linear range of the
analytical'system for each compound.
6.3 Before using any cleanup
procedure, the analyst must process a
series of calibration standards through
the procedure to validate elution
patterns and the absence of
interferences from the reagents.
7. Quality Control.
7.1 Before processing any samples.
the analyst should demonstrate through
the analysis of a distilled water method
blank, that all glassware and reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
chronic laboratory contamination.
7:2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection, Preservation,
and Handling.
8.1 Grab samples must be collected
in glass containers. Conventional
sampling practices should be followed.
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, the sample should be
adjusted to a pH range of 6.0-8.0 with
sodium hydroxide or suifuric acid
8.3 All samples must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Check the ph of the
sample pH with wide-range paper and
adjust to within the range of 5-9 with
sodium hydroide or suifuric acid.
9.2 Add 60 ml methylene chloride to
the sampje bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer the solvent into the separatory
funnel, and extract the sample by
shakingjhe funnel for two minutes with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
minimum of ten 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, but may
include stirring, filtration of the
emulsion through glass wool, or
centrifugation. Collect the methylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium sulfate, and collect it in a 500-ml
Kudema-Danish (K-D) flask equipped
with a 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3, 1979 / Proposed Rules 69493
20-30 ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 clean boiling chips to
the 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 (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-vertial 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
activeiy 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 for at
least 10 minutes while cooling.
9.6 Increase the temperature of the
hot water bath to about 80°C.
Momentarily remove the Snyder column.
add 50 ml of hexane and a new boiling
chip and reattach the Snyder column.
Pour about 1 ml of hexane into the top of
the Snyder column and concentrate the
solvent extract as before. Elapsed time
of concentration should be 5 to 10
minutes. When the apparent volume of
liquid reaches 1 ml, remove the K-D
apparatus and allow it to drain at least
10 minutes while, cooling. Remove the
Snyder column and rinse the flask and
its lower joint into the concentrator tube
with 1-2 ml of hexane, and adjust the
volume to 10 ml A 5-ml syringe is
recommended for this operation.
Stopper the concentrator tube anchstore
refrigerated if further processing will not
tie performed immediately. If the sample
extract requires no further cleanup
proceed with gas chromatographic
analysis. If the sample requires cleanup,
proceed to Section 10.
9.7 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. Record the
sample volume to the nearest 5 ml.
10. Cleanup and Separaton.
10.1 If the entire extract is to be
cleaned up by one of the following two
procedures, it must be concentrated to
about 2 ml. To the concentrator tube in
9.6, add a clean boiling chip and attach
a two-ball micro-Snyder-column. Prewet
the column by adding about 0.5 mi
hexane through the top. Place the K-D
apparatus on a hot water bath [80'Q 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
activeiy chatter but the chambers will
not flood. When the apparent volume of
liquid reaches about 0.5 ml, remove the
K-D apparatus and allow it to drain for
at least 10 minutes while cooling.
Remove the micro-Snyder column, and
rinse its lower joint into the
concentrator tube with 0.2 ml of hexane.
Proceed with one of the following clean-
up procedures.
10.2 Florisil Column Cleanup for
Phthalate Esters
10.2.1 Place 100 g of Florisil into a
500 ml beaker and heat for
approximately 16 hours at 400'C. After
heating transfer to a 500 ml reagent
bottle. Tightly seal and cfibl to room
temperature. When cool add 3 ml of
distilled water which is free of
phthalates and interferences. Mix
thoroughly by shaking or roiling for 10
minutes and let it stand for at least 2
hours. Keep the bottle sealed tightly.
10.2,2 Place lOg of this Florisil
preparation into a 10 mm ID
chromatography column and tap the
column to settle the Florisil. Add 1 cm of
anhydrous sodium sulfate to the top of
the Florisil.
10.2.3 Preelute the column with 40 ml
of hexane. Discard this eluate and just
prior to exposure of the sodium sulfate
layer to the air transfer the 2 ml sample
extract onto the column, using -an
additional 2 ml of hexane complete, the
transfer.
10.2.4 just prior to exposure of the
sodium sulfate layer to the air add 40 ml
hexane and continue the elution of the
column. Discard this hexane eluate.
10.2.5 Next elute the phthalate esters
with 100 ml of 20 percent ethyl ether/80
percent hexane (V/V) into a 500 ml K-D
flask equipped with a 10 ml concentrator
tube. Elute the column at a rate of about
2 ml per minute for all fractions.
Concentrate the collected fraction by
standard K-D technique. No solvent
exchange is necessary. After
concentration and cooling, adjust the
volume of the cleaned up extract to 10
ml in the concentrator tube and analyze
by gas chromatography.
10.3 Alumina Column Cleanup for
Phthalate Esters
10.3.1 Place 100 g of alumina into a
500 ml beaker and heat for
approximately 16 hours at 400* C. After
heating transfer to a 500 ml reagent
bottle. Tightly seal and cool to room
temperature. When cool add 3 ml of
distilled water which is free from
phthalates and interferences. Mix
thoroughly by shaking or rolling for 10
minutes and let it stand for at least 2
hours. Keep the bottle sealed tightly.
10.3.2 Place 10 g of this alumina
preparation into a 10 mm ID
chromatography column and tap the
column to settle the alumina. Add 1 cm
of anhydrous sodium sulfate to the top
of the alumina.
10.3.3 Preelute the column with 40 ml
of hexane. Discard this eluate and just
prior to exposure of the sodium sulfate
layer to the air, transfer the 2 ml sample
extract onto the column, using an
additional 2 ml of hexane to complete
the transfer.
10.3.4 Just prior to exposure of the
sodium sulfate layer to the air add 35 ml
hexane and continue to elution of the
column. Discard this hexane eluate.
10.3.5 Next elute the column with 140
ml of 20 percent ethyl ether/80 percent
hexane (V/V) into a 500 mi K-D flask
equipped with a 10 ml concentrator
tube. Elute the column at a rate of about
2 ml per minute for all fractions.
Concentrate the collected fraction by
standard K-D technique. No solvent
exchange is necessary. After
concentration and cooling adjust the
volume of the cleaned up extract to 10
ml in the concentrator tube and analyze
by gas chromatography.
11. Gas Chromatography,
11.1 Table I summarizes some
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be |
achieved by this method. Examples of
the separations achieved by the primary
column are shown in Figures 1 and 2.
Calibrate the system daily with a
minimum of three injections of
calibration standards.
11.2 Inject 2-5 >tl of the sample
extract using the solvent-flush
technique. Smaller (1.0 jil) volumes can
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 pi, and the resulting
peak size, in area units.
11.3 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
11.4 If the peak area measurement is
prevented by the presence of
interferences, further cleanup is
required.
12. Calculations.
12.1 Determine the concentration of
individual compounds according to the
formula:
ConountnOan. i
(VJ(VJ
-------
69494 Federal Register / Vol. 44, No. 233 / Monday, December 3. 1979 / Proposed Rules
Where:
A—Calibration factor for chroma tographic
system, in nanograma material per area
unit.
B-Peak size in injection of sample extract, in
area units
V,-Volume of extract injected !jd)
V,-Volume of total extract (jil)
V,» Volumeof water extracted (ml)
12.2 Report results in micrograms per
liter without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
13. Accuracy and Precision.
13.1 The U.S. EPA Environmental
Monitoring and Support Laboratory in
Cincinnati is in the process of
conducting a Intel-laboratory method
study to determine the accuracy and
precision of this test procedure.
Bibliography
"Development and Application of Test
Procedures for Specific Organic Toxic
Substances in Wastewatera. Category 1-
Phtaalates." Report for EPA Contract 88-
03-2806 (In preparation).
fonndon Sma
tn*v) Datacton
Compound _____^
Cot1 Cot 2 EC' FIO
Oknwft* polhalata— 2.03 0.05 Q.11 19
Diathyl pMnalatt 2.92 1-27 9.13 31
m n tiiityl pMtiaUM ».86 ISO 0.02 14
Bmy< buy* pnmatta— '8.J4 "5.11 0.02 IS
•«.« "io.s ao4 20
•112 1"«.0 0.11 3t
• Sxmlcoooit toonzo iMih ctnttd wtth i 5% SP-2290/
1.95% SP-2401 ptcMd in • tSO em long x 4 mm 10 glM*
cokmM Mth arrlw gw M (0 ml/mm flow r*M CoKimn wm-
!>•>•*«• I* 180X «KMP4 wt>«r» • mmn* 220-C Undw
aw** oonoWom RT. at AWrtn it S.49 mm. « iag a 10 ml dual vqfuma ol Kw t Mai aamMa «-
tract, and asaumng a GC injactton of 5 mcroMara.
8ILUMQ COOC MaO-01-K
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3,1979 / Proposed Rules
69495
COLUMN: 1.5% SP-2250*
1.35%.SP.2401 ON SUP3.COPORT
TEMPERATURE: 1S04C.
DETECTOR: ELECTRON CAPTURE
ua
UJ *-
— O.
o. ,
e
C
0 24 S 3 10 12
RETENTION TIME-MINUTES
Figure 1. Gas chro;matogram of phthalates
COLUMN: T.5% SP-22Sa*
1.35% SP-2401 ON SUPaCOPORT
TEMPEHATURE: 180"C.
DETECTOR: ELECTRON CAPTURE
UJ
a. i—
/N* rs
2 i£
04 a
ea
-u
»
H-
3
O
c
c
0 4 8 12 1618
RETByTION T1ME-411NUTES
Rgure 2. Gas chromatogram of phthalates
ULUMQ CODE M60-01-C
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Federal Register / Vol. 44, iNo. 233 / Monday. December 3; 1979 / Proposed Rules 69501
OrganochJorina Pesticides and PC3's—
Method 608
1. Scope and Application.
1.1 This method covers the
determination of certain organochlorine
pesticides and polycnlorinated
biphenyls (PCBa). The following
parameters may be determined-by this
method:
Stontjvo.
•4HC.
**«-
*8HC_
30298
30340
M'-OOE-
4,«'-OOT.
OM
39310
30300
Endcwuflw!
EMnutaiN—
Endnufen SuMa
Efldm.
gndrto AMMlyd*
crta
H*pttditor Sparid*.
pca-ioi»_
PC8-12Z1-
PC8-1232-
PC8-1242--
PC8-1246X.
pce-i2S4_
PC8-1«0_
34981
34360
34351
39380
34306
30410
30420
30400
34071
30408
304«0
: . 39600
30504
__ 30600
\2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
.monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest If the user is
attempting to screen samples for any or
all of the compounds above, he most
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table I represent sensitivities that
can be achieved in wastewaters in the
absence of interferences.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons.
2. Summary of Method.
2.1 A 1-liter sample of waste-water is
extracted with methylene chloride using
separatory funnel techniques. The
extract is dried and concentrated to a
volume of 10 ml or less.
Chromatographic conditions are
described which allow for the accurate
measurement of the compounds in the
extract
ZJZ If interferences are encountered,
the method provides selected general
purpose cleanup procedures to aid the
analyst in their elimination.
3. Interferences.
3.1 Solvents, reagents, glassware,
and other sample processing hardware
may yield discrete artifacts and/or
elevated baselines causing
misinterpretation of gas chroma tograma.
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 and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextraded from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being, sampled. While
general cleanup techniques are provided
as part of this method, unique samples
may require additional cleanup
approaches to achieve the sensitivities
stated in Table 1.
3.3 Glassware must be scrupulously
clean. Clean all glassware as soon as
possible after use by rinsing-with the
last solvent used. This should be
followed by detergent washing in hot
water. 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 15 to 30
minutes. Some high boiling materials.
such as PCBs, may not be eliminated by
this treatment. Volumetric ware should
not be heated in a muffle furnace.
Glassware should be sealed/ stored in a
clean environment immediately after
drying or cooling to prevent any
accumulation of dust or other
contaminants. Store inverted or capped
with aluminum foil.
3.4 Interferences by phthalate esters
can pose a major problem in pesticide
analysis. These materials eiute in the
15% and 50% fractions of the Florisil
cleanup. They usually can be minimized
by avoiding contact with any plastic
materials. The contamination from
phthalate esters can be completely
eliminated with the use of a
microcoulometric or electrolytic
conductivity detector.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or 1-quart volume. French
or Boston Round design is
recommended. The container must be
washed and solvent rinsed before use to
minimize interferences.
4.1.2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon. Foil may be
substituted if sample is not corrosive.
4.1.3 Compositing equipment-
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum of 250 ml. Sample containers
must be kept refrigerated during
sampling. No tygon or robber tubing
may be used in the system.
4.2 Separatory funnel—2000 ml, with
Teflon stopcock.
4.3 Drying column—20 mm ID pyrex
chromatographic column with coarse
frit
4.4 Kuderna-Danish (K-D)
Apparatus
4.4.1 Concentrator tube—10 ml,
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked at 1.0 and 10.0 ml level. Ground
glass stopper (size 19/22 joint) is used to
prevent evaporation oi extracts.
4.4.2 Evaporative flask—500 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-0012).
4.4.3 Snyder column—three-ball
macro (Kontea K503000-0121 or
equivalent).
4.4.4 Boiling chips—extracted,
approximately 10/40 mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (±2"C). The bath
should be used in a hood.
4.3 Gas chroma tograpn—Analytical
system complete with gas
chromatograpb suitable for on-cohimn
injection and all required acessories
including electron capture or halogen-
specific detector, column supplies,
recorder, gases, syringes. A data system
for measuring peak areas is
recommended.
4.7 Chromatographic column—Pyrex
400 mm X 25 mm OD, with coarse
fritted plate and Teflon stopcock
(Kontes K-42054-213 or equivalent).
5. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid (1+1}—(ACS) Mix
equal volumes of cone. HaSO« with
distilled water.
5.2 Methylene chloride—Pesticide
quality or equivalent.
5.3 Sodium Sulfate—(ACS) Granular,
anhydrous (purified by heating at 400*C
for 4 hrs. in a shallow tray).
5.4 Stock standards—Prepare stock
standard solutions at a concentration of
1.00 Mg/Ml by dissolving 0.100 grams of
assayed reference material in pesticide
quality isooctane or other appropriate
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them.
-------
69502 Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules
5.5 Boiling chips—Hengar granules
(Hengar Co.; Fisher Co.) or equivalent
5,6 Mercury—triple distilled.
5.7 Aluminum oxide—basic or
neutral, active.
5.3 Hexane—pesticide residue
analysis grade.
5.9 Isooctane (2.2,4-trimethyl
pentane)—pesticide residue analysis
grade.
5.10 Acetone—pesticide residue
analysis grade.
5.11 Diethyi ether—Nanograde.
redistilled in glass if necessary.
5.11.1 Must be free of peroxides as
indicated by EM Quant test strips (Test
strips are available from EM
Laboratories. Inc., 500 Executive Blvd..
Elmsford, N.Y., 10523).
5.1.2 Procedures recommended for
removal of peroxides are provided with
the test strips. After~cleanup 20 ml ethyl
alcohol preservative must be added to
each liter of ether.
5.12 Florisil—PR grade {60/100
mesh); purchase activated at 1250T and
store in glass containers with glass
stoppers or foil-lined screw caps. Before
use activate each batch at least 16 hours
at 130"C in a foil covered glass
container.
6. Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can.
be calculated from Table I as 100 >ig/l in
the final extract, for example, prepare.
standards at 10 ug/1. 50 /ig/1,100 jug/1.
500 pg/L eta, so that injections of 1-5 pd
of each calibration standard will define
the linearity of the detector in the
working range.
6.2 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table L
By injecting calibration standards,
establish the sensitivity limit of the
detector and the linear range of the
analytical system for each compound.
6.3 The cleanup procedure in Section
10 utilizes Florisil chromatography.
Florisil froni different batches or sources
may vary in absorption capacity. To
standardize the amount of Florisil which
is used, the use of lauric acid value
(Mills, 1968) is suggested. The
referenced procedure determines the
adsorption from bexane solution of
lauric acid (mg) per gram Florisil. The
amount of Florisil to be used for each
column is calculated by dividing this
factor into 110 and multiplying by 20
grams.
6.4 Before using any cleanup
procedure, the analyst must process a
series of calibration standards through
the procedure to validate elution
patterns and the absence of
Interferences from the reagents.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate through
the analysis of a distilled water method
blank, that all glassware and reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
chronic laboratory contamination.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate Jhe precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed tc
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection, Preservation.
and Handling.
8.1 Grab samples must be collected
in glass containers. Conventional
sampling practices should be followed.
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, the .sample should be
adjusted to a pH range of 6.0-6.0 with
sodium hydroxide or sulfuric acid.
8.3 All samples must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Check the pH of the
sample with wide-range pH paper and
adjust to within the range of 5-9 with
sodium hydroxide or sulfuric acid.
9.2 Add 60 ml methylene chloride to
the sample bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer the solvent into the separatory
funnel, and extract the sample by
shaking the runnel for two minutes with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
minimum of ten minutes. If the emulsion
interface between layers is more than
one-third the size of the solvent layer,
the analyst must enploy mechanical
techniques to complete the phase
separation. The optimum technique
depends upon the sample, but may
include stirring, filtration of the
emulsion through glass wool, or
centrifugation. Collect the msthylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3—i inches of anhydrous
sodium sulfate.'and collect it in a 500-ml
Kudema-Danish (K-D) flask equipped
with a 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30 ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 clean boiling chips to
the flask and attach a three-bail 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 (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-and allow it to drain for at
least 10 minutes while cooling.
9.6 Increase the temperature of the
hot water bath to about 80°C.
Momentarily remove the Snyder column,
add 50 ml of hexane and a new boiling
chip and reattach the Snyder column.
Pour about 1 ml of hexane into the top of
the Snyder column and concentrate the
solvent extract as before, the elapsed
time of concentration should be 5 to 10
minutes. When the apparent volume of
liquid reaches 1 ml, remove the K-D
apparatus and allow it to drain at least
10 minutes while cooling. Remove the
Snyder column and rinse the flask and
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules 69503
its lower joint into the concentrator tube
with 1-2 ml of hexane, and adjust the
volume to 10 ml. A 5-ml syringe is
recommended for this operation.
stopper the concentrator tube and store
refrigerated if further processing will not
be performed immediately. If the sample
extract requires no further cleanup,
proceed with gas chromatographic
analysis. If the sample requires cleanup.
proceed to Section 10.
9.7 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. Record the
sample volume to the nearest 5 ml.
10. Cleanup and Separation.
10.1 Cleanup procedures are used to
extend the sensitivity of a method by
minimizing or eliminating interferences
that mask or otherwise disfigure the gas
chromatographic response to the
pesticides and PCB's. The Florisil
column allows for a select fractionation
of the compounds and will eliminate
polar materials. Elemental sulfur
interferes with the electron capture gas
chromatography of certain pesticides
but can be removed by the techniques
described below.
10.2 Florisil Column Cleanup
10.2.1 Add a weight of Florisil,
(nominally 21g,) predetermined by
calibration (6.3, 6.4), to a
chromatographic column. Settle the
Florisil by tapping the column. Add
sodium sulfate to the top of the Florisil
to form a layer 1-2 cm deep. Add 60 ml
of hexane to wet and rinse the sodium
sulfate and Florisil. Just prior to
exposure of the sodium sulfate to air,
stop the elution of the hexane by closing
the stopcock on the chromatography
column. Discard the eluate.
10.2.2 Adjust the sample extract
volume to 10 ml and transfer it from the
K-D concentrator tube to the Florisil
column. Rinse the tube twice with 1-2'
ml hexane, adding each rinse to the
column.
10.2.3 Place a 500 ml K-D flask and
clean concentrator tube under the
chromatography column. Drain the
column into the flask until the sodium
sulfate layer is nearly exposed. Elute the
column with 200 ml of 6% ethyl ether in
hexane (Fraction 1) using a drip rate of
about 5 ml/min. Remove the K-D flask
and set aside for later concentration.
Elute the column again, using 200 mi of
15% ethyl ether in nexane (Fraction 2),
into a second K-D flask. Perform the
third elution using 200 ml of 50% ethyl in
hexane (Fraction 3). The elution patterns
for the pesticides and PCB's are shown
in Table II.
10.2.4 Concentrate the eluates by
standard K-D techniques (9.5),
substituting hexane for the glassware
rinses and using the water bath at about
85° C. Adjust final volume to 10 ml with
hexane. Analyze by gas
chromatography.
10.3 Elemental sulfur will usually
elute entirely in Fraction 1. To remove
sulfur interference from this fraction or
the original extract pipet 1.00 ml of the
concentrated extract into a clean
concentrator tube or Teflon-sealed vial.
Add 1-3 drops of mercury and seal.
Agitate the contents of the vial for 15-30
seconds. Place the vial hi an upright
position on a reciprocal laboratory
shaker and shake for 2 hours. Analyze
by gas chromatography.
11. Gas Chromatography.
11.1 Table I summarizes some
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method. Examples of
the separations achieved by these
columns are shown in Figures 1 through
10. Calibrate the system daily with a
minimvnn of three injections of
calibration standards.
11.2 Inject 2-5 /il of the sample
extract using the solvent-Hush
technique. Smaller (1.0 jil) volumes can
be injected if automatic devices-are
employed. Record the volume injected to
the nearest 0.05 jil, and the resulting
peak size, in area units.
11.3 If the peak area exceeds the
linear range of the system, dilute the
extract-and reanalyze.
11.4 If the peak area measurement is
prevented by the presence of
interferences, further cleanup is
required.
12. Calculations.
12.1 Determine the concentration of
individual compounds according to the
formula:
Concentraoon, ^a/l- (A)(BXV'>
(VJ(VJ
Where:
A=Calibration factor for chromatographic
system, in nanograms material per area
unit.
8=Peak size in injection of sample extract, in
area units
Vj=Volume of extract injected (>il)
V,=Volume of total extract (u,l]
V,=Volume of water extracted (ml)
12.2 Report results in micrograms per
liter without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
13. Accuracy and Precision.
13.1 The U.S. EPA Environmental
Monitoring and Support Laboratory in
Cincinnati is in the process of
conducting an interiaboratory method
study to determine the accuracy and
precision of this test procedure.
Bibliography
1. "Development and Application of Te»t
Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 10-
Pesticides and PCB's." Report for EPA
Contract 68-03-2606.
2. Mills, P. A., "Variation of Florisil Activity:
Simple Method for Measuring Absorbent
Capacity and Its Use in Standardizing
Florisil Columns," Journal of the
Association of Official Analytical
Chemists, 51, 29 (1968).
lata* \.—Gas Chramatognphy of Ptstladis ana
PCB's
Parameter
Col. 1 ' Col.2 *
.Detection
Ab*in-
a-8HC . _._
hJlWC , ,.,.„.,„ ..,...,
*9HC
jjiwr.
Qil
n
o
4.10
1.82
i.sr
2JO
2.13
(«)
9.08
7.15
11.75
723
9.20
8^8
10.70
8.10
9 JO
3.35
5.00
C)
C)
C)
C)
C)
(')
C)
n
0.003
0.002
0.004
0.004
0.002
0.04
0.012
0.006
0.018
0.006
0.009
0.01
0.03
0.009
0.023
0.002
0.004
0.40
0.04
0.10
0.10
o.os
0.08
0.08
0.15
" Supeteoport 100/120 mesh coated with 1.5% SP-22SO/
1.95% SP-2401 pocked in « 180 cm long x 4 mm IO glass
column vMtl) 5* Methane/95% Argon earner gM n 60 ml/ mm
flow raw. Column temperature is JOO'C.
'Supetcoport 100/120 mesh coated wim3%OV-l inaieo
cm long x 4 mm 10 glass column witd 5% Mettwe/95%
Argon earner gu at 60 ml/min flow rate. Column temperature
'Detection Kmn 'a calculated from the minimum detectable
GC response being equal to five times the GC background
nose, assuming s 10 ml final volume o« the 1 liter sample
extract and assuming a <3C injection of 5 rncrentsrs.
• Multiple peak response. See Figures 2-10.
-------
69504 Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
ft**** tnd PCBt tMng ftofM Column
Chromftogrtpfty
*yr
2 (1* 3 (M
1O pet) pet) pet)
AWrtn too .
«-8HC 100 .
6-8HC
100
CMORkM '00 _^~_
4,4--oof JZZZZILZIir M .Z!"ZZ!""Z
40*'-DOT 100
OMdrtn 0 TOO
EndOflUlfin I -»«..»..Mw««H».««_nr 37 64 .T_.....Tt.....
EndotulMnll 0 7 tl
EndowiNMi MMO. 0 0 10t
Sndhn 4 M
Endrin ttfttlHffto ri_.. ^ 0 98 38
H*etteNor „ 100 .
McplMNer •poxfcl* ™—-. .-. 100.
ToK*oft4n» .„... M .
PCd-1018. 97 .
PC8-1221 97 .
PC8-124&.
PW-1254
PC8-1J60—
•From •Dottopnunt and AppDetfon o( T««t PraeMurM
tor SptcMo Orgvie Tade SubCMMM m WMMMMI*. CM»
yory iO^M«o«ta and PCS'!. Report tor EPA CenMet »-
03-280*.-
INUUNO COM MW-01-M
-------
COLUMN: 1.5% SP-2250*
1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 200'C.
DETECTOR: ELECTRON CAPTURE
iu
o
o
O S3 u;
o= <
O u.
4 8 12 16
RETENTION TIME-MINUTES
Figure 1. Gas chromatogram of pesticides
COLUMN: 1.5% SP-2250*
1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
S?
!
90
I
5
CO
o
O.
B>
O
8*
°
4 8 12
RETENTION TIME-MINUTES
16
Figure 2. Gas chromatogram of chlordane
O
•a
o
(0
g
(I.
n
-------
COLUMN: 1.5% SP-2250+
1.95?'. SP-2401 ON SUPELCOPORT
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
10 14 18 22
RETENTION TIME-MINUTES
26
Figure 3. Gas chromatogram of toxaphene
COLUMN: 1.5% SP-2250+ 1.95% SP-2401 ON SUPELCOPORT
EMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18
RETENTION TIME-MINUTES
Figure 4. Gas chromatogram of PCB-1016
22
u»
3.
I
8
o
CJ
CO
o
IB
a
CO
I
a
i
01
M
(O
o
•o
o
a>
A
a.
E.
(D
(O
-------
COLUMN: 1.5% SP-2250* 1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 160*C.
DETECTOR: aECTRON CAPTURE
6 10 14 18
RETENTION TIME-MINUTES
COLUMN: 1.5% SP-2250. 1.95'; SP-2401 ON SUPELCOPORT
TEMPERATURE: 160'C.
DETECTOR: ELECTRON CAPTURE
z
o
9
IX
to
(D
3
cr
6 10 14 18
RETENTION TIME-MINUTES
22
24
22
Figure 6. Gas chromatogram of PCB-1232
Figure 5. Gas chromatogram of PC8-1221
O
CO
(ft
Q-
I
a>
ce
O>
CO
-------
COLUMN: 1.SK SP-2250+ 1.95?'. SP-2401 ON SUPELCOPORT
TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18
RETENTION TIME-MINUTES
Figure 7. Gas chromatogram of PCS-1242
22
COLUMN: 1.5% SP-2250* 1.9S?/. SP-2401 ON SUPaCOPORT
TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18
RETENTION TIME-MINUTES
Figure 8. Gas chromatogram of PCB-1248
22
26
9-
I
I
55
to
I
ID
2.
E.
2
-------
COLUMN: 1.5% SP-2250 +
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
SP-2401 ON SUPELCOPORT
COLUMN: 1.5% SP-2250* 1.95X SP-2401 ON SUPELCOPORT
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
2.
90
"2
o
o>
I
3
cr
a
_L
-L
JL
_L
10 14 18 22
RETENTION TIME-MINUTES
26
Figure 10. Gas chromatogram of PCB-1260
6 10 14
RETENTION TIME-MINUTES
18
22
Figure 9. Gas chromatogram of PCB-1254
BILLING CODE *MO-Ot-C
o.
ya
a>
-------
69510
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
Nitroaromatics and Isophorone—
Method 609
1, Scope and Application.
1.1 This method covers the
determination of certain nitroaromatics
and isophorone. The/following
parameters may be determined by this
method:
Slorsl No.
Parameter
Isophorone
Nitrobenzene
2.4-Omrtrotoluene.
2.6-OinttrotokMne.
SAW
34S(1
34826
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used;to meet the
monitoring requirements of the National
Pollutant Discharege Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compunds of interest. If the user is
attempting to screen samples for any or
all of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the leve! of
interferences rather than instrumental
limitations. The limits of detection listed
in Table I represent sensitivities that
can be achieved in wastewaters in the
absence of interferences.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons.
2. Summary of Method.
2.1 A 1-liter sample of wastewater is
extracted with methylene chloride using
separatory funnel techniques. The
extract is dried and exchanged to
toluene while being concentrated to 1.0
ml. Isophorone and nitrobenzene are
measured by flame ionization gas
chromatography. The dinitrotoluenes
are measured by electron capture GC.
2.2 If interferences are encountered.
the method provides a general purpose
cleanup procedure to aid the analyst in
their elimination.
3. Interferences.
3.1 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 running method blanks.
Specific selection of reagents and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextracted from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex r
municipality being sampled. While
general clean-up techniques are
provided as part of this method, unique
samples may require additional-cleanup
approaches to achieve the sensitivities
stated in Table I.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or 1-quart volume. French
or Boston Round design is •
recommended. The container must be
washed and solvent rinsed before use to
minimize interferences.
4.1.2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon. Foil may be
substituted if sample is not corrosive.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum of 250 ml. Sample containers
must be kept refrigerated during_
sampling. No tygon or rubber tubing
may be used in the system.
4,2 Separatory funnel—2000 ml, with
Teflon stopcock.
4,3 Drying column—20 mm ID pyrex
chromatographic column with coarse
frit.
4,4 Kuderna-Danish (K-D)
Apparatus
4,4.1 Concentrator tube—10 ml.
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked. Ground glass stopper (size 19/
22 joint) is used to prevent evaporation
of extracts.
4 4.2 Evaporative flask—500 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-0012).
4,4.3 Snyder column—three-ball
macro (Kontes K503000-0121 or
equivalent).
4,4.4 Snyder column—two-ball micro
(Kontes K-569001-0219 or equivalent).
4.4.5 Boiling chips-solvent extracted.
approximately 10/40 mesh.
4,5 Water bath—Heated; with
concentric ring cover, capable of
temperaWe control (±2"C). The bath
should be used in a hood.
4,6 Gas chromatograph—AnalyticaJ
system complete with gas
chromatograph suitable for on-column
injection and all required accessories
including both electron capture and
flame ionization detectors, column
supplies, recorder, gases, syringes. A
data system for measuring peak areas is
recommended.
4.7 Chromatography column—400
mm long x 10 mm ID, with coarse fritted
plate on bottom and Teflon stopcock.
5. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid (1 + 1)—(ACS) Mix
equal volumes of cone. HiSO* with
distilled water
5.2 Methylene chloride—Pesticide
quality or equivalent.
5.3 Sodium sulfate—(ACS) Granular,
^anhydrous (purified by heating at 400°C
for 4 hrs. in a shallow tray).
5.4 Stock standards—Prepare stock
standard solutions at a concentration of
1.00 p.g/ul by dissolving 0.100 grams of
assayed reference material in pesticide
quality isooctane or other appropriate
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them.
^5.5 Acetone. Haxane, Methanol,
Toluene—pesticide quality or
equivalent.
5.6 Florisil—PR grade (60/100 mesh);
purchase activated at 1250T and store
in glass containers with glass stoppers
or foil-lined screw caps. Before use,
activate each batch overnight at 200"C~
in glass containers loosely covered with
foil.
6. Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest.
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 ;±g/l
in the final extract, for example, prepare
standards at 10 u-g/T, 50 u.g/1,100 jig/1.
500 u.g/1, etc. so that injections of 1-5 ^1
of each calibration standard will define
the linearity of the detector in the
working range.
6.2 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table I.
By injecting calibration standards.
establish the sensitivity limit of the
detector and the linear range of the
analytical system for each compound.
6.3 Before using any cleanup
procedure, the analyst must process a
series of calibration standards through
the procedure to validate elution
patterns and the absence of
interferences from the reagents.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate through
the analysis of a distilled water method
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules ~ 69511
blank, that ail glassware and reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
chronic laboratory contamination.
7.2 .Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the
chromatogram. confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection, Preservation.
and Handling.
8.1 Crab samples must be collected
in glass containers. Conventional
sampling practices should be followed,
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be-iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, the sample should be
adjusted to a pH range of 6.0-6.0 with
sodium hydroxide or sulfuric acid.
8.3 All samples must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Check the pH of the
sample with wide-range pH paper and
adjust to within the range of 5-9 with
sodium hydroxide or sulfuric acid.
9.2 Add 60 ml methylene chloride to
the sample bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer tl}e solvent into the separatory
funnel, and extract the sample by
shaking the funnel for two minutes with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
minimum of ten minutes. If the emulsion
interface between layers is more, than
one-third the size of the solvent layer,
the analyst must employ mechanical
techiques to complete the phase
separation. The optimum technique
depends upon the sample, but may
include stirring, filtration of the
emulsion through glass wool, or
centrifugation. Collect the methylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9,4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium sulfate, and collect it in a 500-ml
Kuderna-Danish (K-D) flask equipped
with a 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30 ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 dean boiling chips to
the 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 (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 and allow it to drain for at
least 10 minutes while cooling. Remove
the Snyder column and rinse the flask
and its lower joint.into the concentrator
tube with 1-2 ml of methylene chloride.
A 5-ml syringe is recommended for this
operation.
9.8 Add 1.0 ml toluene to the
concentrator tube, and a clean boiling
chip. Attach a two-ball micro-Snyder
column. Prewet the micro-Snyder
column by adding about 0.5 ml of
methylene chloride to the.top. Place this
micro-K-D apparatus on a water bath
(60-65°C) so that the concentrator tube
is partially immersed in the hot water.
Adjust the vertical position of the
apparatus and water temperature as
required to complete the concentration
in 5 to 10 minutes. At the proper rate of
distillation the balls will actively chatter
but the chambers will not flood. When
the apparent volume of liquid reaches
0.5 ml. remove the K-D apparatus and
allow it to drain for at least 10 minutes
while cooling. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with a small volume
of toluene. Adjust the final volume to 1.0
ml and stopper the concentrator tube.
and store refrigerated if further
processing will not be performed
immediately. Unless the sample is
known to require cleanup, proceed with
gas chromatographic analysis.
9.7 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. Record the
sample volume to the nearest 5 ml.
10. Cleanup and Separation.
10.1 Prepare a slurry of lOg of
activated Florisil in 10% methylene
chloride in hexane (V/V). Ose it to pack
a 10 mm ID chroma tographyl column.
gently tapping the column toisettle the
Florisil. Add 1 cm anhydrous'sodium
sulfate to the top of the Florisil.
10.1.1 Just prior to exposure of the
sodium sulfate layer to the air transfer
the 1 ml sample extract onto the column
using an additional 2 ml of toluene to
complete the transfer.
10.1.2 Just prior to exposure of the
sodium sulfate layer to the air, add 30 ml
10% methylene chloride in hexane and
continue the elution of the column.
Elution of the column should be at a rate
of about 2 ml per minute. Discard the
eluate from this fraction.
10.1.3 Next elute the column with 30
ml of 10% acetone/90% methylene
chloride (V/V) into a 500 ml K-D flask
equipped with a 10 ml concentrator
tube. Concentrate the collected fraction
by the K-D technique prescribed in 9.5
and 9.6, including the solvent exchange
to 1 ml toluene. This fraction should
contain the nitroaromatics and
isophorone.
10.1.4 Analyze by gas
chromatography.
11. Gas Chromatography.
11.1 Isophorone and nitrobenzene are
analyzed by injection of a portion of the
extract into a gas chromatograph with a
flame ionization detector. The
dinitrotoluenes are analyzed oy a
separate injection into an electron
capture gas chromatograph. Table I
summarizes some recommended gas
chromatographic column materials and
operating conditions for the instruments.
Included in this table are estimated
retention times and sensitivities that
should be achieved by this method.
Examples of the separations achieved
by the primary column are shown in
Figures 1 and 2. Calibrate the system
daily with a minimum of three injections
of calibration standards.
11.2 Inject 2-5 p.1 of the sample
extract using the solvent-flush
technique. Smaller (1.0 /il) volumes can
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 pi, and the resulting
peak size, in area units.
-------
69512 Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
11.3 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
11.4 If the peak area measurement is
prevented by the presence of
interferences, further cle'anup is
required.
12. Calculations.
12.1 Determine the concentration of
individual compounds according to the
formula:.
Where;
A =»Calibration factor for chromatographic
system, in nanograms material per area
unit.
B» Peak size in injection of sample extract, in
area units.
V,» Volume of extract injected (p.1).
Vt=Volume of total extract (jil).
V,»Volume of water extracted (ml).
12.2 Report results in micrograms per
liter without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
13. Accuracy and Precision.
The U.S. EPA Environmental
Monitoring and Support Laboratory in
Cincinnati is in the process of
conducting an in terla bora lory method
study to determine the accuracy and
precision of this test procedure.
Bibliography
"Development and Application of Test
Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 4-
Nitroaromaties-and Isophorone," Report for
EPA Contract No. 68-03-2624 (In
preparation).
Tabte L —Gas Chtomatognphy ol NitroaromaOcs
and laophtmno
Reunion Sma Detection torn.
Compound' (mm.) (jig/1)'
Coi 1' Cot 2» EC FID
laopnorona. 4 49 5.72 5
3.31 4.31 S
5.35 «.54 0.0«
ZWDtnilroioluana 3.52 4.75 0.06
'GaaOvom Q 80/100 maar) coatad witn 1.99% OF-1/
1.5% OV-17 packad in a 1 x V4" 00 glaaa column. FIO
«na)y»» for» and NB raquna nitrogan c«w gas *t 44 rat/mio
and SVC column Mmpmnun. EC vnlyra tar ft* ONT«
nqum 10% McttwiM/90% Argon earner ga« «t 44 ml/mm
flew r*l» and 14S*C column Mmpwaiuni.
'G^Chrom Q SO/100 m**n ocwtwj with 3% OV-101
p*ck«dlnilO> x 14" OO glas* column. FIO tntiysm o< IP and
NB raquna ntrogtn carrw ga» at 44 mt/mn flow raw and
100*C column Mmparatunt. EC anarywa for tna ONTj require*
10% Methana/fM% Argon camar- gas at 44 ml/mm flow rata
and 150'C column Mmparatuni.
•Oataetton limit • calcuMad from th* minimum datactabM
QC nnponaa bamg aqual to fiv* wnaa *• GC background
nma, assuming a 10 ml final votum* of tna 1 Mar Mmpta
axtraot. and aaaummg a GC irnaction of 5 microtWara.
BMXMO COM «fMO-01-M
-------
COLUMN: I.Sfi OV-17 + 1.95% QF-1 ON GAS CHROM Q
TEMPERATURE: 85°C.
DETECTOR: FUME ION12AT10N
L
02468
RETENTION TIME-MINUTES
Figure 1. Gas chromatogram of nitrobenzene and
isophorone
BILLING CODE S5SO-01-C
COLUMN: 1.5% OV-17+ 1.95% QF-1 ON GAS CHROM Q
TEMPERATURE: 14S'C.
DETECTOR: aECTRON CAPTURE
2 46 8 10
RETENTION TIME-MINUTES
*
iz
o
I
s?
CO
I
•o
D.
I
Figure 2. Gas chromatogram of dinitrotoluenes
-------
69514
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
Poiynudaar Aromatic Hydrocarbons-
Method 610
1. Scope and Application.
1.1 This method covers the
determination of certain polynuclear
aromatic hydrocarbons (PAH). The
following parameters may be
determined by this method:
8«nzoyran«
8mo
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules 69515
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them.
5.5 Acetonitrile—Spectral quality.
5.6 Silica gel—100/120 mesh
desiccant (Davison Chemical grade 923
or equivalent). Before use, activate for at
least 16 hours at 130* C in a foil covered
glass container.
6. Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest,
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 fig/1 in
the final extract for example, prepare
standards at 10 p.g/1 50 fig/1,100 fig/1,
500 fig/1, etc. so that injections of 1-5 >il
of each calibration standard will define
the linearity of the detector in the
working range.
6.2 Assemble the necessary HPLC or
gas chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table I
or II. By injecting calibration standards,
establish the sensitivity limit of the
detectors and the linear range of the
analytical systems for each compound.
6.3 Before using'any cleanup
procedure, the analyst must process a
series of calibration standards through
the procedure to validate elution
patterns and the absence of
interferences from the reagents.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate through
the analysis of a distilled water method
blank, that all glassware and reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
laboratory contamination.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt extists over the
identification of a peak on the
chromatogram, confirmatory techniques
such as fraction collection and GC-mass
spectroscopy should be used.
8. Sample Collection, Preservation,
and Handling.
8.1 Grab sample's must be collected
in glass containers. Conventional
sampling practices should be followed,
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, adjust the sample to a pH
range of 6.0-8.0 with sodium hydroxide
or sulfuric acid and add 35 mg sodium
thiosulfate per part per million of free
chlorine per liter.
8.3 All samples must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Check the pH of the
sample with wide-range pH paper and
adjust to within the range of 5-9 with
sodium hydroxide or sulfuric acid.
9.2 Add 60 ml methylene chloride to
the sample bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer the solvent into the separatory
funnel, and extract the sample by
shaking the funnel for two minutes with
periodic venting^to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
munimum of ten minutes. If the emulsion
inteface between layers is more than
one-third the size1 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 of the
emulsion through glass wool, or
centrifugation. Collect the methylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-rnl volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium sulfate, and collect it in a 500-ml
Kuderna-Oanish (K-D) flask equipped
with a 10-ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30-ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 clean boiling chips to
the 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 (60-85* C) so that the
concentrator tube is partially immersed
in the hot water, and the entire lower
rounded surface of the flask is bathed hi
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 apparatus volumn
of liquid reaches 1-ml, remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling. Remove
the Snyder column and rinse the flask
and its lower joint into the concentrator
tube with 1-2-ml of methylene chloride.
A 5-ml syringe is recommended for this
operation. Stopper the concentrator tube
and store refrigerated if further
processing will not be performed
immediately.
9.6 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000-ml graduated cylinder. Record the
sample volume to the nearest 5-mL
9.7 If the sample requires cleanup
before chromatographic analysis,
proceed to Section 10. If the sample does
not require cleanup, or if the need for
cleanup is unkown, analyze an aliquot
of the extract according to Section 11 or
Section 12.
10. Cleanup and Separation.
10.1 Before the silica gel cleanup
technique can be utilized, the extract
solvent must be exchanged to
cyclohexane. Add a 1-10-ml aliquot of
sample extract (in methylene chloride)
and a boiling chip to a clean K-D
concentrator tube. Add 4-ml
cyclohexane and attach a micro-Snyder
column. Prewet the micro-Snyder
column by adding 0.5-ml methylene
chloride to the top. Place the micro-K-D
apparatus on a boiling (100° C) 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
the liquid reaches 0,5-ml, remove K-D
-------
69516 Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
apparatus and allow it to drain for at
least 10 minutes while cooling. Remove
the micro-Snyder column and rinse its
lower joint into the concentrator tube
with a minimum of cyclohexane. Adjust
the extract volume to about 2-ml.
10.2 Silica Gel Column Cleanup for
PAHs.
10-2.1 Prepare a slurry of lOg
activated silical gel in methylene
chloride and place this in a 10 mm ID
chromatography column. Gently tap the
column to settle the silica gel and elute
the methylene chloride. Add 1-2 cm of
anhydrous sodium sulfate to the top of
the silica gel.
10.2.2 Preelute the column with 40-ml
pentane. 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 of
cyclohexane to complete the transfer.
10L2.3 Just prior to exposure of the
sodium sulfate layer to the air, add 25-
ml pentane and continue elution of the
column. Discard the pentane eluate.
10.2.4 Elute the column with 25-ml of
40% methylene chloride/60% pentane
and collect the eluate in a 500-ml K-D
flask equipped with a 10-ml
concentrator tube. Elution of the column
should be at a rate of about 2 ml/min.
10.2,5 Concentrate the collected
fraction to less than 10-ml by K-D
techniques as in 9.5, using pentane to
rinse the walls of the glassware. Proceed
with HPLC or gas chromatographic
analysis.
11. High Performance Liquid
Chromatography HPLC.
11.1 To the extract in the
concentrator tube, add 4 ml acetonitrile
and a new boiling chip, then attach a
micro-Snyder column. Increase the
temperature of the hot water bath to 95-
100* C. Concentrate the solvent as
above. After cooling, remove the micro-
Snyder column and rinse its lower joint
into the concentrator tube with about 0.2
ml acetonitrile. Adjust the extract
volume to 1.0 ml.
11.2 Table I summarizes the
recommended HPLC column materials
and operating conditions for the
instrument. Included in this table are
estimated retention times and
sensitivities that should be achieved by
this method. An example of the
separation achieved by this column is
shown in Figure 1. Calibrate the system
daily with a minimum of three injections
of calibration standards.
11.3 Inject 2-5 ui of the sample
extract with a high pressure syringe or
sample injection loop. Record the
volume injected to the nearest 0.05 jil,
and the resulting peak size, in area
units.
11.4 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
11.5 If the peak area measurement is
prevented by the pressure of
interference, further cleanup is required.
11.8 The UV detector is
recommended for the determination of
napthalene and acenaphthylene and the
fluorescene detector is recommended for
the remaining PAHs.
12. Gas Chromatography.
12.1 The gas chromatographic
procedure will not resolve certain
isomeric pairs as indicated in Table DL
The liquid chromatographic procedure
(Section 11] must be used for these
materials.
12.2 To achieve maximum sensitivity
with this method, the extract must be
concentrated to 1.0 ml. Add a clean
boiling^chip to the methylene chloride
extract in the concentrator tube. Attach
a two-ball micro-Snyder column. Prewet
the micro-Snyder column by adding
about 0.5 ml of methylene chloride to the
top, Place this micro-K-D apparatus on a
hot water bath [60-65* C) so that the
concentrator tube is partially immersed
in the hot water. Adjust the vertical
position of the apparatus and water
temperature as required to complete the
concentration in 5 to 10 minutes. At the
proper rate of distillation the balls will
actively chatter but the chambers will
not flood. When the apparent volume of
liquid reaches 0.5 ml, remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling. Remove
the micro-Snyder column and rinse its
lower joint into the concentrator tube
with a small volume of methylene
chloride. Adjust the final volume to 1.0
ml and stopper the concentrator tube.
12.3 Table H describes the
recommended gas chromatographic
column material and operating
conditions for the instrument. Included
in this table are estimated retention
times that should be achieved by this
method. Calibrate the gas
chromatographic system daily with a
minimum of three injections of
calibration standards.
12.4 Inject 2-5 ul of the sample
extract using the solvent-flush
technique. Smaller (1.0 >il} volumes can
be Injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 u-1, and the resulting
peak size, in ara units.
12.5 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
12.6 If the peak ara measurement is
prevented by the presence of
interferences, further cleanup is
required.
13. Calculations.
13.1 Determine the concentration of
individual compounds according to the
formula:
Concentration, ug/1 —
(VJ(VJ
Where:
A=Calibration factor for chromatographic
system, in nanograms material per area
unit
B=-Peak size in injection of sample extract in
area units
V) =a Volume of extract injected (pi)
Vt=Volume of total extract foil)
V,=Volume of water extracted (ml)
13.2 Report results in micrograms per
liter without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
14. Accuracy and Precision,
14.1- The U.S. EPA Environmental
Monitoring and Support Laboratory in
Cincinnati is in the process of
conducting an interlaboratory method
study to determine the accuracy and
precision of this test procedure.
Bibliography
"Development and Application of Test
Procedures for Specific Organic Toxic
Substances in Wastewaters, Category 9-
PAHs." Report for EPA Contract 8S-03-26Z4
(In preparation).
Tabta \.—High Performance Liquid Chromatography
otPAH's
Compound ' Retention Detection Hfnt (pg/l)'
time (mint
UV
N«phm«U>n»
Acenaphthylene
Acenapnthene
PIlMVMOA .
Phenanthrene
Anthracene
Ruoraflthene. „,
Pyrene......... ™.™
SenZO(a)aiiUWACOnem...
Chrysene *.__
Benzo(b)fluoranthene ._
Senzo(kjfluoranthene._
Benzo(a)pyrene ..._
Dibenzce
1 liter sample extract, and assuming an HPLC injection of
Table IL— Gas Chromatography of PAHs
Compound1
Retention
Time (mm)
Naphthalene..
Acenapnthytene..
Acanaphthene
Ruor«ne_____
4.5
10.4
10.3
12.6
-------
Federal Register 7 Vol. 44. No. 233 / Monday. December 3, 1979 / Proposed Rules
69517
TaM* It—Ota Chromatograpty
Compound'
FMwtfon
•nm*(irin)
Phenanthrene,
Aiwv
FtuaranOMM.
Pyran* —
CXryMM
OiM
1SJ
194
20.»
».e
24.7
29.4
364
3M
'QC oondWonc QmmMorta W-AW-OCM* 100/120 nwMl
CMMd «Wi 3% OV-17. pKted in « V x 2 mm 10 gtew
column, «Mt nttogwi cwMr ga « 40 ml/mbr tow n«.
Cotann Kmptnkirt IM* n*u a 100* C far 4 n*Mm Him
programmed « r/mlnuM to > Ik* how « 280- 0.
Haloethen—Method 611
1. Scope and Application
1.1 This method covers the
determination of certain haloethen. The
following parameters may be
determined by this method:
Bfe<2-cNanMfty« Vim.
-8rameplMnyl
4-CNo
STOfVTHot
34273
3427S
34213
34638
34641
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of finding the specific
compounds of interest If the user is
attempting to screen samples for any or
ail of the4 compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
usually dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection listed
in Table I represent sensitivities, that
can be achieved in wastewaters in the
absence of interferences.
1.4 This method is recommended for
use only by experienced residue
analysts or under the close supervision
of such qualified persons.
2. Summary of Method
2.1 A 1-liter sample of wastewater is
extracted with methylene chloride using
sepaxatory funnel techniques. The
extract is dried and concentrated to a
volume of 10 ml or less.
Chromatographic conditions utilizing a
halide specific detector are described
which allow for the accurate
measurement of the compounds in the
extract.
12 If interferences are encountered,
the method provides a selected general
COLUMN: HC-OOS S1L-X g
MOBILE PHASE' 40% TO 100% ACETONITR1LE IN VKATEB 3
DETECTOR: FLUORESCBICS S
8 12 16 20 24 28 32 38 40
RETENTION TIME-MINUTES
Rgure 1. Liquid chromatogram of polynuclear aromatics
-------
69518 Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 /Proposed Rules
purpose cleanup procedure to aid the
analyst in their elimination.
3. Interferences.
3.1 Solvents, reagents, glassware, and
other sample processing hardware may
yield discrete artificats 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 running
method blanks. Specific selection of
reagents and purification of solvents by
distillation in all-glass systems may be
required.
3.2 Interferences coextracted from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled. While
general clean-up techniques are
provided as part of this method, unique
samples may require additional cleanup
approaches to achieve the sensitivities
stated in Table I.
3.3 Oichlorobenzenes are known to
coelute with haloethers under some gas
chromatographic conditions. If these
materials are present together in a
Sample, it may be necessary to analyze
the extract with two different column
packings to completely resolve all of the
compounds.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or 1-quart volume. French
or Boston Round design is
recommended. The container must be
washed and solvent rinsed before tise to
minimize interferences.
4.1.2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon. Foil may be
substituted if sample in not corrosive.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum of 250 ml. Sample containers
must be kept refrigerated during
sampling. No tygon or rubber tubing
may be used in the system.
4.2 Separatory funnel—2000 ml, with
Teflon stopcock.
4.3 Drying column—20 mm ID pyrex
chromatographic column with coarse
frit.
4.4 Kuderna-Danish (K-D)
Apparatus
4.4.1 Concentrator tube—10 ml,
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked. Ground glass stopper (size l%z
joint) is used to prevent ecaporation of
extracts.
4.4.2 Evaporative flask—500 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-O012).
4.4.3 Snyder column—three ball
macro (Kontes K503000-0121 or
equivalent).
4.4.4 Snyder column—two-ball micro
(Kontes K-569001-0219 or equivalent).
4.4.S Boiling chips—solvent
extracted, approximately *%o mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (±Z'C], The bath
should be used in a hood.
4.0 Gas chromatograph—Analytical
system complete with gas
chromatograph suitable for on-column
injection and all required accessories
including haiide specific detector,
column supplies, recorder, gases,
syringes. A data system for measuring
peak areas is recommended.
4.7 Chromatographic Column—400
mm long x 19 mm ID with coarse fritted
plate on bottom and Teflon stopcock
(Kontes K-420540-0224 or equivalent).
5. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—{ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid (1 +1}—(ACS) Mix
equal volumes of cone. H,SO< with
distilled water.
5.2 Methylene chloride—Pesticide
quality or equivalent
. 5.3 Sodium Sulfate—(ACS) Granular^
anhydrous (purified by heating at 400*C
for 4 hrs. in a shallow tray).
5°A Stock standards—Prepare stock
standard solutions at a concentration of
1.00 ;ig/ud by dissolving 0.100 grams of
assayed reference material in pesticide
quality acetone or other appropriate
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them.
5.5 Florisil—PR Grade (60/100
mesh); purchase activated at 1250T and
store in the dark in glass containers with
glass stoppers or foil-lined screw caps.
Before use, activate each batch
overnight at 130*C in a foil-covered glass
container.
5.6 Hexane, Petroleum ether (boiling
range 30-60°C)—pesticide quality or
equivalent.
5.7 Diethyl Ether—Nanograde,
redistilled in glass, if necessary.
5.7.1 Must be free of peroxides as
indicated by EM Quant test strips. (Test
strips are available from EM
Laboratories, Inc., 500 Executive Blvd.,
Elmsford, N.Y. 10523.)
5.7.2 Procedures recommended for
removal of peroxides are provided with
the test strips. After cleanup 20 ml ethyl
alcohol preservative must be added to
each liter of ether.
6. Calibration.
8.1 Prepare calibration standards
that contain the compounds of interest,
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 pg/1 in
the final extract for example, prepare
standards at 10 fig/1, 50 fig/l 100 jtg/1.
SCO ju.g/1. etc. so that injections of 1-5
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules 69519
identification of a peak on the
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection, Preservation.
and Handling.
8.1 Crab samples must be collected
in glass containers. Conventional
sampling practices should be followed,
except that the bottle must not be
prewashed with sample before
collection. Composite samples should be
collected in refrigerated glass containers
in accordance with the requirements of
the program. Automatic sampling
equipment must be free of tygon and
other potential sources of
contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, the sample should be
adjusted to a pH^nge of 6.0-8.0 with
sodium hydroxide or sulfuric acid.
8.3 All samples; must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Check the pH of the
sample with wide-range pH paper and
adjust to within the range of S-Q with
sodium hydroxide or sulfuric acid.
9.2 Add 60 ml methylene chloride to
(he-sample bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer the solvent into the separatory
funnel, and extract the sample by
shaking the funnel for two minutes with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
minimum, of ten 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, but may
include stirring, filtration of the
emulsion through glass wool, or
centrifugation. Collect the methylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium sulfate, and collect it in a 500-ml
Kuderna-Oanish (K-D) flask equipped
with a. 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30 ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 clean boiling chips to
the 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 (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 1-2 ml, remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling.
Note.—Haloethers have a sufficiently high
volatility that significant losses will occur in
concentration steps if care is not exercised. It
is important to maintain a constant gentie
evaporation rate and not to allow the liquid
volume to fall below 1-2 ml before removing
the K-D from the hot water bath.
9.6 Momentarily remove the Snyder
column, add 50 ml hexane and a new
boiling chip and replace the column.
Raise the temperature of the water bath
to 85-90°C. Concentrate thr extract as in
9.5 except use hexane to prewet the
column. Remove the Snyder column and
rinse the flask and its lower joint into
the concentrator tube with 1-2 ml
hexane. Stopper the concentrator tube
and store refrigerated jf further
processing will not be performed
immediately.
9.7 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. Record the
sample volume to the nearest 5 ml.
9.8 Unless the sample is known to
require cleanup, proceed to analysis by
gas chroma tography.
10. Cleanup and Separation.
10.1 Florisil Column Cleanup for
Haloethers.
10.1.1 Adjust the sample extract
volume to 10 ml.
10.1.2 Place a charge (nominally 20 g
but determined in Section 6.3) of
activated Florisil in a 19 mm ID
chroma tography column. After settling
the Florisil by tapping column, add
about one-half inch layer of anhydrous
granular sodium sulfate to the top.
10.1.3 Pre-elute the column, after
cooling, with 50-60 ml of petroleum
ether. Discard the eluate and just prior
to exposure of the sulfate layer to air,
quantitatively transfer the sample
extract into the column by decantation
and subsequent petroleum ether
washings. Discard the eluate. Just prior
to exposure of the sodium sulfate layer
to the air, begin eluting the column with
300 ml of 6% ethyl ether/94% petroleum
ether. Adjust the elution rate to
approximately 5 ml/min and collect the
eluate in a 500 ml K-D flask equipped
with- a 10 ml concentrator tube. This
fraction should contain all of the
haloethers.
10.1.4 Concentrate the fraction by K-
D as in 9.5 except prewet the Snyder
column with hexane. When the
apparatus is cqpl, remove the column
and rinse the flask and its lower joint
into the concentrator tube with 1-2 ml
hexane. Analyze by gas
chromatography.
11. Gas Chromatography.
11.1 Table I summarizes some
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method.,Examples of
the separations achieved by these
columns are shown in Figures 1 and 2.
Calibrate the system daily with a
minimum- of three injections of
calibration standards.
11.2 Inject 2-5 uJ of ths sample
extract using the solvent-flush
technique. Smaller (1.0 fil) volumes can
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 p,l, and the.resulting
peak size, in area units.
11.3 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
11.4 ' If the peak area measurement is
prevented by the presence of
interferences, further cleanup is
required.
12. Calculations.
12.1 Determine the concentration of
individual compounds according to the
formula:
Concentrate.^/'- W™™
Where:
A = Calibration factor for chromatographic
system, in nanograms material per area-
unit.
B = Peak size tn injection of sample extract, in
area units
V, = volume of extract injected (jxl)
V, = volume of total extract (^1)
V, = volume of water extracted (ml)
-------
69520 Federal Register / Vol. 44. No. 233 / Monday. December 3, 1979 / Proposed Rules
12.2 Report results in micrograms per
liter without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported, •
13. Accuracy and Precision. The U.S.
EPA Environmental Monitoring and
Support Laboratory in Cincinnati is in
the process of conducting an
interiaboratory method study to
determine the accuracy and precision of
this test procedure.
Bibliography
1. "Development and Application of Test
Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 2-
Haloethers." Report for EPA Contract 88-03-
2833 (In preparation).
2. Mills, P. AM "Variation of Florisil
Activity. Simple Method for Measuring
Absorbent Capacity and Its Use in
Standardizing Florisil Columns." Journal of
the Association of Official Analytical
Chemists. SI. 29 (1968).
Tub* I—Gt$ Chfomuognphy ofH»to9th*»
(mm.) OMKdon
Compound fen*
(ug/L)'
Cot i' CoU2'
Stl(2-GhloioMpR]py4 «nv..» 8.41 9.70 0.9
a*e-cNom**iQ torn 9J2 9.08 0.5
Blxa 2.1 mm O gttn column wtti unn>
Ngn purity Mkm camir/gH
-------
COLUMN: 3!i SP-1000 ON SUPELCOPORT
PROGRAM: 60*C-2 MINUTES 8"/MINUTE TO 230*C.
DETECTOR: HALL ELECTROLYTIC CONDUCTIVITY
COLUMN: TENAX GC
PROGRAM: 150*C.-4 MINUTES IS'/MINUTE TO 310*C.
DETECTOR: HALL ELECTROLYTIC CONDUCTIVITY
ee
UJ
5
UJ
2"
a.
O
UJ g
2
UJ
O
i
o
—t
9
CM
DT
5
i
A
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I I
UJ
UJ
JJJ
s
K1"
UJ
i
0
5
CSI
£
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V
11 1 11
) 2 4 6 8 10
1LI
<
£1
UJ
*
sf
g
UJ
o
C£
O
^
o
rsi
i2
ca
1
Jl
t A
12 14
cc
UJ
£
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1
.j
2
UJ
E
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o
X
o
1
/v_
t i
16 18
UJ
S jg
uj 5
-j Q
2 <
UJ
j£
j^
2
1
a,
.,
n
i
Ju
s.
1 L
20 22 24
RETENTION TIME-MINUTES
Figure 1. Gas chromatogram of haloethers
p
I
s
i?
8
I
_1_
04s 12 16 20
RETENTION TIME-MINUTES
Figure 2. Gas chromatogram of haloethers
24
-------
69522 Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
Chlorinated Hydrocarbons—Method 612
\. Scope and Application.
1.1 This method covers the
determination of certain chlorinated
hydrocarbons. The following parameters
may be determined by this method.
STOnETNo.
34388
HotachlorocyclopwiUiMn*...
VMxKMonXwnm*
HoxacMaroDutadiwi*..
1 .2-OfchkyoMnzwM .....
\ ,3-OteNorolMremw -----------
1 , 4-OJcWorob«n»n» --------------
2-cntoron»pMh»)«n« -------------
39700
34391
34396-
34636
34551
34566
94571
1*561
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutant Discharge Elimination System
(NPDES). As such, it presupposes a high
expectation of rinding the specific
compounds of interest. If the user is
attempting to screen samples for any or
ail of the compounds above, he must
develop independent protocols for the
verification of identity.
1.3 The sensitivity of this method is
ususally dependent upon the level of
interferences rather than instrumental
limitations. The limits of detection li-ted
in Table I represent sensitivities that
can be achieved in waste-waters ir the
absence of inteferences.
1.4 This methodis recommended for
use only by experienced resid- c.
analysts or under the close f pervision
of such qualified persons.
2. Summary of Method.
2.1 A 1-liter sample of wastewater is
extracted with methylene chloride using
separatory funnel techniques. The
extract is dried by passing through a
sodium sulfate column and concentrated
to a volume of l&ml or less.
Chromatographic conditions are
described which allow-for the accurate
measurement of the compounds in the
extract.
2.2 If inteferences are encountered
or expected, the method provides a
selected general purpose cleanup
procedure to aid the analyst in their
elimination.
3. Interferences.
3.1 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
inteferences under the conditions of the
analysis by running method blanks.
Specific selection of reagents and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextracted from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled. While
general clean-up techniques are
provided as part of this method, unique
samples-may require, additional cleanup
approaches to achieve the sensitivities
states in Table 1.
4, Apparatus and-Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter or 1-quart volume. French
or Boston Round design is
recommended. The container must be
washed and solvent rinsed before use to
minimize interferences.
4.1 2 Bottle caps—Threaded to screw
on to the sample bottles. Caps must be
lined with Teflon. Foil may be
substituted if sample is not corrosive
and the foil is found to be interference
free.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum of 250 ml. Sample containers
must be kept refrigerated during
sampling. No tygon or. rubber tubing
may be used in the system.
4.2 Separatory funnel—2000 ml, with
Teflon stopcock.
4.3 Drying column—20 mm ID pyrex
chromatographic column with coarse
frit.
4.4 Kuderna-Danish (K-D)
Apparatus
• 4.4.1 Concentrator tube—10 ml,
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked. Ground glass stopper (size 19/
22 joint) is used to prevent evaporation
of extracts.
4.4.2 Evaporative flask—500 ml
(Kontes K-57001-0500 or equivalent).
Attach to concentrator tube with
springs. (Kontes K-662750-0012).
4.4.3 Snyder column—three-ball
macro (Kontes K503000-0121 or
equivalent);
4.4.4 Snyder column—two-ball micro
(Kontes K-569001-0219 or equivalent).
4.4.5 Boiling chips—solvent
extracted, approximately 10/40 mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (±2° C). The bath
should be used in a hood.
4.8 Gas chrqmatograph—Analytical
system complete with gas
chromatograph suitable for on-column
injection and all required accessories
including electron capture detector,
column supplies, recorder, gases,
syringes. A data system for measuring
peak areas is recommended.
4.7 Chromatography column—300
mm long x 10 mm ID with coarse fritted
disc at bottom and Teflon stopcock.
5. Reagents.
5.1 Preservatives:
5.1.1 Sodium hydroxide—(ACS) 10 N
in distilled water.
5.1.2 Sulfuric acid—(ACS) Mix equal
volumes-of cone. H,SO» with distilled
water.
5.2 Methylene chloride, Hexane and
Petroleum ether (boiling range 30-
80°C)—Pesticide quality orequivalenL
5.3 Sodium sulfate—(ACS) Granular,
anhydrous (purified by heating at 400*C
for4 hrs. in a shallow tray).
5.4 Stock standards—Prepare stock
standard solutions at a concentration of
1.00 /ig/ul by dissolving 0.100 grams of
assayed reference material in pesticide
quality isooctane or other appropriate
solvent and diluting to volume in a 100
ml ground glass stoppered volumetric
flask. The stock solution is transferred
to ground glass stoppered reagent
bottles, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them.
5.5 Florisil—PR grade (60/100 mesh);
purchase activated at 1250T and store
in the dark in glass containers with glass
stoppers or foil-lined screw caps. Before
use, activate each batch at 130'C in foil-
covered glass containers.
8. Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest,
either singly or mixed together. The
standards should be prepared at
concentrations covering two or more
orders of magnitude that will completely
bracket the working range of the
chromatographic system. If the
sensitivity of the detection system can
be calculated from Table I as 100 fxg/1
in the final extract, for example, prepare
standards at 10 jig/1. 50 /ig/1,100 jxg/1.
500 fig/1, etc. so that injections of 1-5 p.1
of each calibration standard will define
the linearity of the detector in the
working range.
6.2 Assemble the necessary gas
chromatographic apparatus and
establish operating parameters
equivalent to those indicated in Table I.
By injecting calibration standards,
establish the sensitivity limit of the
detector and the linear range of the
analytical system for each compound.
8.3 The cleanup procedure in Section
10 utilizes Florisil chromatography.
Florisil from different batches or sources
may vary in absorption capacity. To
standardize the amount of Florisil which
is used, the use of lauric acid value
(Mills, 1968) is suggested. The
referenced procedure determines the
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules 69523
adsorption from hexane solution of
lauric acid (mgj per gram Florisil. The
amount of Florisil to be used for each
column is calculated by dividing this
ratio by 110 and multiplying by 20
grams.
6.4 Before using any cleanup
procedure, the analyst must process a
series of calibration standards through
the procedure to validate elution
patterns and the absence of
interferences from the reagents.
7. Quality Control.
7.1 Before processing any samples,
the analyst should demonstrate through
the analysis of a distilled water method
blank, that all glassware and reagents
are interference-free. Each time a set of
samples is extracted or there is a change
in reagents, a method blank should be
processed as a safeguard against
chronic laboratory contamination.
7.2 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
Where doubt exists over the
identification of a peak on the
chromatogram, confirmatory techniques
such as mass spectroscopy should be
used.
8. Sample Collection. Preservation.
and Handling.
8.1 Grab samples must be collected
in glass containers, leaving a minimum
headspace. Conventional sampling
practices should be followed, except
that the bottle must not be prewashed
with sample before collection.
Composite samples should be collected
in refrigerated glass containers in
accordance with the requirements of the
program. Automatic sampling equipment
must be free of tygon and other potential
sources of contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, the sample should be
adjusted to a pH range of 6.0-8.0 -with
sodium hydroxide or sulfuric acid.
8.3 All samples should be extracted
immediately and must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction.
9.1 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory runnel. Check the pH of the
sample with wide-range pH paper and
adjust to within the range of 5-9 with
sodium hydroxide or sulfuric acid.
9.2 Add 60 ml methylene chloride to
the sample bottle, seal, and shake 30
seconds to rinse the inner walls.
Transfer the solvent into the separatory
funnel, and extract the sample by
shaking the funnel for two minutes with
periodic venting to release vapor
pressure. Allow the organic layer to
separate from the water phase for a
minimum of ten 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, but may
include stirring, filtration of the
emulsion through glass wool, or
centrifugation. Collect the methylene
chloride extract in a 250-ml Erlenmeyer
flask.
9.3 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.4 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium suifate, and collect it in a 500-ml
Kuderna-Danish (K-D) flask equipped
with a 10 ml concentrator tube. Rinse
the Erlenmeyer flask and column with
20-30 ml methylene chloride to complete
the quantitative transfer.
9.5 Add 1-2 clean boiling chips to the
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 (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 1-2 ml, remove the K-D
apparatus and allow it to drain for at
least 10 minutes while cooling.
Note.—The dichlorobenzenes have a
sufficiently high volatility that significant
losses may occur in concentration steps if
care is not exercised. It is important to
maintain a constant gentle evaporation rate
and not to allow the liquid volume to fall
below 1-2 ml before removing the K-D from
the hot water bath.
9.6 Momentarily remove the Snyder
column, add 50 ml hexane and a new
boiling chip and -jplace the column.
Raise the temperature of the water bath
to 85-90" C. Concentrate the extract as
in 9.5, except using hexane to prewet the
column. Remove the Snyder column and
rinse.the flask and its lower joint into
the concentrator tube with 1-2 ml of
hexanei A 5-ml syringe is recommended
for this operation. Stopper the
concentrator tube and store refrigerated
if further processing will not be
performed immediately.
9.7 Determine the original sample-
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1000 ml graduated cylinder. Record the
sample volume to the nearest 5 ml.
9.8 Unless the sample is known to
require cleanup, proceed to analysis by
gas chroma tography.
10. Cleanup and Separation.
10.1 Florisil column cleanup for
chlorinated Hydro-carbons.
10.1.1 Adjust the sample extract to
10ml.
10.1.2 Place a 12 gram charge of
activated Florisil (see 6.3) in a 10 mm ID
chromatography column. After settling
the Florisil by tapping the column, add a
1-2 cm layer of anhydrous granular
sodium suifate to the top.
10.1.3 Pre-elute the column, after
cooling, with 100 ml of petroleum ether.
Discard the eluate and just prior to
exposure of the suifate layer to air,
quantitatively transfer the sample
extract into the column'by decantation
and subsequent petroleum ether
washings. Discard the eluate. Just prior
to exposure of the sodium suifate layer
to the air, begin eluting the column with
200 ml petroleum ether and collect the
eluate hi a 500 ml K-D flask equipped
with a 10 ml concentrator tube. This
fraction should contain all of the
chlorinated hydrocarbons.
10.1.4 Concentrate the fraction by K-
D as in 9.5 except prewet the column
with hexane. When the apparatus is
cool, remove the Snyder column and
rinse the flask and its lower joint into
the concentrator tube with 1-2 ml
hexane. Analyze by gas
chromatography.
11. Gas Chromatography.
11.1 Table I summarizes the
recommended gas chromatographic
column materials and operating
conditions for the instrument. Included
in this table are estimated retention
times and sensitivities that should be
achieved by this method. Examples of
the separations- achieved by this column
are shown in Figures 1 and 2. Calibrate
the system daily with a minimum of
three injections of calibration standards.
11.2 Inject 2-5 ul of the sample
extract using the solvent-flush
technique. Smaller (1.0 ul) volumes can
-------
69524 Federal Register / Vol. 44. No. 233./ Monday, December 3. 1979 / Proposed Rules
be injected if automatic devices are
employed. Record the volume injected to
the nearest 0.05 ul, and the resulting
peak size, in area units.
11.3 If the peak area exceeds the
linear range of the system, dilute the
extract and reanalyze.
11.4 If the peak area measurement is
prevented by the presence of
interferences, further cleanup is
required.
12. Calculations.
12.1 Determine the concentration of
individual compounds according to the
formula:
Concentrwon, «/.. 'AMBi'V')
(VJ(VJ
Where:
A = Calibration factor for chromatographic
system, in nanograms material per area
unit.
8 = Peak size in injection of sample extract, in
area units
V, = Volume of extract injected (ul)
V,=Volume of total extract (ul]
V, = Volume of water extracted (ml]
12.2 Report results in micrograms per
liter without correction, for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
13. Accuracy and Precision. The U.S.
EPA Environmental Monitoring and
Support Laboratory in Cincinnati is in
the process of conducting an
interlaboratory method study to
determine the accuracy and precision of
this test procedure.
Bibliography
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 63-03-
2625 (In preparation).
2. Mills. P. A., "Variation of Flonsil
Activity; Simple Method for Measuring
Absorbent Capacity and Its Use in\
Standardizing Florisil Columns," Journal of
the Association of Official Analytical
Chemists, 51.29 (1968).
Tatoto L—Gas Chromatograptty of Cfilormatad
Hydrocarbons
Retention
time (mm.) Detection
Compound col 1 ' limit (ng/0 *
1 ,3-dKhkxoberttene
1 ,4^Jn:hlorooenzene
1 ,2-dichlorobenzene
Hexachkxobutadiens
1 ,2.4-«tehtorceeraene „
2-chloronapnthalena
Hexacnlorobenzene
4.0
4.3
48
5J
11 8
12.4
•1.5
•2.5
•70
O.OOA
0.01 a
0.001
0012
0.001
0.006
O.OOT
0.015
0001
'Ow CtifonrQ 30/100 meed coated vnlri 1.5% OV-1/
1 5% OV-22S packed m a 1 3 m long x 2 mm !O gfus
column with 5V Methane/95% Argon earner gat at 30 ml/
mm flow rate. Column temperature a 75" C except where '
indicate*. 160* C. Under these condmone R.T. of Alarm a 18.8
minutes at 160* C.
' Detection limit is calculated from the minimum detectaola
GC response of the electron capture detector being, equal to
five om« the GC background none, assuming a 10 TH final
volume of the 1 liter sample extract, and assuming a QC «v
lection of 5 microttters.
BILLING COOE SWO-01-M
-------
COLUMN: 1.5%OV-1
TEMPERATURE: 75*C.
DETECTOR: aECTRON CAPTURE
1.5% OV-225 ON GAS CHROM Q
03
i
o
CM
2
Q
0.
§
O
O
x
COLUMN: 1.5%OV-H
1.5% OV-225 ON GAS CHROM Q
TEMPERATURE: 160°C.
DETECTOR: ELECTRON CAPTURE
II
LU
LU
o
§
<
X
LU
J.
I 1
8 12 16
RETENTION TIME-MINUTES
20
8 12 16
RETENTION TIME-MINUTES
I
SO
o
i
Figure 2. Gas chromatogram of chlorinated hydrocarbons
Figure 1. Gas chromatogram of chlorinated hydrocarbons
s
Qu
0)
a
a>
o
(D
a"
CD
CO
VI
CO
o
•o
o
CO
(B
a.
to"
tfl
en
CO
-------
69532
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
Purgeables—Method 624
\. Scope and Application.
l.l This method is designed to
determine volatile organic materi-als that
are amenable to the purge and trap
method. The parameters listed in'Table
1 may be determined by this method.
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutants Discharge Elimination System
(NPDES).
1.3 The detection limit of this method
is usually dependent upon the level_of
interferences rather than instrumental
limitations. The limits listed in Table 2
represent sensitivities that can be
achieved in wastewaters.
1.4 The GC/MS parts of this method
are recommended for use only by
persons experienced in GC/MS analysis
or under the close supervision of such
qualified persons.
1.5 The trapping and chromatographic
procedures described do not apply to
the very volatile pollutant,
dichlorodifluoromethane. An alternative
three stage trap containing charcoal is to
be used if this compound is to be
analyzed. See EPA Method 601 and
Reference 1. Primary ion for quantitative
analysis of this compound is 101. The
secondary ions are 85, 87, and 103.
1.6 Although this method can be
used for measuring acrolein and
acrylonitrile, the purging efficiencies are
low and erratic. For a more reliable
quantitative analysis of these
compounds, use direct aqueous injection
(Ref. 4-6) or EPA Method 603, Acrolein
and Acrylonitrile, EMSL, Cincinnati,
Ohio.
2. Summary of Method.
21. A sample of wastewater is
purged with a stream of inert gas. The
gas is bubbled through a 5 ml water
sample contained in a specially
designed purging chamber. The volatile
organics are efficiently transferred from
the aqueous phase into the gaseous
phase where they are passed through a
sorbent bed designed to trap out the
organic volatiles. After purging is
complete, the trap is backflushed while
being rapidly heated in order to
thermally desorb the components into
the inlet of a gas chromatograph. 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 spectrometer operating
parameters are given.
3. Interferences.
3.1 Interferences coextracted from
the samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled. Impurities
in the purge gas and 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 interferences under the conditions
of the analysis by running method
blanks. Method blanks are run by
charging the purging device with
organic-free water and analyzing it in a
normal manner. The use of non-TFE
plastic tubing, non-TFE thread sealants.
or flow controllers"with rubber
components in the purging device should
be avoided.
3.2 Samples can be contaminated by
diffusion of volatile organics
(particularly methylene chloride)
through the septum seal into the sample
during shipment and storage. A field
blank prepared from organic-free 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 level and low level
samples are sequentially analyzed. To
reduce cross contamination, it is
recommended that the purging device
and sample syringe be rinsed out twice,
between samples, with organic-free
water. Whenever an. unusually
concentrated sample is encountered, it
should be followed by an analysis of
organic-free water to check for cross-
contamination. For samples containing
Urge amounts of water'soluble
materials, suspended solids,'high boiling
compounds, or high organohalide levels,
it may be necessary to wash out the
purging device with a soap solution,
rinse with distilled water, and then dry
in a 105°C oven between analyses.
4, Apparatus and Materials.
4.1. Sampling equipment, for discrete
sampling.
4.1.1 Vial, with cap—40 ml capacity
screw cap (Pierce #13075 or equivalent).
Detergent wash and dry vial at 105°C for
one hour before use.
4.1.2 Septum—Teflon-faced silicone
(Pierce #12722 or equivalent). Detergent
wash and dry at 105°C for one hour
before use.
4.2 Purge and trap device—The
purge and trap equipment consists of
three separate pieces of apparatus: a
purging device, a trap, and a desorber.
The complete device is available
commercially from several vendors or
can be constructed in the laboratory
according to the specifications of Bellar
and Lichtenberg (Ref. 2,3). The sorbent
trap consists of V» in. O.D. (0.105 in. ID.)
x 25 cm long stainless steel tubing
packed with 15 cm of Tenax-GC (60-80
mesh) and 8 cm of Davison Type-15
silica gel (36-60 mesh). See figures 1
through 4. Ten centimeter traps may be
used providing that the recoveries are
comparable to the 25 cm traps.
4.3 Gas chromatograph—Analytical
system complete with a temperature
programmable gas chromatograph
suitable for on-column injection and all
required accessories including an
analytical column.
4.3.1 Column 1—An 8 ft. stainless
steel column (Vs in. OD.x 0.90 to 0.105 in.
ID) packed with 1% SP-1000 coated on
60/80 mesh Carbopack B preceded by a
5-cm precolumn packed with 155 SP-1000
coated on 60/80 mesh Chromosorb W. A
glass column (1A in OD x 2 mm IDT may
be substituted. The precolumn is
necessary only during conditioning.
4.3.2 Column 2—An 8 ft. stainless
steel column (Vs in OD x 0.09 to 0.105 in.
ID) packed with 0.2% Carbowax 1500
coated on 80/80 mesh Carbopack C
preceded by a 1 ft. stainless steel
column (Va in. OD x 0.09 to 0.105 in. ID)
packed with 3% Carbowax 1500 coated
on 60/80 mesh Chromosorb W. A glass
column (V* in. OD x 2 mm ID) may be
substituted. The precolumn is necessary
only during conditioning.
4.4 Syringes—glass, 5-ml hypodermic
with Luer-Lok tip (3 each).
4.5 Micro syringes—10, 25,100 /j.1.
4.6 2-way syringe valve with Luer
ends (3 each; Teflon or Kel-F).
4.7 Syringe—5 ml gas-tight with shut-
off valve.
4.8 8-inch, 20-gauge syringe needle—
One per each 5-ml syringe.
4.9 Mass Spectrometer—capable of
scanning from 20-260 in six seconds or
less at 70 volts (nominal), and producing
a recognizable mass spectrum at unit
resolution from 50 ng of DFTPP when
injected through the GC inlet. The mass
spectrometer must-be interfaced with a
gas chromatograph equipped with an
all-glass, on-column injector system
designed for packed column analysis.
All sections of the transfer lines must be
glass or glass-lined and deactivated. Use
Sylon-CT, Supelco, (or equivalent) to
deactivate. The GC/MS interface can
utilize any separator that gives
recognizable mass spectra (background
corrected) and acceptable calibration
points at the limit of detection specified
for each compound in Table 2.
4.10 A computer system should be
interfaced to the mass spectrometer to
allow acquisition of continuous mass
scans for the duration of the
chromatographic program. The computer
system should also be equipped with
mass storage devices for saving all data
from GC-MS runs. There must be
-------
Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 /Proposed Rules 69533
computer software available to allow
searching any GC/MS run for specific
ions and plotting the intensity of the
ions with respect to time or scan
number. The ability to integrate the area
under a specific ion plot peak is
essential for quantification.
5. Reagents.
5.1 Sodium thiosulfate—(ACS)
Granular.
5.2 Trap Materials
5.2.1 Porous polymer packing 60/80
mesh chromatographic grade Tenax GC
(2,6-diphenylene oxide).
5.2.3 Silica gel-(35-60 mesh)—
5.2.2 Three percent OV-1 on
Chromosorb-W 60/80 mesh. Davison,
grade-15 or equivalent.
5.3 Activated carbon—Filtrasorb-200
(Calgon Corp.) or equivalent.
5.4 Organic-free water
5.4.1 Organic-free water is defined
as water free of interference when
employed in the purge and trap
procedure described herein. It is
generated by passing tap water or well
water through a carbon filter bed
containing about 1 ib. of activated
carbon.
5.4.2 A water system (Millipore
Super-Q or equivalent) may be used to
generate organic-free deionized water.
5.4.3 • 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 one hour. While
still hot, transfer the water to a narrow
mouth screw cap bottle equipped with a
Teflon seal.
5.5 Stock standards (2 mg/ml)—
Prepare stock standard solutions 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 NIQSH/MESA
approved toxic gas respirator should be
worn when the analyst handles high
concentrations of such materials.
5.5.1 Place about 9.8 ml of methanol
into a 10 ml ground glass stoppered
volumetric flask. Allow the flask to
stand, unstoppered, for about 10 minutes
or until all alcohol wetted surfaces have
dried. Tare the flask to the nearest 0.1
mg.
5.5.2 Add the assayed reference
material:
5.5.2.1 Liquids—using a 100 jil
syringe, immediately add 2 to 3 drops of
assayed reference material to the flask,
then reweigh. Be sure that the drops fall
directly into the alcohol without
contacting the neck of the flask.
5.5.2.2 Gases—To prepare standards
of bromomethane, chioroethane,
chloromethane, and 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
methyl alcohol menicus. Slowly inject
the reference standard into the neck of
the flask (the heavy gas will rapidly
dissolve into the methyl alcohol).
5.5.3 Reweigh the flask, dilute to
volume, stopper, then mix by inverting
the flask several times. Transfer the
standard solution to a 15-ml screw-cap
bottle equipped with a Teflon cap liner.
5.5.4 Calculate the concentration in
mg per ml (equivalent to jxg per )d) from
the net gain in weight.
5.5.5 Store stock standards at 4° C.
Prepare fresh standards every second
day for the four gases and 2-
chloroethylvinyl ether. All other
standards must be replaced with fresh
standards each week.
5.6 Surrogate Standard Dosing
Solution—From stock standard solutions
prepared as above, add a volume to give
1000 jug each of bromochloromethane,
2-bromo-l-chloropropane, and 1,4-
dichlorobutane to 40 ml of organic-free
water contained in a 50-ml volumetric
flask, mix and dilute to volume. Prepare
a fresh surrogate standard dosing
solution weekly. Dose the surrogate
standard mixture into every 5-ml sample
and reference standard analyzed.
6. Calibration.
6.1 Using, the stock standards,
prepare secondary dilution standards of
the compounds of interest, either singly
or mixed together in methanoi. The
standards should be at concentrations.
such that the aqueous standards
prepared in 6.2 will bracket the working
range of the chromatographic system. If
the limit of detection listed in Table 2 is
10 fig/1, for example, prepare secondary
methanolic standards at 100 fj.g/1, and
500 fig/1, so that aqueous standards
prepared from thee secondary
calibration standards, and the primary
standards, will define the linearity of the
detector in the working range.
6.2 Using both the primary and
secondary dilution standards, prepare
calibration standards,by carefully
adding 20.0 ^il of the standard in
methanol to 100, 500, or 1000 ml of
organic-free water. A 25 fil syringe
(Hamilton 7JD2N or equivalent) should be
used for this operation. These aqueous
standards must be prepared fresh daily.
6.3 Assemble the necessary gas
chromatographic and mass spectrometer
apparatus and establish operating
parameters equivalent to those
indicated in Table 2. By injecting
secondary dilution standards, establish
the linear range of the analytical system
for each compound and demonstrate
that the analytical system meets the
limit of detection requirements in Table
2.
8.4 Assemble the necessary purge
and trap device. Pack the trap as shown
in Figure 2 and condition overnight at a
nominal 180° C by backflushing with an
inert gas flow of at least 20 ml/min.
Daily, prior to use, condition the traps
for 10 minutes by backflushing at 180° C.
Analyze aqueous calibration standards
(6.2) according to the purge and trap
procedure in Section 9. Compare the
responses to those obtained by injection
of standards (6.3), to determine the
analytical precision. The analytical
precision of the analysis of aqueous
standards must be comparable to data
presented by Bellar and Lichtenberg
(1978, Ref. 1) before reliable sample
analysis may begin.
6.5 Internal Standard Method — The
internal standard approach is
acceptable for the purgeable organics.
The utilization of the internal standard
method requires the periodic
determination of response factors (RF)
which are defined in equation 1.
Eq. (1) RF
Where:
A, is (he integrated area or peak height of
the characteristic ion for the priority pollutant
standard.
Au is the integrated area or peak height of
the characteristic ion for the internal
standard.
Cu is the amount of the internal standard in
W
C, is the amount of the pollutant standard
in fig.
The relative response ratio for each
pollutant should be known for at least
two concentration values — 50 ng
injected to approximate 10 ^ig/1 and 500
ng to approximate the 100 ug/1 level.
Those compounds that do not respond
at either of these levels may be run at
concentrations appropriate to their
response. The response factor (RF) must
be determined over all concentration
ranges of standard (C,) which are being
determined. (Generally, the amount of
internal standard added to each extract
is the same so that C& remains
constant.) This should be done by
preparing a calibration curve where the
response factor (RF) is plotted against
the standard concentration (C,). Use a
minimum of three concentrations over
the range of interest. Once this
calibration curve has been determined,
it should be verified daily by injecting at
least one standard solution containing
internal standard. If significant drift has
occurred, a new calibration curve must
be constructed.
Note. — EPA, through its contractors and
certain of its Regional Laboratories, is
currently evaluating selected compounds for
-------
69534
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
use as internal standards in the analysis of
organic* by purge and trap.
6.6 The external standard method
can also be used at the discretion of the
analyst. Prepare a master calibration
curve using a ipjniHum* of three
standard solutions of each of the
compounds that are to be measured. Plot
concentrations versus integrated areas
or peak heights (selected characteristic
ion for GC/MS). One point on each.
curve should approach the method
detection limit After the master set of
instrument calibration curves have been
established, they should be verified
daily by injecting at least one standard
solution. If significant drift has occurred.
a new calibration curve must be
constructed.
7. Quality Control.
7.1 Before'processing any samples,
the analyst should daily demonstrate,
through the analysis of an organic-free
water method blank, that the entire
analytical system is interference-free.
72 Standard quality assurance
practices should be used with this
method. Field replicates should be
collected to validate the precision of the
sampling technique. Laboratory
replicates should be analyzed to
validate the precision of the-analysis.
Fortified samples should be analyzed to
validate the accuracy of the analysis.
7.3 The analyst should maintain
constant surveillance of both the
performance of the analytical system
and the effectiveness of the method in
dealing with each sample matrix by
determining the precision of the method
in blank water and spiking each 5-ml
sample, standard, and blank with
surrogate halocarbons.
7.3.1 Determine the precision of the
method by dosing blank water with the
compounds selected as surrogate
standards—bromochlorome thane, 2-
bromo-1-chloropropane, and 1,4-
dichlorobutane—and running replicate
analyses. Calculate the recovery and its
standard deviation. These compounds
represent early, middle, and late eluters
over the range of the pollutant
compounds.
7.3.2 The sample matrix can affect.
the purging efficiencies of individual
compounds; therefore, each sample.must
be dosed with the surrogate standards
and analyzed in a manner identical to
the internal standards in blank water. If
the recovery of the surrogate standard
shows a deviation greater than two
standard deviations (7.3.1), repeat the
dosed sample analyses. If the deviation
is again greater .than two standard
deviations, dose another aliquot of the
same sample with the compounds of
interest at approximately two times the
measured values and analyze. Calculate
the recovery for the individual
compounds using these data.
8. Sample Collection. Preservation,
and Handling.
8.1 Grab samples must be collected
hi glass-containers having a total
volume greater than 20 ml Fill the
sample bottles in such a manner that no
air bubbles pass through the sample as
the bottle is being filled. Seal the bottle*
so that no air bubbles are entrapped in
it Maintain the hermetic seal on the
sample bottle until time of analysis.
8.2 The sample must be iced or
refrigerated from the time of collection
until extraction. If the sample contains
residual chlorine, add sodium:
thiosulfate- preservative (10 ^g/40 ml] to
the empty sample bottles just prior to
shipping to the sample site, fill with
sample just to overflowing, seal the
bottle, and shake vigorously for 1
minute.
8.3 All samples must be analyzed
within 7 days of collection.
9. Sample Extraction and Gas
Chromatography.
9.1 Remove standards and samples
from cold storage (approximately an
hour prior to an analysis) and bring to
room temperature by placing in a warm
water bath at 20-25'C.
9.2 Adjust the purge gas (nitrogen or
helium) flow rate to 40 ml/min. Attach
the trap inlet to the purging device, and
set the device to the purge mode. Open
the syringe valve located on the purging
device sample introduction needle.
9.3 Remove the plunger from a 5 ml
syringe and attach a closed syringe
valve. Open the sample bottle (or
standard) and carefully pour the sample
into the syringe barrel until it overflows.
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.
Since this process of taking an-aliquot
destroys the validity of the sample for
future analysis, the analyst should fill a
second syringe at this, time to protect
against possible loss of data. Add 5.0 p.1
of the surrogate spiking solution (7.3)
through the valve bore, then close the
valve.
9.4 Attach the syringe-valve
assembly to-the syringe valve on the
purging device. Open the syringe valve
and inject the sample into the purging
chamber.
9.5 Close both valves and purge the
sample for 12.0 ±.05 minutes.
9.8 After the 12-minute purge time,
attach the trap to the'chromatograph,
and adjust the device to the desorb
mode. 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 20 to 60 ml/
min for 4 minutes. If rapid heating
cannot be achieved, 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 initial program
temperature of 45'C.
9.7 While the trap is being desorbed
into the gas chromatograph, empty the
purging chamber using the sample
introduction syringe. Wash the chamber
with two 5-ml flushes of organic-free
water. After the purging device has been
emptied, continue to allow the purge gas
to vent through the chamber until the frit
is dry, and ready for the next sample.
9.8 After desorbing the sample for
four minutes, recondition the trap by
returning the purge and trap device to
the purge mode. Watt 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 seven
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. (Note:
If this bake out step is omitted, the
amount of water entering the GC/MS
system will progressively increase
causing deterioration of and potential
shut down' of the system.)
9.9 The analysis of blanks is most
important in the purge and trap
technique since the purging device and
the trap can be contaminated by
residues from very concentrated
samples or by vapors in the laboratory.
Prepare blanks by filling a sample bottle
with organic-free water that has been
prepared by passing distilled water
through a pretested activated carbon
column. Blanks should be sealed, stored
at 4'C, and analyzed with each group of
samples.
10. Gas Chromatography—Mass
Spectrometry.
10.1 Table 2 summarizes the
recommended gas chromatographic
column materials and operting
conditions for the instrument. Included
in this tab.e are estimated retention
times and sensitivities that should be
achieved by this method. An example of
the separation achieved by Column 1 is
shown hi Figure 5.
10.2 GC-MS Determination—
Suggested analytical conditions for
determination of the pollutants
amenable to purge and trap, using the
Tekmar LCS-1 and GC/MS are given
below. Operating conditions vary from
one system to another; therefore, each
analyst must optimize the conditions for
each purge and trap and GC/MS system.
10.3 Purge Parameters.
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
69535
Sample size—S.O ml.
Purge gas—Helium, high purity grade.
Purge time—12 minutes.
Purge flow—40 ml/rain.
Trap dimensions—V4 in. O.D. (0.105 in.
I.D.)x25 on long.
Trap sorbent—Tenax-GC. 60/80 mesh (15
cm), plus Type 15 silica gel, 35/60 mesh (8
cm).
Desorption flow—20 ml/min.
Desorption time—4 min.
Desorption temperature—180* C-
10.4 Mass Spectrometer Parameters.
Electron energy—70 volts (nominaj).
Mass range—20*27, 33-260 amu.
Scan time—6 seconds or less.
10.5 Calibration of the gas-
chroma tography-mass spectrometry
(GC-MS system—Evaluate the system
performance each day that it is to be
used for the analysis of samples or
blanks by examining the mass spectrum
ofDFTPPorSFB.
10.5.1 To use DFTPP, remove the
analytical column and substitute a
column more appropriate to the boiling
point of the reference compound (e.g. 3%
SP-2250 on Supelcoport). Inject a
solution containing 50 ng DFTPP and
check to insure that the performance
criteria listed in Table 3 are met
10.5.2 To use BFB, infect a solution
containing 20 ng BFB and check to
insure that the performance criteria
listed in Table 4 are met.
10.5.3 If the system performance
criteria are not met for either test, the
analyst must retune the spectrometer
and repeat the performance check. The
performance criteria must lie met before
any samples or standards may be
analyzed.
10.8 Analyze an internal or external
calibration standard to develop
response factors for each compound.
11. Qualitative and Quantitative
Determination.
11.1 To qualitatively identify a
compound, obtain an Extracted Ion
Current Profile (EICP] for the primary
ion and at least two other ions (if
available) listed in Table 5. The criteria
below must be met for a qualitative
identification.
11.1.1 The characteristic ions for the
compound must be found to maximize in
the same or within one spectrum of each
other.
11.1.2 The retention time at the
experimental mass spectrum must be
within ±60 seconds of the retention
time of the authentic compound.
11.1.3 The ratios of the three EICP
peak heights must agree within ±20%
with the ratios of the relative intensities
for these ions in a reference mass
spectrum. The reference mass spectrum
can be obtained from either a standard
analyzed through the GC-MS system or
from a reference library.
11.1.4 Structural isomers that have
very similar mass spectra can be
explicitly identified only if the resolution
between the isomers in a standard mix
is acceptable. Acceptable resolution is
achieved if the valley height between
isomers is less than 25% of the sum of
the two peak heights. Otherwise,
structural isomers are identified as
isomeric pairs.
11.2 The primary ion listed in Table 5
is to be used to quantify each
compound. If the sample produces an
interference for the primary ion, use a
secondary ion to quantify.
11.3 For low concentrations, or direct
aqueous injection of acryionitrile and
acrolein, the characteristic masses listed
for the compounds in Table 5 may be
used for selected ion monitoring (SIM).
SIM is the use of a mass spectrometer as
a substance selective detector by
measuring the mass spectrometric
response at one or several characteristic
masses in real time.
11.4 Internal Standard Method
Calculations—By adding a constant
known amount of internal standard (C*
in fig) to every sample extract the
concentration of the pollutant (C0) in
fig/1 in the sample is calculated using
equation 2,
Eq. «C,- _
(AJ(F»F)(VJ
Where:
V, is the volume of the original sample in
liters, and the other terms are defined as
in Section 6 .5. To quantify, add the
internal standard to the 5.0-ml sample no
more than a few minutes before purging
to minimize the possibility of losses due
to evaporation, adsorption, or chemical
reaction. Calculate the concentration by
using the previous equations with the
appropriate response factor taken from
the calibration curve.
11.5 Extenral Standard Method
Calculations—The concentration of the
unknown can be calculated from the
slope and intercept of the multiple point
calibration curve. The unknown
concentration can be determined using
equation 3.
(A)
Eq. (3) mlcroorarna per liter »ng/ml- -—
(VJ
•Where:
A—Mass of compound from calibration curve
(ng/5 ml).
V.» volume of water purged (5 ml).
11.6 An alternate external standard
approach for purgeables utilizes a single
point calibration. Prepare and analyze a
reference standard that closely
approximates the response for each
component in a sample. Calculate the
concentration in the sample using
Equation 4.
(AKB)
Eq. 4 microgram* per liter—
Where:
A=»area of the unknown
B=»concentration of standard 0*g/l)
C«area of the standard.
11.7 Report ail results to two
significant figures. When duplicate and
spiked samples are analyzed, all data
obtained should be reported. Report
results in micrograms per liter without
correction for recovery data.
12. References.
1. "The Analysis of Haiogenated Chemical
Indicators of Industrial Contamination in
Water by the Purge and Trap Method." U.S.
EPA. Environmental Monitoring and Support
Laboratory, Cincinnati, OH. 45268, Dec. 1978.
2. "Symposium on Measurement of Organic
Pollutants in Water and Wastewater," ASTM
Special Publication. 1979 (In Press).
3. "Determining Volatile Organics at
Microgram-per-Uter Levels by Gas
Chromatography," T. A. Bellar and J. J.
Lichtenberg, Jour. AWWA, 88. 739-744, Dec.
1974.
4. ASTM Annual Standards—Water, part
31, Method 02908 "Standard Recommended
Practice for Measuring Water by Aqueous-
Injection Gas Chromatography."
5. ASTM Annual Standards—Water, part
31, Method D3371 "Tentative Method of Teat
for Nitriles in Aqueous Solution of Gas Liquid
Chroma tograph."
6. "Direct Analysis of Water Samples for
Organic Pollutants-with Gas
Chroma tography-Mass Spectrometry,"
Harris. L. E., Budde, W. L, and Eichelberger,
J. W. Aaal. Chem^ 46,1912 (1974).
Bibliography
1. "Sampling and Analysis Procedures for
Screening of Industrial Effluents for Priority
Pollutants," March 1977 (revised April 1977).
USEPA. Effluent Guidelines Division.
Washington, D.C. 20460.
2. "Proceedings: Seminar on Analytical
Methods for Priority Pollutants," Volume 1—
Denver, Colorado. November 1977; Volume
2—Savannah, Georgia, May 1978: Volume 3—
Norfolk. Virginia. March 1979; USEPA.
Effluent Guidelines Division. Washington.
D.C. 20460.
PmnwMr
STORETNo.
AcroMrt
Aerytonrtri*.
eromonwtMn
C*rt>on Tmd
CDk
2-CMoroctfiyMnyl «txr
DlbromocMoronwthtn*—
1.1-OlcNeraMMM.
34210
34215
34238
34413
32101
32104
32102
34301
34311
34879
32108
34418
34106
34488
-------
S953S
•••1-^-M.i
TaW« 1 — Continued
1 ,2-Ochloroathana
1.1-OicniOfO«man»
tran»- 1 ,2-D»cftioro«man
' .2-Oicflloropnipana ...
co- 1 .3-Otalilofoorop«o«
tran». 1 ,3-Ocrtcfopropai
£thyl_«n.ana
MathyMrw chtorida
•
na
1 . 1 .22-T«trachk>K_Mh-r-»
Tetrachlofetnana
1,1.1-TnchlonMIIWM
1 . 1 -Mnchttroatfian*
Trtcnloroathana
TncNoroouoromelhana'.
Tafc-B_»
vln^H chlunda — r— I.HW...L
•«••••-•
cuiuer o, ia
/» / WO]
sosea Rule
S
^^-OrrpPK^^tnatonAbundv>ca ^.i.-BFBKwttonaarttoAtonOincvCrit**
34531
34501
34546
34541
34561
34561
34371
34423
34516
34475
34508
34511
39180
34488
34010
38175
Tabto 2.— Gas Chromatograpty of Organic* 6?
Putyeantt Trap-
Compound
cttorammhana
Qromom«nan«
wyl emend* _____
cnlofoemana
nwthylan* crtcnda.
inehJaofluoromeiftana..
1.1-tfcNorootnana
Sromocntoromatnana
(Ss>
U-<»chloro«in*i«
traos-l.2-
*CftlorO«tn«n» _____
chtofofofm ....,„_._„...._
1 .2-dhXkiR.Htww
1 . 1 . i -tncttoroflttun* _
cartxjn i«tncnK>n»
IxonxxlicNaranwman*.
i.2-<*chkxopropane
Wna-1,3-
Oicftlofopropane
MchlomMhjn-. ,
1.1--Wchloro«thana_.
oa-1 .3-dBhlonjpiepana'
2-chkxoathylvinvl «har
2^Nuiiiu»|.
cftoreprapvi* (SS}_
bromolorm ......
1.1A2-
UHracnionoatnana
1,*-*cftloroout-n»(SS» _
tokMna .... _
cNorooanzvna '""
atnyibanzBoa
•emwn
•crvtonitnl.
Rwandan HIM
(mautta)
Cot.1'
150
2.17
2.67
3.33
525
7.18
7.92
1.46
0-30
10.08
10.68
11.40
1260
13.02
13.65
14.92
15.22
1580
16.46
16_52
16.53
16.00
19-23
21.6Z
2147
iiTi"
Cot.2'
210
2.50
2.57
2-S2-
4.03
5.14
5.25
8.31 _
6.48
6.81
7.70
8.2»-
9.28
9.4S
10.36
11.30
11.7O
11JW
1288
1266
12.68
^95
13.71
1182 _
15.41
17.70
17.4*
18.13 _
18.53
20.57
25.08
Urn. of
-atac-on'
IPO/I)
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
"1 1 1.1
10
10
10
to
10
10
•100
MOO
M*»» ton abundttwtt crttartt
51 30 to 60 pet o< mtst 1S
'0 __, U»» than 2 pet o« m«ii
127 40 to ao pet at man is
• su
18. 75
16*. 95
l«9.
«. 98
Mau
97 Laattnanl po of maw 1S8. 173
is» 8a»« pMk. too pet ralakv* 17*
199 ., **_"*"? 175"~ -
iw. — . 5io9oeto
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3, 1979 / Proposed Rules
69537
OPTIONAL
FOAIU
(TRAP
V4 IN.
0. 0. EXIT
c
d
1
-EXIT '/a IN.
0. 0.
~-14MM 0. D.
INLET 5i IN.
0. D.
SAMPLE INLET
2-WAY SYRINGE VALVE
•17O.1. 20 GAUGE SYRINGE NEEDLE
. 0. D. RUBBER SEPTUM
IQfiMfl GLASS FRIT
MEDIUM POROSITY
1
^_10MM. 0. D.
•"*)
j V.
5
S
H
7
—INLET
'/i IN. 0. D.
cr
/
/
1/16 IN. O.D.
y'STAJIVLtii iltt1
i
^PSI
13X MOLECULAR
SIEVE PURGE
GAS FILTER
' PURGE GAS
* FLOW
fc CONTROL
Figure 1. Purging device
PACKING PROCEDURE
GLASS KUM
WOOL 5MM
GRADE 15
SILICA GEL
8 CM
TENAX 15 CM
3% OV-1 1CIW
GLASS 5MM
WOOL
TRAP INLET
CONSTRUCTION
COMPRESSION
FITTING NUT
ANO FERRULES
14FT. 7-n/FOOT
RESISTANCE WIRE
WRAPPED SOLID
THERMOCOUPLE/
CONTROLLER
SENSOR
CJL / TUBING 25CM
0.105 IN. I.O.
CL
J-£
0.125 IN. O.D.
STAINLESS STEEL
Figure 2. Trap packings and construction to include
deaorb capability
-------
63538 Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
PURGE GAS . .
FLOW CONTROL '
LIQUID INJECTION PORTS
13X MOLECULAR
SIEVE FILTER %
/,
-COLUMN OVEN
\ nnnn I CONFIRMATORY COLUMN
jti U LI U^-y-^Q OETECTOR
pJ¥irir-f ^-ANALYTICAL COLUMN
* OPTIONAL 4-PORT COLUMN
SaECTlON VALVE
SB-PORT TRAP INLET
WIRE CHEATER
" CONTROL
PURGING
DEVICE
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 80° C.
Figure 3. Schematic of purge and trap device - purge mode
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
LIQUID INJECTION-PORTS
PURGE GAS
FLOW CONTROL^
13X MOLECULAR
SIEVE FILTER
-COLUMN OVEN
n n n n I CONFIRMATORY COLUMN
TO DETECTOR
-ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SaECTlON VALVE
6-PORT TRAP INLET
VALVE J RESISTANCE WIRE H£ATER
^ CONTROL
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 80°C.
PURGING
DEVICE
Figure 4. Schematic of purge and trap device - desorfr mode
-------
LU
-COLUMN: 1% SP-1000 ON CARBOPACK-B
PROGRAM: 45'C-3 MINUTES, 8VMINUTE TO 220°C
3?
8-
2.
UJ
8 10 12 14
RETENTION TIME - MINUTES
Figure 5. Gas chromatogram of volatile organics by purge and trap
28' 30
I
B-
a
I
8
BIU-INQ CODE 65VO-01-C
n
(B
a.
I
(D
-------
60540
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
Base/Neutrals. Acids, and Pesticides—
Method 625
1. Scope and Application.
1.1 This method covers the
determination of a number of organic
compounds that are solvent extractable
and amenable to gas chroma tography.
The parameters listed hi Tables 1, 2 and
3 may be determined by this method.
1.2 This method is applicable to the
determination of these compounds in
municipal and industrial discharges. It is
designed to be used to meet the
monitoring requirements of the National
Pollutants Discharge Elimination System
(NPDES].
,1.3 The detection limit of this method
is usually dependent upon the level of
interferences rather than instrumental
limitations. The limits listed in Tables 4,
5, and 6 represent the minimum quantity
that must be injected into the system to
get confirmation by the mass
spectrometric method described below.
1.4 The GC/MS parts of this method
are recommended for use only by
analysts experienced with GC/MS or
under the close supervision of such
qualified persons.
2. Summary of Method.
2.1 A1 to 2 liter sample of
wastewater is extracted with methylene
chloride using separatory funnel or
continuous extraction techniques. If
emulsions are a problem, continuous
extraction techniques should be used.
The extract is dried over sodium sulfate
and concentrated to a volume of 1 ml
using a Kuderna-Danish (K-D)
evaporator. Chromatographic conditions
are described which allow for the
separation of the compounds in the
extract.
2.2 Quantitative analysis is performed
by GC/MS using either the internal
standard or external standard
technique.
3. Interferences.
3.1 Solvents, reagents, glassware, and
other sample processing hardware may
yield discrete artifacts and/or elevated
baselines causing misinterpretation of
chromatograms. 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 and
purification of solvents by distillation in
all-glass systems may be required.
3.2 Interferences coextracted from the
samples will vary considerably from
source to source, depending upon the
diversity of the industrial complex or
municipality being sampled.
3.3 The recommended analytical
procedure may not have sufficient
resolution to differentiate between
certain isomeric pairs. These are
anthracene and phenanthrene, chrysene
and benzo(a)anthracene, and
benzo(b)fluoranthene and
benzo(k)fluoranthene. The GC retention
time and mass spectral data are not
sufficiently unique to make an
unambiguous distinction between these
compounds. Alternative techniques
should be used to identify and quantify
these specific compounds. See
Reference 1.
4. Apparatus and Materials.
4.1 Sampling equipment, for discrete
or composite sampling.
4.1.1 Grab sample bottle—amber
glass, 1-liter to 1-gallon volume. French
or Boston Round design is
recommended. The container must be
washed and solvent rinsed before use to
minimize interferences.
4.1.2 Bottle caps—Threaded to fit
sample bottles. Caps must be lined with
Teflon. Aluminum foil may be
substituted if sample is not corrosive.
4.1.3 Compositing equipment—
Automatic or manual compositing
system. Must incorporate glass sample
containers for the collection of a
minimum of 1000 ml. Sample containers
must be kept refrigerated during
sampling. No plastic or rubber tubing
other than Teflon may be used in the
system.
4.2 Separatory funnel—2000 ml, with
Teflon stopcock (Ace Glass 7228-T-72
or equivalent).
4.3 Drying column—A 20 mm ID
pyrex chromatographic column
equipped with coarse glass frit or glass
wool plug.
4.4 Kuderna-Danish (K-D)
Apparatus
4.4,1 Concentrator tube—10 ml,
graduated (Kontes K-570050-1025 or
equivalent). Calibration must be
checked. Ground glass stopper (size 19/
22 joint) is used to prevent evaporation
of extracts.
4.4.2 Evaporative flask—500 ml
(Kontes K-STOOl-^SOO or equivalent).
Attach to concentrator tube with
springs.. {Kontes K-662750-0012).
4.4.3 Snyder column—three-ball
macro (Kontes K503000-0232 or
equivalent).
4.4.4 Snyder,column—two-ball micro
(Kontes K-569002-0219 or equivalent).
4.4.5 Boiling chips-extracted,
approximately 10/40 mesh.
4.5 Water bath—Heated, with
concentric ring cover, capable of
temperature control (±2' C). The bath
shouhi be used in a hood.
4,8 Gas chromatograph—Analytical
system complete with gas
chromatograph capable of on-column
injection and all required accessories'
including column supplies, gases, etc.
4.8.1 Column 1—For Base/Neutral
and Pesticides a 8-foot glass column (V*
in OD x 2 mm ID) packed with 3% SP-
2250 coated on 100/120 Supelcoport (or
equivalent).
4.6.2 Column 2—For Acids, a 6-foot
glass column (Vi in OD x 2 mm ID)
packed with 1% SP-1240 DA coated on
100/120 mesh Supelcoport (or
equivalent).
4.7 Mass Spectrometers-Capable of
scanning from 35 to 450 a.m.u. every 7
seconds or less at 70 volts (nominal) and
producing a recognizable mass spectrum
at unit resolution from 50 ng of DFTPP
when the sample is introduced through
the GC inlet (Reference 2). The mass
spectrometer must be interfaced with a
gas chromatograph equipped with an
injector system designed for splitless
injection and glass capillary columns or
an injector system designed for on-
column injection with all-glass packed
columns. All sections of the transfer
lines must be glass or glass-lined and
must be deactivated. (Use Sylon-CT,
Supelco, Inc., or eqviivalent to
deactivate.)
Note.—Systems utilizing a jet separator for
the GC effluent are recommended since
membrane separators may lose sensitivity for
light molecules and glass fnt separators may
inhibit the elution of polynuclear aromatics.
Any of these separators may be used
provided that it gives recognizable mass
spectra and acceptable calibration points at
the limit of detection specified for each
individual compound listed in Tables 4, 5,
and 8.
4.8 A computer system must be
interfaced to the mass spectrometer to
allow acquisition of continuous mass
scans for the duration of the
chromatographic program. The computer
system should also be equipped with
mass storage devices for saving all data
from GC-MS runs. There must be
computer software available to allow
searching any GC-MS run for specific
ions and plotting the int'ehsity of the
ions with respect to time or scan
number. The ability to integrate the area
under any specific ion plot peak is
essential for quantification.
4.9 Continuous liquid-liquid
extractors—Teflon or glass connecting
joints and stopcocks, no lubrication.
(Hershberg-Wolf Extractor—Ace Glass
Co., Vineland, N.J. P/N 6841-10 or
equivalent).
5. Reagents.
5.1 Sodium hydroxide—(ACS) 6N in
distilled water.
5.2 Sulfuric acid—(ACS) 6N in
distilled water.
5.3 Sodium sulfate—(ACS) granular
anhydrous (rinsed with methylene
chloride (20 ml/g) and conditioned at
400° C for 4 hrs.).
-------
Federal Register / Vol. 44, No. 233 / Monday. December 3. 1979 / Proposed Rules 69541
5.4 Methylene chloride—Pesticide
quality or equivalent.
5.5 Stock standards—Obtain stock
standard solutions at a concentration of
1.00 fig/p.1. For example, dissolve 0.100
grams of assayed reference material in
pesticide quality isooctane or other
appropriate solvent and dilute to volume
in a 100 ml ground glass stoppered
volumetric flask. The stock solution is
transferred to 15 ml Teflon lined screw
cap vials, stored in a refrigerator, and
checked frequently for signs of
degradation or evaporation, especially
just prior to preparing working
standards from them. Protect PNA
standards from light.
6t Calibration.
6.1 Prepare calibration standards
that contain the compounds of interest,
either singly, or mixed together. The
standards should be prepared at
concentrations that will bracket the
working range of the chromatographic
system (two or more orders of
magnitude are suggested). If the limit of
detection (Tables 4, 5, or 6) can be
calculated as 20 ng injected, for
example, prepare standards at 1
10 Aig/nd- 100 MJ/nd- etc. so that
injections of 1-5 /U of the calibration
standards will define the linearity of the
detector in the working range.
6.2 Assemble the necessary gas
chromatographic apparatus and'
establish operating parameters
equivalent to those indicated in Tables
4, 5, and 6. By injecting calibration
standards, establish the linear range of
the analytical system and demonstrate
that the analytical system meets the
limits of detection requirements of
Tables 4, 5, and 6.. If the sample gives
peak areas above the working range,
dilute and reanalyze.
6.3 Internal Standard Method—The'
internal standard approach is
acceptable for all of the semivolatile
organics. The utilization of the internal
standard method requires the periodic
determination of response factors (RF)
which are defined in equation 1.
Eq. 1 RF=(A,Ci,)/(Al.C,)
Where:
A, is the integrated area or peak height of the
characteristic ion for the pollutant
standard.
At, is the integrated area or peak height of the
characteristic ion for the internal
standard.
Ci, is the amount (jtg) of the internal
standard.
C, is the amount (jug) of the pollutant
standard.
6.3 The relative response ratio for
the pollutants should be known for at
least two concentration values—20 ng
injected to approximate 10 ^g/1 and 200
ng injected to approximate the 100 fj.g/1
level. (Assuming 1 ml final volume and a
2 pi injection). Those compounds that 'do
not respond at either of these levels may
be run at concentrations appropriate to
their response.
The response factor (RF) should be
determined over all concentration
ranges of standard (C,) which are being
determined. (Generally, the amount of
internal standard added to each extract
is the same (20 fig) so that Q, remains
constant.) This should be done by
preparing a calibration curve where the
response factor (RE) is plotted against
the standard concentration (C,), using a
minimum of three concentrations over
the range of interest Once this
calibration-curve has been determined,
it should be verified) daily by injecting at
least one standard solution containing
internal standard. If significant drift has
occurred, a new calibration curve must
be constructed. To quantify, add the
internal standard to the concentrated
sample extract no more-than a few
minutes before injecting into the GC/MS
to minimize the possibility of losses due
to evaporation, adsorption, or chemical
reaction. Calculate the concentration by
using the previous equations with the
appropriate response factor taken from
the calibration curve. Either deuterated
or fluorinated compounds can be used
as internal standards and surrogate
standards. Naphthalene-cU, anthracene-
dio, pyridine-dg, aniline-dj, nitrobenzene-
ds. 1-fluoronaphthalene, 2-
fluoronaphthalene, 2-fluorobiphenyl,
2,2'-difluorobiphenyl, and 1,2,3,4,5-
pentafluorobiphenyl have been used or
suggested as appropriate internal
standards /surrogates for the base-
neutral compounds. Phenol-d«,
pentafluorophenol, 2-perfluoromethyl
phenol, and 2-fluorophenol have been
used or suggested for the acid
compounds. Compounds used as
internal standards are not to be used as
surrogate standards. The internal
standard must be different from the
surrogate standards.
6.5 The external standard method
can also be used at the discretion of the
analyst. Prepare a master calibration
curve using a minimum of three
standard solutions of each of the
compounds that are to be measured. Plot
concentrations versus integrated areas
or peak heights (selected characteristic
ion for GC/MS). One point on each
curve should approach the limit of
detection (Tables 4, 5, and 6). After the
master set of instrument calibration
curves have been established, they
should be verified daily by injecting at
least-one standard solution. If significant
drift has occurred, a new calibration
curve must be constructed.
7. Quality Control.
7.1 Before processing any samples,
demonstrate through the analysis of a
method blank, that all glassware and
reagents are interference-free. Each time
a set of samples is extracted or there is
a change in reagents, a method blank
should be processed as a safeguard
against chronic laboratory
contamination.
7.2 . Standard quality assurance
practices should be used with this
method. Field replicates should be
collected and analyzed to determine the
precision of the sampling technique.
Laboratory replicates should be
analyzed to determine the precision of
the analysis. Fortified samples should be
analyzed to determine the accuracy of
the analysis. Field blanks should be
analyzed to check for contamination
introduced during sampling and
transportation.
8. Sample Collection, Preservation,
and Handling,
8.1 Grab samples must be collected in
glass containers. Conventional sampling
practices should be followed, except
that the bottle must not be prerinsed
with sample before collection.
Composite samples should be collected
in refrigerated glass containers in
accordance with" the requirements of the
program. Automatic sampling equipment
must be ffee of tygon and other potential
sources of contamination.
8.2 The samples must be iced or
refrigerated from the time of collection
until extraction. Chemical preservatives
should not be used in the field unless
more than 24 hours will elapse before
delivery to the laboratory. If the samples
will not be extracted within 48 hours of
collection, they must be preserved as
follows:
8.2.1 If the sample contains residual
chlorine, add 35 mg of sodium
thiosulfate per 1 ppm of free chlorine per
liter of sample.
8.2.2 Adjust the pH of the water
sample to a pH of 7 to 10 using sodium
hydroxide or sulfuric acid. Record the
volume of acid or base used.
8.3 All samples must be extracted
within 7 days and completely analyzed
within 30 days of collection.
9. Sample Extraction (Base/Neutrals.
Acids, and Pesticides).
9.1 Samples may be extracted by
separatory funnel techniques or with a
continuous extractor as described in
Section 10. Where emulsions prevent
acceptable solvent recovery with the
separatory funnel technique, the analyst
must use the continuous extractor.
9.2 The details of the extraction
technique should be adjusted according
to the sample volume. The technique
described below assumes a sample
-------
69542 Federal Register / Vol. 44. No. 233 / Monday, December 3. 1979 / Proposed Rules
volume of 1000 ml. For volumes
approximating 2-liters, the volume of
extraction solvent should be adjusted to
250,100, and 100 ml for the serial
extraction of the base neutrals, and 200,
100, and 100 ml for the acids.
9.3 Mark the water meniscus on the
side of the sample bottle for later
determination of sample volume. Pour
the entire sample into a two-liter
separatory funnel. Adjust the pH of the
sample with 6N NaOH to 11 or greater.
Use multirange pH paper for the
measurements. Proceed to Section 10 if
continuous extraction is used.
9.4 Add 60 ml methylene chloride to
the sample bottle, cap, and shake 30
seconds to rinse the walls. Transfer the
solvent into the separatory funnel, and
extract the sample by shaking the funnel
for two minutes with periodic venting to
release excess vapor pressure. Allow
the organic layer to separate from the
water phase for a minimum of ten
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, but may include stirring,
filtration of the emulsion through glass
wool, or centrifugation. (If the emulsion
cannot be broken, that is, recovery is
less than 80% of the added solvent
corrected for the water solubility of
methylene chloride, transfer the sample,
solvent and emulsion into a continuous
extractor and proceed as described in
Section 10). Collect the methylene
chloride extract in a-250-ml Erlemneyer
flask.
9.5 Add a second 60-ml volume of
methylene chloride to the sample bottle
and complete the extraction procedure a
second time, combining the extracts in
the Erlenmeyer flask.
9.6 Perform a third extraction in the
same manner. Pour the combined
extract through a drying column
containing 3-4 inches of anhydrous
sodium sulfate, and collect it in a 500 ml
K-D flask equipped with 10 ml
concentrator tube. Rinse the Erlenmeyer
with 20 to 40 ml of methylene chloride.
Pour this through the drying column.
Seal, label as base/neutral fraction, and
proceed with the acid extraction. If the
extract must be stored overnight before
analysis by GC/MS, it may be
transferred to a 2 ml serum vial
equipped with a Teflon-lined rubber
septum and crimp cap.
9.7 Acid (Phenols) Extraction—Adjust
the pH of the water, previously
extracted for base-neutrals, with 6N
H»SO« to 2 or below. Serially extract
with 60, 60 and 60 ml portions of
distilled-m-glass methylene chloride.
Collect and combine the extracts in a
250-ml Erlenmeyer flask then dry by
passing through a column of anhydrous
sodium sulfate. Rinse the Erlenmeyer
with 20 to 40 ml of methylene chloride
and pour through the drying column.
Seal, label acid fraction and prepare for
concentration.
9.8 Concentrate the extracts (Base/
Neutrals and Acids) in a 500 ml K-D
flask equipped with a 10 ml concentrator
tube.
9.9 Add 1 to 2 clean boiling chips to
the flask and attach a three-ball macro-
Snyder column. Prewet the Snyder
column by adding about 1 ml methylene
chloride through the top. Place the K-D
apparatus on a warm water bath (60 to
65° C | so that the concentrator tube is
partially immersed in the water, and the
entire lower rounded surface of the flask
is bathed with water vapor. Adjust the
vertical position of the apparatus and
the water temperature as required to
complete the concentration in 15 to 20
minutes. At the proper rate of
distillation the balls of the column
actively chatter but the chambers do not
flood. When the liquid has reached an
apparent volume 1 ml. remove the K-D
apparatus and allow the solvent to drain
for at least 10 minutes while cooling.
Remove the Snyder column and rinse
the flask and its lower joint into the
concentrator tube with 1 to 2 ml of
methylene chloride. A 5-ml syringe is
recommended for this operation.
9.10 Add a dean boiling chip and
attach a two-ball micro-Snyder column
to the concentrator tube in 9.8. Prewet
the column by adding about 0.5 ml
methylene chloride through the top.
Place the K-D apparatus on a warm
water bath (60 to 65°C) so that the
conceintrator tube is partially immersed
in the 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 actively chatter but the
chambers do not flood. When the liquid
reaches an apparent volume of about 0.5
ml, remove the K-D from the water bath
and allow the solvent to drain and cool
for at least 10 minutes. Remove the
micro-Snyder column and rinse its lower
joint into the concentrator tube with
approximately 0.2 ml of methylene
chloride. Adjust the final volume to 1.0
ml, seal, and label as acid fraction.
9.11 Determine the original sample
volume by refilling the sample bottle to
the mark and transferring the liquid to a
1,000-ml graduated cylinder. Record the
sample volume to the nearest 5 ml
10. Emulsions/Continuous Extraction.
10.1 Place 100 to 150 ml of methylene
chloride in the extractor'and 200-500 ml
methylene chloride in the distilling flask.
10.2 Add the aqueous sample (pH 11
or greater) to the extractor. Add blank
water as necessary to operate the
extractor and extract for 24 hours.
Remove the distilling flask and pour the
contents through a drying column
containing 7 to 10 cm of anhydrous
sodium sulfate. Collect the methylene
chloride in a 500 ml K-D evaporator
flask quipped with a 10 ml concentrator
tube. Seal, label as the base/neutral
fraction, and concentrate as per sections
9.8 to 9.10.
10.3 Adjust the pH of the sample in
the continuous extractor to 2 or below
using 6N sulfuric acid. Charge a clean
distilling flask with 500 ml of methylene
chloride. Extract for 24 hours. Remove
the distilling flask and pour the contents
through a drying column containing 7 to
10 cm of anhydrous sodium sulfate.
Collect the methylene chloride layer on
a K-D evaporator flask equipped with a,
10 ml concentrator tube. Label as the
acid fraction. Concentrate as per
sections 9.8 to 9.10.
11. Calibration of the GC-MS System.
11.1 At the beginning of each day,
the mass calibration of the GC-MS
system must be checked and adjusted if
necessary to meet DFTPP specifications
(11.3). Each day base-neutrals are
measured, the column performance
specification (12.1) with benzidine must
be met Each day the acids are
measured, the column performance
specification (13.1) with
pentachlorophenol must be met DFTPP
can be mixed in solution with either of
these compounds to complete two
specifications with one injection, if
desired.
11.2 To perform the mass calibration
test of the GC-MS system, the following
instrumental parameters are required:
Electron energy—70 volts (nominal).
Mass range—35 to 450 a.m.u.
Scan time—7 seconds or less.
11.3 GC-MS system calibration-
Evaluate the system performance each
day that it is to be used for the analysis
of samples or blanks by examining the
mass spectrum of DFTPP. Inject a
solution containing 50 ug DFTPP and
check to insure that performance criteria
listed in Table 10 are met. If the system
performance criteria are not met, the
analyst must retune the spectrometer
and repeat the performance check. The
performance criteria must be met before
any samples or standards may be
analyzed.
12. Gas Chromatography-Mass
Spectrometry of Base/Neutral Fraction.
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3. 1979 / Proposed Rules 69543
12.1 At the beginning of each day
that base/neutral analyses are to be
performed, inject 100 nanograms of
benzidine either separately or as part of
a standard mixture that may also
contain 50 ng of DFTPP. The tailing
factor for oenzidine should be less than
3. Calculation of the tailing factor is
given in Reference 2 and described in
Figures.
12.2 Establish chromatographic
conditions equivalent to those in Tables
4 and 5. Included in these tables are
estimated retention times and
sensitivities that can be achieved by this
method. Examples of the separations
achieved by these columns are shown in
Figures 1 and 3 through 7.
12.3 Program the GC/MS to operate
in the Extracted Ion Current Profile
(EICP) mode, and collect EICP for the
three ions listed in Tables 7 and 8 for
each compound being measured.
Operating in this mode, calibrate the
system response for each compound as
described in Section 6, using either the
internal or external standard procedure.
12.4 If the internal standard
approach is being used, the analyst may
not add the standard to sample extracts
until immediately before injection into
the instrument Mix thoroughly.
12.5 Inject 2 to 5 jil of the sample
extract. The solvent-flush technique is
preferred. If external calibration is
employed, record the volume injected to
the nearest 0.05 pi. If the response for
any ion exceeds the linear range of the
system, dilute the extract and reanalyze.
12.6 Qualitative and quantitative
measurements are made as described in
Section 14. When the extracts are not
being used for analysis, store them in
vials with unpierced septa in the dark at
14° C.
13. Cas Chromatography/Mass
Spectromatry of Acid Fraction.
13.1 At the beginning of each day
that acid fraction analyses are to be
performed, inject 50 nanograms of
pentachlorophenoi either separately or
as part of a standard mixture that may
also contain DFTPP. The tailing factor
for pentachlorophenoi should be less
than 5. Calculation of the tailing factor is
given hi Reference 2 and described in
Figure 8.
13.2 Establish chromatographic
conditions equivalent to those in Table
6. Included in this table are estimated
retention times and sensitivities that can
be achieved by this method. An example
of the separation achieved by the
column is shown in Figure 2.
13.3 Program the GC/MS to operate
in the Extracted Ion Current Profile
mode, and collect EICP for the three ions
listed in Table 9 for each phenol being
measured. Operating in this mode,
calibrate the system response for each
compound as described in Section 8
using either the internal or external
standard procedure.
13.4 If the internal standard
approach is being used, the analyst may
not add the standard to sample extracts
until immediately before injection into
the instrument. Mix thoroughly.
13.5 Inject 2. to 5 jul of the sample
extract. The solvent-flush technique is
preferred. If external standard
calibration is employed, record the
volume injected to the nearest 0.05 pi. If
the response for any ion exceeds the
linear range of the system, dilute the
extract and reanalyze.
13.6 Qualitative and quantitative
measurements are made as described in
Section 14. When the extracts are not
being used for analysis, store them in
vials with unpierced septa in the dark at
4'C.
14. Qualitative and Quantitative
Determination.
14.1 To qualitatively identify a
compound, obtain an Extracted Ion
Current Profile (EICP) for the primary
ion and the two other ions listed in
Tables 7, 8, or 9. The criteria below must
be met for a qualitative identification.
14.1.1 The characteristic ions for the
compound must be found to maximize in
the same or within one spectrum of each
other."
14.1.2 The retention time at the
experimental mass spectrum must be
within ±60 seconds of the retention
time of the authentic compound.
14.1.3 The ratios of the three EICP
peak heights must agree within ±20%
with the ratios of the relative intensities-
for these ions in a reference mass
spectrum. The reference mass spectrum
can be obtained from either a standard
analyzed through the GC-MS system or
from a reference library.
14.1.4 Structural isomers that have
very similar mass spectra can be
explicitly identified only if the resolution
between the isomers in a "standard mix .
is acceptable. Acceptable resolution is
achieved if the valley height between
isomers is less than 25% of the sum of
the two peak heights. Otherwise,
structural isomers are identified as
isomeric pairs.
14.2 In samples that contain an
inordinate number of interferences the
chemical ionization (CI) mass spectrum
may make identification easier. In
Tables 7 and 8 characteristic CI ions for
most of the compounds are given. The
use of Chemical ionization MS to support
El is encouraged but not required.
14.3 When a compound has been
identified, the quantification of that
compound will be based on the
integrated area from the specific ion plot
of the first listed characteristic ion in
Tables 7, 8 and 9. If the sample produces
an interference for the first listed ion,
use a secondary ion to quantify.
Quantification will be done by the
external or internal standard method.
14.4 Internal Standard—By adding a
constant known amount of internal
standard (Q, in fig) to every sample
extract, the concentration of pollutant
(C0) is pg/1 in the sample is calculated
using equation 2.
Eq. 2
(A,) (CJ
(Au)
(Vo)
Where: V. is the volume of the original
sample in liters, and the other terms are
denned as in Section 6.3.
14.5 External Standard—The
concentration of the unknown can be
calculated from the slope and intercept
of the calibration curve. The unknown
concentration can be determined using
equation 3.
Eq. 3
Micrograms/liter = ng/ml
(A)(Vt)
where:
A=mass of compound from calibration curve
(ng).
Y!=volume of extract injected (/il).
Vt» volume of total extract (jil).
V,« volume of water extracted (ml).
14.6 Report all results to two
significant figures. Report results in
micrograms per liter (Base/Neutrals and
Acids) without correction for recovery
data. When duplicate and spiked
samples are analyzed, all data obtained
should be reported.
14.7 In order to minimize
unnecessary GC-MS analysis of method
blanks and field blanks, the field blank
may be screened on a FID-GC equipped
with the appropriate SP-2250 or SP-1240
DA columns.
15. References
-------
69544 Federal Register / Vol. 44. No. 233 / Monday. December 3. 1979 / Proposed Rules
1. Method 610, Polynuclear Aromatic
Hydrocarbons, EMSL, Cincinnati, Ohio
45288, 1979.
2. "Reference Compound to Calibrate Ion
Abundance Measurement in Cas
Chromatography — Mass Spectrometry
Systems." }. W, Eicheiberger, L. E. Harris
and W. L Budda, Anal. Cheat. 47, 995-1000
(1975).
Bibliography
1. "Sampling and Analysis Procedures for
Screening of Industrial Effluents for Priority
Pollutants," March 1977 (revised April
1977). USEPA, Effluent Guidelines Division.
Washington, D.C. 20460.
2. "Proceedings.— Seminar on Analytical
Methods for Priority Pollutants":
Volume 1 — Denver, Colorado, November
1977
Volume 2 — Savannah, Georgia. May 1978
Volume 3 — Norfolk, Virginia, March 1979
USEPA, Effluent Guidelines Division.
Washington. D.C. 20460.
T»»il*A.—uasa-NeutralExtractati/es
Compound STORET
No.
Acenaphthene ..... .. - • . ,. - , 34205
AcannpntfiyWne. , 34200
Anlhrac«n«. . ........ ,.., , ,.. 345?O
Benzo(a)anthracana 34526
8erzo(l()fluorantnene 34242
Ctan7o(g,n i)p*ry*tr» ?4f?1
3***tfcjiri* 39120
«^2^NQfO«tiy<)«ih«r •U77t
8m<2-chloroethoxy)melhane 34278
8o<2-ettiylhexyl)phthalate _ _ 39100
Bw(2-chloroiaopropyl)ether 34283
4-Bromophenyl phenyl ether. _._„._ 34636
Butyl benzyl phthalate 34292
2-CNoronaphdlalene _... .. 34581
4OHorophenyt phenyl ether _. 34641
0*enzo-n-r'"C>Vl*™~« tUM
N-NitroaodlpnenyuuiMW ._ - ,-m, ,„..-- 344.1$
Phenantnrene 34461
2.3.7,8-Tetrachlorodibenzo^dioxin 34675
1 ,2.4-Tnchlorobenzene 34551
Table) i-Aca Bmctatles
Compound STORET
No.
4-CMoro-3-melhylphenol . . 34452
2.4-Otahlorophenol 34601
Table t— Ada Extractabtos— Continued
Compound STORET
Na
2 4-Otmethylphenol ... 34606
2,4-OWtwpfienol 34619
2-MethyM,64Mlrophenol. 34657
Z4.6-Titahtoroprienol 34621
Table *.— Pesticide Extractablea
Compound STOH6T
Na
Aldrm___ . _ _ .... _ 39330
a-8HC _. , , ,„._ 39337
d-BHC 39340
g-SHC . .__ _._ 34259
4,4'-OOO....-_. —.. _._._. ... „._ . 39310
44-npr ........... „ . -men
Endoaulfan 9 _ - 34359
Cnrt0ftulfan Surfatt V351
Ervjrtn • 19300
En*nAJdehyde 34366
HeptacNor Epoiode 39420
PTB.inm iuj7i
PCS-1221 - .. . 39488
PCB.1M? .,. , „ „ 34497
pea. 1242 39494
PCB-12S4 39504
Table 4— Gas Chromatography of Base/ Neutral
ExtractatHes
Reten- Unit of detection
Compound don ##
(Ma) ng injected w/1
1,3-Oichkxobenzene 74 20 10
1 4^*4t4yc£ianz9n4 73 20 10
Hexachloroethane 8.4 20 to
as<2-cMoro«inyl)etrier 8.4' 20 10
1.2-Ocnlorobertzene „ 8.4 20 10
Nitrobenzene 11.1 20 10
1.2,4-TricrHorobenzene 11.6 20 10
laophorone 119 20 10
Naphthalene 12.1 20 10
BislZ-chloroethoxy) methane 12-2 20 10
Hexachlorocyclopentadiene 13.9 20 10
2-Chloroniphtnajene 15.9 20 10
AcanaphWene „. 178 20 10
Dimethyl phtfiatate _ 183 20 10
Fluorene.... ....,,,_ 19.5 20 10
4-CNoropnenyl phenyl ether ._ _ 19.5 20 10"
2,4-Oinrtrotohjene 19 fl 20 10
1,2-Oiphenyl hydrazme' . 20.1 20 10
Dlethyl phthalate 20.1 20 10
4.8romoptwnyl pheny* «th«r._ „ 21 2 20 10
Anthracene .. ._ 22.8 20 10
Di-n-butyl phthalate ..._ _ 24 7 20 10
Ruorantnene _ „ _ 28.5 20 10
Pyrene 273 20 10
Senzidine,.... . . 28.8 20 10
Butyl benzyl phthalaie 29.9 20 10
Sis(2-ethythaxy() pnthalate.. .306 20 10
Chryaena „„ 31.5 20 10
Table 4— Gas Chromatography of Base/Neutral
Extractabtes — Continued
Reten- Limit of detection
Compound ton ##
time #
(mm.) ng intected i>9/1
Senzofalanthrecene 31 5 20 to
nu 3rt .
Toxaphene . _ ... 25 to 34 ... .
PT.fJ.1J49 ,,, . JO !(>.•?? .....„, ,
•6 toot glass column (V, m. OO x 2 mm IO) packed with
3% SP-2250 coated on 100/120 mesh Supelcoport Camer
gas: hetum at 30 ml per nun. Temperature program: Isother-
mal for 4 minutes at 50* C, then a* per minute to 270* HoM
at 270* C for 30 minutes. If desired, capillary or SCOT col-
umns may be used.
#This is a minimum level at which the entire analytical
system must give mass spectral confirmation. (Nanograms in-
lected is based on a 4 jj" injection of a one-Mar sample that
has been extracted and concentrated to a volume ol 1.0 mL
Table 6.— Gas Chromatography of Acid Extractables
Reten- Unit of
Compound ten time* detection*
(mm) — — —
ng injected ng/l
2-Chkxophenol 59 50 25
2-Nitrophenol 64 50 25
Phenol 80 50 25
2.4-OlmettrytptienoL _ 9.4 50 25
2,4-Dichlorophenol 98 50 25
2.4.6-Trlchlorophenof 11.8 50 25
2,4-Omitrophenol _ „ 159 500 250
-------
Federal Register / Vol. 44. No. 233 / Monday, December 3.. 1979 / Proposed Rules 69545
Tabl» 6.— Gas Ctirom'atography of Acid
ExtractaUes— Continued
Reien- Limit of
Compound tion lime* detection*
ng injected ng/l
2-MethyM 6-dmrtrophenol 162 500
Pentachlorophenol 175 50
4-Nltropnenol . — 20.3 50
250
25
25
•6 foot^glass column (V* m. OO x 2 mm ID) Packed with
1% SP-1240 DA coated on 100/120 mesh Supelcoport Cam-
ar gas: helium at 30 ml per min. Temperature program: 2 mm
isothermal at 70', then 8' per mm to 200' C.Jf desired, capil-
lary or SCOT columns may be used.
#Thw is a minimum level at which the entire analytical
system must give mass spectral confirmation. (Nanograms in-
lected is based on a 2 nl miection of a one liter sample that
has been extracted and concentrated to 1.0 ml.)
Table 1.— Base/Neutral Sxtractables Characteristic Ions
1 2-Dichiorobenzene -
HexaChtOTObUtadlene „ „ „.,.
Naphthalene —> •
Hexachlorocyclopentadiene
2-Chloronaphthalene
Fluorine
2,4-Oinitrotoluane .
1 2'0iphenylhydrazine l . « ....
Dietnylphthalate • -...—
N-Nitrosodlptwiylamirw'
Anthracene i ...
Ottxityt phthalata .... .... •« ...
Pyrene ...
Banzidine . .-
ChfVSene . ._. „ „
Characteristic ions
Electron impact
146
146
117
93
146
45
130
92
77
225
180
128
93
237
162
152
154
63
165
168
204
165
77
149
168
284
243
178
178
149
202
202
184
149
149
228
223
252
149
252
252
252
276
278
276
42
45
188
148
148
201
S3
148
77
42
95
123
223
182
129
95
235
164
151
153
194
63
16S
206
69
93
177
168
142
250
179
179
ISO
101
101
92.
91 . .
167
228
229
254
253
253
253
139
139
138
74
49
322
94
113
113
199
95
113
79
101
138
65
227
145
127
123
272
127
153
152
164
121
167
141 . ....
163
105
150
167
249
141
178
176
104
100
100
185
279
229
226
126
125
125
125
277
279
277
44.
51
320
80
Chemical lonczation
(methane)
148
148
199
S3
146
77
139
124
223
181
129
65
235
163
152
154
151
33
166
183
185
177
169
284
249
178
178
149
203
203
185
149
149
228
228
252
252
252
27B
278
276
59
189
143
148
201
1 07
148
135
167
152
225
183
157
107
237
191
153
155
163
211
167
211
213
223
170
286
251
179
179
205
231
231
213
299
229
229
253
253
253
277
279
277
217 ...
150
150
203
109
150
137
178
164
227
209
169
137
239
203
181
183
164
223
195
223
225
251
198
288
277
207
207
279
243
243
225
327
257
257
281
281
281
305
307
305
1 Detected as azobenzene.
'Detected as diphenytamine.
'Suggested internal standard.
-------
69546
Federal Register / Vol. 44. No. 233 / Monday. December 3.1979 / Proposed Rules
T«bt« •.-**«#**•«•
font
Compound
CtwraoMMto ien* •MeMn knpwt
a-8HC_
9-8HC..
d-SHC-
andoaultinl_
andoaulfan !)„
4,4'-OOT
«nd
-------
COLUMN: 3% SP-2250 ON SUPELCOPORT
PROGRAM: BO°C, 4 MIN. 8'PER MIN TO 270*C
DETECTOR: MASS SPECTROMETER
2,4-DINITROTOLUENE-' N NITBOSO OIPHENVIAMINE
10
15
20 25 30
RETENTION TIME-MINUTES
35
40
4$
I
<
r-
•z
o
o
I
o
•i
to
Figure 1. Gas chromatogram of base/neutral fraction
•o
o
I
a
a
-------
COLUMN: IK SP-1240DA ON SUPELCOPORT
PROGRAM: 70°C-2 MIN. 8*/MIN TO 200°C.
DETECTOR: MASS SPECTROMETER.
£
(B
s.
I
CL
to
a
(D
O
(D
g
cr
co
t-»
CO
8 10 12 14
RETENTION TIME-MINUTES
16
18
20'
22
Figure 2. Gas chromatogram of acid fraction
CO
OB
CL
CD
CO
-------
Federal Register / Vol. 44, No. 233 / Monday, December 3. 1979 / Proposed Rules 69549
COLUMN: 3% SP-2250 ON SUP&COPORT
PROGRAM: 50'C-4 MIN, 8«/MlNUTE TO 270'C
DETECTOR: MASS SPECTROMETER
10 15 20
RETENTION TIME-MINUTES
25
30
Figure 3. Gas chromatogram of pesticide fraction
-------
69550
Federal Register / Vol. 44, No. 233 / Monday, December 3, 1979 / Proposed Rules
COLUMN: 3% SP-2250 ON SUPELCOPORT
PROGRAM: 50*C, 4 WIN. 8"PER WIN TO 270*C
DETECTOR: MASS SPECTROMETER
PEAKS GIVING THE THREE
CHARACTERISTIC IONS
20
25
30
RETENTION TIME-MINUTES
Figure 4. Gas chromatogram of chlordane
-------
COLUMN: 354 SP-2250 ON SUPELCOPORT
PROGRAM: 50*C. 4 MIN. 8°PER WIN TO 270*C
DETECTOR: MASS SPECTROMETER
COLUMN: 3% SP-2250 ON SUPaCOPORT
PROGRAM: 50°C. 4 MIN, 8*PER MIN TO 27CPC
DETECTOR: MASS SPECTROMETER
25 3d
RETENTION TIME-MINUTES
35
Figure 5. Gas chromatogram of toxaphene
A ffl/e 224 PRESENT
B m/e 260 PRESENT
C m/e 294 PRESENT
25 3T
RETENTION TIME-MINUTES
35
Figure 6. Gas chromatogram of Arochlor 1248
S
a.
I
n>
-------
COLUMWc 3X SP-2250 ON SUPaCOPERT
PROGRAM: 50*0. 4 WIN, 8* PER MIN TO 270"C
DETECTOR: MASS SPECTROMETER
A m/e 294 PRESENT
B rn/e 330 PRESENT
C m/e 362 PRESENT
20 25 30
RETENTION TlME-MINUTtS
Figure 7. Gas chromatogram of Arochlor 1254
BILLING CODE 6560-01-C
TAILING FACTOR
Example calculation: Peak Height = DE = 100mm
10% Peak Height = BD = 10 mm
Peak Width at 105; Peak Height - AC = 23 mm
AB=tT! mm
12
Therefore: Tailing Factor- — =1.1
11
Figure 1 . Tailing factor calculation
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