Friday
October 26, 1984
Part VIII
Environmental
Protection Agency
40 CFR Part 136
Guidelines Establishing Test Procedures
for the Analysis of Pollutants Under the
Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
ENVIRONMENTAL PROTECTION
AGENCY
40 CFR Part 136
[FRL-2636-6]
Guidelines Establishing Test
Procedures for the Analysis of
Pollutants Under the Clean Water Act
Note: This reprint incorporates typographi-
cal corrections which were published in the
Federal Register of Friday, [anuary 4, 1985 on
page 695.
AGENCY: Environmental Protection
Agency (EPA).
ACTION: Final Rule and Interim Final
Rule with Request for Comments.
SUMMARY: EPA is amending its
Guidelines Establishing Test Procedures
for the Analysis of Pollutants, as
proposed on December 3.1979. EPA is
also reprinting existing test procedures
for the convenience of readers.
However, only those test procedures
which are new or revised are being
promulgated for purposes of judicial
review. The purpose of this amendment
is tcpestablish:
• New test procedures (including
quality control requirements) for the
analysis of priority toxic organic
polhitants;
• A new test procedure for the
measure of carbonaceous biochemical
oxygen demand (CBOD);
• A new test procedure based upon
inductively coupled plasma optical
emission spectroscopy for the analysis
of most of the regulated trace metal
pollutants: and.
• Mandatory container materials,
preservations, and holding times for
samples of the parameters covered by
this regulation.
The quality control requirements
establish control limits for acceptable
analytical performance. However, the
specific control limits in the test
procedures for the priority toxic organic
pollutants are being promulgated as an
interim rule with a request for
comments. Comments should be limited
to the calculation of the numerical
warning limits for the revised quality
control sections.
In accordance with the Clean Water
Act (CWA), these procedures will be
required for filing applications for
National Pollution Discharge
Elimination System (NPDES) permits
and for State certifications. These test
procedures will also be used for
compliance monitoring and to express
pollutant quantities, characteristics, or
properties in effluent limitations
guidelines and standards and in
pretreatment standards set forth at 40
CFR Parts 402 through 699 (unless
otherwise specifically noted or defined
in those parts).
DATES: In accordance with 40 CFR
100.01 (45 FR 26048), this regulation shall
be considered issued for purposes of
judicial review at 1:00 p.m. eastern time,
November 7,1984. These regulations
shall become effective for all methods
except CBOD5 on January 22,1985. The
regulation relating to CBODs [40 CFR
§ 136.3(a) Table IB, parameter 14] will
be effective November 23.1984.
Comments on the interim final rule for
specific control limits [40 CFR § 136.3(a)
Table 1C, footnote 7, and Table ID.
footnote 7] will be accepted until
December 24,1984.
Under section 509(b)(l) of the Clean
Water Act, judicial review of this
regulation can be obtained only by filing
a petition for review in the United States
Court of Appeals within 90 days after
these regulations are considered issued
for purposes of judicial review (see
NRDC v. EPA. 673 F.2d 402, D.C. Cir..
1982). Under Section 509(b)(2) of the
Clean Water Act, the requirements of
this regulation may not be challenged
later in civil or criminal proceedings
brought by EPA to enforce these
requirements.
The information collection
requirements contained in 40 CFR
136.3(e), has not been approved by the
Office of Management and Budget
(OMB) and is not effective until OMB
approves it. The incorporation by
reference of certain publications listed
in the regulation is approved by the
Director of the Federal Register as of
January 22,1985.
ADDRESSES: Comments on the
calculation of control limits should be
labelled as "Section 304(h): Control
Limit Calculations" and submitted to:
Dr. Robert B. Medz, Water and Waste
Management Monitoring Research
Division, Office of Research and
Development (RD-680), U.S.
Environmental Protection Agency, 401 M
Street, SW., Washington, D.C. 20460.
Telephone Number: (202) 382-5788.
Most of the public record for this
rulemaking will be available for
inspection from 8:00 A.M. to 4:00 P.M. in
EPA's Public Information Reference
Unit, Room 2404 (rear of the EPA
Library), 401 M Street, SW.,
Washington. D.C. 20460. The remainder
of the record (primarily materials
describing interlaboratory studies) will
be available at the Environmental
Monitoring and Support Laboratory at
the Andrew W. Breidenbach
Environmental Research Center, 26
West St. Clair Street, Cincinnati. Ohio
45268, from 8:00 a.m. to 4:00 p.m.,
Monday through Friday.
The EPA information regulation (40
CFR Part 2) allows the Agency to cha
a reasonable fee for copying.
FOR FURTHER INFORMATION CONTAC1.
Dr. Robert B. Medz. at the address listed
above or call (202) 382-5788.
SUPPLEMENTARY INFORMATION:
Outline of Preamble Discussion
I. Authority
II. History and Background
A. Structure and History of 40 CFR Part 136
B. Consent Decree and the Priority
Pollutants
III. Summary of the Proposed Amendment
A. Purposes
B. GC, HPLC. and GC/MS Test Procedures
C. Quality Control and Quality Assurance
D. Other Table I Organic Parameters
E. ICP Test Procedure
F. CBODi Test Procedure
IV. Highlights of Final Test Procedures
A. Restructuring of Table I
B. GC. HPLC, and GC/MS Test Procedures
C. ICP Test Procedure
0. CBOOs Test Procedure
E. Table II: Required Containers.
Preservation Techniques, and Holding
Times
F Incorporation by Reference
V. Public Participation and Response to Most
Significant Comments
A. GC. HPLC. and GC/MS Test Procedures
1. Policy on Applicability of Test
Procedures
2. Flexibility and Analysts' Professional
Judgment
3. Quality Control and Quality Assuranj
B. ICP Method '
C. CBOD, Method
D. Table II: Required Containers,
Preservation Techniques, and Holding
Times
E. Cost Estimates
F. Publication of Full Texts of Test
Procedures
G. Consistency of Analytical Methods
Approved Under Different Acts
VI. Economic Analyses
VII. Effective Dates
I. Authority
Today's amendment was proposed on
December 3,1970 (44 FR 69464). It is
being promulgated under the authority
of sections 301, 304(h) and 501(a) of the
CWA, 33 U.S.C. 1251 et seq. Section 301
forbids anyone to discharge any
pollutant into navigable waters except
pursuant to an NPDES permit issued
under the CWA. Permits are issued
under § 402, which is referenced in
Section 304(h). Subsection 304(h)
requires the Administrator to
"promulgate guidelines establishing test
procedures for the analysis of pollutants
that shall include the factors which must
be provided in any certification
pursuant to section 401 of the Act or
permit application pursuant to sectiojj
402 of the Act." Section 501(a)
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authorizes the Administrator "to
prescribe such regulations as are
necessary to carry out his functions
under the Act."
The Administrator has also made
these test methods applicable to
monitoring and reporting of NPDES
permits (40 CFR Part 122. Subsections
122.21. 122.41. 122.44. and 123.25), and
implementation of the pretreatment
standards issued under section 307 of
the CWA (40 CFR Part 403. Subsections
403.10, and 403.12).
II. History and Background
A. Structure and History of 40 CFR Part
136
The Administrator first approved test
procedures for the analysis of
wastewater pollutants on October 16.
1973. and first amended the list of
approved test procedures on December
1, 1976 (See 38 FR 28758, October 16.
1973 and 41 FR 52780. December 1. 1976).
The full texts of the approved test
procedures incorporated by reference in
the regulation are considered to be part
of the regulatory language. Most of the
test procedures were cited from the
following compilations of consensus test
procedures:
• "EPA Manual of Methods for the
Chemical Analysis of Water and
Wastes,"
• "Standard Methods for the
Examination of Water and
Wastewater,"
• "American Society for Testing and
Materials (ASTM) Annual Book of
Standards. Part 31, Water."
• "Official Methods of Analysis of the
Association of Official Analytical
Chemists (AOAC)."
• "Methods for Determination of
Inorganic Substances in Water and
Fluvial Sediments of the U.S. Geological
Survey."
Additional test procedures were
incorporated from other standards
groups, such as the American National
Standards Institute (ANSI), or from the
open literature. Several test procedures.
such as those for the analysis of
benzidine. were incorporated from
specific EPA sources.
Test procedures have previously been
approved for about 115 different
parameters. Those procedures apply to
the analysis of inorganic (metal, non-
metal, mineral), nutrient, demand,
residue, radiological, organic.
bacteriological, and physical
parameters. For any given parameter,
the regulations generally approved
several different analytical methods.
The December 1.1976 amendments to 40
CFR Part 136 approved certain test
procedures which were identified in
tabular form (Table I). The discharge
parameters to be measured were
presented alphabetically. Each
parameter was followed by a brief test
procedure description and by page
numbers of the incorporated references.
This unambiguously identified the
approved test procedure.
An equivalency program is provided
in 40 CFR Part 136. Under this program
the Administrator may approve
alternate test procedures developed and
proposed by dischargers or other
persons. If dischargers or other persons
wish to use such alternate test
procedures, they must apply to the State
or Regional EPA permitting office (for
limited approval) and to the Director of
the Environmental Monitoring and
Support Laboratory in Cincinnati (for
nationwide approval). The equivalency
provisions are included in these
guidelines to encourage the development
of new analytical methods, and to give
analysts a number of options for
resolving analytical problems that may
be unique to specific wastewaters.
Finally, there may be discharges from
some particular industries which need to
be regulated on the basis of parameters
or test procedures which have not been
proposed and approved within the scope
of the test procedures guidelines under
40 CFR Part 136. EPA may include such
parameters and alternate test
procedures within the rule-making for
these industries in accordance with the
provisions prescribed at 40 CFR 401.13.
"Test Procedures for Measurements."
Such test procedures may be integrated
into the text of future amendments and
revisions of 40 CFR Part 136.
The following provides a brief
regulatory history of 40 CFR Part 136
prior to the current amendment.
• First proposal: 38 FR 17318 (at 40
CFR Part 130, later redesignated as Part
136), June 29,1973.
• First promulgation: 38 FR 28750 (at
40 CFR Part 136), October 16, 1973.
• First amendment proposal: 40 FR
24535, June 9, 1975.
• First amendment promulgation: 40
FR 52780. December 1.1976.
• Second amendment proposal: 44 FR
69464, December 3,1979.
• Correction Document, second
amendment proposal: 44 FR 75031,
December 18,1979.
• Comment period reopened, second
amendment proposal: 46 FR 3033,
January 13,1981.
• Equivalent Alternate Test
Procedure Approvals:
Chemical Oxygen Demand: 43 FR 9341.
March 7.1978.
pH and Ammonia: 43 FR 38618, August
29,1978.
Nitrite Nitrogen: 44 FR 25505, May 1,
1979.
Manganese: 44 FR 34193. June 14,1979.
Chemical Oxygen Demand: 45 FR 26811,
April 21,1980.
Copper and Zinc: 45 FR 36166, May 29,
1980.
Iron: 45 FR 43459. June 27, 1980.
Residual Chlorine: 46 FR 58489,
December 2.1981.
Many reviewers of the proposed
amendments requested that certain
documents upon which the procedures
were based be made available for
review. In response, the Administrator
sent 38 supporting documents to EPA's
Regional Offices and to the EPA
Headquarters Library in Washington.
D.C., for inspection by the public. The
closing date for comments was also
extended from February 1.1980. to April
28.1980, to permit adequate time for
public inspection of the record.
The Agency also started a series of
formal inter-laboratory collaborative
validation studies (each comprising of 15
to 20 laboratories) for the trace organic
priority pollutant test procedures and
the trace metal ICP test procedure.
These were designed to establish
expected inter-labaoratory precision
and accuracy characteristics of the test
procedures.
Late in 1980, representatives of the
Chemical Manufacturers Association
(CMA). the American Petroleum
Institute (API), and EPA met informally
to discuss the reliability of some of the
proposed test procedures. On January 5.
1981. these representatives met again to
more formally discuss these concerns.
CMA and API felt the test procedures
for the toxic organic priority pollutants
should not be promulgated as final until
the inter-laboratory validation studies
had been completed. The comment
period was reopened on January 12,
1981, to allow all interested persons to
inspect the official transcript of the
January 5,1981 meeting, and if needed.
to amend their earlier comments on the
proposed regulation. The extended
comment period closed on February 2.
1981.
B. Consent Decree and the Priority
Pollutants
In 1976, the Agency entered into a
consent decree in the District Court of
the District of Columbia (Natural
Resources Defense Council, Inc., et al. v.
Train. 8 ERC 2120 (D.D.C. 1976), as
modified 12 ERC 1833 (D.D.C. 1979), and
by the Court's Orders of October 26.
1982. August 2. 1983. January 6,1984.
and July 5,1984), and the decree
requires EPA to study the occurrence of
65 categories of pollutants in industrial
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wastewaters, and to limit the discharge
of those pollutants in effluent by issuing
effluent guidelines based on the "best
available control technology
economically achievable" (BAT),
pretreatment standards for new and
existing sources, and new source
performance standards. (Note: Within
those 65 categories. 114 specific organic
toxic pollutants and 15 inorganic
pollutants were identified for a total of
129 specific toxic pollutants studied by
EPA. Bis-chloromethyl ether (46 FR
10723, Feb. 4,1981) and
dichlorofluoromethane and
trichlorofluoromethane (46 FR 46103.
Jan. 8.1981) have since been removed
from the list, leaving 126 toxic pollutants
now listed as "priority" toxic
pollutants). The Agency began
development of test procedures for
measuring these pollutants in complex
industrial wastewater matrices.
Paragraph 4(c) of the Consent Decree
also required the EPA to establish and
implement a program to identify and
study pollutants other than the priority
pollutants. At a minimum, EPA was to
conmer those additional pollutants
listeij in Appendix C of the Settlement
Agreement. Those additional pollutants
were not included in the December 3,
1979 proposal of the regulation. In
studying Methods 1624 and 1625. EPA
has evaluated applying the methods to
those additional pollutants. A separate
notice in today's Federal Register
proposes to extend the scope of those
methods to include the paragraph 4(c)
pollutants.
The 1976 Test Procedures Guidelines
(41 FR 56780, December 1.1976),
provided approved test procedures,
selected from the various consensus
standards, for 14 of the 15 inorganic
priority toxic pollutants. The exception
was asbestos, for which no adequate
procedure was then available. The 1976
Guidelines also provided approved test
procedures, similarly selected, for
several chlorinated organic compounds
(including PCBs, pesticides, benzidine,
and pentachlorophenol). However,
neither those procedures nor existing
consensus standards were adequate to
meet the testing requirements for all of
the 114 priority toxic organic pollutants.
To fill this gap, the Agency embarked
on an extensive program to develop
additional test procedures to implement
sections 301, 304(h) and 402. By 1979,
these test procedures had been
developed to a stage that represented
the state-of-the-art analysis of the trace
organic priority pollutants in industrial
wastewater discharges. On December 3,
1979 the Agency proposed these
methods, together with a test procedure
for analysis of trace metals by
inductively coupled plasma optical
emission (ICP) and a test procedure for
determining the carbonaceous
biochemical oxygen demand of
municipal wastewaters, as amendments
to 40 CFR Part 136.
III. Summary of the Proposed Regulation
A. Purposes
On December 3,1979, the Agency
proposed to revise the "Guidelines
Establishing Test Procedures for the
Analysis of Pollutants." The primary
purposes of this proposal were:
(1) To amend Table I. List of
Approved Test Procedures, by adding
the priority toxic organic pollutant
parameters and approved alternate test
procedures for their analysis;
(2) To add an approved test procedure
for a new parameter, "carbonaceous
biochemical oxygen --demand" (CBOD),
which is important to secondary
biological treatment technology for
municipal wastewaters;
(3) To approve .an additional state-of-
the-art test procedure based on the
inductively coupled plasma optical
emission (ICP) principle for the analysis
of most of the trace metal parameters
which were already covered in Table I;
and,
(4) To remove Table I, footnote 1,
which recommended sample container
materials, preservation procedures, and
maximum holding times, and to
specifically list those elements as
mandatory requirements in a new Table
II. "Required Containers, Preservation
Techniques, and Holding Times."
Several test procedures were included
for each of the priority toxic organic
pollutants. This allowed several
analytical options for most analyses.
The CBOD parameter was included for
analysis of a new specific measure of
oxygen demand. The ICP test procedure
was included to provide an additional
and more rapid tool for trace metal
analysis. Mandatory sample container
materials, preservation techniques, and
maximum holding times were included
because these have been found to be
critical to assure NPDES data quality.
The test procedures for the toxic
organic priority pollutants were
developed by the Agency in response to
the mandates of the Consent Decree. At
the time of proposal, the test procedures
had been subjected to intensive single
laboratory developmental testing. They
were considered to be the best state-of-
the-art test procedures available for the
routine analysis of treated wastewaters
for organic pollutants. They also
appeared to be applicable to the
analysis of untreated wastewaters.
Multi-laboratory validations of these
test procedures had been planned but
had not yet been started. The Agency
decided to propose the test procedures
for priority toxic organic pollutants
before completion of the inter-
laboratory validation studies because:
• Even without inter-laboratory
validation these were (and are) the most
tested and intensively validated test
procedures available for the analysis of
the toxic organic priority pollutants in
industrial and municipal wastewater
discharges,
• Many permits were expiring and
permit renewals would require some
provision for priority pollutant analysis.
• The new round of BAT effluent
guidelines regulations would include
limits on priority toxic organic pollutant
discharges, and.
• The priority toxic organic pollutants
would need to be reported under the
permits regulations at 40 CFR Parts 122
and 123, and by pretreatment
regulations at 40 CFR Part 403.
The following discussion covers the
provisions of the proposed regulation in
more detail.
B. GC, HPLC. and GC/MS Test
Procedures
Since 1976 the text of the regulation
has listed pollutant parameters
alphabetically in Table I. "List of
Approved Test Procedures,"—either al
specific compounds or entities such as
"Benzidine" or as classes of compounds
or entities such as "Pesticides." The
individual parameters within such
classes, when identified, were entered
alphabetically within the class.
Approved test procedures were then
identified by test procedure descriptors
and by page numbers in specifically
identified references. In the case of
"Pesticides" and several other organic
chemical classes, the Table refers to the
full text of the approved test procedures
in order to clearly identify the scope of
the test procedures.
In 1979 the Agency proposed to
consolidate all of these organic chemical
parameters under a new class entry,
"Organic Compounds", and to identify
alphabetically all of the specific organic
compounds which were included within
the scope of the various approved test
procedures. All of the test procedures
and organic compounds which were
approved in 1976 continued to be
approved, and were re-printed
(unmodified) in the proposal only
because Table I was being restructured.
The new organic chemical entries in
Table I were the 114 (now 111) priority
toxic organic pollutants and the
proposed test procedures for their
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analysis. Twelve of the proposed test
procedures were based on gas
chromatography (GC) and/or high-
pressure liquid chromatography (HPLC).
Three of the proposed alternate test
procedures were based on gas
chromatography with mass
spectrometric detection (GC/MS).
Chromatography is the technique by
which compounds in mixtures are
separated by interaction between a
mobile and a stationary phase. The
stationary phase is placed in a tube
called a column, and the mixture is
moved through the column by the mobile
phase under the influence of a pressure
differential across the tube. Different
compounds are carried through the
column at different speeds by the mobile
phase. The order in which the
compounds exit the column (elute) is
determined by the chemical and
physical properties of the compounds
and of the mobile and stationary phases.
In each method the stationary and
mobile phases are selected to give the
most effective separation of the
compounds of interest.
In gas chromatography (GC), the
mobil'e phase is a gas. and the stationary
phasfr'is usually a high molecular weight
liquid; coated on an inert support or on
the ciSlumn wall of a very small
diameter tube (an open tubular column).
In liquid chromatography (LC) the
mobile phase is a solvent, and the
stationary phase is usually a selective
adsorbant. As compounds in the mixture
elute [exit) from the column, their
presence is indicated by various
detectors. One detector is the mass
spectrometer, hence the term GC/MS.
Following detection, the compounds can
be identified and then quantified by a
computer or by the analyst.
Separations by chromatographic
techniques are not always complete.
Several compounds within a mixture
which have similar chemical and/or
physical properties may simultaneously
elute from the column, along with the
compounds of interest. These are known
as interfering compounds or interferents.
Where compounds are easily
identified, conventional detectors can
often make better quantitative
assessments than GC/MS can.
However, in many cases, the
conventional detectors used in
chromatography are not able to
discriminate between the compounds of
interest and the interferents. In such
instances, a mass spectrometer usually
is able to discriminate between the
compounds of interest and the
interferents. Thus, it would be the
preferred detection system where
interferents are expected or many
compounds must be identified. An
alternative solution is to use a second
column containing a different stationary
phase. This aids in the identification of
the compounds of interest by providing
additional qualitative identification
when conventional chromatography
detectors are used.
The low cost of the conventional GC
detectors, relative to MS. makes the GC
option particularly attractive for routine
monitoring of small numbers of
pollutants. On the other hand, the GC/
MS test procedures allow for the
simultaneous or rapid sequential
measurement of large numbers of
different organic pollutants. They also
provide certain structural information
that can be used to minimize
interferences that would mask
compound identification by the less
specific conventional GC detectors.
EPA divided priority toxic organic
pollutants into 12 categories, based on
their physical and chemical properties
and chemical structures. A GC or HPLC
test procedure was then developed for
each category, with the expectation that
the pollutants within each category
could be measured by a single
procedure. These procedures were to be
routinely used where the pollutants to
be measured were known to have a high
probability of occurrence. GC and HPLC
could also be used for qualitative
identifications of unknown materials.
although the proposed GC/MS test
procedures were more suitable for this
purpose. In most cases, several GC or
HPLC configuations of inlet, column.
operating conditions, and detectors
were recommended with each
procedure. Each test procedure
stipulated that, if it were used to screen
samples for priority pollutants, an
analyst needed to verify any compound
identified with an independent
analytical protocol. The GC/MS was
suggested as such a protocol.
In the proposed tests of the organic
toxic priority pollutant test procedures.
it was not EPA's intent to require
separate samples for each test
procedure. Subsequent comments have
indicated that this was not clear from
the test procedure texts or in the
proposed sample preservation and
holding time requirements in Table II.
C. Quality Control and Quality
Assurance
Quality control (QC) includes all of
the means taken by an analyst or an
analytical laboratory to make certain
that the total measurement system,
including the analyst's performance and
matrix problems, are calibrated
correctly or accounted for. and remain
in calibration or accounted for in all
ensuing analyses. Quality assurance
(QA) includes all the means taken
within or beyond the laboratory to make
certain that the measurement systems in
different laboratories in a monitoring
network remain in calibration with a
common external standard, and hence
with each other. QA/QC seeks to assure
that analyses of the same substances
taken by different analysts at different
times and places are of the same quality
and are comparable within known
statistical confidence limits. EPA
proposed that the QC within the GC and
HPLC test procedures require the use of
field replicates to validate the precision
of sampling techniques. Laboratory
replicates and fortified samples were
also proposed in 1979 to validate the
precision and accuracy of analyses.
Since EPA's studies in this area were
not yet finalized at the time of the
proposal, additional quality control
guidance was described in general terms
and proposed as necessary to enable
evaluation of the performance of test
procedures.
Similar GC configuration and quality
control guidance was proposed for the
GC/MS test procedures, except that the
GC/MS could be used in a screening
application without a mandatory
confirmation protocol. However, a
separate, more intensive quality control
procedure was proposed and described
in detail, as it might be applicable to the
GC/MS test procedures.
D. Other Table I Organic Parameters
Proposed Table I was restructured to
include the previously designated
organic parameters benzidine,
pentachlorophenol. Aldrin. 6-BHC. y-
BHC. chlordane. 4,4'-DDD. 4,4'-DDE. 4.4'-
DDT, Dieldrin. Endosulfan. Endrin, and
Heptachlor as specific entries in the
organic compounds subsection. The
previously approved test procedures (41
FR 52780,'Dec. 1, 1976) for most of the
organic pollutants incorporated within
Table I had been entered without
changes under the broad parameters
"chlorinated organic compounds (except
pesticides)" and "pesticides." These
broad parameters included test
procedures for chlorinated organic
solvents, chlorinated hydrocarbon
pesticides, carbamate pesticides.
triazine pesticides, phosphate
pesticides, and chlorinated phenoxy
carboxylic acid pesticides. Approved
test procedures for these parameters
have been available on request from
EPA's Environmental Monitoring and
Support Laboratory in Cincinnati, Ohio
(EMSL-CI).
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E. ICP Test Procedure (ICP)
The ICP test procedure was proposed
as an additional option for the analysis
of 23 trace elements: aluminum, arsenic,
barium, beryllium, boron, cadmium,
calcium, chromium, cobalt, copper, iron,
lead, magnesium, manganese.
molybdenum, nickel, potassium.
selenium, silicon, silver, sodium,
vanadium, and zinc. It was listed in
Table I, under Metals. Lithium and
strontium were also included within the
scope of the ICP method although the
consent decree had not listed them as
high priority toxic parameters. This
proposal did not affect the previously
approved atomic absorption (AA),
voltametric, gravimetric, flame
photometric, and colorimetric test
procedures referenced in Table I for
these elements.
The ICP test is a rapid, multielement
procedure, representing state-of-the-art.
It was proposed because of its time- and
cost-effectiveness, and because
available data showed it to be
equivalent to the other approved
procedures for analyzing the designated
meflls in wastewater discharges.
F. CBOD* Test Procedure
The CBODs test procedure was a new
parameter proposed in Table I. It
responded to many requests for an
approved procedure to measure the
carbonaceous component of oxygen
demand. It was a draft version of the
consensus method now included as
Method 507 paragraph e(6) in the 15th
Edition of "Standard Methods for the
Analysis of Water and Wastewater."
The CBODs is a different measure of
oxygen demand than the total BODs
(Standard Method 507). Thus it cannot
be used to analyze oxygen demand
when an NPDES permit calls for BOD»
to be measured. The CBODs procedure
uses a nitrogen biochemical oxygen
demand inhibitor. This inhibitor allows
oxygen to be consumed only by
organisms that require carbon as their
nutrient source. In the presence of such
an inhibitor, the nitrogen compounds
remain refractory to biochemical
degradation, since the activity of
nitrifying organisms is suppressed.
IV. Highlights of Final Test Procedures
A. Restructuring of Table I
Users familiar with the former text of
section 136 will first notice the
reorganization of Table I. which lists
pollutant parameters for which
approved analytical methods exist, and
indicates the approved method(s)
available for each parameter. In the 1976
regulations, the parameters in Table I of
§ 136.3(a) were organized around broad
categories, such as bacteriological test
procedures and test procedures for
metallic or residue parameters. These
broader categories were then entered
alphabetically into Table I.
Analyses for an additional 111 organic
parameters have now become essential.
However, entering all those parameters
into a single list of approved methods
became unwieldy, especially with the
proliferation of footnotes to the table.
Therefore, to make Table I easier to use.
it has been restructured for this final
rulemaking into five sub-tables:
• Table IA. List of Approved
Biological Test Procedures.
• Table IB. List of Approved Inorganic
Test Procedures.
• Table 1C. List of Approved Test
Procedures for Non-Pesticide Organic
Compounds.
• Table ID. List of Approved Test
Procedures for Pesticides.
• Table IE. List of Approved
Radiological Test Procedures.
Throughout Table I. EPA has updated
numerous references to consensus
methods, e.g., from the 14th to the 15th
edition of "Standard Methods." These
changes are technical amendments
without substantive effect and are
promulgated as final amendments. EPA
has determined that notice and public
procedure on these updates are
unnecessary and contrary to public
interest. See 5 U.S.C. 553(b)(3)(B).
These technical amendments should
not affect any on-going enforcement
actions or other regulatory actions on
analyses performed with earlier
methods. Today's amendments do not
approve consensus methods adopted
since the proposal when they contain
substantive revisions to the previously
approved methods. Instead, the Agency
has retained its approval of the prior
method. Examples discussed below
include the retained approval of the 14th
edition Standard Method for phenols
(4AAP), and the limited approval of the
U.S.G.S. method for fecal streptococci
which, as approved, is identical to the
previous U.S.G.S. method.
Table IA includes bacteriological test
parameters which were approved in the
1976 Guidelines. Approved methods for
their analysis are now listed in a new
format: they are not substantively
changed. Previously cited references
have been updated. With the exception
of the U.S.G.S. test procedures for fecal
streptococci, no changes have been
made in the test procedures. As noted
only editorial changes have been made
in the texts of the other test procedures
in these updated references.
A new EPA reference is now
approved for several bacteriological test
parameters. The updated USGS fecal
streptococci test procedure is approved
only if the dissolution of the nutrient
medium is conducted in a boiling wai
bath. This is because dissolution on b
hot plate or over a open flame (which
appears to be permitted in the updated
reference) can lead to scorching or to
other alterations in the nutrient medium.
Table IB includes all of the inorganic
and physical parameters that were in
the 1976 Guidelines. The previously
cited references have been updated.
The ICP test procedure is now
approved as an additional alternate test
procedure for the analysis of 25 Table IB
trace element parameters. Antimony
and thallium are now included within
the scope of the ICP test procedure in
response to information made available
in comments which were received. The
only new parameter which has been
added to Table IB is the Carbonaceous
Biochemical Oxygen Demand (CBODs
parameter.
Table 1C includes 97 organic, non-
pesticide chemical parameters. Test
procedures for 21 of these parameters
were pproved in the 1976 Guidelines
and continue to be approved,
unchanged, in Table 1C. Ninety-five of
the Table 1C parameters (including 19
parameters approved since 1976) are
priority toxic organic pollutants for
which new test procedures were t
proposed. The new test procedures 4
essentially the same as those propose.
with the exception that. (1) where
possible, they have been made more
flexible in response to comments and (2)
quality assurance and quality control
standards have been defined. Two
newly-modified GC/MS test procedures.
Methods 1624 and 1625, which are
variants of Methods 624 and 625. have
been added to Table 1C. They differ
from Methods 624 and 625 by utilizing
stable, isotopically labeled analogs of
the analytes as internal standards for
GC/MS analysis. This allows the
analyst to accurately correct for
variability in analyte recovery
efficiency, responding to a criticism
raised by commenters.
With the exception of the test
procedures for benzyl chloride and
epichlorohydrin. all test procedures in
Table 1C prescribe quality control limits.
The actual specific control limits are the
sole elements of this regulation which
are promulgated as an "interim final"
action. This is because the data base
and calculations of these limits have
been developed since proposal.
However, the idea of specific
mandatory, acceptability criteria wa
fully proposed, favorably commente
on. and finally accepted. Thus the
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
regulatory framework for the specific
limits is being promulgated as final.
Table ID contains the 67 parameters
included under the general "pesticides"
parameter in the 1976 Guidelines.
Although most pesticides are organic
compounds, they have been listed
separately in Table ID rather than with
the other organic parameters in Table 1C
because of the wide association
between this subset of organic
compounds and their end use. Sixteen of
the 67 parameters are priority pollutants.
Three additional pesticides were
identified as priority pollutants under
the consent decree. Table ID therefore
now identifies 70 specific pesticides, of
which 19 are priority pollutants.
Methods 608 and 625. which were
proposed for the priority organic toxic
pollutants, were revised to incorporate
substantive comments. All other
references in Table ID have been
updated, but the updated references do
not require any substantive changes
from previously approved test
procedures.
Table IE now includes the five
radiological test procedures approved in
the 1976 Guidelines. All references have
been updated, and an EPA reference has
beea^dded. There are no substantive
textual changes in these updated test
procedures.
B. GC. HPLC. and GC/MS Test
Procedures
Analyses for organics depend upon a
variety of chromatographic techniques.
See subsection III-B above. EPA
proposed and is approving two HPLC
methods (605 and 610). 10 GC methods.
and three GC/MS methods (613. 624,
and 625). In addition. EPA has
responded to critiques of Methods 624
and 625 by approving two GC/MS/
isotope dilution variants (1624 and 1625).
Each method is accompanied by a
specific set of quality assurance (QA)
procedures. The QA process relies on
specific control limits calculated for
each parameter for which the method
can be used. The control limits indicate
the outer range of precision and
accuracy found in an extensive inter-
laboratory study. The limits represent
the minimum threshold of quality
expected of competent laboratories: 95
percent confidence level per compound
for the 600 series and the 99 percent
confidence level across the set of
compounds for the 1624 and 1625
methods. Most analyses should have far
better precision and accuracy. The
calculations of specific numerical
control limits for the calibration and
quality control sections of the GC,
HPLC, and GC/MS test procedures is
interim final. This means that thev are
legally effective, but that EPA will
accept comments on their calculation.
All other parts of these test procedures
are finally approved for the analysis of
the parameters which are indicated in
Tables 1C and ID.
Each method is approved for specific
organic compounds. In general. GC
Methods 601-603 and GC/MS Methods
624 and 1624 are approved for the
analyses of the purgeable priority
pollutants. GC Methods 604 and 606-612
and GC/MS Methods 625 and 1625 are
approved for the analysis of the non-
purgeable, volatile priority pollutants.
including, for Method 625 only, the
priority pesticide pollutants. Method 625
is also approved for screening samples
for 2.3.7.8-TCDD (2.3.7.8-
tetrachlorodibenzo-p-dioxin). but only
GC/MS Method 613 is approved for final
qualitative confirmation or
quantification of 2.3,7,8-TCDD in
samples. HPLC Methods 605 and 610 are
also approved for the analysis of the
nonpurgeable volatiles (the benzidines
and polynuclear aromatic
hydrocarbons). Methods 1624 and 1625
are approved for use interchangeably
with the other test procedures which are
being approved for the analysis of the
priority toxic organic pollutants. Their
, most significant difference from
Methods 624 and 625 is the requirement
that, where available, stable.
isotopically-labeled analogs of the
priority pollutants are to be used as
method internal standards. Since
Methods 624 and 625 do permit
flexibility in the selection of internal
calibration standards and surrogate
standards. Methods 1624 and 1625 are.
in essence, acceptable variants
permitted by Methods 624 and 625. They
improve on Methods 624 and 625 and
are generally preferable. However,
Methods 624 and 625 are also being
approved because they are widely
available, slightly less expensive, and
they are of use when interference and
recovery efficiency are not expected to
be problems.
In general, both GC/MS and non-MS
test procedures have been approved for
each of the priority toxic pollutants.
Most of the revisions of the proposed
test procedures were made either for
clarification or to give the analyst more
flexibility to practice professional
judgment. These procedures now
contain a section on safety, cautioning
analysts of the potential hazards
associated with exposure to the
chemical reagents required by the test
procedures, or to the toxic chemicals
being analyzed. Recommended and
mandatory quality assurance practices
are also given in each of the test
procedures.
Methods 601-604, 606-609. 611-613.
624. 625. 1624. and 1625 include
specifications for performing the tests.
These specifications are based on a
required primary GC column and
specified detector. A primary HPLC
column and specified detector are
required for Methods 605 and 610 and
specifications are provided. The primary
column is also used to identify the
pollutant. A secondary column and
detector are also defined, but not
required, for non-MS Methods 601-604
and 606-611. The secondary column and
detector can be used for confirmation of
priority pollutants identified by the
primary column for unfamiliar (non-
routine) samples (see sections 1.2 of the
methods). The GC/MS test procedures
are suggested as the confirmatory test
for identifications made by Methods 605
and 612. and may also be used as the
confirmatory test for identifications
made by Methods 601-604 and 606-611.
For example, an unfamiliar sample
which would be likely to need
confirmation would be a single sample
taken for an NPDES application. See 40
CFR 122.21. In contrast, routine
monitoring, such as'that for discharge
monitoring reports, would be less likely
to require a secondary column for
confirmation since the sample is more
likely to be familiar to the analyst.
Methods 606. 609. 611 and 612 all use
essentially the same procedure for
sampling, sample extraction, and
concentration. Thus a single sample may
be used to measure the parameters
within the scope of these methods.
Sample container materials,
preservation techniques, and holding
times are critical to the procedures and
are specifically defined (Methods 601-
613, 624. 625, 1624 and 1625). The design
and operation of the purge-and-trap
device in Methods 601-603. 624 and
1624. and the sample extraction
procedures of Methods 604-613. 625 and
1625 are precisely defined as well.
In response to public comments.
substantive revisions were made to
allow more flexibility in the remaining
parts of Methods 601-613, 624. 625,1624
and 1625. In Methods 604-613, after the
sample has been extracted, the analysts
are now free to choose a technique to
concentrate the extract. The same
flexibility is provided for selecting the
GC or HPLC configurations (column
packings, operating conditions, and
detectors). When analysts use
concentration techniques or
chromatographic configurations other
than those described in the test
procedures, their approaches must meet
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8 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
the performance criteria defined in the
section of the procedures dealing with
calibration and quality control.
The most difficult task in finalizing the
methods for organic analyses was
defining the relationship between
desirable flexibility in the methods and
necessary quality control. The proposal
specifically solicited comments on both
issues and many comments were
received on each. The final methods
resolve the issue by allowing far greater
method flexibility, but by establishing
specific control limits as a mandatory
part of the quality control procedure.
The proposal noted, and comments
confirmed, that method flexibility
should be inherent in the methods.
Historically, rigid protocols have been a
problem in organics analyses. For
example, an analyst may be using a
method, other than mass spectrometry,
to identify a few specific components
out of the several million known to
exist. This requires that interferences be
overcome and "canned" approaches
may not effectively address
interferences, particularly where
matrices are variable or diverse. Thus,
the £ood and Drug Administration
(FDA) and AOAC and other method
standardization organizations have
usually provided optional "clean-up"
procedures for organics, for example,
permitting analysts to use Florisil clean-
up for pesticides. Further, the analyst
may be interested in measuring only a
few compounds, while the proposed
method may be designed to measure
large categories of compounds. For
example, a particular industry may be
regulated only for the compound that
elutes from the gas chromatography
after a long program temperature run.
An inflexible method might require the
analyst to go through the entire
temperature run to look for a single peak
that elutes late in the chromatogram.
This may be needlessly inefficient. For
such reasons, EPA has decided to permit
flexibility in chromatographic
conditions.
Commentors also raised concerns
about inflexibility in sample
preparation. They objected to the
Kuderna-Danish glassware
concentration technique being the only
approved approach for concentrating
extracts. In fact, if the analysts are
measuring only the less volatile
compounds in a method category, it may
not be necessary to require a rigid
procedure for concentration. In this
case, it may be appropriate to allow
other procedures for concentrating
extracts.
After considering these issues, the
Agency has decided to allow limited
flexibility within the methods.
Specifically, chromatographic
conditions, including column packings
and detectors can be varied. This
approach allows continued technical
development of the methods. Thus EPA
avoided a rigid prescription of
technology that would soon be obsolete
due to the rapid advances occurring in
chromatography. However, the primary
objective underlying this flexibility is to
enhance precision and accuracy for
each analysis. Flexibility should not be
permitted if the altered technique would
be less precise or less accurate than the
standard approved analytical method.
Thus, a corollary of increased flexibility
was an increased need for a rigorous
and unambiguous quality control
procedure.
These basic decisions had become
clear by the time of the second.
reopened comment period. The
comments received in the second
comment period again supported the
issue of quality control and requested
that the criteria be specified more
clearly. Another general comment was
that the criteria should wait for the
results of the inter-laboratory method
validation studies and be based upon
those results. Today's rulemaking
reflects these comments, while
specifying that EPA will accept further
comments, limited specifically to the
calculation of control limits from that
new data base.
The quality control procedures now
take two different forms. First, there is a
"start-up test" to establish the
laboratory's basic ability to set up and
operate the analytical equipment and
procedure. The purpose of the start-up
test is two-fold; it establishes that
analytical equipment has been properly
set up. and it demonstrates the basic
ability of the analyst to recognize the
compounds of interest. It is required
every time the method is changed. It
requires the analysis of four spiked
distilled water samples. The analyst
compares his measures of precision and
accuracy to establish criteria developed
from the inter-laboratory method
validation studies. Because of the basic
threshold nature of the start-up test, the
methods allow the test to be performed
with reagent water.
If the analyst fails the criteria for
accuracy or precision in the start-up
test, the analyst is to repeat the test for
any compound that fails a criterion. If
the analyst is measuring, for example.
eight compounds at once using Method
601. and fails the criteria for three of
them, the analyst is required only to
repeat the three that failed provided the
method is not changed. It is not very
difficult to meet the criteria for any
individual compound. However, when
one is analyzing for numerous
compounds there is an accumulation.
failure probabilities: that is, an I
increased likelihood that one of sevei
parameters will fail for "statistical"
reasons. Thus EPA allows a "second
pass" opportunity to meet the criteria.
as long as the method is not changed.
Exhibit 1, below, offers some guidance
as to when analysts may want to skip
the "second pass" opportunity based on
an excessive number of test criteria
failures occurring on the first pass. An
excessive number of failures should not
occur if the system is operating properly.
Thus, such a number of failed criteria
may suggest poor operation to the
analyst. In this case, the first pass
criteria failures suggested the
compound(s) tested would fail a second
round. The analyst may wish to simply
adjust the system and reinitiate the
start-up test.
If the method is changed as a result of
the initial test, the startup begins again.
For example, if the start-up test
indicates zero recovery of vinly chloride
and a check reveals that the instrument
trap was installed backwards, the
operator must correct the problem and
reinitiate the test for all compounds,
since the method was just modified.
The second form of quality control is
contained in the ongoing quality con»«^-
program. Laboratories are required \
analyze blank samples (e.g.. reagent
water) daily, and to analyze spiked
wastewater samples periodically. Ten
percent of all samples are to be spiked
(five percent for Methods 624 and 625).
The resulting accuracy of recovery must
be compared to the established
accuracy criteria for the method
developed from the results of the inter-
laboratory method studies.
If an analyst fails one or more
accuracy criteria with the spiked
wastewater, the analyst must analyze a
check sample (e.g.. spiked reagent
water). The purpose of analyzing the
check sample is to establish whether the
inaccuracy is caused by matrix effects
or by the laboratory operating
improperly (i.e., out of control). Again.
accuracy results are compared to the
established accuracy criteria. The
criteria for acceptable accuracy in these
methods are based upon accuracy
derived from testing reagent water. Use
of check samples rather than spiked
wastewater to verify the accuracy
criteria for a laboratory is consistent
with the fact that one set of regression
equations in the inter-laboratory method
study is derived from reagent water
That set of regression equations is t
basis quality control criteria.
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The decision to rely on spiked
wastewater samples for the initial test is
an alternative to requiring that analyses
be conducted on ten percent spiked
reagent water samples (to verify
laboratory control) and ten percent of
spiked wastewater samples (to verify
matrix effects). Accordingly, the need to
also analyze a check sample is reduced
to a second-tier requirement which is
only mandated if accuracy criteria are
not met with spiked wastewater.
The limits that are in the methods
have been derived on a compound-by-
compound and method-by-method basis.
They are derived directly from the inter-
laboratory method validation studies.
The formal inter-laboratory validation
studies for Methods 601-602, 604-613,
624, 625, and 1625 have been completed
with 15 to 20 laboratories. These fifteen
methods have been revised to include
methods performance results derived
from these studies.
Two methods (603 and 1624) have not
been subject to an inter-laboratory
validation study. A formal inter-
laboratory validation study for Method
603 has not been completed due to an
error in the draft method. Although the
error was corrected. EPA was not able
to gierform an inter-laboratory validation
study on the same scale as performed
for the other methods. However, one
commercial laboratory did validate the
method and that validation was verified
by EPA's laboratory. In addition, the
method is similar to Methods 601 and
602 and the results from the validation
are similar. EPA believes that the
validation of 603 is adequate to
establish that the method is appropriate.
Therefore, Method 603 is being
promulgated with warning limits based
upon the best data now available.
Method 1624 was not formally
validated through an inter-laboratory
study. The specifications for Method
1624 were developed from Method 624
which was formally validated. In
informal multi-laboratory and single-
laboratory studies. Method 1624 has
been shown to yield slightly better
performance on treated effluents than
Method 624, but this improvement is
insufficient to warrant a separate inter-
laboratory validation study.
The multi-laboratory validation
studies were designed according to the
method of W.J. Youden (Youden, W.J.,
"Statistical Technique for Collaborative
Tests," Statistical Manual of the
Association of Official Analytical
Chemists. 1975) in which pairs of
samples having slightly different spiked
concentrations of the compound of
interest are analyzed. Each
collaborating analyst analyzes a sample
only once and reports a single value. By
having the analyst perform the analysis
as he would have done for a normal
routine sample, the Youden design helps
to avoid accidental manipulation of data
that can sometimes occur in a
laboratory doing replicate
determinations.
Each Youden sample pair for a given
parameter is prepared so that the
concentration of the pollutant of interest
in one-half of the pair is similar to, but
measureably different from, the
concentration of the pollutant in the
other half. Three Youden pairs were
analyzed for each of the parameters.
The mean values of each of the three
pairs were designed to spread over a
usable and realistic range of
concentrations. The lowest
concentration pair was prepared so that
the concentration would be above the
minimum detection concentration for the
method.
The Youden pairs, prepared as
concentrates, were spiked into six
different water matrices: distilled water,
municipal drinking water, a surface
water vulnerable to synthetic chemical
contaminants, and usually, three
different industrial wastewaters from
industries that normally would be
regulated for the priority pollutants
under study. The data were reduced to
four statistical relationships related to
the overall study: (1) Multi-laboratory
mean recovery for each sample, (2)
accuracy expressed as relative error or
bias (the difference between the multi-
laboratory mean recovery and the true
value divided by the true value), (3) the
multi-laboratory standard deviation of
the spike recovery for each sample, and
(4) the multi-laboratory relative
standard deviation. In addition, two
statistics were reduced from the raw
data relating to the single-analyst
performance: (1) Single-analyst standard
deviation, and (2) single-analyst relative
standard deviation.
The single-analyst standard
deviations were calculated for each of
the sample pairs according to the
method of Youden by (1) calculating the
difference for recoveries from each
sample pair reported by each analyst,
(2) calculating the average value of
these differences across the entire study,
(3) calculating a "sum-of-the-squares"
by adding the square of the differences
between each difference and mean
difference, (4) dividing the "sum-of-the-
squares" by the degrees of freedom to
give the single-analyst variance, and (5)
taking the square root of the variance to
give the single-analyst standard
deviation.
Fifteen to twenty-five percent of the
data generated in the multi-laboratory
validation studies were discarded as
outliers, i.e., data too far from the vast
majority of data to be acceptable.
Outliers were determined based on
widely accepted statistical tests
prescribed by ASTM and AOAC.
There is an apparent linear
relationship between the mean
recovered spike values and the true
spike values, overall standard deviation,
and single-analyst standard deviation.
These linear relationships have been
expressed as regression equations over
the concentration ranges studied in each
matrix. Six different regression
equations are derived for each of the six
matrices for any given compound. In
most cases the variations of the six lines
do not appear to be statistically
significant at the 5% significance level.
The conclusions were reached for each
water type by using the F-distribution to
compare variance statistics of waste
waters with those of distilled water.
Mean recoveries were compared
between wastewater and distilled water
using paired t-test statistics.
EPA is aware that there are limits to
the strength of these analyses. These
comparisons assume independence
among the observations and this was
not exactly the case since the "spike"
was made up of mixtures of all of the
compounds under consideration in each
method and hence there was an
interdependence among compounds.
Despite these limitations, the tests still
provide strong evidence that water type
generally had no statistically significant
effect on the method's performance.
The multi-laboratory tests support an
important conclusion. If a laboratory
performs well with the methods using
distilled water, it should be able to
obtain good results with surface waters
and industrial wastewaters. Based upon
this conclusion, the multi-laboratory
regression equations for accuracy and
single-analyst overall precision for
distilled or reagent water have been
incorporated into the quality assurance
and quality control provisions of the
final texts of Methods 601, 602, 604-613,
624, and 625 to define method
performance. The regression equations
for the other matrices are also included
in the texts of the methods.
The multi-laboratory validation of
Method 1625 was performed at a single
concentration in a reagent water matrix.
Specifications were derived for linearity
of calibration, for calibration
verification, for retention time precision,
for compound recovery from a reagent
water matrix, and for precision and
accuracy of analysis by isotope dilution
and internal standard techniques. All
specifications derived from the study
are applied at the same level at which
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10 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
they were tested, and sample matrices
which show labeled compound
recoveries significantly different from
recoveries of these compounds from
reagent water are diluted with reagent
water to bring these recoveries into the
expected range.
It is also important to note that the
studies provide a strong basis for setting
control limits which represent a range of
acceptability. The studies show that
most laboratories will do far better.
especially on a single-operator, single-
laboratory basis. Other performance
studies, completed since the inter-
laboratory analyses, incorporate too
much flexibility to be directly analogous
to EPA's collaborative test of the
methods. However, they appear to
confirm the assumption that most
laboratories will exceed the minimum
standards and indicate that method
variability will be well within the range
of the control limits.
The final specifications derived for all
of the organics methods (except 603)
were the result of a statistical analysis
of the data from the multi-laboratory
studiae. These specifications adopt
initirfprecision and accuracy for all
methods. For start-up calibration
verification, they specify control limits
for Methods 601. 602, 624.1624, 625 and
1625. For on-going accuracy, they
specify control limits for recovery of
pollutant spikes for Methods 601-613,
624, and 625, and for recovery of labeled
compound spikes for Methods 1624 and
1625. The methods allow for
simultaneous testing of all the
parameters listed in each method.
In theory, a problem could arise from
simultaneous tests for numerous
compounds. The control limits have
been calculated to allow only a 5%
likelihood that a result that exceeds the
limits for each compound is merely a
statistical fluctuation (rather than actual
error). However, the chance of
"statistical error" rises with the number
of compounds being tested.
EPA has corrected for this possibility
in several ways. First, most users will
not apply each analysis to all
parameters simultaneously; thus they
will have a greater chance of passing all
test criteria. Second, in order to allow
for simultaneous testing of all
parameters in a given method, the
specifications for accuracy and
precision have either been broadened.
or a re-test has been allowed, or both.
The technique of using a re-test was
chosen because a one-test-only
specification which allowed for
simultaneous testing of a large number
of parameters would be so broad as to
have little meaning. The provision for a
re-test preserved a meaningful
specification while allowing for
simultaneous testing of all parameters. If
a laboratory fails the re-test as well as
the initial test, the likelihood of
"statistical error" is extremely low (5%
times 5%. i.e.. .0025 for a given
compound). Third, when a re-test is
required, it need only be performed on
the particular compounds which failed
the initial test. Finally, the control
criteria for Methods 1624 and 1625—
those most likely to be simultaneously
used on many compounds—were
determined based on the 99% confidence
level.
As a voluntary guide to laboratories
practicing a given method, the following
Exhibit 1 gives suggested numbers of
first pass test criteria failures which are
unlikely if the laboratory is satisfying
the probability based quality control
specifications. It assumes all parameters
in a given method are tested
simultaneously. The Exhibit indicates
the maximum number of parameters for
which each method can be used
simultaneously. The two right-hand
columns dicate a certain number of
unacceptable results. If the analyst finds
that number, or a greater number, of
unacceptable results, he may conclude
that the entire analysis is flawed. If so. it
may be more efficient to repeat the
entire analysis than to re-examine only
the compounds which exceed the
control limits.
EXHIBIT 1.—SUGGESTED MAXIMUM NUMBER OF
TEST CRITERIA FAILURES WHICH JUSTIFY
REPEATING ENTIRE ANALYSIS
Number of
simuKane-
oo»
Number of test cntena
failure*
I I
: Start-up | On-going'
601
602
603/605
604
606
607
606
609
610
611
612
613
624
625
1624
1625
29
7
2
11
6
3
25
4
16
5
9
1
31
61
86
151
7
3
2
3
2
6
3
5
3
4
2
7
11
12
7
4
2
2
3
2
2
4
2
3
2
3
g
7
7
5
1 BaMd on twee the numb* of parameters being tMMd
sine* both accuracy and pncMion are baing evaluated.
' Bated on the number of paramtMra baing Mated.
Section 8 of each method defines
acceptable analytical performance limits
for the GC, HPLC, and GC/MS test
procedures (Methods 601-613.624. 625.
1624. and 1625). These acceptable
performance limits are also specified in
Footnote 7 to Table 1C, "List of
Approved Test Procedures for Non-
Pesticide Organic Compounds," and
Footnote 7 to Table ID. "List of
Approved Test Procedures for
Pesticides." System performance is
acceptable only when the average
recoveries and standard deviations of
spikes of the pollutants of interest into
reagent water meet these performance
standards. Where large numbers of
parameters are being analyzed (see
Exhibit 1 above), there is an increased
chance that at least one parameter will
fail for either average recovery or
standard deviation limits based purely
on chance. Where such failure occurs,
the spiking and recoveries must be
repeated, but only for the failed
parameters. Repeated failure confirms a
general problem with the analytical
measurement system. When such failed
recoveries are experienced the system is
judged to be out-of-control for the failed
parameter. Thus, the results for the
failed parameters in unspiked samples
are suspect and cannot be reported to
show regulatory compliance.
The acceptance criteria for spikes into
samples for each parameter were
calculated to include both an allowance
for error in prior measurement of the
background and another allowance for
error in prior measurement of spike
concentrations. The calculation
assumed a spike-to-background ratio of
- to 1. Thus such error will be accounted
: T to the extent the analysts' spike-toj"
background ratio approaches 5 to 1. Ira
many cases this allows analysts a
greater margin of error than should
actually be expected. This is because
the calculation assumes that two prior
errors are cumulative, ignoring the
degree to which they actually cancel
each other out.
Today's final test procedures
represent an effort to provide the
maximum uniformity that is practical for
a wide cross-section of classes of
chemical compounds. They will be
continually Devaluated for their general
applicability to complex wastewater
matrices.
The substantive revisions made in the
GC, HPLC, and GC/MS methods in
response to comments are discussed in
the public participation section of this
preamble. Three of the most significant
changes include: (1) Addition of a
confirmatory column to Method 602; (2)
deletion (from 613) of the gas
chromatographic/electron capture (GC/
EC) test procedure for screening for
2.3.7,8-TCDD, and (3) revision of
Methods 613 and 625 to show that
Method 625 may be used whenever
screening for 2,3,7.8-TCDD is required.
The full text of the approved GC, HPLC
and GC/MS test procedures are being!
printed in Appendix A of this regulatiol
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
11
The GC, HPLC. and GC/MS test
procedures are now cited in the
regulations in the new Table 1C, "List of
Approved Test Procedures for Non-
Pesticide Organic Compounds." and
Table ID, "List of Approved Test
Procedures for Pesticides."
C. ICP Test Procedure
The ICP test procedure is cited in the
regulation as an additional analytical
option for trace metal analysis in the
new Table IB, "List of Approved
Inorganic Test Procedures."
The ICP test procedure, Method 200.7,
has been changed only slightly from the
version proposed on December 3,1979.
EPA proposed that lithium and
strontium be analyzed using the ICP test
procedure, since these parameters could
be analyzed using this method. Because
EPA did not propose or develop
accuracy or precision criteria for these
parameters, EPA is unable to approve
the ICP test procedure for them. EPA is
considering the ICP and other
alternative test procedures in a separate
rulemaking. In light of additional
information received in the public
comments showing good recoveries for
antimony and adequate recoveries for
thallium by the proposed test procedure,
both of these metals have been added to
the scope of the ICP test procedure. Also
in response to public comments the
detection limit for silica has been
doubled and the wavelengths of the
metal are now given to the third
decimal. In section 3 of the ICP test
procedure a new definition for "Quality
Control Sample" has been provided for
clarification, and a new section on
safety has been added to alert the
analyst to the hazards of the toxic
reagents and pollutants involved. Other
revisions made in response to comments
are discussed in the public participation
section of this preamble. The full text of
the ICP procedure is printed as
Appendix C to this regulation.
D. CBODs Test Procedure
The final test procedure for CBODs is
essentially the same as that proposed.
See Section III-D, above. EPA's
proposed test procedure was taken from
a draft Standard Methods test procedure
for CBODs.
The final method language is the same
as the language now included in the 15th
edition of Standard Methods. This has
required minor changes from the
wording of the proposal, but no
substantive changes were required.
E. Table II: Required Containers.
Preservation Techniques, and Holding
Times
Table II in Section 136.3(e) now
restricts the materials of which sample
containers can be made, and specifies
the procedures by which samples are to
be preserved. Table II also limits the
maximum time for which samples may
be held from the the time of sampling
until they are analyzed. Table II has
been restructured in this final regulation
to correlate with the parameters in the
new Tables IA, IB, 1C. ID, and IE in
Section 136.3(a). Table II allows cross-
reference between the container.
preservative, and holding times and the
individual parameters in Tables IA, to
IE.
In response to comments, several
changes were made in Table II of the
final regulations for prescribed
container materials, preservation
requirements, and holding times of
wastewater samples. Where supported
by comments, changes were made
primarily in holding times. In response
to comments, EPA has adopted the
requirement that some samples be
analyzed immediately, to avoid sample
degradation. This would be as soon as
the sample is collected and labelled.
generally within 15 minutes. Longer
holding times are generally not
appropriate where the sample may
quickly degrade. However, a longer time
period may be justified under the
variance procedure. Exhibits 3 and 4,
below, show that for organic compounds
and pesticides, the holding times were
generally extended from 30 days after
extraction to 40 days after extraction.
Changes were also made to enable a
single sample to be used for analyses of
extractable organics and of pesticides.
This was a step towards the goal of
uniformity, sought by EPA and by the
commenters.
Table II as promulgated also allows a
variance to holding times under
§ 136.3(e). Analysts may exceed the
holding times if they have data on file to
show that the specific types of samples
are stable for a longer time and if they
receive a variance from the Regional
Administrator.
No changes were made for container
materials, preservation requirements, or
holding times in final Table II from the
proposed requirements for the biological
parameters listed in Table IA, or the
radiological parameters listed in Table
IE. Changes which were made in Table
II for inorganic parameters listed in
Table IB, organic parameters listed in
Table 1C, and pesticide parameters
listed in Table ID are summarized in the
following Exhibits 2, 3, and 4, of this
preamble. Proposed and final container
materials, preservation requirements.
and holding times in Exhibits in 2, 3, and
4 are given only for the affected
pollutant parameters in Tables IB, 1C
and ID of the regulation.
EXHIBIT 2.—CHANGES MADE IN TABLE II FOR TABLE IB PARAMETERS
Parameter
CT tved obe
Requirement
Holding time .
Holding time
Preservative
Preservative....
Hokfina time
Holding time
Holding time ..
Change
From (proposed)
NaiSiOi
48 hours.
0.05% KaCriOr
None
1 hour
P and G.
14 days
14 days
7 days
28 days ....
Cod to 4 "C
To (final)
Analyze immediately.
Ascorbic acid.
Add: Remove surfide as cadmium sulftde.
Analyze immediately.
24 hours.
Delete.
Add: HCI or H,SO, to pH<2.
Analyze immediately.
Add: Store in dark.
G only.
7 days.
7 days.
48 hours.
Add: NaOH topH>9.
7 days.
Analyze immediately.
None required.
-------
12
Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations
EXHIBIT 3.—CHANGES MADE IN TABLE II FOR TABLE 1C PARAMETERS
Panvnaiar
Phenols '
f^BnrWfew i
HaHjtthar* '
TCOD '
RaQunMnanf
("I
Praaarvatfra
rr^MrvBtiva
HoMina toma
Holding torn*
•• jafcu*
Holding tima
Holding tima
• IMIiUB
n.jj^u .j^
rrasarMaBVa
Holding Mm
Pi «•»•»»«
Holding Mm
' Tha foNowino optonrt praaarvatton may ba uaad whan tha astanakad cataooriaa ara to oa i
' Droppad a» « pygnataf-saa Punjaatta Hatocaftona and Cntonnaiad Hydrocarbom.
EXHIBIT 4.— CHANGES MADE IN TAG
Cnanga
From (proposad)
(1
3 days
H.SO.IO
pH<2 30 days altar attraction
NfcSiOi
30 days aflar mraclion
NfcSiOi
30 days altar extraction
NajSiOi
30 days altar axtracfeon
30 days attar axtracVon
30 days aftar airtraction
NfcSiO,
30 days altar airtraction
To«man
(l
Add: HO to pH<2. Samptoa not faosxxg
days.
14 days.
OaMad.
40 days altar attraction.
Add: pH 2-7 it 1 24iphanylhydr
prassnt adjust pH to 4.0±0.2.
OaMad.
40 days altar artracton.
Ad* Stora « dark.
40 days altar sxtrackon. adjust pH
DaMad.
40 days altar attraction.
OaMad.
40 day* aftar attraction.
Add: Stora in dark.
40 days aftar attraction.
40 days attar aJdracMon.
DalaMd.
40 days aftar «x1rsclion.
40 days aflar axtracfeon.
7-10 (or
nalyzad in a smgM sampla: cool to 4 'C. idd 0.008* N.AO,. stora n dark, adjust pH 6.0-9.0.
LE II FOR TABLE ID PARAMETERS
Paramalaf
RaQumfnant
Prosaivatnra
Holding tima
Changa
From (nnxiosaJ)
NaiSiOt
30 days altar airtracaon
To(ftnal)
OaMad.
Add:pHS-9.
40 days altar axtnctton.
F. Incorporation by Reference
The analytical methods approved are
lengthy and detailed. Many are readily
available to the public. Thus, § 136.3(a)
has'been revised to show that the full
texts of the test procedures taken from
the various references in Tables IA, IB,
1C. ID and IE are "incorporated by
reference" into the regulatory language
of these Guidelines in accordance with
the regulations of the Office of the
Federal Register, and with the approval
of the Director of the Office of the
Federal Register. Methods which are not
readily available are printed in full as
appendices to this notice.
As a convenience to the users,
§ 136.3(b) has also been added. This
cross-references Tables LA, IB. 1C, ID,
and IE and the parameters therein. It
cites specific references, the sources
from which they may be readily
acquired, and indicates, where
available, approximate costs of the
references.
The full texts of the test procedures
cited are available for inspection only at
the Office of the Federal Register
Information Center, Room 8301,1100 L
Street, NW.. Washington, D.C. Full texts
of non-copyrighted and copyrighted test
procedures are available from the
sources indicated in § 136.3(b).
To accommodate this new paragraph
(b), paragraphs (b) and (c) in the existing
regulation have been redesignated as
paragraphs (c) and (d), respectively.
V. Public Participation and Response to
Most Significant Comments
Two hundred and twenty letters,
many with attachments of data and
other information, were received in
response to the Administrator's
December 1979 request for comments.
Ten additional letters were received in
response to the reopened comment
period in January 1981. Letters were
received from industries, Federal and
State agencies, industrial and trade
associations, universities, testing
laboratories, research institutes,
engineering and consulting firms, local
government agencies, standards and
professional organizations, and from
one private citizen.
To facilitate analysis, comments
within the letters were classified into
three categories—policy, technical, and
general, and then into thirteen sub-
categories. The comments were then
reviewed by a technical group of EPA
analytical experts and by an EPA policy
review group. The record contains a
response to the comments, organized on
the basis of these subcategories. The
final rulemaking today incorporates
changes based upon this review. This
section highlights significant and generic
comments and EPA responses.
In the following paragraphs of this
section only issues of major concern will
be discussed. Many lesser technical
questions or minor issues which were
raised are discussed in the "Response to
Comments" document contained in the
record.
A. GC, HPLC, and GC/MS Test
Procedures
1. Policy on Applicability of Test
Procedures
EPA requested comments on the
general applicability of the proposed
methods. To present the best state-of-
the-art test procedures for analyzing
trace organic pollutants in industrial
wastewaters, EPA specifically requested
commenters to share their experiences
and data relative to sensitivity,
precision, accuracy, and detection limits
of the proposed GC, HPLC, and GC/MS
test procedures.
Comment: Several commenters felt
that the EPA should not publish some of
these test procedures (specifically, the
GC, HPLC. and the GC/MS) as
regulations until they had been fully
validated by multi-laboratory studies.1
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 13
Response: No data or information
were provided to the EPA in the
comments that would permit revisions
of the texts of the proposed test
procedures to better represent the state-
of-the-art, to enhance general
applicability, or to preclude interference
problems. Consequently. EPA believes
these GC, HPLC, and GC/MS test
procedures to be the best state-of-the-art
test procedures currently available for
the analysis of wastewater discharges
for the priority toxic organic pollutants.
The multi-laboratory validation
studies for Methods 601-602, 604-613,
624, and 625 have been completed and
support the general applicability of
these test procedures to the analysis of
treated wastewater effluents. Since the
front-end chemistry of the GC/MS test
procedures, i.e., sample preservation,
holding times, and purge and trap or
methylene chloride extraction, are
similar for the GC and the GC/MS test
procedures, it would be expected that
the precision and accuracy of these test
procedures will be substantially the
same over the concentration ranges
which give linear detector responses.
This has been substantiated by the
validation studies.
Comment: Several commenters felt
that GC. HPLC, and GC/MS test
procedures did not meet rigorous
criteria, such as those of the consensus
standards organizations, had not been
precisely written, and did not meet the
state-of-the-art.
Response: The Agency recognizes that
these test procedures include some
state-of-the-art techniques which have
not been as widely used as the
consensus methods which were
approved and promulgated in 1973 and
1976. But for Methods 601-602, 604-613,
624, 625.1624, and 1625. method
performance is now supported by the
multi-laboratory validation studies.
These 16 test procedures are now
backed by data which meet criteria at
least as rigorous as that employed by
consensus standards organizations.
Only single-laboratory data are
available for 4 parameters (2-
chloroethylvinyl ether, 1,2-
Dichloropropane. cis-1,3-
Dichloropropene, and trans-1,3-
Dichloropropene) in Method 601, 2
parameters (acrolein and acrylonitrile)
in Method 603,1 parameter
(Hexachlorocyclopentadiene) in Method
612, and 5 parameters (Bromomethane,
2-chloroethylvinyl ether, 1,2-
Dichloropropane. cis-1,3-
Dichloropropene, and trans-1,3-
dichloropropene) in Method 624. It was
necessary to resort to single-laboratory
testing due to the long-term instability of
these parameters in the Youden pair
samples used in the multi-laboratory
validation studies. In many instances
these single-laboratory evaluated test
procedures have been more
exhaustively tested for their
applicability to industrial wastewaters
than the currently approved test
procedures in these guidelines which are
cited from consensus sources.
The procedures promulgated today, as
written, most precisely define the
current state-of-the-art for the analysis
of trace organic compounds in
wastewater discharges. No better
methods are now available for general
use.
Comment: Several commenters said
that these test procedures had not been
demonstrated by the Agency to be
generally applicable to all wastewater
effluents.
Response: These test procedures were
developed by single laboratories to be
applicable under various conditions of
preservation and holding times, to
samples taken from wastewaters which
were shown in various surveys to have
the compounds of interest. Their general
applicability to multiple industrial
wastewater effluents is now
corroborated by the multi-laboratory
validation studies, which specifically
searched for matrix effects. In specific
cases of interference, analysts may seek
leave to use alternative procedures
under 40 CFR 136.4.
Comment: Several commenters
suggested that these test procedures be
issued as "interim test procedures"
without regulatory impact within the
limits of their analytical uncertainties.
Response: This comment was made
prior to the completion of the multi-
laboratory validation studies.
Acceptable performance for these
methods is now verified for analysis of
industrial wastewater effluents.
Mandatory quality control within the
text of the test procedures further
strengthens their credible applicability
to effluent analysis. When spike or
surrogate recoveries are poor (do not
fall within the acceptable performance
levels for the parameters of concern),
the method will not have produced data
acceptable for NPDES reporting
purposes such as permit applications
and discharge monitoring reports
demonstrating compliance.
Comment: Several commenters felt
that the test procedures should be
continually reevaluated to ensure they
are kept abreast of the state-of-the-art.
They requested that the test procedures
be re-submitted for comment and peer
review after the 20-laboratory validation
tests were completed.
Response: The Agency decision to
conduct the 20-laboratory validation
studies was made in the spirit expressed
by these commenters. The studies have
been completed, and their results are
incorporated in the revised texts of
these test procedures. These results
corroborate the general applicability of
the validated test procedures to multiple
wastewater effluents. EPA may amend
40 CFR Part 136, if comments or future
results of these studies suggest that this
is necessary. EPA is accepting comment
on the actual calculation of specific
control limits.
2. Flexibility and Analysts' Professional
Judgment
EPA requested commenters'
experiences and opinions on the
flexibility that should be incorporated
within the GC, GC/MS. and HPLC
procedures. Optimum flexibility would
render the test procedures most
generally applicable to industrial
discharges, without seriously
compromising data quality.
After careful study, EPA has found
most of the comments received in
response to this request to have merit
and has reacted positively to them.
Comment: Several commenters
requested that test procedures be
revised to allow the analyst maximum
leeway to exercise professional
judgment to adapt the test procedures to
the sample at hand. They felt that
defining test procedures in terms of
"good laboratory practices" and
analytical criteria would allow the
incorporation of technological advances
not permitted in rigidly structured
procedures. The commenters
specifically requested flexibility in GC
temperature programming and
extraction solvent stripping procedures.
Response: Where technically justified.
the proposed test procedures have been
relaxed to allow flexibility for analysts
to exercise professional judgment. This
flexibility is being incorporated within
the text of the GC and HPLC procedures,
in recognition of the rapid advances
occuring in the state-of-the-art for
packed and open tubular columns. A
primary configuration of GC or HPLC
components is described within the test
procedure. However, to optimize the
applicability of the chromatographic
technique to analytical requirements
unique to specific discharges, the
analyst is allowed to use professional
judgment in selecting packed or open
tubular columns, operating temperature
programs, carrier gas or solvent flow
rates, and detectors. Analysts may also
use their discretion in selecting cleanup
procedures. EPA has also relaxed the
strict protocol for sample extract
concentration. In the proposed test
-------
14 Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
procedures, sample extracts were
concentrated by means of a Kuderna-
Danish concentrator, which consists of a
concentrator tube, an evaporative flask,
and a Snyder column. Many studies
comparing the Kuderna-Danish
concentration technique to a variety of
others have provided no statistically
conclusive support for the rigid protocol
for most materials tested. Accordingly.
within the scope of the GC, HPLC, and
GC/MS test procedures, analysts are
permitted some discretion in selection of
concentration techniques.
Comment: Other commenters said
these analytical criteria for the test
procedures should be in terms of
mandatory verification and quality
control procedures.
Response: The EPA agrees. With the
added flexibility in test procedures for
analyst discretion, an added
responsibility is borne by the analyst for
mandatory quality control and
validation procedures. These
requirements have long been recognized
as elements of good analytical
laboratory practices. Accordingly,
Methods 601-613. 624. and 625 now
incorporate verification and quality
control and the criteria for reportable
analytical results for regulatory
purposes.
Comment: Many of the commenters
were concerned with the costs
associated with rigidly prescribed
methods, especially where required use
of several such methods would call for
significant capital expenditures in highly
specialized equipment.
Response: The flexibility which has
been incorporated into the test
procedures is expected to encourage
rapid incorporation of technical
advances, and to allow the dischargers
to select the most cost-effective
analytical options when acquiring major
analytical equipment. However, any
modifications of Methods 601-613, 624,
625,1624, and 1625, which are not
expressly permitted in the text of the
methods as given in Appendix A to this
part, will be considered major
modifications and must be approved as
alternate test procedures under the
provisions of § § 136.4 and 136.5.
Sample preservation and holding
times have also been standardized to
allow the same sample to be used for
more than one method. This flexibility
allows monitoring costs to be minimized
and existing equipment to be fully
utilized. Additionally, contract
laboratory services are available when
the analytical load of a discharger does
not justify purchase of specialized
analytical equipment (See cost
discussion below).
General Response to Comments on
Flexibility and Analysts' Professional
Judgment: Flexibility is permitted only in
discretionary elements of the
chromatographic test procedures.
Changes in non-discretionary elements
of the test procedures are outside the
scope of the flexibility provision.
Change may be made only when
conditions of analytical equivalency are
demonstrated within the provisions of
§§ 136.4 and 136.5 of this regulation. For
all discretionary changes which analysts
may make, their laboratory records
should corroborate that the quality of
the data generated meets all stated
performance criteria of precision and
accuracy. (See 40 CFR 122.21,122.41.
122.44 and 403.12).
3. Quality Control and Quality
Assurance
This issue is integrally related to the
preceding discussion of allowable
flexibility The availability of rigorous
quality control criteria is what allows
the flexibility now built into the
methods. Conversely, the allowance for
modification based on professional
judgment makes it essential that there
be a procedure for assuring that those
modifications do not undermine the
quality of reported data.
EPA proposed, within the test
procedures, that method blanks should
be processed each time a set of samples
was to be extracted, or whenever there
was a change in reagents. This was to
safeguard against chronic laboratory
contamination. EPA also proposed that
standard quality assurance practices be
used. The practices proposed included
field replicates to validate the precision
of the sampling technique, laboratory
replicates to validate the precision of
the analysis, and special (or fortified)
samples to validate the accuracy of the
analysis. If doubt existed in the
identification of any peak in a gas
chromatogram, the proposed quality
assurance required a confirmation by
use of a technique such as MS. In
addition. Appendix III of the proposal
required an intensive quality control
plan for the GC/MS test procedures. The
Agency requested comments on the
proposed levels of quality assurance/
quality control (QA/QC) and on any
additional levels that should be
specified in the test procedures or left to
the discretion of the analyst.
Comments: Most commenters,
including several large industrial
associations, restricted their comments
to the operational details of the
suggested programs, thus implying
acceptance of the concept of the
mandatory quality control procedures.
Others specifically urged adoption of
such procedures.
Some commenters addressed the issl
of appropriate costs and number of
samples that should be analyzed for
continuing QA/QC. One corporation
suggested that 10%-20% of cost was
reasonable, with one control sample for
10 samples analyzed. A trade
association stated the QA/QC should
not exceed 20% of the analytical costs.
Another corporation suggested 10%
spiked samples and duplicate analysis
of all purgeable samples. A permitting
authority suggested that 15%-20% QA/
QC was reasonable.
Some commenters specified that
mandatory QC should be an integral
part of all of the proposed organic test
procedures. There were no objections to
the required analysis of four spiked
reagent water samples, as proposed in
Appendix III.
Response: The comments have
confirmed EPA's belief that a
reasonable level of mandatory quality
assurance should be incorporated in all
of the test procedures for organic
priority pollutants to assure that data of
known and acceptable quality are
produced.
The analyst must demonstrate correct
use of the method, reagents, and
equipment before analyzing samples.
Users of Methods 601-613, 624, 625.
and 1625 must demonstrate their abili
to generate data of acceptable quality
by analyzing four spiked aliquots of
reagent water containing each
parameter of interest, and meeting
performance specifications based upon
the multiple-laboratory methods studies
completed by the Agency. For a limited
number of parameters discussed earlier,
the performance specifications are
based upon other laboratory test data.
Reagent water is used to ensure that
there are no matrix effects in this
preliminary test.
On-going quality assurance for
Methods 601-612 consists of mandatory
spiking of 10% of all samples (spiking
with the compounds being measured)
and comparing the degree and precision
of spike recovery with the test
procedure criteria established in the
multiple-laboratory methods studies.
This will lead to a QA/QC cost in the
range of 10% of the analytical costs. For
Method 613, samples must be spiked
with isotopically labeled 2,3,7,8-TCDD
internal standards. For Methods 624 and
625 the requirement is for a 5%
mandatory spiking of samples. See
Section IV B above.
Users of these methods are required
to spike all samples with surrogates.
These QA/QC procedures are
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 15
essentially the same as those proposed
except that criteria are now specified in
more detail and specific control limits
are based upon multiple-laboratory
studies. If the recovery of any parameter
falls outside the control limits, the
laboratory performance for that
parameter is inconsistent with control,
and the analytical results for that
parameter in the unspiked sample are
suspect and may not be reported for
regulatory compliance purposes.
If the performance criteria for the GC,
HPLC. and GC/MS test procedures
which have been reprinted in full text in
Appendix A of this part cannot be
attained in a specific wastewater
matrix, the analyst may consult the
Director of EPA's Environmental
Monitoring and Support Laboratory in
Cincinnati. Ohio, for technical
assistance. If the method must be
modified beyond its allowable flexibility
to meet the performance criteria, the •
procedures of § 136.4(b) are available.
Although these requirements for
quality assurance and control are
minimum EPA standards, analysts are
expected to perform significantly better
in almost all cases. They are also
encouraged to participate in additional
performance testing to assure continued
analytical proficiency.
B. ICP Method
EPA proposed the ICP instrumental
test procedure as an alternative to the
approved test procedures for elemental
analysis (such as the AA and
colorimetric test procedures). In
particular, comments were solicited on
the general applicability of the ICP test
procedure to industrial discharges.
Comment: Some commenters thought
that the detection limits given in the test
procedures were unrealistic. They
suggested that industrial samples would
need to be concentrated before such
limits could be achieved.
Response: The detection limits given
in Table 1 of the ICP test procedure
were taken from an EPA publication
(EPA-600/4-7&-017, "Inductively
Coupled Plasma—Atomic Emission
Spectroscopy—Prominent Lines"). These
detection limits are intended as guides
for instrumental limits and are published
for informational purposes. The method
performance in terms of precision and
accuracy were determined at different
concentrations of analytes in distilled
water in a multi-laboratory test. These
are reported in Table 4 in the ICP test
procedure and are the performance
standards which are required by section
12 of the test procedure for instrument
quality assurance.
Comment: Some commenters thought
that the ICP test procedure was not
equivalent to the AA test procedure, and
especially to the furnace AA test
procedure.
Response: The approved furnace AA
test procedure can be used to measure
concentrations lower than the ICP
technique can measure. Data filed with
the EPA show the equivalency of the
AA and ICP test procedures across the
concentration ranges that are common
to the two procedures. The common
ranges include the concentrations which
are of regulatory concern. These
comparisons were made over a number
of industrial discharges and a broad
concentration range.
Comment: Several commenters asked
for the approval of the direct current
plasma (DCP) atomic emission
spectrometric method as equivalent to
the ICP and AA test procedures, and
requested its inclusion in Table IB as an
approved test procedure for elemental
analysis.
Response: EPA did not have data to
show the applicability of the DCP
technique to wastewater analysis and
accordingly did-not propose it as an
alternate test procedure for elemental
analysis. Data showing applicability to
vvastewaters were not made available
by the commenters, although data were
presented to show the applicability of
DCP to sample matrices other than
wastewater. Because of this incomplete
data base. EPA was unable to include
DCP in the list of approved test
procedures.
Recently, a DCP manufacturer has
provided such data to EPA for approval
thorugh the equivalency provisions of
§§ 136.4 and 136.5, and'EPA is currently
considering approval of the DCP test
procedure for the analysis of trace
elements in wastewater.
Comment: Several commenters had
difficulty understanding the guidance
given for background corrections and for
high dissolved solids (salt buildup)
interferences.
Response: When using the ICP
procedure it is important to make
background corrections. The analyst
must determine and incorporate into the
analysis the valid correction factors for
spectral interferences caused by high
dissolved solids concentrations. Section
2.1 of the ICP test procedure has been
revised to make this clear. Additional
guidance has also been given to correct
for salt buildup at the tip of the
nebulizer.
Comment: Some commenters thought
that the suggested combinations of
elements in stock solutions were
ambiguous.
Response: The mixed standard
solutions suggested by the EPA in the
test procedure were found to be
compatible mixtures. Other
combinations may be used within the
scope of the test procedure, as long as
precipitations and other chemical
reactions do not occur. The test
procedure text has been revised to make
this clear.
Comment: Some commenters felt that
other wavelengths of light which were
also emitted in the characteristic
spectrum of the element should be
permitted.
Response: Other wavelengths may be
used when those given in the test
procedure are obscured. In using them.
the analyst must demonstrate that they
are free from interfering wavelengths or
that interference results can be
corrected.
Comment: Several commenters
presented data and suggested that
antimony and thallium should be
included within the scope of the ICP test
procedure.
Response: Antimony and thallium are
priority pollutants and the EPA agrees
they should be included within the
scooe of the ICP test procedure since the
suuir.ued data show these metals can
be recovered in excess of 90% from
spiked samples using the ICP test
procedure.
Comment: Several commenters
requested sources from which verified
outside samples could be obtained.
Response: Verified outside check
samples are necessary for testing the
analytical system. Upon request, the
Director of EPA's Environmental
Monitoring and Support Laboratory in
Cincinnati. Ohio, will either supply such
samples or provide information on
where quality control check samples can
be obtained.
Comment: Several commenters found
errors in the tables of reference
wavelengths.
Response: Suggested corrections of
errors in the tables and various other
parts of the ICP test procedure text have
been made.
Comment: Several commenters
questioned the estimated costs for
analyses made by the ICP test
procedure, relative to other approved
test procedures.
Response: Cost estimates for analysis
by the ICP test procedure were given
only as general comparisons with the
costs of the other alternate test
procedures in Table IB, which are still
approved for trace metal analyses. The
analyst may exercise professional
judgment in selection of the approved
alternate test procedure from Table IB
which best meets his analytical needs. If
the ICP instrument system is available
to the analyst, it may well be the most
-------
16 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
cost-effective approved alternate test
procedure, especially if large numbers of
samples containing several of the ICP-
approved metals are to be analyzed.
Otherwise, the AA or one of the other
approved alternate test procedures may
prove to be more cost-effective. The
estimates given for the ICP analyses are
average contract charges experienced
by the EPA for multi-element analyses
of large lots of samples.
C. (CBOEk) Method
The Agency proposed the CBODs test
procedure in response to requests from
environmental analysis laboratories to
measure the carbonaceous BOD of
wastewater without the complications
caused by the nitrogenous oxygen
demand.
Comments: EPA requested comments
and additional data on the control of
nitrification in BOD measurements.
Many letters were received, several
accompanied by extensive test data. A
majority of the letters expressed
confusion over the use of the CBODs
versus the traditional 5-day BOD test.
None of the commenters questioned the
test procedure.
Response: The CBODs method
(Parameter No. 14, Table IB) is being
added to the list of approved
measurements. This method is new, and
should not to be confused with the
traditional 5-day BOD test which is
listed in Table IB as Parameter No. 9.
The nitrogen inhibitor is not a
procedural option, rather CBODs must
be designated as the pollutant or
effluent limitation that is measured to
report the CBOD5 parameter.
A discharger whose permit requires
reporting the traditional 5-day
biochemical oxygen demand (Parameter
No. 9, Table IB) may not use a nitrogen
inhibitor in the procedure for reporting
results. Only when a discharger's permit
explicitly states that CBODs is the
parameter whose monitoring is required
may the permittee report data using the
CBODs nitrogen inhibited analytical
method.
D. Table II: Required Containers,
Preservation, and Holding Times
EPA proposed mandatory
preservation techniques, container
materials, and holding times for
industrial wastewater samples. It also
proposed a mechanism by which a
permit holder or analytical laboratory
could obtain a variance from these
requirements if justified by data. The
permittee or analyst was also limited to
shorter holding periods, if this was
necessary to maintain sample stability.
In the proposed guidelines, these were
paragraph 136.3(d) and footnote 4 to
Table II. These provisions are now
contained in paragraph 136.3(e) and
footnote 3 of Table II.
Comment: Some commenters thought
that Table II was too detailed and
should not be included in the text of the
regulation. They suggested that EPA
make it available under a separate
cover.
Response: The EPA disagrees. EPA
continues to believe that the integrity of
the sample is critical to the quality of
monitoring data. It is best presented and
publicized as a requirement through
inclusion in the regulation.
Comment: Several commenters felt
that preservation techniques, holding
times, and container materials should be
presented as recommendations only.
They thought the diversity of
wastewater samples and sampling
requirements could best be judged by
the professional taking the sample.
Response: EPA agrees that the
judgment of the professional taking the
sample is critical to the integrity of the
sample. However, after the sample is
taken, sample integrity is maintained
and verified only by control of container
materials, preservation procedures, and
holding times. Only with verifiable
sample integrity can analytical data be
correlated to the sample source. EPA's
evaluation of NPDES monitoring
practices has confirmed that inadequate
sample preservation and failure to
observe recommended holding times are
major analytical problem areas. For
these reasons EPA has included Table II
in the mandatory text of this regulation.
Comment: Other commenters said that
there should be more standardization of
the container, holding time, and
preservation requirements between
parameters in order to minimize the
number of samples that needed to be
taken.
Response: EPA agrees that this is
desirable. It is, however, secondary to
quality assurance. Table II requirements
have now been standardized to a
significant degree. This minimizes the
number of preserved samples that need
to be taken.
Comment: Several commenters felt
that there were stable wastewater
sample types that would not require
adherence to the mandatory
requirements of Table II.
Response: When a permit applicant
knows that Table II requirements do not
apply to his wastewater—and he has
data to substantiate this conclusion—
the variance provisions of paragraph
136.3(e) and footnote 3 of Table II allow
less stringent requirements to prevail.
Such variances must be obtained from
the Regional Administrator under these
provisions.
General Response: Many commenters
provided data or information relating
specific requirements of Table II. The!
changes made in Table II in response 1
these comments were summarized
earlier in Exhibits 2. 3. and 4 in section
4(e) of this preamble.
E. Cost Estimates for Methods
EPA is concerned with the costs of
analyses required by the new
regulations, because of their overall
importance to programs under the Clean
Water Act. In the December 3.1979.
Federal Register, the Agency solicited
comments on these costs and listed
EPA's typical costs for Carbonaceous
BOD. ICP, GC, HPLC, and the GC/MS
analyses.
Comments: Several comments from
the public addressed issues such as the
high cost of routine analysis, the
appropriateness of the figures presented
by EPA, and the variations in cost
experienced, depending upon lot size,
throughput, and complexity of the
analyses performed. Nearly all of the
comments received were directed
toward organic analyses and the related
requirements for capital equipment.
Several comments also seemed to rely
on the erroneous assumption that the
proposed methods would require
separate samples for each procedure.
Response: EPA noted a wide range j
unit costs cited, based largely on
volume. These regulations do not require
that each permittee perform its own
analyses. When the small volume of
analytical loads makes unit costs less
economical, the permittee may use
commercial laboratories, where
analytical work loads can be
maintained at levels which optimize
costs.
EXHIBIT 5.—EPA CURRENT EXPENDITURES FOR
WATER AND WASTEWATER ANALYSIS FOR
PRIORITY POLLUTANTS
! COM
113 Organic Compounds by GC/MS (indudas i
li«M t laboratory Minks) i
Each Rapfcata Simp* tor 114 Orgsncs by GC/ I
MS
Each Priority Petulant Spfead Sampla tor 114
Organcs by GC/MS (with data calculations)
13 Elamantal Compounds by AA-Flama or Flams-
ICP—Up to 21 Salactad Elamarm
Cyarida. EPA Standard Mathod Cyanida Amand-
ataM to CNorinalion or Fraa Cyanida..
i mat- I
GC (Avaraga of costs tor mathods 601-612)..
'seoo.oo
'900.00
'600.00
'130.00
•30.00
'4.50
'332.00
•100.00
'Baaad upon actual pncas par lampM offarad to tha
Aganey tor 200-sampla bid lots.
'Baaad upon actual pncas oftarad to tha Aganey tor 750
ICP and 6500 AA sampla bid tots.
'Bissd upon actual pncas oftarad par sampla to tha
Aganey tor 250-sampta bid tots.
'Baaad upon actual pncas par sampla tor $400 tarr
(900 volets*. 4SOO asmi-volatla). avaragad tor Ihraa I
tracts with tha Aganey.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 17
The Agency has compiled the data
presented in Exhibit 5 from actual
(lowest) contract bids, obtained through
a competitive bidding process. As
shown in the footnotes, they were based
upon lots of specific sizes and on sample
throughputs of approximately 200
samples per month for GC/MS and
transmission electron microscope (for
asbestos), and 100 per month for
elemental analyses. The table costs
were constructed on the basis of survey
analysis. Permittees may expect
somewhat lower costs if they are only
analyzing for certain specific
parameters.
A commenter making a commercial
quotation places the cost of a quality
assured analysis of the 111 currently
designated organic toxic pollutants at
S500 per sample. However, comparison
of the Agency's and other commenters'
cost data has generally been impossible.
since the Agency has not known the
factors on which the various
commenters' cost estimates were based.
The Agency has estimated analytical
costs in other regulations which
implement various NPDES monitoring
requirements of the Clean Water Act.
Analytical costs were estimated in the
preambles of the Consolidated Permits
Regulations (45 FR 33290, May 19,1980)
and NPDES Regulations (44 FR 34408,
June 14,1979), which are consistent with
the estimates given in Exhibit 5.
Analytical cost estimates consistent
with those given above have also been
included in promulgations of effluent
limitations guidelines and pretreatment
standards for individual industrial
categories.
While EPA agrees with many
commenters that less costly procedures
would be highly desirable, the proposed
methods appear to be the most cost-
effective currently available. These cost
estimates include 4 spiked reagent water
analyses and 10% spiked sample
analyses. Ongoing research by this
Agency and others should produce less
expensive approaches that, with testing,
will prove equivalent or better,
especially for analyzing trace organic
toxic pollutants in industrial and
municipal wastewater.
To encourage development of new,
improved, and less costly analytical
technologies, the Agency, by
establishing the Equivalency Program
(see § 136.4 and 136.5), has provided a
mechanism for their rapid approval and
use of new procedures.
F. Publication of Full Texts of Test
Procedures
Comments: Several commenters
thought that it was inappropriate for the
EPA to publish the full texts of test
procedures within these guidelines.
They felt that the Agency had reversed
its past policy to "incorporate test
procedures by reference" into these
regulations.
Response: When test procedures are
readily available from other sources.
EPA will incorporate them by reference.
However, test procedures for routine
analysis of trace organic pollutants, with
the exception of a few pesticidal organic
chemicals, were unavailable to meet the
analytical requirements for controlling
priority organic pollutants. EPA
developed the proposed test procedures
through a high-priority research effort.
For the convenience of the public and to
facilitate timely comment, EPA
published the full texts of the draft
procedures in the preamble of the
proposed regulation. In order to continue
the convenience, and to ensure that the
final methods are as widely distributed
as the proposed methods, EPA is
including, as an appendix to this
amendment, the full text of the
approved, Agency developed, test
procedures for. analysis of the priority
pollutants. It is possible that in the
future full text of these test procedures
will become available from other
sources.
In general, the text of approved test
procedures will not be published in the
Federal Register when they are readily
available from other sources. EPA will
continue to "incorporate approved test
procedures by reference" in these
guidelines as it has in the past. Methods
are clearly incorporated by reference in
§ 136.3(a) and a new § 136".3(b) has been
added to clearly identify the sources of
these references. This should resolve the
commenter's concerns.
G. Consistency of Analytical Methods
Approved Under Different Acts
Comments: Several commenters were
concerned that EPA was requiring the
regulated community to use two
different test procedures for analyzing
similar or identical compounds, one set
under the Safe Drinking Water Act, and
another in these Guidelines under the
Clean Water Act. They concluded that
the cost of compliance with these
various requirements unduly burdened
the "rate payer" by what appeared to be
duplication.
Response: EPA makes every effort to
ensure that the analytical methods
prescribed under its regulations are
proliferated only when necessary.
Regulations issued under different Acts
are applicable to the matrices defined
by those Acts. Where analytical
methods are prescribed in these
regulations, consideration must be given
to the concentration ranges that must be
measured and the interferents to be
expected.
Interferents may be unique to a matrix
or may be common to several matrices
covered by these Acts and regulations.
Whenever possible, within the
information and data available to the
Agency, the same methods are approved
for measuring the same parameters in
the different matrices. There are many
instances where this is not possible. For
example, for drinking water analyses
both a purge and trap procedure
(Method 501.1) and an alternate solvent
extraction procedure (Method 501.2) are
approved for the analysis of the
trihalomethanes (THM). In drinking
water, interferents for THM analysis are
minimal. If the drinking water extraction
procedure were applied to wastewater
samples for the purgeable compounds,
extracts which result would present
much more formidable analytical
problems than would have been the
case if the purge and trap procedure had
been used. Therefore, it is not possible
lr' approve the drinking water THM test
. '-ndures for the analysis of THMs
a.r' . related puraeable pollutants in
wastewater samples. On the other hand.
Method 601 can easily be used for
analysis of THMs in drinking water.
VI. Regulatory Analyses
(a) Under Executive Order 12291, EPA
must judge whether a regulation is
"major" and therefore subject to the
requirement of a "Regulatory Impact
Analysis." This regulation is not major
for the following reasons: (1) It only
prescribes analytical methods and
sample handling requirements that
ensure a uniform measure of pollutants
across all wastewater discharges within
minimum acceptance criteria. It does not
require that analyses actually be made.
The purpose is to ensure that the quality
of environmental monitoring data meets
certain minimum standards.
(2) The impact of this regulation will
be far less than $100 million.
(a) The regulation affects unit
monitoring costs for other regulatory
programs, e.g., effluent guidelines
regulations and the implementation
regulations of the National Pollutant
Discharge Elimination System (NPDES),
and the pretreatment programs.
However, it does not impose those costs.
In fact, the monitoring costs for other
programs are considered in each other
rule-making. This is appropriate because
total (rather than unit) monitoring costs
are determined by the monitoring
provisions of those other regulations.
(b) This regulation has deliberately
provided approval of several analytical
options for most compounds. This often
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18 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
allows NPDES permittees to select the
option that is best suited to their
particular monitoring requirements and
that will minimize their monitoring
costs. In addition, the approval of the
CBODs analysis will facilitate
modification of CWA permits to allow
treatment works and their control
authorities to focus on the best measure
of oxygen demand, thereby achieving
treatment economies that will reduce
costs of treatment significantly.
(c) Further, through the equivalency
provisions, these test procedure
guidelines have been designed to
encourage the development of
innovative analytical methods by the
private sector and to encourage the
competitive viability of the instrument
manufacturing industry. The
equivalency provision also allows
individual dischargers to gain approval
of analytical systems of their own
design that may futher reduce their total
monitoring costs.
(3) The impact of compliance with
these regulations will not be
concentrated on any particular sectors
of American industry.
This regulation was submitted to the
Office of Management and Budget
(OMB) for review as required by
Executive Order 12291. Any comments
from OMB to EPA and any EPA
response to those comments will be
available for public inspection at the
Public Information Reference Unit.
Room M2904 (EPA Library—Rear), TM-
213. Environmental Protection Agency,
401 M Street. SW., Washington. D.C.
20460. Phone: (202) 382-5926, Office
Hours 8:00 a.m. to 4:30 p.m.
(b) Under the Regulatory Flexibility
Act. 5 U.S.C. 601 et seq., EPA is required
to determine whether a regulation will
significantly affect a substantial number
of small entities so as to require a
regulatory analysis. The regulation
requires no new reports beyond those
already now required. The analytical
techniques approved here either can be
handled by small facilities, or are
widely available by contract at a
reasonable price. Therefore, in
accordance with 5 U.S.C. 605(b), I
hereby certify that this rule will not
have a significant adverse economic
impact on a substantial number of small
facilities.
(c) The equivalency information
p. jvision in this rule has been submitted
for approval to the Office of
Management and Budget (OMB) under
the Paperwork Reduction Act of 1980. 44
U.S.C. 3501 et seq. It is not effective until
OMB approves it and a technical
amendment to that effect will be
published in the Federal Register.
VII. Effective Dates
The effective date of this rule is
January 24,1985 for all methods except
the method for CBOD5. This date. 90
days hence, was chosen to allow
analysts sufficient time to learn of the
new methods and to implement the
necessary changes in laboratory
practices. After January 22,1985, data
reported to EPA must be generated
using the methods approved under 40
CFR Part 136, including these methods
added by today's amendment.
An effective date of 30 days,
November 23,1984, was chosen for using
the method for CBOD5. This is the only
method for a new parameter. Treatment
works and permitting authorities are
eager to begin measurement and
analysis for this new parameter because
it may result in achieving treatment
economies and reduce costs
significantly. Although data reported to
permitting authorities or EPA on this
parameter must be generated using this
method after 30 days, there is no
requirement that any treatment works
monitor for this parameter or use the
new method until a treatment works
applies for and receives a permit
modification. Permits may be modified
after EPA's amendment to the secondary
treatment rule is effective. The amended
secondary treatment rule should be
effective before or on the same day that
today's rule approving the CBOD5
method is effective. There is no reason
why EPA should postpone the effective
date of this rule for more than 30 days
and possibly delay use of this method
and parameter for persons whose permit
can be changed quickly.
The Director of the Federal Register
has approved all materials which are
"incorporated by reference" in the text
of the regulation to be incorporated.
They are incorporated by reference on
the effective dates given above.
List of Subjects in 40 CFR Part 136
Water Pollution Control;
Incorporation by reference.
Dated: September 26.1984.
William O. Ruckelshaus.
Administrator.
Regulation
For the reasons set out in the
Preamble, Part 136, Chapter 1,
Subchapter D of Title 40, Code of
Federal Regulations, is amended as set
forth below. Only new provisions are
being promulgated at this time;
unchanged provisions are reprinted for
the purpose of clarity.
1. The authority citation for Part 136 is
revised to read as follows:
Authority: Sections 301. 304(h). 307 and
501(a). Pub. L. 95-217. 91 Slat. 1566. et seq. (33
U.S.C. 1251, et seq.) (the Federal Water
Pollution Control Act Amendments of 1972 ;
amended by the Clean Water Act of 1977).
2. In paragraphs 136.1 (a) and (c).
reference to the Federal Water Pollution
Control Act Amendments of 1972
(FWPCA) is replaced by reference to the
Clean Water Act of 1977. and
applicability under paragraph 136.l(a) is
extended to Parts 122-125 of Title 40. As
revised, paragraphs (a) and (c) of § 136.1
read as follows:
§ 136.1 Applicability.
(a) An application submitted to the
Administrator, or to a State having an
approved NPDES program for a permit
under § 402 of the Clean Water Act of
1977, as amended (CWA), and/or to
reports required to be submitted under
NPDES permits or other requests for
quantitative or qualitative effluent data
under Parts 122 to 125 of Title 40, and,
* * * * .
(c) Certifications issued by States
pursuant to § 401 of the CWA. as
amended.
3. In § 136.2. paragraph (a) is revised
to reference the Clean Water Act of
1977. and paragraphs (f). (g), and (h) are
removed, and a new paragraph (f) is
added defining "Detection limit." and
paragraphs (g) and (h) are reserved, as i
follows:
§136.2 Definition*.
As used in this part, the term:
(a) "Act" means the Clean Water Act
of 1977, Pub. L. 95-217. 91 Slat. 1566. et
seq. (33 U.S.C. 1251 et seq.) (The Federal
Water Pollution Control Act
Amendments of 1972 as amended by the
Clean Water Act of 1977).
*****
(f) "Detection limit" means the
minimum concentration of an analyte
(substance) that can be measured and
reported with a 99% confidence that the
analyte concentration is greater than
zero as determined by the procedure set
forth at Appendix B of this Part.
(g) [Reserved.]
(h) [Reserved.]
4. In § 136.3. Table I is restructured
into five new tables by transferring the
biological parameters formerly
designated as parameters 4, 5. 6, 7 and 8
to new Table IA, entitled "List of
Approved Biological Test Procedures,"
by adding an additional EPA reference
to the approved test procedures, and by
updating the references to Standard
Methods and USGS: by transfering the
inorganic parameters formerly
designated as parameters 1-3,10-13.15-1
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
19
93, 96-98, and 104-115 to new Table IB,
entitled "List of Approved Inorganic
Test Procedures", adding two new
inorganic parameters, Carbonaceous
Biochemical Oxygen Demand (CBOD5)
and Nitrate-Nitrite, including an
additional test procedure based upon
the inductively coupled plasma
technique in Table IB for 25 of the metal
parameter designations, by including 10
methods approved under the
equivalency provisions of §i 136.4[d)
and 136.5[e), and updating references to
EPA, Standard Methods, ASTM, AOAC
and USGS test procedures; by deleting
former parameter 14 (Chlorinated
organic compounds) and by entering the
individual chlorinated organic
compounds into new Table 1C, entitled.
"List of Approved Test Procedures for
Non-Pesticide Organic Compounds",
transferring old parameters 9
(Benzidine) and 94 (Pentachlorophenol)
to Table 1C. by including the 78
additional proposed non-pesticidal
organic parameters and by adding 17
new test procedures in Table 1C; by
deleting former parameter 95
(Pesticides) and by entering the 68
individual pesticides into new Table ID,
entitled "List of Approved Test
Procedures for Pesticides", by including
the 2 additional proposed pesticide
parameters, and the two new test
procedures in Table ID; and by
transferring the former radiological
parameters 99-103 to new Table IE,
entitled "Approved Radiological Test
Procedures", adding an EPA reference to
the approved test procedures, and
updating the Standard Methods, ASTM
and USGS references. As revised, Table
I reads as follows:
§ 136.3 Identification of Test Procedures.
TABLE IA.—LIST OF APPROVED BIOLOGICAL TEST PROCEDURES
I
Parameter and units Method '
Bacteria: I
EPA"
Reference (Method Number or Pagel
Standard
Methods tsth
Ed.
I
ASTM !
I
USGS
1. Conform (fecal) number per 100 ml ! MPN. 5 tube. 3 dilution!; or. membrane filter (MR 4. single steo ... p. 132 980C !
p. 124 1 909C , 8-0050-77.
2. Coliform (fecal) in presence of chlorine number per 100 ml MPN, 5 tube. 3 dilution | p.132
3. Colilorm (total, number per 100 ml , MPN. 5 lube. 3 dilution: or. MF • single step or two step p • u ...
908C
-I
908A I
| i p. :J j 909A , ! 8-0025-77.
4. Coliform (total) in presence of chlonne, number per 100 ml ; MPN, 5 tube, dilution; or MF * with enrichment | p • 4 : 908A , ,
! c •' 909(A+A.5c)...., |
5 Fecal streptococci, number per 100 ml ! MPN. 5 tube, 3 dilution; MF «; or, plate count p. 139 i 910A , ,
. p. 136 9108 1 , 80055-77.8
| p. 143 910C | j
Table IA Notes
1 The method must be specified when results are reported.
' "Microbiological Methods for Monitoring the Environment, Water and Wales. 1978", EPA-600/8-78-017, U.S. Environmental Protection Agency.
3 Greeson, P.E.. el a/.. Methods tor Collection and Analysis of Aquatic Biological and Microbiological Samples. "U.S. Geological Survey. Techniques of Water-Resources Investigations.
Book 5. Chapter A4. Laboratory Analysis. 1977.
• 0.45 um membrane filter or other pore size certified by the manufacturer to fully retain organisms to be cultivated, and free of extractables which could interfere with their growth and
development.
s Approved only if dissolution of the KF Streptococcus Agar (Section 5.1. USGS Method 8-0055-77) is made in a boiling water bath to avoid scorching of the medium.
TABLE IB.—LIST OF APPROVED INORGANIC TEST PROCEDURES
, Reference (method No. or page)
Parameter, units, and method
or phenolphthatein end point.
2. Alkalinity, as CaCOi, mg/L Electrometric or color-
imetnc:
3. Aluminum— Total 3 mg/L: Digestion 3 followed by:
4. Ammonia (as N). mg/L Manual distillation ° (at
pH 9.5):
Followed by .
5. Antimony— Total 3, mg/L: Digestion 3 followed by:
6. Arsenic— Total 3, mg/L
Hydride
7. Barium— Total 3. mg/L: Digestion 9 followed by:
8. Beryllium— Total 3. mg/L: Digestion » followed by:
AA furnace
EPA 1979
305.1
310 1
310.2
202.1
2022
350.2
350.2
3502
350 3
350.1
204.1
2042
206.5
206.3
2062
2064
208 1
208 2
210 1
210.2
Standard methods 15th
Ed.
402(4. d)
403
303C
304
306B
417A
41 7B
4170
417F
303A
304
303E
304
3078
303C
304
303C
304
ASTM
D1067-70(E)
01067(8)
01426-79(A|...
01426-79(0)
D1426-79(C)
02972-78(8).
02972-78(A)
03645-78
USGS'
1-1030-78
1-2030-78
1-3051-78
1-3520-78
1-4523-78
1-3062-78
1-3060-78
1-3084-78
1-3095-78
Other
P 548'
Method 200.7. •
P. 553. «
t
Method 2007*
Method 2007'
Method 2007*
-------
20 Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
TABLE IB.—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued
Parameter, units, and method
9. Biochemical oxygen demand (BOOO. mg/L
Winkler (Azide modification)
Or etectroda method
EPA 1979
405 1
Reference (nwtltod No. or page)
Standard methods 15th ASTM USQS , j ^
i Method 200 7 *
3096 , j
507 ; ' 1-1578-78 j P 17 •
, ; : P. 548.'
320.1..
10 Boron—Tot*. mg/L: !
CotonmatK (curcumm) or 212.3...
inductively coupled plasma
11 Bromide. mg/L Titnmetnc
12. Cadmium—Total". mg/L: Digestion1 followed .
by: i
AA direct aspiration , 213.2...
AAtumace 213.2...
Inductively coupled plasma |
VoRametry" or |
Cotonmetric (Dithnone) i
13. Calcium—Total' mg/L: Digestion9 followed by: i
Atomic absorption I 215.1...
Inductively coupled plasma i
Or EDTA Mratlon 215.2...
14 Carbonaceous Biochemical oxygen demand I
(CBOOO. mg/L: Winklar (Ante moditicalion) or |
electrode method with nitrification inhibitor. j
15. Chemical rwygen demand (COO). mg/L: !
Titnmetnc cotonmetnc j 410.1...
Manual or ; 410.2...
i 410.3...
Automated | 410.4...
Spectropholometnc
16 Chloride. mg/L:
Titnmetnc (ewer nitrate) or •
Mercuric nitrate \ 325.3
Cotonmetnc (femcyanide) manual or j
Automated , 325.1 or 325.2
17 Chlorine Total residual. mg/L: '
Titrimalnc^mperometnc" i 330.1
Starch end point I 330.2
lodomatncor : 330.3
OPO-FAS | 330.4
Spectropnotometnc. DPD; or i 330.5
Electrode i
18. Chromium VI dissolved. mg/L 0.45 micron flltra- i
tion with: I
Extraction and atomic absorption, or ; 218.4
Cotohmetnc (Diphenylcarbande)
19. Chromium—Total3. mg/L :
Digestion1 (optional extraction) followed by 218.3
AA direct aspiration ; 218.1
AAtumace ! 218.2
Inductively coupled plasma I
Or cotonmetnc (Diphenytcarbazide) ;
20. Cobalt—Total«. mg/L Digestion1 followed by: '•
AA direct aspiration I 219.1
AA furnace, or I 219.2
inductively coupled plasma i
21. Color, platinum Cobalt units or dominant wave- i
length hue. luminance, purity
Cotonmetnc. ADMI j 110.1
Platinum cobalt: or , 110.2
Spectropholometnc , 110.3
22. Copper—Total3. mg/L: Digestion' followed by: j
AA direct aspiration I 220.1
AA furnace I 220.2
Inductively coupled plasma ,
Cotonmetnc (Neocuprome) t
Bicincnoninate i
23 Cyanide—Total mg/L:
Manual disMMUon with MgCI, , 335.2
Followed by trtnmetnc i 335.2
Manual or j 335.2
Automated15 spectropnotometrtc , 335.3
24. Cyanide amenable to chtonnation, mg/L Manual i 335.1
distillation with MgCI,: Followed by titnmemc.
manual or automated ' * Spectropnotometnc. :
25. Fluoride—Total. mg/L: ;
Manual damnation' ;
Followed by manual or , 340.2
Automated electrode I
SPADNS , 340.1
Or automated complexone ! 340.3
26. Gold—Total1. mg/L Digestion5 followed by: ;
A« direct aspiration i 231.1
Or AA furnace 231.2
27 Hardness—Total as CaCo,. mg/L: i
Automated cotonmetnc 130.1
EDTA titration j 130.2
Inductively coupled plasma ,
Or atomic absorption (sum , 215.1 +
of Ca and Mg as their respective carbonates) > 242.1
; 404A i I 1-3112-78 ,
i i ; Method 200.7 <
, D1246-77IC) , 1-1125-78 1 P S44."
303A or 3038 j 03557-78 (A or B) i 1-3135-78 or 1-3136-78 ..... Pg. 557 '
304 i i P 37 •
! , Method 200.7.'
D3557-78(C) i
3108
i 303A | D511-77(C) '. 13152-78..
. 311C D511-77(B)..
507(5.8.6) :
Method 200.7«
508A....
D1252-78 , I-3560-78 P 550' and
i I-3562-78 P 17« and
1-3561-78 i (">.)
I 407A : 0512-67(8) 1-1183-78
: 4078 l D512-«7(A) i 1-1184-78 P. 554.'
i 0512-67(C) , 1-1187-78
! 4070 ! 1-2187-78 ;
. 408C , D1253-76(A)
4088
. 408A 01253-76(8)
4080
408E
3038 : 1-1232-78 ..
1-1230-78..
303A or 3038
304
01687-7/10)
312A D1687-77IA)
, I-3236-78 P. 557 «
Method 200.7.'
303A or 3038 03558-77 (A or B) I-3240-78 or I-3239-78
304
P 37. •
. Method 200.7.'
2040
204A
2048
1-1250-78 .
303A or 3038 \ D1688-77 (D or E) 1-3271-78 or 1-3270-78.... P. 557« and P. 37"
304 ,
Method 200.7.'
3138 D1688-77(A)
I")
4120
4128 P 22.'
412C D2036-75(A) ,
. 4120 i D2036-75(A) , I-3300-78
412F 02036-75(8)
413A
4138
, 413C
413E
01179-72(8)
i D1179-72(A)
i 1-4327-78..
303A
304
3148 01126-67(8) 1-1338-78
Method 200.7«
303A I 1-3153-784
i 1-3448-78..
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 21
TABLE IB.—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued
Parameter, units, and method
28. Hydrogen ion (pH). pH units:
29. Indium— Total3. mg/L Digestion1 followed by:
30. Iron— Total3. mg/L:
AA direct aspiration
31. KjeKJahl nitrogen— Total (as N), mg/L
32. Lead— Total3. mg/L Digestion3 followed by:
33. Magnesium— Total 3, mg/L Digestion 3 followed
by:
34. Manganese— Total >. mg/L Digestion ' followed
by:
35. Mercury— Total s. mg/L
36. Molybdenum— Total 3, mg/L: Digestion 3 fol-
lowed by:
37. Nickel— Total 3, mg/L Digestion3 followed by:
38. Nitrate (as N). mg/L
39. Nitrate-nitrite (as N), mg/L
Or automated; or
40. Nitrite (as N). mg/L
41. Oil and grease— Total recoverable, mg/L Gravi-
metric (extraction).
42. Organic carbon— Total (TOO). mg/L Combustion
or oxidation.
43. Organic nitrogen (as N), mg/L Total KjeWahl N
minus ammonia N.
44. Ortnopnosphate (as P). mg/L Ascorbic acid
metfxx), automated
45. Osmium— Total ', mg/L Digestion ' followed by:
46. Oxygen, dissolved, mg/L
47. Palladium— Total *, mg/L Digestion3 followed
IV
t"V AA h
48. Phenols. mg/L
49. Phosphorus (elemental), mg/L Gas-liquid cnro-
matography.
Reference (method No. or page)
EPA 1979
150.1
235.1
235 2
236 1
2362
351 3
351 3
351 3
351 3
351 1
351 2
351 4
239 1
239.2
242.1
243 1
243.2
245 1
245.2
246 1
2462
249 1
249.2
352 1
See parameters 39 and
40.
353 3
353.2
353 1
354 1
413 1
415 1
See parameters 31 and 4 .
365 1
3652
365 3
252 1
252.2
3602
360 1
253 1
2532
420 1
420 1
4202
Standard methods 15th
Ed.
423
303A
304
303A or 303B
3038
304
3158
420A or 8
4170
4178
41 7E
303A or 303B
304
316B
303A
3188
303A or 3038
304
3198
303F
303C
304
303A or 303B
304
321B
See parameters 39 and
40.
418C
418F
419
503A
505
420A
424G
424F
303C
304
4218
421F
ASTM
01293-78IA) or 01293-
78(B).
01068-77
(Cor 0)
D1068-77(A)
03590-77 ..
03559-78 (A or B)
03559-78(0)
0511-77(8)
D51t-77(A)
0858-77 (B or C)
D858-77(A)
03223-79
01886-77 (C or 0)
0092-71
See parameters 39 and
40.
03867-79(8)
D3867-79(A)
01254-67
D2579-78(A) or 02579-
78(8).
03590-77 minus 01426-
79(A).
0515-78(A)
01589-60(A)
D1 783-70 (A or B)
USGS'
1-1586-78
1-3381-78
1-4551-78
1-4552-78
1-3399-78
1-3447-78
1-3454-78
1-3462-78
1-3490-78 .
1-3499-78
See parameters 39 and
40.
1-4545-78
M540-78
See parameters 31 and 4 .
1-4601-78
1-1575-78
M576-78
Other
,,..,
P. 557.'
Method 200.7.'
("•]
P. 552.'
P. 557.'
Method 200.7.«
P. 557.'
Method 200.7.4
P. 557.'
Method 200.7.'
P. 564.'
18.
P. 559.'
Method 200.7.'
Method 200.7.'
P. 554.'
P. 28."
19.
P. 551 ' and P. 4.'°
PP. 552-53.'
P. 561.'
P. 550.'
P. S27."
P. S28."
26.
26.
21.
-------
22 Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE IB.—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued
Reference (method No. or page)
Parameter, units, and method
EPA 1979
Standard methods 15m
Ed.
ASTM
USGS'
Other
50 Phosphorus—Toial. mg/L:
Persulfate digestion 365.2 ! 424C (III) i ; p
Followed by manual or | 365.2 or 365.3 424F ! D515-78IA) |
Automated ascorbic aod 365.1 , 424G : I-4600-78..
Reduction; or Mrm-aulomaMd Block digester I 365.4 ; , i M603-7S..
51 Platinum—Total'. mg/L: Digestion J followed by: i !
AA direct aspiration ,255.1 303A i..
Or AA lumac* j 255.2 304 ]..
52. Potassium—Total '. mg/L. Digestion ' followed I ! I
by: ' i i
Atomic absorption 258.1 ; 303A , 1-3630-78 p. 560.'
Inductively coupled plasma ; ! Matted 200.7 <
Or flame photometric I i 322B , 01428-64(A| I
53. Residua—total. mg/L: Gravimetric. 103-105'C j 160.3 i 209A ! I 1-3750-78 !
54. Residue—MtarabM. mg/L: Grsvimetnc, 180'C j 160.1 209B j 1-1750-78
55. Residue—nonWteradle. (TSS). mg/L: Grew-j 160.2 , 2090 : 1-3765-78
matnc. 103-105'C post washing ol residue.
56 Residue—settteaole. mg/L. Volumetric (Imhofl 160.5 , 209F
cone) or gravimetric. j :
57 Residue—volet*. mg/L: Gravimetric. 550'C 160.4 i 209E i 1-3753-78
58 Rhodium—Total'. mg/L: Digestion' followed
by:
AA direct aspiration i 265.1 303A....
Or AA furnace | 267.2 i 304
59. Ruthenium—Total'. mg/L: Digestion ' lolkMred I
by:
AA direct aspiration 267.1 303A....
Or AA furnace i 267.2 304
60. Selenxn—Total3 mg/L: Digestion " followed by: ;
AA fumance , 270.2 304
Inductively coupled plasma j : i Method 200 7«
Ornydnde I 270.3 303E ! , D3859-79 j I-3667-78
61. Mica—Dissolved, mg/L: 0.45 micron filtration: '
Followed by manual or , 370.1 «5C ; D859-68(B) | 1-1700-78
Automated cotonmetnc (Molvbdosilicate). or j , ! I-2700-78
inductively coupled plasma i.... i i ! Method 200.7.«
62. Silver—Total " mg/L: Digestion ' followed by: i I
AA direct aspiration 272.1 303A or 303B ! 1-3720-78 P. 557» and p. 37 •
AA furnace, or 272.1 304 1
Inductively coupled plasma ! j ; Method 200.74
63 Sodium—Total *. mg/L: Digestion ' followed by:
Atomic absorption ! 273.1 303A , : I-3735-78 P 581.'
Inductively coupled plasma , : ! Method 200.7.«
Or flame photometric : D1428-64(A)
64. Specific conductance, mnos/cm: Wheatslone 120.1 205 D1125-77(A) I 1-1780-78 P 547.'
DndQO- ' ' i
65. Sullate (as SO.). mg/L: i !
Automated methytthymol blue : 375.2 i i 1-2822-78
Gravimetric, or ', 375.3 «6A or 4Z6B D516-68IA) i PP 562-63.'
Turtudimemc \ 375.4 ', 426C j D516-68(B) j
66. SulMe (as S), mg/L: ' '
Titnmetnc (lOOSne) or 376.1 ; 427D j I I-3840-78
Cokximetnc (methylene blue) , 376.2 427C , |
67. Sulfite las SO,), mg/L: Titrimetnc (iodine wdale)..[ 377.1 ; 428F j D1339-78IC)...
68 Surfactants. mg/L. Cokximetric (methylene blue)..! 425.1 '. 512A ; D2330-68(A).
69 Temperature. 'C.: Thermometnc j 170.1 ; 212 ! ! («).
70. Thallium—Total." mg/L: Digestiona followed by: i ] i j
AA direct aspiration i 279.1 j 303A |
AA furnace, or | 279.2 j 304 ; I |
Inductively coupled plasma i j ! Method 200 7«
71 Tin—Total.' mg/L: Digestion ' followed by: !
AA direct aspiration or I 282.1 303A j I-3850-78..
AA furnace 282.2 304
72. Titanium—Total.' mg/L: Digestion > followed by: j
AA direct aspiration or.. 283.1 j 303C....
AA furnace 283.2 i 304 ,
73. Turbidity. NTU: Nepnetometnc 180.1 j 214A 01889-71 | I-3860-78 ,
74 Vanadium—Total.3 mg/L. Digestion' followed i ' j
by: i ! i
AA direct aspiration j 286.1 ; 303C ; j
AA furnace 266.2 1 304 j I
Inductively coupled plasma I i | ! Method 200.7.<
Or colonmeiric (Gallic aod) ', : ; 03373-75
75. Zinc—Total.1 mg/L Digestion1 followed by: i ; :
AA direct aspnUonb 289.1 303A or 303B | 01691-77(0) 1-3900-78 i P. 5S7.«
AA furnace 289.2 ; 304 : D1691-77(C) i P. 37."
Inductively coupled plasma ! I i , Method 200.7 <
Or cotonmetnc (Zinconi
1979.
T*MQ !• MBtAfl
> "Methods for Analysis of Inorganic Substances m Water and Fluvial Sediments." US. Department of the Interior. U.S. Geological Survey, Open-File Report 78-679. or "Methods for
nmnatnn of Inorganic Substances m Water and Fluvial Sediments." N.W. Skougstad. »/«/ U.S. Geological Survey. Techniques of Water-Resources Investigation. Book 5. Chapter A1.
1 "Official Methods of Analysis of the Association of Official Analytical Chemists" methods manual. 13th ed. (1980).
1 For the determination ol total metals the sample is not filtered before processing. A digestion procedure is required to soli
plena. Two digeston procedures are given m "Methods for Chemical Analysis of Water and Wastes. 1979." One (} 4 1.3). i
idedi
al and to destroy possMa orgarnc-metal
._., . r . . las. 1979." One (}4.1.3). is a vigorous dneation usmg nitric aod. A less vigorous digestion
using nitric and nvdrochotooc acids ({ 4.1 4) is preferred: however, the analyst should be cautioned that this rnM digestion may not suffice tor all sample types. Particularly, if a ce4onmeinc
procedure is to be employed, it is necessary to ensure that all or
preferred making certain that at no time does the sample go "
graphite furnace technique, inductively coupled plasma, as wel
digestion and in all cases the method wnte-up should be consulted for specific instructions and/or cautions.
that all organc-metallic bonds be broken so that the metal is n a reactive stale. In those srtuasons, the vigorous digeston is
pie go to oryneee. Samples containing large amounts of organic materials would also benefit by this vigorous digestion. Use
as well as dMermnMons for certain elements such as arsenic, the noble mettle, mercury, illinium, and trtanwn require e m
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 23
Not*: II the digestion procedure lor direct aspiration or graphite furnace atomic absorption analysis included in one ol the other approved references is different than the above, the EPA
procedure must be used.
Dissolved metals are defined as those constituents which will pass through a 0.45 micron membrane filter. Following filtration of the sample, the referenced procedure for total metals must
be followed. Sample digestion ot the filtrate lor dissolved metals, or digestion of the original sample solution for total metals may be omitted for AA (direct aspiration or grapnite furnace) and
ICP analyses provided the sample has a low COO and the filtrate meets the following criteria:
(a) Is visibly transparent
(b) Has no perceptible odor, and
(c) Is tree of paniculate or suspended matter following acidification.
' The full text ot Method 200.7. "Inductively Coupled Plasma Atomic Emission Spectrometnc Method for Trace Element Analysis ol Water and Wastes." is given at Appendix C of this Part
136.
1 Manual distillation is not required if comparability data on representative effluent samples are on company file to show that this preliminary distillation step is not necessary: however.
manual distillation will be required to resolve any controversies.
' Ammonia. Automated Electrode Method. Industrial Method Number 379-75WE. dated February 19. 1976. Technicon AutoAnalyzer II. Technicon Industrial Systems. Tanytown. New York
10591.
' Carbonaceous biochemical oxygon demand (CBODO must not be confused with the traditional BOD* test which measures "total BOD". The addition ol the nitrification inhibitor is not a
procedural option, but must be included to report the CBOD, parameter. A discharger whose permit requires reporting the traditional C80D> may not use a nitrification inhibitor in the procedure
for reporting the results. Only when a discharger's permit specifically states CBOD, is required can the permittee report data obtained using the nitrification inhibitor.
' American National Standard on Photographic Processing Effluents. Apr. 2. 1975. Available from ANSI. 1430 Broadway. New York. NY 10018.
* The use of normal and differential pulse voltage ramps to increase sensitivity and resolution is acceptable.
10 Chemical Oxygen Demand. Method 8000. Hach Handbook ol Water Analysis. 1979. Hach Chemical Company, P.O. Box 389. Loveland. Colorado 80537.
1' COD Method. Oceanography International Corporation. 512 West Loop. P.O. Box 2980. College Station. Texas 77840.
11 The Dack titratran method will bo used to resolve controversy.
13 National Council ol the Paper Industry lor Air and Stream Improvement. Inc.. Technical Bulletin 253. December 1971.
" Copper. Bicinchonmate Method. Method 8506. Hach Handbook of Water Analysis. 1979. Hach Chemical Company. P.O. Sox 389. Loveland. Colorado 80537.
15 After the manual distillation is completed, the auto-analyzer manifolds in EPA Methods 335.03 (Cyanide) or 420.2 (phenols) are simplified by connecting the re-sample line directly to the
sampler. When using the manifold setup shown in Method 335. the buffer 6.2 should be replaced with the buffer 7.6 found in Method 335.2.
"Hydrogen Ion (pH) Automated Electrode Method. Industrial Method Number 378-75WA. October 1976. Technicon Auto-Analyzer II. Technicon Industrial Systems. Tanytown. New York
10591.
» Iron. 1.10-Phenanthroline Method. Method 8008. 1980, Hach Chemical Company. P.O. Box 389. Loveland. Colorado 80537.
"Manganese. Penodate Oxidation Method. Method 8034, Hach Handbook ot Wastewater Analysis. 1979, pages 2-113 and 2-117, Hach Chemical Company, Loveland. Colorado 80537
" Nitrogen, Nitrite. Method 8507. Hach Chemical Company. P.O. Box 389, Loveland. Colorado 80537.
"Goertitz. D.. Brown. E.. "Methods for Analysis of Organic Substances in Water." U.S. Geological Survey. Techniques of Water-Resources Investigations. Book 5. Chapter A3. p.4 (1972).
21R.F. Addison and P..G. Ackman. "Direct Determination of Elemental Phosphorus by Gas-Liquid Chromatography," Journal of Chromatography. Vol. 47. No. 3. pp. 421-426. 1970.
" Recommended methods for the analysis of silver in industrial wastewaters at concentrations ol 1 mg/L and above are inadequate where silver exists as an inorganic halide. Silver
halides such as the bromide and chloride are relatively insoluble in reagents such as nitric acid but are readily soluble in an aqueous buffer ol sodium thiosulfate and sodium hydroxide to a pH
of 12. Therefore, lor levels ot silver above 1 mg/L. 20 ml ol sample should be diluted to 100 mL by adding 40 mL each of 2 M Na,S,O, and 2M NaOH. Standards should be prepared in me
same manner. For levels of silver below 1 mg/L the recommended method is satisfactory.
" Stevens. H.H.. Ficke. J.F.. and Smoot. G.F.. "Water Temperature-Influential Factors. Field Measurement and Data Presentation." U.S. Geological Survey. Techniques of Water-Resources
Investigations. Book 1. Chapter 01. 1975.
"Zinc Zincon Method. Method 8009. Hach Handbook of Water Analysis. 1979. pages 2-231 and 2-333. Hach Chemical Company, Loveland. Colorado 80537.
" "Selected Analytical Methods Approved and Cited by the United States Environmental Protection Agency." Supplement to the Fifteenth Edition ol Standard Methods tor the Examination
ot Water and Wastewater (I9BI1.
" The approved method is that cited in Standard Methods for the Examination ot Water and Wastewater, 14th Edition. The colorimetric reaction is conducted at a pH of 10.0 ± 0.2. The
approved methods are given on pp. 576-81 ot the 14th Edition: Method 510A for distillation. Method 510B for the manual colorimetric procedure, or Method 510C for the manual
spectrophotometric procedure.
"ORION Research Instruction Manual. Residual Chlorine Electrode Model 97-70. 1977. Onion Research Incorporates. -;;j Memorial Drive. Cambridge. Massachusetts 02138.
TABLE 1C.—LIST OF APPROVED TEST PROCEDURES FOR NON-PESTICIDE CYANIC COMPOUNDS
Parameter'
EPA Method Number ' '
GC
GC/MS
HPLC
Other
32 Chrysene
4e. CIS- ,J u oprope
48. Diathvl ohtnalate
610
610
603
603
610
602
610
610
610
610
610
606
611
611
606
601
601
601
611
601
604
601. 602
601
601
601
601
612
604
611
610
610
601
601 602 612
601. 602. 612
601. 602, 612
601
601
601
601
601
604
601
601
601
606
625 1625
625. 1625
•624 1624
•624, 1624
625. 1625
624 1624
'625 1625
625. 1625
625, 1625
625. 1625
625 1625
625 1625
625 1625
625. 1625
625 1625
625 1625
624 1624
624. 1624
624. 1624
625 1625
624. 1624
625. 1625
624 1624
624 1624
624 1624
624 1 624
624 1624
625 1625
625 1625
625 1625
625. 1625
625. 1625
624 1624
624, 625 1625
624. 625, 1625
625, 1624, 1625
625 1625
624, 1624
624 1624
624. 1624
624. 1624
625, 1625
624. 1624
624. 1624
624, 1624
625. 1625
610
610
1
610
605
610
610
610
610
610
.
L
.
610
610
605
Note 3 p 130'
Note 6, p.
S102.
Note 3. p. 130;
Note 3, p 130-
Note 3 p 130'
-------
24 Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
TABLE 1C.—LIST OF APPROVED TEST PROCEDURES FOR NON-PESTICIDE ORGANIC COMPOUNDS—Continued
Parameter'
30 Oinndwl nhHuKU
51 Di-obutyt rtuhalale
52 Dtavoctyl phthefets
M •» 4.riMinvtf^tfM^
54 2 4-Dfni8'otokjene
55 2 B-Oinifrololuene
rm f oir+iirwrihwrkvi
57 EttiyttMnMOft
58 RuOTaVlttWfM
~1 Itt&vf+^-fg^
61 HtKKMorotiulMMnc
0? HOTayhtpfPO cine int>divl '
GC
604
608
608
608
604
609
809
602
610
610
612
612
612
612
610
609
601
604
610
609
604
604
607
607
607
611
608
606
608
806
608
608
608
604
810
604
610
601
601
602
612
601
601
601
601
604
601
GC/MS
625, 1625
625. 1625
625.1625
625.1625
625. 1825
625. 1625
625. 1625
624. 1624
625. 1625
625. 1625
625.1625
625. 1625
•625,1625
625. 1625
625, 1625
625. 1825
624. 1624
625. 1625
625. 1625
625. 1825
625, 1625
625. 1625
625. 1625
•625. 1625
•625. 1825
625. 1625
1
l
t
625. 1625
625. 1625
625. 1625
625. 1625
"613
624, 1624
624. 1824
624. 1624
625. 1625
624. 1624
624. 1624
624. 1624
624.
625. 162S
624, 1624
Table 1C Notes
ven at Appendix A. "Test Procedures for Analysis of Organic Pollutants," of this F
•e test procedures is gwen at Appendix B. "Definition and Procedure for the Determ
orophenol and Pesticides m Water and Wastawater." U.S. Environmental Protect
rytonrtrte. However, when they are known to be present the preferred method for tr
w compounds.
A of thia Part 136) rn accordance with procaduraa aach in section 8.2 of each of thaaa Methods. Additionally, each
lament to tha Fiftaan
xuracy witn Mathodi
lafioratory, on an on
iv. i mwa w , wi lai i
HPLC
610
610
610
[
i
625.
625,
625.
625.
625.
625,
625.
610
610
art 136. Ttv
nation of th
ion Agancy.
•aa two coi
lhay ara km
601-613. 624. 825. 1624. and 1
•going ba*at muat «p*a and an
OMrl
NoM 3. p. 130:
NoM 6. p.
S102.
NoM 3. p. 130:
NOM 3, p. 43;
NOM 3. p. 43:
NoM 3. p. 43;
NOM 3. p. 43:
NOM 3, p. 43.
NoM 3. p. 43:
NOM 3. p. 43;
NoM 3. p. 140;
Hota 3, p. 130;
NOM 3. p. 130;
NOM 3. p. 130:
Nota 3, p. 130:
a (Mndardoad ta«t
i Mathod OaMctwn
SapMmbar. 1978.
npoundaia Mathod
Mm to ba praaant.
tor fn» EKamnmon
625 (Saa Appandnt
Myia 10% (5% lor
ina'racovaiy of any pararnaiar laHa outaida lha warning limit*.' tha analyacal raauM tor thai paramatar in trw~unap*mf sarnpM ara~ auapact and cannot ~ba raporiad » damonattaM raguiatwy
coinplianca.
Nota.—Tnaaa warning KmiM am promukjaMd u an "Marim final acton with a raquaat for commanta."
TABLE ID.—LIST OF APPROVED TEST PROCEDURES FOR PESTICIDES '
Paramatar w)/L)
Mathod
EPA»
Standard i
15th Ed
I ASTM
Othar
1. AWrin
, GC
6061
509A
03086
Nota 3.
p.
7;
NOW 4. p. 30.
625 i
...i NoM 3. p. 83: NoM 6. p. S68.
....( NoM 3. p. 94; NoM 6, p. S18.
GC/MS
2. Amatryn ; GC....
3. Ammocarb , TLC..
4. Alraton i GC I j MOM 3. p. 83: NoM 6. p. 368
5. Atraima GC j , i NoM 3. p. 83; NoM 6, p. S88.
6. Aanpnoa mathyl i GC | j , MOM 3. p. 25; NoM 6. p. SSI.
7. Barban TLC ; ! l NOM 3, p. 104; NoM 6. p. S64.
8. a-BHC ! GC i 608 i 509A l D3088 I NoM 3. p. 7.
, GC/MS j «625i | .... i
9. 0-BHC iGC
; GC/MS
10. 4-BHC : GC
606 I
625 ,
608 :
03086 I
03086 I
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
25
TABLE ID.—LIST OF APPROVED TEST PROCEDURES FOR PESTICIDES '—Continued
!
Parameter M9/U Method
i
j GC/MS
' GC/MS
14 Carbophenothion GC
. GC-MS
GC-MS
19 4 4'-DDE - GC
GC/MS
20 4 4'-DDT GC
! GC/MS
27 Dicotol • GC
GC/MS
30 Oisutfoton GC
31 Diuron j TLC
32 Endosullan 1 . 1 GC
| GC/MS
33 Endosulfan II - GC....
; GC/MS
GC/MS
35 Endrin GC
GC/MS
GC/MS
39 Fenuron-TCA TLC
GC/MS ....
42 Isodrin GC/MS
GC
43 Linuron TLC
50 Monuron-TCA 1 TLC
51 Neburon TLC
53 Parathion ethyl GC..
54 PCNB — - GC
55 Perthane j GC
56 Prometon I GC
59 Propham TLC
62 Siduron . TLC
65 Swep TLC
66 2 4 5-T — GC
GC/MS ..
EPA'-'
!625
608
625
608
625
608
625
608
625
608
625
608
625
608
-25
-.08
•525
o08
625
608
'625
608
625
608
625
608
625
608
625
Standard
Methods
15th Ed
509A
509A
509A
5098
509A
509A
. .
509A
509A
509A
509A
. SOSA
509A
509A
509A
509A
509A
509A
509A
509A
509A
509A
509B
509B
509A
509A
A"-"-
03086
03086
03086
D3086
03086
03086
03086
03086
03086
03086
D3086
03086
...
03086
D3086
Other
Note 3 p 7
Note 3 p 94* Note 6 p S60
Note 4 p 30- Note 6 p S73
Note 3 p 7
Note 3 p ?• Note 4 p 30
Note 3 p 7- Note 4 p 30
S51.
Note 3 p 1 1 5
Note 4 p 30- Note 6 p S73
Note 3 p 7
Note 4 p 30' Note 6 p S73
Note 3 p • Note 6 p S51
Note 3 p 7
Note 3 p 7
Note 3 p 7- Note 4 p 30
Note 4 p 30- Note 6 p S73
Note 3 p 104- Note 6 p S64
Note 3 p 104- Note 6 p S64
Note 3 p 7' Note 4 p 30
Note 3 o 7' Note 4 p 30' Note 6 p
S73.
Note 4 p 30: Note 6 p S73
Note 3. p. 104; Note 6 p S64.
Note 3 p 25- Note 4 p 3Q- Note 6 p
S51.
Note 3 p 94* Note 6 p S60
Note 3 p 7' Note 4 p 30
Note 3 p 94* Note 6 p S60
Note 3 p 7
Note 3 p 104* Note 6 p S64
Note 3 p 104- Note 6 p S64
Note 3. p. 104; Note 6 p S64.
Note 3 p 25* Note 4 p 30
Note 3 p 25
Note 3 p 7
Note 3. p. 83; Note 6. p. S68.
Note 3. p. 83; Note 6, p. S68
Note 3. p. 83; Note 6, p. 566.
Note 3. p. 104; Note 6, p. S64.
Note 3, p. 94; Note 6 p S60
Note 3 p 83* Note 6 p S66
Note 3. p. 104; Note 6. p. S64.
Note 3. p. 83; Note 6. p. S68
Note 3 p 7
Note 3 p. 104; Note 6 p S64.
Note 3 p 11 5; Note 4 p 35
Note 3 p 1 1 5
Note 3 p 83* Note 6 p S66
Note 3. p. 7; Note 4. p. 30.
Note 3 p 7
Table ID Notts
1 Pesticides are listed in this table by common name for the convenience of the reader. Additional pesticides may be found under Table 1C, where entries are listed by chemical name.
' The full text of methods 608 and 625 are given at Appendix A. "Test Procedures for Analysis of Organic Pollutants." of this Part 136. The standardized test procedure to be used to
determine the method detection limit (MOL) for these test procedures 19 given at Appendix B. "Definition and Procedure for the Determination of the Method Detection Limit", of this Part 136.
3 "Methods for Benzidine. Chlorinated Organic Compounds, Pentachlorophenol and Pesticides in Water and Wastewater," U.S. Environmental Protection Agency. September. 1978. This
EPA publication includes thin-layer chromatography (TLC) methods.
4 "Methods for Analysis of Organic Substances in Water." U.S. Geological Survey. Techniques of Water-Resources Investigations. Book 5. Chapter A3 (1972).
9 The method may be extended to include a-BHC. 6-BHC. endosulfan I, endosulfan II. and endnn. However, when they are known to exist. Method 608 is the preferred method.
• "Selected Analytical Methods Approved and Cited by the United States Environmental Protection Agency." Supplement to the Fifteenth Edition of Standard Methods tor the Examination
of Water and Wastewater (1981).
' Each analyst must make an initial, one-time, demonstration of their ability to generate acceptable precision and accuracy with Methods 608 and 625 (See Appendix A of this Part 136) in
accordance with procedures given in Section 8.2 of each of these methods. Additionally, each laboratory, on an on-going basis, must spike and analyze 10% of all samples analyzed with
-------
26 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 60B of 5S ol •» samples analyzed with Method 62S to monitor and evaluate laboratoiY data quaMy m accordance with Sections 8.3 and 8.4 ol these method!. When the recovery ol
any parameter fan* outside the warning units, the analytical remits lor that parameter m the unspfced sample are suspect and cannot be reported to demonstrate regulatory compliance
Not*.—These warning limits are promulgated as an 'interim final action with a request for comments."
TABLE IE.—LIST OF APPROVED RADIOLOGICAL TEST PROCEDURES
: , Reference (method No or page)
1
2
3
4
5
Alpha-Total, ff' pe< liter
Alpha-Counting error, p'
Aipna-Counnng error. D'
Beta-Counting error, p"
lal Radium- Total, p" p*
(b) •'•'*«. ff-' per liter ..
Parameter and units
1 per liter
1 per liter
Methods
Proportional or scintillation counter
Proportional or scintillation counter
Proportional counter
Proportional counter
Scintillation counter
EPA i
900.0
App0ndtx B .
900.0
'. Appendix B...
9030
: 903.1
Standard
Methods
15th Ed.
.: 703
703
703
703
705
706
ASTM
01943-6*
D 1943-66
D 1890-66
01890-66
D2460-70
03454-79
: PP
' P
PP
p.
p.
USGS'
. 75 and 78 '
79.
. 75 and 78. '
79
81
Table IE Notes
' Prescribed Procedures lor Measurement ol Radoactnnty in Drinking Water. EPA-600/4-80-032 (1980 update), US Environmental Protection Agency. August 1980
- Fishman. MJ and Brown. Eugene. "Selected Methods of the US. Geological Survey ot Analysis of Wastewaters." US. Geological Survey. Open-File Report 76-177 (1976)
' The method found on p 75 measures only the dissolved portion while the method on p 78 measures only the suspended portion Therefore, the two results must be added to obtain me
5. In § 136.3. paragraph (a) is revised
to show that the full text of approved
test procedures have been incorporated
by reference, into the regulation to read
as follows:
§ 136.3 Identification of test procedures.
(a) Parameters or pollutants, for which
methods are approved, are listed
together with test procedure
descriptions and references in Tables
IA. IB. 1C. ID. and IE. The full text of the
referenced test procedures are
incorporated by reference into Tables
IA. IB. 1C, ID, and IE. The references and
the sources from which they are
available are given in paragraph (b) of
this section. These test procedures are
incorporated as they exist on the day of
approval and a notice of any change in
these test procedures will be published
in the Federal Register. The discharge
parameter values for which reports are
required must be determined by one of
the standard analytical test procedures
incorporated by reference and described
in Tables IA. IB. 1C. ID. and IE. or by
any alternate test procedure which has
been approved by the Administrator
under the provisions of paragraph (d) of
this section and sections 136.4 and 136.5
of this Part 136. Under certain
circumstances (§§ 136.3 (b) or (c) or 40
CFR Part 401.13) other test procedures
may be used that may be more
advantageous when such other test
procedures have been previously
approved by the Regional Administrator
of the Region in which the discharge will
occur, and providing the Director of the
State in which such discharge will occur
does not object to the use of such
alternate test procedure.
6. In § 136.3. paragraphs (b) and (c)
are redesignated as (c) and (d) and a
new paragraph (b) is added to itemize
the references which are "incorporated
by reference" and to identify the sources
from which they may be obtained. As
added, the new paragraph (b) reads as
follows:
§ 136.3 Identification of test procedures.
(b) The full texts of the methods from
the following references which are cited
in Tables IA, IB. 1C. ID. and IE are
incorporated by reference into this |
regulation and may be obtained from the
sources identified. All costs cited are
subject to change and must be verified
from the indicated sources.
REFERENCES, SOURCES. AND COSTS
Table
Parameters
Reference, source and cost
IA—EPA .
IA—Standard Methods
IB-Standard
ID—Standard Methods
IE—Standard Methods
18—Standard Methods
IB—Other (Standard Methods Supplement)
1C—Other (Standard Methods Supplement)
ID—Other (Standard Methods Supplement)
IA—U S Geological Survey (USGS)
IB—EPA
IB—ASTM
ID—ASTM
IE-ASTM
•• 1-5
1-10. 12-46. 50-75
1. 8, 11. 12, 15. 17-20. 26, 28. 32. 33, 35. 40. 41.
'• 44. 46. 48. 52-54, 64, 66. 67. 69. 70
1-5
. 48
11.47
13. 56
2-7, 13. 14 16. 21-23. 25. 29-31 37. 38. 39. 41-
< 45. 47. 49. 50, 51, 56-63. 65. 68
1. 3, 5
[ 1-13, 15-48. 50-75
1. 2. 4. 6. 8. 11-13, 15-17. 19. 20, 22-25. 27. 28.
, 30-35. 37-40. 42-44. 46. 48. 50. 52, 60. 61, 63-
I 65. 67, 66. 73-75
i 1. 8-11. 15. 18-20. 27. 32, 33, 35. 40. 41, 46. 55.
I 69
! 1-5
"Microbiological Methods for Monitonng the Environment. Water and wastes."
United States Environmental Protection Agency. EPA-600/8-78-017 1978
Available from: ORD Publications. CERI. US Environmental Protection Agency.
Cincinnati, Ohio 45268.
Standard Utttmto lor trtt ExtmntUon of Wtltr tnd WttUwtttr. Joint Editonal
Board. American Public Health Association. American Water Works Association.
and Water PoHutjon Control Federation 15th Edition. 1981. Available from
American Public Health Association. 1015 Fifteenth Street. N.W.. Washington.
DC 20036. Cost: SSO.OO including the Supplement to the Fifteenth Edition
Ibid, 14th Edition.
"Selected Analytical Methods approved and Cited by the United States Environ-
mental Protection Agency," Supplement to the 15th Edition ot Sttndtrd Mftfi-
ods tor tht extmntUon ol Wtltr tna Wttttwtttr (1961). Available from:
American Public Health Association. 1015 Fifteenth Street. NW. Washington.
DC 20036 Cost Included with the 15th Edrtnn ot SunatrO Methods lor tnt
eiainrmton ol Wtttr tna Wtsttwtttr
"Methods for Collection and Analysis of Aquatic Bioiog«al and Microbiologwal
Samples." edited by PE. Greeson. T A. Ehlke, G A Irwm. B.W. bum. and K V
Slack: U.S. Geological Survey. Techniques of Water-Resources Investigation
(USGS TWRI). Book 5. Chapter A4 (1977) Revised edition. 332 pages.
Available from: U.S. Geological Survey. Branch ol Distribution. 1200 South Eads
Street Arlington. VA 22202. (Authorized agent ot the Superintendent of Docu-
ments. Government Printing Office.) Cost: S9.2S. Prices are subject to change
"Methods lor Chemical Analysis ot Water and Wastes". EPA-600/4-79-020
United States Ermronrnental Protection Agency. March. 1979. Available from
ORD Publications. CERI. U.S. Environmental Protection Agency Cincinnati. Ohio
45268
"Annual Book ol Standards. Part 31. Water", American Society lor Testing and
Materials. I960. Available from: American Society for Testing and Mateneis.
1918 Race Street Philadelphia. PA 19103. Con available from publisher
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 27
REFERENCES, SOURCES, AND COSTS—Continued
Table
IB USGS
18 Other (ANSI)
IB — Other
IB— Other
IB Other
IB — Other
IS other (USGS)
IB— Other (USGS)
ID-Other (USGS)
1C— EPA
ID— EPA
1C— Other (EPA)
IE EPA
IE USGS ••
2. 3. 4. 6-13. 15. 1
44. 46. 50. 52-55
2 4 9 12 15 16
52. 62-65. 75.
9 12 1 5. 20. 22 2
3 5-8 10 12 13
52. 60-63, 70. 74
21
4
15
15
17
22
28
30
34
40
75
49
69 ....
42
1. 11. 14. 17-20. 2
46. 52. 66. 69
1-12. 14-55. 57-97
1, 8-11. 15. 18.
7. 13. 22. 24, 27.
1-8 11-13. 15-24,
56-70
1-5
1-5
Parameters
6. 18-23. 25. 27. 28. 30-40. 43.
57.60-66. 71. 73. 75.
19 22 30-35 38. 42-44 46 50
3 38 62 75
9 20 22 27 30 32-34 36 37
. 75.
3, 25. 28. 29. 35, 37. 40-42. 44,
19. 20, 28, 32-36. 40. 41. 69
56. 66, 76-83. 88. 89. 91. 93
26. 28. 30-33. 35. 38-41. 43-54
Reference, source and cost
"Methods (or determination of inorganic substances in water and fluvial sedi-
ments." N.W. Skougstad and others, editors: USGS— TWHI Book 5, Chapter A1.
1979. $10.00. Available from: U.S. Geological Survey. Branch of Oistnbution.
1200 South Eads Street. Arlington. VA 22202. (Authorized agent of the Super-
intendent of Documents, Government Pnnting Office). Pnces are subiect to
change.
methods manual. 13th Edition (1980). Price.- $78.00. Available from: The
Association of Official Analytical Chemists. 1111 N. 19th St.. Suite 210.
Arlington, VA 22209.
1975. Available from: American National Standards Institute. 1430 Broadway.
New York. New York 10018.
procedure. Method 200.7. is pnnted in Appendix C of this Part 136.
"An Investigation of Improved Procedures for Measurement of Mill Effluent and
Receiving Water Color." NCASI Technical Bulletin No. 253. OecemOer. 1971.
Available from: National Council of the Paper Industry for Air and Stream
Improvements. Inc., 260 Madison Avenue. Cost available from publisher.
Ammonia. Automated Electrode Method. Industnal Method Number 379-75WE
dated February 19. 1976. Technicon AutoAnalyzer n. Method and price available
from Technicon Industrial Systems. Tarrytown. New York 10591.
1979. Method and price available from Hacn Chemical Company. P.O. Box 389.
Loveland, Colorado 80537.
QIC Chemical Oxygen Demand Method. Method and pnce available from Ocean-
ography International Corporation. 512 West Loop. P.O. Box 2980. College
Station, Texas 77840.
ORION Research Instruction Manual. Residual Chlorine Electrode Model 97-70
1977. Method and price available from Orion Research Incorporated, 840
Memorial Drive. Cambridge, Massachusetts 02138.
Analysis, 1979. Method and pnce available from Hacn Chemical Company, P.O.
Box 389. Loveland. Colorado 80537.
75WA, October 1976, Technicon AutoAnalyzer II. Method and Price available
from Technicon Industnal Systems. Tarrytown. New York 10591.
available from Hacn Chemical Company, P.O. Box 389. Loveland. Colorado
80537.
Periodate Oxidation Method for Manganese. Method 8034, Hach Handbook for
Water Analysis, 1979. Method and Price available from Hacn Chemical Compa-
ny. P.O. Box 389. Loveland. Colorado 80537.
Nitrite Nitrogen, Hach Method 8507. Method and pnce available from Hach
Chemical Company, P.O. Box 389. Loveland. Colorado 80537.
Zincon Method tor Zinc Method 8009 Hach Handbook for Water Analysis 1979
Method and price available from Hach Chemical Company, P.O. Box 389.
Loveland. Colorado 80537.
by R.F. Addison and R.G. Ackman. Journal of Chromatography. Volume 47. No.
3. pp. 421-426, 1970. Available in most public libraries. Back volumes of the
Journal of Chromatography are available from Elsevter/ North-Holland. Inc..
Journal Information Centre. 52 Vanderbilt Avenue, New York. NY 10164. Cost
available from publisher.
by H.H. Stevens, Jr., J. Ficke. and G.F. Smoot: USGS-TWfll Book 1. Chapter
D1. 1975. 65 pages, $1.60. Available from: U.S. Geological Survey, Branch of
Distribution, 1200 South Eads Street. Arlington. VA 22202. Prices are subject lo
change.
"Methods for analysis of organic substances in water." by 0. F. Goetlitz and
Eugene Brown: USGS-TWRI. Book 5. Chapter A3. 1972. 40 pages. $.90.
Available from: U.S. Geological Survey. Branch of Distribution. 1200 South Eads
Street Arlington. VA 22202. Prices are subject to change.
The full texts of Methods 601-613. 624. 625. 1624. and 1625 are printed in
appendix A of this Pan 136. The full text for determining the method detection
limit when using the test procedures is given in Appendix B of this Part 136.
"Methods for Benzidine. Chlorinated Organic Compounds. Pentachloropnenol and
Laboratory. United States Environmental Protection Agency. Cincinnati, Ohio
1978. Available from: ORD Publications. CERI. U.S. Environmental Protection
Agency, Cincinnati. Ohio 45268.
"Prescnbed Procedures for Measurement of Radioactivity in Drinking Water "
EPA-600/4-80-032 (1980 Update), United States Environmental Protection
Agency. 1980. Available from: ORD Publications. CERI. U.S. Environmental
Protection Agency. Cincinnati. Ohio 45268.
"Selected Methods of the U.S. Geological Survey of Analysis of Wastewaters " by
M.J. Fishman and Eugene Brown; U.S. Geological Survey Open File Report 76-
77 (1976). $13.50. Available from: U.S. Geological Survey, Branch Distribution.
1200 South Eads Street. Arlington. VA 22202.
The full texts of all the test procedures 7. In section 136.3 a new paragraph (e) § 136.3 Identification of test procedures.
cited are available for inspection at the
Office of the Federal Register
Information Center, Room 8301,1110 L
Street, N.W., Washington, D.C. 20408.
is added together with a new Table II
entitled, "Table II, Required Containers,
Preservation Techniques, and Holding
Times," to read as follows:
(e) Sample preservation procedures,
container materials, and maximum
allowable holding times for parameters
cited in Tables IA, IB. 1C, ID, and IE are
-------
28
Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations
prescribed in Table II. Any person may
apply for a variance from the prescribed
preservation techniques, container
materials, and maximum holding times
applicable to samples taken from a
specific discharge. Applications for
variances may be made by letters to the
Regional Administrator in the Region in
which the discharge will occur.
Sufficient data should be provided to
assure such variance does not adversely
affect the integrity of the sample. Such
data will be forwarded by the Regional
Administrator to the Director of the
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio for
technical review and recommendations
for action on the variance application.
Upon receipt of the recommendations
from the Director of the Environmental
Monitoring and Support Laboratory, the
Regional Administrator may grant a
variance applicable to the specific
discharge to the applicant. A decision tc
approve or deny a variance will be
made within 90 days of receipt of the
application by the Regional
Administrator.
TABLE II.—REQUIRED CONTAINERS, PRESERVATION TECHNIQUES, AND HOLDING TIMES
Parameter No /name
Container'
Preservation«
Maximum holding lime *
Table IA—Bacterial Tests:
1-4. Coliiorm, fecal and total : P. G
5. Fsca) streptococci ! P. G
Table IB—Inorganic Tests: :
1. Acidify , P G
2. Alkalinity P. G
4. Ammonia i p. G
9. Biochemical oxygen demand I P. G
11. Bromide i P. G
14. Biochemical oxygen demand, carbonaceous ! P, G
15. Chemical oxygen demand I P. G
16. Chloride I P. G
17. Chlorine, total residual : P. G
21. Color i P. G
23-24. Cyanide, total and amenable to chkxination j P, G
25. Fluoride I P
27. Hardness ! P. G
28. Hydrogen on (pH) \ P. G
31. 43. Kfektahl and organic nitrogen ; P. G
Metals:'
18. Chromium VI i P. G
35. Mercury , P. G
3. 5-«. 10. 12. 13. 19. 20. 22. 26. 29. 30. 32-34. 38. 37. 45, 47. 51, 52, 58- I P, G
60. 82. 63. 70-72. 74, 75. Metals, except chromium VI and mercury
38. Nitrate P. G
39. Nrtrate-nitnte , P. G
40. Nitrite i P. G
41. Oil and grease i G
42. Organic carbon P. G
44. Orthophoephate : P. G
46. Oxygen. Dissolved Probe G Bottle and top..
47. Winner i do
48. Phenols I G only
49. Phosphorus (elemental) I G
50. Phosphorus, total P. G
53. Residue, total P. G
54. Residue. Filterable : P, G
55. Residue. Nonfilterable (TSS) P. G
56. Residue. Samaeble P. G
57. Residue, volatile , P. G
61. SHca P
64. Specific conductance , P, G
65. Sultate P. G
66. SulMe f, G
Cool. 4'C, 0008% Nai&Cs'
. ...do
Cool. 4-C
do
Cool, 4-C. H,SO, to pH<2
Cool, 4'C
None required
Cool. 4-C
Cool. 4'C. H,SO. to pH<2
None required
do
Cool. 4-C
Cool. 4-C. NaOH to pH> 12. 0 6g ascorbic acid '
: None required
HNCs to pH<2. H,SO. to pH<2
None required
Cool. 4'C. H,SO. to pH<2
Cool. 4-C
HNO, 10 pH<2...
do
Cool, 4-c
Cool. 4-C. HiSO. topH<2
Cool, 4-C
Cool. 4'C. H,SO. to pH<2
Cool. 4-C. HCI or H,SO. to pH<2
Filter immediately. Cool, 4'C
None required
Fix on site and store in dark
Cool. 4'C. H,SOi to pH<2
Cool. 4'C
Cool. 4-C. H,SO. to pH<2
Cool, 4-C
do
do
do
do
..do..
.do.
..do..
67. SurMe • P. G
68. Surfactants P .G
69. Temperature P, G
73. Turbidity P. G
Table 1C—Organic Tests.'
13. 18-20. 22. 24-28. 34-37. 39-43. 45-47. 56. 66, 88. 89. 92-95. 97 ' G. Telflon-lined septum .
PurgeabM Hatocarbons.
6. 57. 90. PurgeeMe aromatic hydrocarbons do
3. 4. AcreMm and acryloratnle *>
23. 30. 44, 49. 53. 67, 70. 71. 83. 85. 96. Phenols " G. Teflon-lined cap
7. 38. BenzKlmes"
u. 17. 48. 50-52. Phfhalate esters"
..do..
...do..
72-74 Nitrosammes"•" do....
76-82. PCBs" acrytonitnle do....
54, 55. 65. 69. Nitroaromatics and isopnorone" do....
1. 2. 5. 8-12, 32. 33. 58. 59. 64. 68. 84. 66 Polynuclear aromatic do
hydrocarbons.''
15, 16. 21, 31. 75. Haloethers" , do....
29. 35-37. 60-63. 91. Chlonnated hydrocarbons '' do ...
87. TCDD '' 00 -
Table to—Pesticides Tests:
1-70. Pesticides" do....
Table IE—Radmogwal Tests:
1-5. Aloha, beta and radnjm P, G
Cool. 4'C add zinc acetate plus sodium hydroxide to
PH>9.
None required [[[
Cool. 4-C [[[
None required [[[
Cool, 4-C [[[
Cool. 4'C. 0008% Na,S,0,.» .........................................
Cool. 4-C. 0.008% NaAO,5. HC1 to pH2«
Cool. 4-C. 0 008% Na,S>Oi'; Adjust PH 10 4-5 >
Cool. 4'C. 0.008% Na,SiO,s
do
Cool, 4-C
Cool. 4'C. store in dark. 0.008% Na,S,O, '•
Cool. 4-C
Cod. 4-C. 0008% NaiSXV store m dark
. ..Jo
Cool. 4-C. 0.008% Na,S,(V
Cool. 4-C
Coot. 4-C, 0008% Na,S.O,>
Cool. 4'C. pH 5-9"
6 hours
Do
14 days.
Do.
28 days.
48 hours
. 28 days
48 hours
28 days.
Do
Analyze immediately
. 48 hours
14 days.'
28 days.
6 months.
Analyze immediately
28 days.
24 hours
. 28 days.
6 months.
. 48 hours
28 days.
48 hours.
28 days
Do
48 hours
Analyze immediately
8 hours.
28 days.
48 hours.
28 days.
7 days.
48 hours.
7 days.
48 hours.
7 days.
28 days
Do.
Do.
7 days.
Analyze immediately
48 hours.
Analyze
48 hours
14 days.
Do
Do
7 days until extraction.
40 days after
extraction.
7 days until extraction '
7 days until extraction:
40 days after
extraction
Do.
Do.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
29
'Sample preservation should be performed immediately upon sample collection. For composite chemical samples each aliquot should be preserved at the time of collection. When use ol
an automated sampler makes it impossible to preserve each aliquot then chemical samples may be preserved by maintaining at 4'C until compositing and sample splitting is completed.
'When any sample is to be shipped by common carrier or sent through the United States Mails, it must comply with the Department ol Transportation Hazardous Materials Regulations (49
CFR Part 172). The person ottering such material tor transportation is responsible lor ensuring such compliance. For the preservation requirements of Table II. the Office ol Hazardous
Materials. Materials Transportation Bureau. Department of Transportation has determined that the Hazardous Materials Regulations do not apply to the following materials: Hydrochloric aod
(HO) in water solutions at concentrations of 0.04% by weight or less (pH about 1.96 or greater): Nitric acid (HNCM in water solutions at concentrations of 0.15% by weight or less (pH about
1.62 or greater); Sulfuhc acid (H,SO.) in water solutions at concentrations of 0.35% by weight or less (pH about 1.15 or greater); and Sodium hydronde (NaOH) in water solutions at
concentrations of 0.080% by weight or less (pH about 12.30 or less).
•Samples should be analyzed as soon as possible after collection. The times listed are the maximum times that samples may be held before analysis and still be considered valid. Samples
may be held for longer periods only it the permittee, or monnonng laboratory, has data on file to show that the specific types of samples under study are stable tor the longer time, and has
received a variance from the Regional Administrator under § 136 3(e) Some samples may not be stable for the maximum time period given in the table. A permittee, or monitoring laboratory, is
obligated to hold the sample for a shorter time if knowledge exists to show mat this is necessary to maintain sample stability. See § 138.3(e) tor details.
'Should only be used in the presence of residual chlorine.
•Maximum holding time a 24 hours when sulfide is present. Optionally all samples may be tested with lead acetate paper before pH adjustments in order to determine if sufflde is present.
It sulfide is present, it can be removed by the addition of cadmium nitrate powder until a negative spot test is obtained. The sample is filtered and then NaOH is added to pH 12.
'Samples should be filtered immediately on-ste before adding preservative tor dissolved metals.
'Guidance applies to samples to be analyzed by GC. 1C, or GC/MS for specific compounds.
•Sample receiving no pH adjustment must be analyzed within seven days of sampling.
"The pH adjustment is not required if acrolein will not be measured. Samples for acrolem receiving no pH adjustment must be analyzed within 3 days of sampling.
"When the extractable analytes ot concern fall within a single chemical category, the specified preservative and maximum nolding times should be observed tor optimum safeguard of
sample integrity. When the analytes ol concern tall within two or more chemical categories, the sample may be preserved by cooling to 4'C. reducing residual chlorine with 0.008% sodium
thiosulfate. stonng in the dart, and adjusting the pH to 6-9; samples preserved in this manner may be held for seven days before extraction and tor forty days after extraction. Exceptions to
this optional preservation and holding time procedure are noted in footnote 5 (re the requirement for thiosultate reduction ot residual chlorine), and footnotes 12, 13 (re the analysis ot
benzidine).
"If 1.2-diphenylhydrazine is likely to be present, adjust the pH of the sample to 4.0±0.2 to prevent rearrangement to benzidine.
"Extracts may be stored up to 7 days before analysis if storage is conducted under an inert (oxidant-free) atmosphere.
"For the analysis of diphenylnrlrosamine, add 0.008% Na,S,Oi and adjust pH to 7-10 with NaOH within 24 hours of sampling.
'•The pH adjustment may be performed upon receipt at the laboratory and may be omitted if the samples are extracted within 72 hours of collection. For the analysis ot aldrin. add 0.008%
Na.S.0,.
8. Appendices A. B, and C are added to
Part 136 to read as follows:
APPENDIX A TO PART 136—METHODS
FOR ORGANIC CHEMICAL ANALYSIS OF
MUNICIPAL AND INDUSTRIAL
WASTEWATER.
Method 601—Purgeable Halocarbons
1. Scope and Application
1.1 This method covers the determination
of 29 purgeable halocarbons.
The following parameters may be
determined by this method:
Parameter
Carbon tetrachloride
Chloroethane
2-Chloroethytvinyl ether
1 2-Dichlorobenzene
1.3-Oichlorobenzene
1 2-Dichk>roethane ..
1 1 -Dichtoroethane
trans-1 2-Dichloroethene
Memylene chloride
112 2-Tetrachtoroetnane
1 1 1-Trichloroethane
1 1 2-Trich!oroethane
Vinyt chloride
STORET
No.
32101
32104
34413
32102
34301
34311
34576
32106
34418
32105
34536
34566
34571
34668
34496
34531
34501
34546
34541
34704
34699
34423
34516
34475
34506
34511
39180
34488
39715
CAS No.
75-27-4
75-25-2
74-83-9
56-23-5
108-90-7
75-00-3
100-75-8
67-66-3
74-87-3
124-48-1
95-50-1
541-73-1
106-46-7
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
10061-01-5
10061-02-6
75-09-2
79-34-5
127-18-4
71-55-6
79-00-5
79-01-6
75-69-4
75-O1 -4
1.2 This is a purge and trap gas
chromatographic (GC) method applicable to
the determination of the compounds listed
above in municipal and industrial discharges
as provided under 40 CFR 136.1. When this
method is used to analyze unfamiliar samples
for any or all of the compounds above,
compound identifications should be
supported by at least one additional
qualitative technique. This method describes
analytical conditions for a second gas
chromatographic column that can be used to
confirm measurements made with the
primary column. Method 624 provides gas
chromatograph/mass spectrometer (GC/MS)
conditions appropriate for the qualitative and
quantitative confirmation of results for most
of the parameters listed above.
1.3 The method detection limit (MDL.
defined in Section 12.1) ' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed.
depending upon-the nature of interfer°r.ces in
the sample matrix.
1.4 Any modification of this metric.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the operation of a purge and
trap system and a gas chromatograph and in
the interpretation of gas chromatograms.
Each analyst must demonstrate the ability to
generate acceptable results with this method
using the procedure described in Section 8.2.
2. Summary of Method
2.1 An inert gas is bubbled through a 5-
mL water sample contained in a specially-
designed purging chamber at ambient
temperature. The halocarbons are efficiently
transferred from the aqueous phase to the
vapor phase. The vapor is swept through a
sorbent trap where the halocarbons are
trapped. After purging is completed, the trap
is heated and backflushed with the inert gas
to desorb the halocarbons onto a gas
chromatographic column. The gas
chromatograph is temperature programmed to
separate the halocarbons which are then
detected with a halide-specific detector.a-3
2.2 The method provides an optional gas
chromatographic column that may be helpful
in resolving the compounds of interest from
interferences that may occur.
3. Interferences
3.1 Impurities in the purge gas and
organic compounds outgassing 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
laboratory reagent blanks as described in
Section 8.1.3. The use of non-Teflon plastic
tubing, non-Teflon thread sealants, or flow
controllers with rubber components in the
purge and trap system should be avoided.
3.2 Samples can be contaminated by
diffusion of volatile organics (particularly
fluorocarbons and methylene chloride)
through the septum seal into the sample
during shipment and storage. A field reagent
blank prepared from reagent water and
carried through the sampling and handling
protocol can serve as a check on such
contamination.
3.3 Contamination by carry-over can
occur whenever high level and low level
samples are sequentially analyzed. To reduce
carry-over, the purging device and sample
syringe must be rinsed with reagent water
between sample analyses. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of reagent 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
detergent solution, rinse it with distilled
• water, and then dry it in a 105"C oven
between analyses. The trap and other parts
of the system are also subject to
contamination; therefore, frequent bakeout
and purging of the entire system may be
required.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined: however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified "for the information of the
analyst.
-------
30
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: carbon tetrachloride,
chloroform, 1.4-dichlorobenzene. and vinyl
chloride. Primary standards of these toxic
compounds should be prepared in a hood. A
NIOSH/MESA approved toxic gas respirator
should be worn when the analyst handles
high concentrations of these toxic compounds
5. Apparatus and Materials
5.1 Sampling equipment, for discrete
sampling.
5.1.1 Vial—25-mL capacity or larger.
equipped with a screw cap with a hole in the
center (Pierce «13075 or equivalent).
Detergent wash, rinse with tap and distilled
water, and dry at 105 *C before use.
5.1.2 Septum—Teflon-faced silicone
(Pierce "12722 or equivalent). Detergent
wash, rinse with tap and distilled water, and
dry at 105 'C for 1 h before use.
5.2 Purge and trap system—The purge and
trap system consists of three separate pieces
of equipment: a purging device, trap, and
desorber. Several complete systems are now
commercially available.
5.2.1 The purging device must be designed
to accept 5-mL samples with a water column
at least 3 cm deep. The gaseous head space
between the water column and the trap must
have a total volume of less than 15 mL. The
purge gas must pass through the water
column as finely divided bubbles with a
diameter of less than 3 mm at the origin. The
purge gas must be introduced no more than 5
mm from the base of the water column. The
purging device illustrated in Figure 1 meets
these design criteria.
5.2.2 The trap must be at least 25 cm long
and have an inside diameter of at least 0.105
in. The trap must be packed to contain the
following minimum lengths of adsorbents: 1.0
cm of methyl silicone coated packing (Section
6.3.3), 7.7 cm of 2.6-diphenylene oxide
polymer (Section 6.3.2), 7.7 cm of silica gel
(Section 6.3.4). 7.7 cm of coconut charcoal
(Section 6.3.1). If it is not necessary to
analyze for dichlorodifluoromethane. the
charcoal can be eliminated, and the polymer
section lengthened to 15 cm. The minimum
specifications for the trap are illustrated in
Figure 2.
5.2.3 The desorber must be capable of
rapidly heating the trap to 180 *C. The
polymer section of the trap should not be
heated higher than 180 *C and the remaining
sections should not exceed 200 *C. The
desorber illustrated in Figure 2 meets these
design criteria.
5.2.4 The purge and trap system may be
assembled as a separate unit or be coupled to
a gas chromatograph as illustrated in Figures
3 and 4.
5.3 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
columns, gases, detector, and strip-chart
recorder. A data system is recommended for
measuring peak areas.
5.3.1 Column 1—8 ft long x 0.1 in. ID
stainless steel or glass, packed with 1% SP-
1000 on Carbopack B (60/80 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 12. Guidelines for the use of alternate
column packings are provided in Section 10.1.
5.3.2 Column 2—6 ft long x 0.1 in. ID
stainless steel or glass, packed with
chemically bonded n-octane on Porasil-C
(100/120 mesh) or equivalent.
5.3.3 Detector—Electrolytic conductivity
or microcoulometric detector. These types of
detectors have proven effective in the
analysis of wastewaters for the parameters
listed in the scope (Section 1.1). The
electrolytic conductivity detector was used to
develop the method performance statements
in Section 12. Guidelines for the use of
alternate detectors are provided in Section
10.1.
5.4 Syringes—5-mL glass hypodermic with
Luerlok tip (two each), if applicable to the
purging device.
5.5 Micro syringes—25-fiL, 0.006 in. ID
needle.
5.6 Syringe valve—2-way, with Luer ends
(three each).
5.7 Syringe—5-mL, gas-tight with shut-off
valve.
5.8 Bottle—15-mL. screw-cap, with Teflon
cap liner.
5.9 Balance—Analytical, capable of
accurately weighing 0.0001 g.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.1.1 Reagent water can ge generated by
passing tap water through a carbon filter bed
containing about 1 Ib of activated carbon
(Filtrasorb-300. Calgon Corp.. or equivalent).
6.1.2 A water purification system
(Millipore Super-Q or equivalent) may be
used to generate reagent water.
6.1.3 Reagent water may also be prepared
by boiling water for 15 min. Subsequently,
while maintaining the temperature at 90 'C.
bubble a contaminant-free inert gas through
the water for 1 h. While still hot, transfer the
water to a narrow mouth screw-cap bottle
and seal with a Teflon-lined septum and cap.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Trap Materials:
6.3.1 Coconut charcoal—6/10 mesh sieved
to 26 mesh. Barnebey Cheney, CA-580-26 lot
# M-2649 or equivalent.
6.3.2 2.6-Diphenylene oxide polymer—
Tenax. (60/80 mesh), chromatographic grade
or equivalent.
6.3.3 Methyl silicone packing—3% OV-1
on Chromosorb-W (60/80 mesh) or
equivalent.
6.3.4 Silica gel—35/60 mesh, Davison.
grade-15 or equivalent.
6.4 Methanol—Pesticide quality or
equivalent.
6.5 Stock standard solutions—Stock
standard solutions may be prepared from
pure standard materials or purchased as
certified solutions. Prepare stock standard
solutions in methanol using assayed liquids
or gases as appropriate. 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.
6.5.1 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 min or until all alcohol wetted
surfaces have dried. Weigh the flask to the
nearest 0.1 mg.
6.5.2 Add the assayed reference material:
6.5.2.1 Liquid—Using a 100 pL syringe.
immediately add two or more 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.
6.5.2.2 Gases—To prepare standards for
any of the six halocarbons that boil below 30
' C (bromomethane. chloroethane,
chloromethane. dichlorodifluoromethane.
trichlorofluoromethane, vinyl chloride), fill a
5-mL valved gas-tight syringe with the
reference standard to the 5.0-mL mark. Lower
the needle to 5 mm above the methanol
meniscus. Slowly introduce the reference
standard above the surface of the liquid (the
heavy gas will rapidly dissolve into the
methanol).
6.5.3 Reweigh. dilute to volume, stopper.
then mix by inverting the flask several times.
Calculate the concentration in fig/pL from
the net gain in weight. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.5.4 Transfer the stock standard solution,
into a Teflon-sealed screw-cap bottle. Store,
with minimal headspace, at -10 to -20 *C
and protect from light.
6.5.5 Prepare fresh standards weekly for
the six gases and 2-chloroethylvinyl ether. All
other standards must be replaced after one
month, or sooner if comparison with check
standards indicates a problem.
6.6 Secondary dilution standards—Using
stock standard solutions, prepare secondary
dilution standards in methanol that contain
the compounds of interest, either singly or
mixed together. The secondary dilution
standards should be prepared at
concentrations such that the aqueous
calibration standards prepared in Sections
7.3.1 or 7.4.1 will bracket the working range of
the analytical system. Secondary dilution
standards should be stored with minimal
headspace and should be checked frequently
for signs of degradation or evaporation,
especially just prior to preparing calibration
standards from them.
6.7 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Assemble a purge and trap system
that meets the specifications in Section 5.2.
Condition the trap overnight at 180 *C by
backflushing with an inert gas flow of at least
20 mL/min. Condition the trap for 10 min
once daily prior to use.
7.2 Connect the purge and trap system to
a gas chromatograph. The gas chromatograph,
must be operated using temperature and flow
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations 31
rale conditions equivalent to those given in
Table 1. Calibrate the purge and trap-gas
chromatographic system using either the
external standard technique (Section 7.3) or
the internal standard technique (Section 7.4).
7.3 External standard calibration
procedure:
7.3.1 Prepare calibration standards at a
miminum of three concentration levels for
each parameter by carefully adding 20.0 fit of
one or more secondary dilution standards to
100. 500, or 1000 mL of reagent water. A 25-^L
syringe with a 0.006 in. ID needle should be
used for this operation. One of the external
standards should be at a concentration near.
but above, the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector. These aqueous
standards can be stored up to 24 h, if held in
sealed vials with zero headspace as
described in Section 9.2. If not so stored, they
must be discarded after 1 h.
7.3.2 Analyze each calibration standard
according to Section 10, and tabulate peak
height or area responses versus the
concentration in the standard. The results
can be used to prepare a calibration curve for
each compound. Alternatively, if the ratio of
response to concentration (calibration factor)
is a constant over the working range (<10%
relative standard deviation, RSD), linearity
through the origin can be assumed and the
average ratio or calibration factor can be
used in place of a calibration curve.
7.4 Internal standard calibration
procedure — To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
The compounds recommended for use as
surrogate spikes in Section 8.7 have been
used successfully as internal standards.
because of their generally unique retention
times.
7.4.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest as described in
Section 7.3.1.
7.4.2 Prepare a spiking solution containing
each of the internal standards using the
procedures described in Sections 6.5 and 6.6.
It is recommended that the secondary
dilution standard be prepared at a
concentration of 15 pg/mL of each internal
standard compound. The addition of 10 pL of
this standard to 5.0 mL of sample or
calibration standard would be equivalent to
7.4.3 Analyze each calibration standard
according to Section 10. adding 10 fiL of
internal standard spiking solution directly to
the syringe (Section 10.4). Tabulate peak
height or area responses against
concentration for each compound and
internal standard, and calculate response
factors (RF) for each compound using
Equation 1.
. Equation 1.
RF =
where:
A,=Response for the parameter to be
measured.
A,, = Response for the internal standard.
C,, = Concentration of the internal
standard.
C,=Concentration of the parameter to be
measured.
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A,/AU, vs. RF.
7.5 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of a
QC check sample.
7.5.1 Prepare the QC check sample as
described in Section 8.2.2.
7.5.2 Analyze the QC check sample
according to Section 10.
7.5.3 For each parameter, compare the
response (Q) with the corresponding
calibration acceptance criteria found in Table
2. If the responses for all parameters of
interest fall within the designated ranges,
analysis of actual samples can begin. If any
individual Q falls outside the range, proceed
according to Section 7.5.4.
Note: The large number of parameters in
Table 2 present a substantial probability that
one or more will not meet the calibration
acceptance criteria when all parameters are
analyzed.
7.5.4 Repeat the test only for those
parameters that failed to meet the calibration
acceptance criteria. If the response for a
parameter does not fall within the range in
this second test, a new calibration curve,
calibration factor, or RF must be prepared for
that parameter according to Section 7,3 or 7.4.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in Section
10.1) to improve the separations or lower the
cost of measurements. Each time such a
modification is made to the method, the
analyst is required to repeat the procedure in
Section 8.2.
8.1.3 Each day. the analyst must analyze a
reagent water blank to demonstrate that
interferences from the analytical system are
under control.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of 10
fig/mL in methanol. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Prepare a QC check sample to
contain 20 /ig/L of each parameter by adding
200 pL of QC check sample concentrate to
100 mL of reagent water.
8.2.3 Analyze four 5-mL aliquots of the
well-mixed QC check sample according to
Section 10.
8.2.4 Calculate the average recovery (X)
in Mg/L, and the standard deviation of the
recovery (s) in ng/L, for each parameter of
interest using the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, then the system performance is
unacceptable for that parameter.
Note: The large number of parameters in
Table 2 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.8 When on?or more of the parameters
tested fail at least one of the acceptance
-------
32
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
criteria, the analyst mutt proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.0.1 Locate and correct the source of
the problem and repeat the test for all
parameter* of interest beginning with Section
8.2.3.
8.2.8.2 Beginning with Section 8.2.3, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2.3.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 20 ug/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.2 Analyze one 5-mL sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8J.1) appropriate for the background
concentrations in the sample. Spike a second
5-mL sample aliquot with 10 uL of the QC
check sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, when T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 20 pg/L. the analyst must use
either the QC acceptance criteria in Table 2,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter (1) Calculate
accuracy (X') using the equation in Table 3.
substituting the spike concentration (T) for C
(2) calculate overall precision (S'J using the
equation in Table 3, substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) ±2.44(100 S'l
T)*.'
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note: The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory. If the entire list of parameters in
Table 2 must be measured in the sample in
Section 8.3, the probability that the analysis
of a QC check standard will be required is
high. In this case the QC check standard
should be routinely analyzed with the spiked
sample.
8.4.1 Prepare the QC check standard by
adding 10 u.L of QC check sample concentrate
(Sections 8.2.1 or 8.3.2) to 5 mL of reagent
water. The QC check standard needs only to
contain the parameters that failed criteria in
the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P.)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (a,). Express the accuracy
assessment as a percent recovery interval
from P-2s, to P+2«^ If p=90% and s,«10%.
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.8 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram. confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
8.7 The analyst should monitor both the
performance of the analytical system and the
effectiveness of the method in dealing with
each sample matrix by spiking each sample,
standard, and reagent water blank with
surrogate halocarbons. A combination of
bromochloromethane, 2-bromo-l-
chloropropane. and 1.4-dichlorobutane is
recommended to encompass the range of the
temperature program used in this method.
From stock standard solutions prepared as in
Section 8.5, add a volume to give 750 fig of
each surrogate to 45 mL of reagent water
contained in a 50-mL volumetric flask, mix
and dilute to volume for a concentration of 15
ng/jiL Add 10 pL of this surrogate spiking
solution directly into the 5-mL syringe with
every sample and reference standard
analyzed. Prepare a fresh surrogate spiking
solution on a weekly basis. If the internal
standard calibration procedure is being used.
the surrogate compounds may be added
directly to the internal standard spiking
solution (Section 7.4.2).
9. Sample Collection. Preservation, and
Handling
9.1 All samples must be iced or
refrigerated from the time of collection until
analysis. If the sample contains free or
combined chlorine, add sodium thiosulfate
preservative (10 mg/40 mL is sufficient for up
to 5 ppm Ck) to the empty sample bottle just
prior to shipping to the sampling site. EPA
Methods 330.4 and 330.5 may be used for
measurement of residual chlorine.'Field test
kits are available for this purpose.
9.2 Grab samples must be collected in
glass containers having a total volume of at
least 25 mL Fill the sample bottle just to
overflowing 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. If preservative
has been added, shake vigorously for 1 min.
Maintain the hermetic seal on the sample
bottle until time of analysis.
9.3 All samples must be analyzed within
14 days of collection.'
10. Procedure
10.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
estimated retention times and MDL that can
be achieved under these conditions. An
example of the separations achieved by
Column 1 is shown in Figure 5. Other packed
columns, chromatographic conditions, or
detectors may be used if the requirements of
Section 8£ are met.
10.2 Calibrate the system daily as
described in Section 7.
10.3 Adjust the purge gas (nitrogen or
helium) flow rate to 40 mL/min. Attach the
trap inlet to the purging device, and set the
purge and trap system to purge (Figure 3).
Open the syringe valve located on the
purging device sample introduction needle.
10.4 Allow the sample to come to ambient
temperature prior to introducing it to the '
syringe. 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 to just short of overflowing. Replace
the syringe plunger and compress the sample.
Open the syringe valve and vent any residual
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations
33
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 10.0 fiL of the
surrogate spiking solution (Section 8.7) and
10.0 >iL of the internal standard spiking
solution (Section 7.4.2), if applicable, through
the valve bore, then close the valve.
10.5 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.
10.6 Close both valves and purge the
sample for 11.0±0.1 min at ambient
temperature.
10.7 After the 11-min purge time, attach
the trap to the chromatograph, adjust the
purge and trap system to the desorb mode
(Figure 4), and begin to temperature program
the gas chromatograph. Introduce the trapped
materials to the GC column by rapidly
heating the trap to 180 *C while backflushing
the trap with an inert gas between 20 and 60
mL/min for 4 min. If rapid heating of the trap
cannot be achieved, the GC column must be
used as a secondary trap by cooling it to
30 *C (subambient temperature, if poor peak
geometry or random retention time problems
persist) instead of the initial program
temperature of 45 *C
10.8 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 reagent water.
10.9 After desorbing the sample for 4 min,
recondition the trap by returning the purge
and trap system to the purge mode. Wait 15 s
then close the syringe valve on the purging
device to begin gas flow through the trap. The
trap temperature should be maintained at
160 *C After approximately 7 min, turn off the
trap heater and open the syringe valve to
stop the gas flow through the trap. When the
trap is cool, the next sample can be analyzed.
10.10 Identify the parameters in the
sample by comparing the retention times of
the peaks in the sample chromatogram with
those of the peaks in standard
chromatograms. The width of the retention
time window used to make identifications
should be based upon measurements of
actual retention time variations of standards
over the course of a day. Three times the
standard deviation of a retention time for a
compound can be used to calculate a
suggested window size: however, the
experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
10.11 If the response for a peak exceeds
the working range of the system, prepare a
dilution of the sample with reagent water
from the aliquot in the second syringe and
reanalyze.
11. Calculations
11.1 Determine the concentration of
individual compounds in the sample.
11.1.1 If the external standard calibration
procedure is used, calculate the
concentration of the parameter being
measured from the peak response using the
calibration curve or calibration factor
determined in Section 7.3.2.
11.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.4.3 and Equation 2
Equation 2.
(A.)(CU)
Concentration (ug/L) =
where:
A.=Response for the parameter to be
measured.
AU=Response for the internal standard.
Cu = Concentration of the internal
standard.
11.2 Report results in /ig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
12. Method Performance
12.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Table 1 were obtained using reagent water.'
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
12.2 This method is recommended for use
in the concentration range from the MDL to
1000 x MDL Direct aqueous injection
techniques should be used to measure
concentration levels above 1000 x MDL
12.3 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 8.0 to 500 pig/L* Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136. Appendix B.
2. Bellar. T.A., and Lichtenberg. J.|.
"Determining Volatile Organics at
Microgram-per-Litre-Levels by Gas
Chromatography," Journal of the American
Water Works Association. 66. 739 (1974).
3. Bellar, T.A., and Lichtenberg. J.J. "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds," Proceedings
from Symposium on Measurement of Organic
Pollutants in Water and Wastewater,
American Society for Testing and Materials,
STP 686, C.E. Van Hall, editor, 1978.
4. "Carcinogens—Working With
Carcinogens," Department of Health.
Education, and Welfare. Public Health
Service, Center for Disease Control, National
Institute for Occupational Safety and Health.
Publication No. 77-206, August 1977.
5. "OSHA Safety and Health Standards.
General Industry" (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2206 (Revised,
January 1976).
8. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication, Committee on Chemical Safety,
3rd Edition, 1979.
7. Provost, L.P., and Elder, R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15, 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
8. "Methods 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric, DPD) for
Chlorine, Total Residual," Methods for
Chemical Analysis of Water and Wastes.
EPA 600/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, March 1979.
9. "EPA Method Validation Study 23,
Method 601 (Purgeable Halocarbons)," Report
for EPA Contract 68-03-2856 (in preparation).
10. "Method Validation Data for EPA
Method 601," Memorandum from B. Potter.
U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory. Cincinnati. Ohio 45268.
November 10,1983.
TABLE 1.—CHROMATOQRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Ptfifftttar
CNoronnCWK . -• .-
vinyl cnterid» - • .. —
CNoRMIhttW
^r * PfcJitamOMiK
trim 1.3 DicfiluimUmii -
R6t0ndon time (min)
Column 1
1.50
^17
£82
2.67
3.33
5.25
7.18
7.93
».30
10.1
Column J
5.26
705
nd
5.28
8.68
10.1
nd
7.72
12.6
9.38
Method dgtacliun
fenit04/L)
0.06
1.16
1.81
0.18
0.52
0.25
nd
0.13
0.07
0.10
465-028 0-85-2
-------
34 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS—Continued
PwAVTMtor
CNorafcvm
1 2-OicMoro0tharTfl
1 1 1-ThcNoroothtM •
1 tJ-TrichtaroathaV*
2-CMoKMtttytvinyl ttfwr
1 i j jg-TtfrachrTjf otthant
Ttttachto'fttihfM'
1 3-Oichlofobtftttnt --- ,, , , — , -
l 2-Oichtefobtftff*» - .
HdsoUon
Column 1
107
11 4
12.6
130
13 7
14 (
15.2
159
16.5
165
16.5
16.0
1f.2
21.6
21 7
24.2
340
34.9
354
«m* 0.1 in. ID lUmlMi MM! of gjatt column wWi Datum camar gaa at 40 m
Column 1 cundMiona.
Column tamparalura haU at 45
Column 2 concMon* PorMC (100/120
tamparalurt Md«50'Clor3m1n*wn programmad (16 •C/mn to 170 'C »nd h«td tor 4 mm
~* - --
no^
•I 40 mL/iMn (tow rut
ID ttHMt MMl or glm column wWi htfun dnw 0M «t 40 mL/nwi Row ntt. Column
TABLE 2.—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 601 •
PmvMMr
RwgctarO
UnMtori
(M/L)
FtengtforX
(M/L)
nathani
Carton Mkachtortda..
Chtorobanzana
Chtoromathana..
152-244
14.7-25.3
11.7-26.3
13.7-26.3
14.4-25.9
15.4-24.6
110-26.0
15.0-25.0
11.9-26.1
13.1-264
14.0-28J)
64-M.1
13.9-26.1
164-2U
1.2-Otehto
124-17.4
144-CU
124-27.2
154-244
Mnyfchtarida
144-164
15.7-24J
15.4-24.6
1SJ-26.7
13.7-26J
4.3
4.7
7.6
5.6
5.0
4.4
8.3
4.5
7.4
U
U
6.1
U
U
6.4
U
74
74
4.0
M
44
44
6.0
6.7
10.7-32.0
5.0-294
3.4-24.5
11.9-2S.3
104-27.4
11.3-2S4
4.5-35.5
12.4-24.0
1.7-464
64-M4
114-144
1JMI4
11.4-27.1
10.1-29.9
64414
7.0-274
42-172
13-159
0-144
41-143
11-150
46-137
14-166
49-113
7-
42-141
47-112
11-147
44-156
22-176
22-176
6.1-294
94-264
7.4-26.1
64-29.9
41-136
3B-146
21-156
26-163
O-ConeMMIon mmuntd ki QC GhHk tHi**. in M/L (84<*jn 7.5.3).
t-SMndard dnHMon of tour raeeMry mMMOTMnliJn |>g/L Otelton 6J.4).
x-Av»r»g« neoKify tor tour r«ca»lty nm«ur»m«n». in ug/L (S«clton 67.4).
P, P,.Pwomt raeoMty nwMurad (Btedon 6.3A Sw«anT4J).
D-MMM; nNuR mu* to gnMtor tt«n mo.
•CrMrt* wm mcuHHtf Miuming • OC duck tmvto conon>»»on ol 20 ofl/L
Not*: The6e criteria an baaed directly
upon the method performance data in Table
3. Where necei»ary, the limiti for recovery
have been broadened to a«6ure applicability
of the limite to concentration* below thoae
U6ed to develop Table 3.
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 601
fMOww
Cvtral praoWon.
-
1.12C-1.02
OJ6C-106
0.76C-177
0.99C-1.04
1.00C-1J3
0.66C-1J3
1.00C
ojsc-oje
0.11X+OXM
0.12X+OJ6
OJ6J+OJ7
0.15X+OJ6
O.ltX-0.02
O.t4jt-0.11
0.13«+0.16
OMC+2.71
OJK^-I.TO
0.1 1X+ 1.10
OJOJU0.97
021«+t41
OJ6Jt+Ol94
OJOS+OJi
0.1SX+6.13
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 35
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 601—Continued
Parameter
Accuracy, as
'
,
recovery. X'
Single analyst
precision. V IMS'
Overall precision.
S' (ug/L)
1.3-0ichlorobenzene '. 0.95C4-0.43 | 0.14X4-2.33
M-OchkxoDeniene 0.93C-0.09 0.15*40.29
1.1-Oicnloroethane 0.95C-1.08 ' 0.08X4-0.17
1.2-Dichloroethane 1.IMC-1.08 0.11X4-0.70
1.1-Dichloroethene 0.98C-0.87 0.21*-0.23
trans-t,2-Dichloroethene 0.97C-0.16 0.11X4-1.46
1,2-Dichloropropane' 1.00C 0.13X
cis-t.S-Oichkxopropene' 1.00C 0.18*
lans-I.S-Oichloropropene' 1.00C 0.18*
Methylene chlonde 0.91C-0.93 0.11X4-0.33
1.1.2.2-Tetrachloroetrwne 0.95C4-0.19 0.MX+ 2.41
Tetrachtoroethene 0.94C + 0.06 0.14*+ 0.38
1.1.1-Tnchtoroethane 0.90C-0.16 0.15*+ 0.04
1.1,2-Trichloroethane 0.86C + 0.30 0.13*-0.14
Triehloroethene 0.87C+0.48 0.13*-0.03
Tnchlorolluoromethane 0.89C-0.07 0.15*+0.67
Vinyl chloride 0.97C-0.36 0.13*+0.65
0.26X 4 2.34
0.20X+041
0 MX+094
0.158^0.94
0.29X-0.40
0.17X+ 1.46
0.23X
032X
0.32X
0.21X+ 1.43
0.23X4.2.79
0.18X + 2.21
0.20X + 0.37
0.19X+0.67
0.23X4-0.30
0.26X4-0.91
0.27X4.0.40
X' = Expected recovery for one or more measurements of a sample containing a concentration of C. in pg/L.
Sg's Expected single analyst standard deviation of measurements at an average concentration found of X. in pg/L.
S' = Expected interlaooratory standard deviation of measurements at an average concentration found of X. in ug/L.
C = True value for the concentration, in ug/L
X=Average recovery found lor measurements of samples containing a concentration of C, in ug/L
•Estimates based upon the performance in a single laboratory.10
BILLING CODE 6560-50-M
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36
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
OPTIONAL
FOAM
TRAP
-BIT X IN.
0.0.
— 14MM 0. 0.
INLET X IN.
0. D.
X IN. _
0. 0. EXIT
10MH GLASS FRIT
MEDIUM POROSITY
SAMPLE INLET
MAY SVMNGE VALVE
-17CM. 20 GAUGE SYRINGE NEEDLE
6MM. 0. D. RUBBER SEPTUM
~1QMM. 0. D.
-»-INLET
X IN. 0. D.
1/11 IN. O.D.
f STAINLESS STEEL
131 MOLECULAR
SIEVE PURGE
GAS FILTER
PURGE GAS
FLOW
CONTROL
Figure 1. Purging device.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
37
PACKING PROCEDURE
CONSTRUCTION
GLASS CMIIFI
WOOL "f E
ACTIVATED , 1
CHARCOAL 7.7C
-
GRADE 15 7
SILICA GEL1'1*'
TENA1 7.7 C
3540V-1
GLASS WOOL1CM
A
1
/I
.
5MM
%
I
\
-
_
TRA
7 ^/FOOT
RESISTANCE
WIRE WRAPPED
SOLID <
(DOUBLE LAYER)
1SCM
7~ /FOOT.
RESISTANCE
WIRE WRAPPED
SOLID
(SINGLE LAYER)
8CM-
INLET
-------
38
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
CARRIER OAS PLOW CONTROL LIQUID IMJfCTION
PRESSURE REGULATOR
fODCVfCTOR
ANAUTICM. COLUHM
\ OPTIONAL 4-PORT COLUMN
SELECTION VALVE
19X MOLfCULAM
SIEVE FILTH
DffVKl
HEATER CONTROL
i.ALLUNOKTWEEN
TRAP AND OC
SHOULD • ttEATED
TOtOX
Figure 3. Purge and trap •ysttfn«purg« mod*.
CARRIER GAS
FLOW CONTROL LIQUID INJECTION PORTS
PRESSURE
K6ULATOR
_
V WIIWBk *« V|^
nOtl CONTROL^
131 MOLECULAR |
SIEVE FILTER
COLUMN OVEN
__CONFIRMATO* COLUMN
> TO DETECTOR
' ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SaECTWN VALVE
INLET
VALVE J RESISTANCE VIRE MEATB|
^CONTROL
PURGING
OCVICE
ALL LINES KTKEN
TRAP AND GC
SHOULD K HEATED
TO M°C.
Figurt 4. Purgt and trap system • dasorb mode.
-------
COLUMN: IX SP-1000 ON CAR80PACX-B
PROGRAM: 45°C FOR 3 WIN. 8*C/MIN TO 220°C
DETECTOR: HALL 700-A aECTROLVTIC CONDUCTIVITY
T «>• a « 5 1}
»- . o o <
18 20 22' 2*
02 4 .6 8 10 12 14 16
RETENTION TIME, MIN.
Figure 6. Gas chromatogram of purgeable halocarbons.
m
a.
(B
09
to
<
o
CD
z
o
a.
Q;
v;
O
o
a
cr
o
50
c_
!B
u
03
a.
50
i
CO
co
-------
40 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 802-Purgeable Aromatic*
/. Scope and Application
1.1 This method covers the determination
of various purgeable aromatics. The following
parameters may be determined by this
method:
PwaiTWMr
BcnzcAV
CMorabtnnnt
1.3-Did*xot>«r««n«
EIHyw^flWo*
TokM««
STOflET
No
34030
34301
34536
34566
3457 1
34371
34010
CAS No
71-43-2
108-90-7
95-50-1
S41-73-1
106-46-7
tOO-41-4
10S-M-3
1.2 This is a purge and trap gas
chromatographic (CC) method applicable to
the determination of the compounds listed
above in municipal and industrial discharges
as provided under 40 CFR 136.1. When this
method is used to analyze unfamiliar samples
for any or all of the compounds above.
compound identifications should be
supported by at least one additional
qualitative technique. This method describes
analytical conditions for a second gas
chromatographic column that can be used to
confirm measurements made with the
primary column. Method 624 provides gas
chromatograph/mass spectrometer (GC/MS)
conditions appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above.
1.3 The method detection limit (MDL,
defined in Section 12.1)' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed.
depending upon the nature of interferences in
the sample matrix.
1.4 Any modification of this method.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the operation of a purge and
trap system and a gas chromatograph and in
the interpretation of gas chromatograms.
Each analyst must demonstrate the ability to
generate acceptable results with this method
using the procedure described in Section 8.2.
2. Summary of Method
2.1 An inert gas is bubbled through a 5-
mL water sample contained in a specially-
designed purging chamber at ambient
temperature. The aromatics are efficiently
transferred from the aqueous phase to the
vapor phase. The vapor is swept through a
sorbent trap where the aromatics are
trapped. After purging is completed, the trap
is heated and backflushed with the inert gas
to desorb the aromatics onto a gas
chromatographic column. The gas
chromatograph is temperature programmed to
separate the aromatics which are then
detected with a photoionixation detector.1'
2.2 The method provides an optional gas
chromatographic column that may be helpful
in resolving the compounds of interest from
interferences that may occur.
3. Interferences
3.1 Impurities in the purge gas and
organic compounds outgassing 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
laboratory reagent blanks as described in
Section 6.1.3. The use of non-Teflon plastic
tubing. non-Teflon thread sealants, or flow
controllers with rubber components in the
purge and trap system 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 field reagent blank prepared
from reagent water and carried through the
sampling and handling protocol can serve as
a check on such contamination.
3.3 Contamination by carry-over can
occur whenever high level and low level
samples are sequentially analyzed. To reduce
carry-over, the purging device and sample
syringe must be rinsed with reagent water
between sample analyses. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of reagent water to check for cross
contamination. For samples containing large
amounts of water-soluble materials.
suspended solids, high boiling compounds or
high aromatic levels, it may be necessary to
wash the purging device with a detergent
solution, rinse it with distilled water, and
then dry it in an oven at 105 *C between
analyses. The trap and other parts of the
system are also subject to contamination:
therefore, frequent bakeout and purging of
the entire system may be required.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified *'for the information of the
analyst.
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: benzene and 1.4-
dichlorobenzene. Primary standards of these
toxic compounds should be prepared in a
hood. A NIOSH/MESA approved toxic gas
respirator should be worn when the analyst
handles high concentrations of these toxic
compounds.
5. Apparatut and Materials
5.1 Sampling equipment, for discrete
sampling.
5.1.1 Vial—25-mL capacity or larger,
equipped with a screw cap with a hole in the
center (Pierce »13075 or equivalent).
Detergent wash, rinse with tap and distillel
water, and dry at 105 *C before use.
5.1.2 Septum—Teflon-faced silicone
(Pierce «12722 or equivalent). Detergent
wash, rinse with lap and distilled water, and
dry at 105 *C for 1 h before use.
5.2 Purge and trap system—The purge and
trap system consists of three separate pieces
of equipment: A purging device, trap, and
desorber. Several complete systems are now
commercially available.
5.2.1 The purging device must be designed
to accept 5-mL samples with a water column
at least 3 cm deep. The gaseous head space
between the water column and the trap must
have a total volume of less than 15 ml. The
purge gas must pass through the water
column as finely divided bubbles with a
diameter of less than 3 mm at the origin. The
purge gas must be introduced no more than 5
mm from the base of the water column. The
purging device illustrated in Figure 1 meets
these design criteria.
5.2.2 The trap must be at least 25 cm long
and have an inside diameter of at least 0.105
in.
5.2.2.1 The trap is packed with 1 cm of
methyl silicone coated packing (Section 6.4.2)
and 23 cm of 2.6-diphenylene oxide polymer
(Section 6.4.1) as shown in Figure 2. This trap
was used to develop the method performance
statements in Section 12.
5.2.2.2 Alternatively, either of the two
traps described in Method 601 may be used,
although water vapor will preclude the
measurement of low concentrations of
benzene.
5.2.3 The desorber must be capable of
rapidly beating the trap to 180 'C. The
polymer section of the trap should not be
heated higher than 180 *C and the remaining
sections should not exceed 200 *C. The
desorber illustrated in Figure 2 meets these
design criteria.
5.2.4 The purge and trap system may be
assembled as a separate unit or be coupled to
a gas chromatograph as illustrated in Figures
3,4. and 5.
5.3 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
columns, gases, detector, and strip-chart
recorder. A data system is recommended for
measuring peak areas.
5.3.1 Column 1—6 ft long x 0.082 in. ID
stainless steel or glass, packed with 5* SP-
1200 and 1.75% Bentone-34 on Supelcoport
(100/120 mesh) or equivalent This column
was used to develop the method performance
statements in Section 12. Guidelines for the
use of alternate column packings are
provided in Section mi.
5.3.2 Column 2—8 ft long x 0.1 in ID
stainless steel or glass, packed with 5% 1,2,3-
Tris(2-cyanoethoxy)propane on Chromosorb
W-AW (80/80 mesh) or equivalent.
5.3.3 Detector—Photoionization detector
(h-Nu Systems, Inc. Model Pl-51-02 or
equivalent). This type of detector has been
proven effective in the analysis of
wastewaters for the parameters listed in the
scope (Section 1.1). and was used to develop
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations 41
the method performance statements in
Section 12. Guidelines for the use of alternate
detectors are provided in Section 10.1.
5.4 Syringes—5-mL glass hypodermic with
Luerlok tip (two each), if applicable to the
purging device.
5.5 Micro syringes—25-^L, 0.006 in. ID
needle.
5.6 Syringe valve—2-way. with Luer ends
(three each).
5.7 Bottle—15-mL. screw-cap, with Teflon
cap liner.
5.8 Balance—Analytical, capable of
accurately weighing 0.0001 g.
ft Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.1.1 Reagent water can be generated by
passing tap water through a carbon filter bed
containing about 1 Ib of activated carbon
(Filtrasorb-300, Calgon Corp., or equivalent).
6.1.2 A water purification system
(Millipore Super-Q or equivalent) may be
used to generate reagent water.
6.1.3 Reagent water may also be prepared
by boiling water for 15 min. Subsequently,
while maintaining the temperature at 90 *C,
bubble a contaminant-free inert gas through
the water for 1 h. While still hot, transfer the
water to a narrow mouth screw-cap bottle
and seal with a Teflon-lined septum and cap.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Hydrochloric acid (1+1)—Add 50 mL
of concentrated HC1 (ACS) to 50 mL of
reagent water.
6.4 Trap Materials:
8.4.1 2,6-Diphenylene oxide polymer—
Tenax, (60/80 mesh), chromatographic grade
or equivalent.
6.4.2 Methyl silicone packing—3% OV-1
on Chromosorb-W (60/80 mesh) or
equivalent.
8.5 Methanol—Pesticide quality or
equivalent.
6.6 Stock standard solutiona—Stock
standard solutions may be prepared from
pure standard materials or purchased as
certified solutions. Prepare stock standard
solutions in methanol using assayed liquids.
Because of the toxicity of benzene and 1,4-
dichlorobenzene, primary dilutions of these
materials should be prepared in a hood. A
NIOSH/MESA approved toxic gas respirator
should be used when the analyst handles
high concentrations of such materials.
6.6.1 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 min or until all alcohol wetted
surfaces have dried. Weigh the flask to the
nearest 0.1 mg.
6.6.2 Using a 100-ftL syringe, immediately
add two or more 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.
6.6.3 Reweigh, dilute to volume, stopper,
then mix by inverting the flask several times.
Calculate the concentration in fig/Ml from
the net gain in weight. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.6.4 Transfer the stock standard solution
into a Teflon-sealed screw-cap bottle. Store
at 4 *C and protect from light.
6.6.5 All standards must be replaced after
one month, or sooner if comparison with
check standards indicates a problem.
6.7 Secondary dilution standards—Using
stock standard solutions, prepare secondary
dilution standards in methanol that contain
the compounds of interest, either singly or
mixed together. The secondary dilution
standards should be prepared at
concentrations such that the aqueous
calibration standards prepared in Sections
7.3.1 or 7.4.1 will bracket the working range of
the analytical system. Secondary solution
standards must be stored with zero
headspace and should be checked frequently
for signs of degradation or evaporation,
especially just prior to preparing calibration
standards from them.
6.8 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Assemble a purge and trap system
that meets the specifications in Section 5.2.
Condition the trap overnight at 180 *C by
backflushing with an inert gas flow of at least
20 mL/min. Condition the trap for 10 min
once daily prior to use.
7.2 Connect the purge and trap system to
a gas chromatograph. The gas chromatograph
must be operated using temperature and flow
rate conditions equivalent to those given in
Table 1. Calibrate the purge and trap-gas
chromatographic system using either the
external standard technique (Section 7.3) or
the internal standard technique (Section 7.4).
7.3 External standard calibration
procedure:
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter by carefully adding 20.0 /tL of
one or more secondary dilution standards to
100, 500, or 1000 mL of reagent water. A 25-
jiL syringe with a 0.006 in. ID needle should
be used for this operation. One of the
external standards should be at a
concentration near, but above, the MDL
(Table 1) and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector. These aqueous standards must be
prepared fresh daily.
7.3.2 Analyze each calibration standard
according to Section 10, and tabulate peak
height or area responses versus the
concentration in the standard. The results
can be used to prepare a calibration curve for
each compound. Alternatively, if the ratio of
response to concentration (calibration factor)
is a constant over the working range (<10%
relative standard deviation. RSO], linearity
through the origin can be assumed and the
average ratio or calibration factor can be
used in place of a calibration curve.
7.4 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that (he measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
The compound, a.a.a.-trifluorotoluene.
recommended as a surrogate spiking
compound in Section 8.7 has been used
successfully as an internal standard.
7.4.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest as described in
Section 7.3.1.
7.4.2 Prepare a spiking solution containing
each of the internal standards using the
procedures described in Section 6.6 and 6.7. It
is recommended that the secondary dilution
standard be prepared at a concentration of 15
ug/mL of each internal standard compound.
The addition of 10 /il of this standard to 5.0
mL of sample or calibration standard would
be equivalent to 30 pg/L.
7.4.3 Analyze each calibration standard
according to Section 10, adding 10 jiL of
internal standard spiking solution directly to
the syringe (Section 10.4). Tabulate peak
height or area responses against
concentration for each compound and
internal standard, and calculate response
factors (RF) for each compound using
Equation 1.
Equation 1.
RF=-
(AJfCiJ
(A*)(C.)
where:
A,=Response for the parameter to be
measured.
A,,=Response for the internal standard.
Q,=Concentration of the internal standard
C,=Concentration of the parameter to be
measured.
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios. A\/Ah, vs. RF.
7.5 The working calibration curve.
calibration factor, or RF must be verified on
each working day by the measurement of a
QC check sample.
7.5.1 Prepare the QC check sample as
described in Section 8.2.2.
7.5.2 Analyze the QC check sample
according to Section 10.
7.5.3 For each parameter, compare the
response (Q) with the corresponding
calibration acceptance criteria found in Table
2. If the responses for all parameters of
interest fall within the designated ranges,
analysis of actual samples can begin. If any
individual Q falls outside the range, a new
calibration curve, calibration factor, or RF
must be prepared for that parameter
according to Section 7.3 or 7.4.
8. Quality Control
ai Each laboratory that uses this method is
required to operate a formal quality control
program. The mimimum requirements of this
program consist of an initial demonstration of
-------
42
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chroma tography, the analyst is
permitted certain options (detailed in Section
10.1) to improve the separations or lower the
cost of measurements. Each time such a
modification is made to the method, the
analyst is required to repeat the procedure in
Section 8.2.
8.1.3 Each day, the analyst must analyze a
reagent water blank to demonstrate that
interferences from the analytical system are
under control.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 6.3.
6.1.5 The laboratory must on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control Thisprocedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analysed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that ia generated This procedure is
described in Section 8.5.
6J To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
BwJ.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of 10
ug/mL in methanol. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency.
Environmental Monitoring and Support
Laboratory in Cincinnati. Ohio, if available. If
not available from that source, the QC check
•ample concentrate mud be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8£.2 Prepare a QC check sample to
contain 20 pg/L of each parameter by adding
200 pL of QC check sample concentrate to
100 mL of reagent water.
&2J Analyze four 5-mL aliquot* of the
well-mixed QC check sample according to
Section 10.
8.2.4 Calculate the average recovery (X)
in M8/L. and 'ne standard deviation of the
recovery (s) in pg/L. for each parameter of
interest using the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 2 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.6 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.6.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.3.
8.2.6.2 Beginning with Section 8.2.3, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 84.3.
8.3 The laboratory must on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample ia being checked against a regulatory-
concentration Umit the spike should be at
that limit orlto 5 times higher than the
background concentration determined in
Section 84.Z whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 20 jig/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.2 Analyze one 5-mL sample aliquot to
determine the background concentration (B)
of each parameter. II necessary, prepare a
new QC check sample concentrate (Section
8A1) appropriate for the background
concentrations in the sample. Spike a second
5-mL sample aliquot with 10 pL of the QC
check sample concentrate and analyse it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, when T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 20 pg/L the analyst must use
either the QC acceptance criteria in Table 2,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter. (1) Calculate
accuracy (X') using the equation in Table 3,
substituting the spike concentration (T) for C;
(2) calculate overall precision (S') using the
equation in Table 3, substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) ± 2.44(100 S'/
T)%.'
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note: The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 10 uL of QC check sample concentrate]
(Sections 8il or 84.2) to 5 mL of reagent
water. The QC check standard needs only to
contain the parameters that failed criteria in
the test in Section 8J.
8.4.2 Analyse the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (PJ as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4 J Compare the percent recovery (PJ
for each parameter with the "^"•«p~"'«"g
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected The analytical result for that
parameter in the unapiked sample is suspect
and may not be reported for regulatory
compliance purposes.
&5 As part of the QC program for the
laboratory, method accuracy for waatewatar
samples must be assessed and records must
be maintained After the analysis of five
•piked wastewater samples as in Section 8J,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (s,). Express the accuracy
assessment as a percent recovery interval
from P-2a, to P+2H. If P-90% and s-10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e,g. after each five to ten new accuracy
measurements).
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations 43
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromalogram. confirmatory techniques such
as gas chromalography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
8.7 The analyst should monitor both the
performance of the analytical system and the
effectiveness of the method in dealing with
each sample matrix by spiking each sample.
standard, and reagent water blank with
surrogate compounds (e.g. a, a, a,-
trifluorotoluene) recommended to encompass
the range of the temperature program used in
this method. From stock standard solutions
prepared as in Section 6.6, add a volume to
give 750 ng of each surrogate to 45 mL of
reagent water contained in a 50-mL
volumetric flask, mix and dilute to volume for
a concentration of 15 mg/jtL. Add 10 \iL of
this surrogate spiking solution directly into
the 5-mL syringe with every sample and
reference standard analyzed. Prepare a fresh
surrogate spiking solution on a weekly basis.
If the internal standard calibration procedure
is being used, the surrogate compounds may
be added directly to the internal standard
spiking solution (Section 7.4.2).
ft Sample Collection, Preservation, and
Handling
9.1 The samples must be iced or
refrigerated from the time of collection until
analysis. If the sample contains free or
combined chlorine, add sodium thiosulfate
preservative (10 mg/40 mL is sufficient for up
to 5 ppm Cla) to the empty sample bottle just
prior to shipping to the sampling site. EPA
Method 330.4 or 330.5 may be used for
measurement of residual chlorine.'Field test
kits are available for this purpose.
9.2 Collect about 500 mL of sample in a
clean container. Adjust the pH of the sample
to about 2 by adding 1 + 1 HC1 while stirring.
Fill the 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.
9.3 All samples must be analyzed within
14 days of collection.1
10. Procedure
10.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
estimated retention times and MDL that can
be achieved under these conditions. An
example of the separations achieved by
Column 1 is shown in Figure 6. Other packed
columns, chromatographic conditions, or
detectors may be used if the requirements of
'Section 8.2 are met.
10.2 Calibrate the system daily as
described in Section 7.
10.3 Adjust (he purge gas (nitrogen or
helium) flow rate to 40 mL/min. Attach the
trap inlet to the purging device, and set the
purge and trap system to purge (Figure 3).
Open the syringe valve located on the
purging device sample introduction needle.
10.4 Allow the sample to come to ambient
temperature prior to introducing it to the
syringe. 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 to just short of overflowing. Replace
the syringe plunger and compress the sample.
Open the syringe valve and vent any residual
air while adjusting the sample volume to 5.0
mL. 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 10.0 \iL of the
surrogate spiking solution (Section 8.7) and
10.0 |iL of the internal standard spiking
solution (Section 7.4.2), if applicable, through
the valve bore, then close the valve.
10.5 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.
10.6 Close both valves and purge the
sample for 12.0±0.1 min at ambient
temperature.
10.7 After the 12-min purge time,
disconnect the purging device from the trap.
Dry the trap by maintaining a flow of 40 mL/
min of dry purge gas through it for 6 min
(Figure 4). If the purging device has no
provision for bypassing the purger for this
step, a dry purger should be inserted into the
device to minimize moisture in the gas.
Attach the trap to the chromatograph, adjust
the purge and trap system to the desorb mode
(Figure 5), and begin to temperature program
the gas chromatograph. Introduce the trapped
materials to the GC column by rapidly
heating the trap to 180 *C while back/lushing
the trap with an inert gas between 20 and 60
mL/min for 4 min. If rapid heating of the trap
cannot be achieved, the GC column must be
used as a secondary trap by cooling it to 30
*C (subambient temperature, if poor peak
geometry and random retention time
problems persist) instead of the initial
program temperature of 50 'C.
10.8 While the trap is being desorbed into
the gas chromatograph column, empty the
purging chamber using the sample
introduction syringe. Wash the chamber with
two 5-mL flushes of reagent water.
10.9 After desorbing the sample for 4 min,
recondition the trap by returning the purge
and trap system to the purge mode. Wait 15 s,
then close the syringe valve on the purging
device to begin gas flow through the trap. The
trap temperature should be maintained at 180
"C. After approximately 7 min, turn off the
trap heater and open the syringe valve to
stop the gas flow through the trap. When the
trap is cool, the next sample can be analyzed.
10.10 Identify the parameters in the
sample by comparing the retention times of
the peaks in the sample chromatogram with
those of the peaks in standard
chromatograms. The width of the retention
time window used to make identifications
should be based upon measurements of
actual retention time variations of standards
over the course of a day. Three times the
standard deviation of a retention lime for a
compound can be used to calculate a
suggested window size; however, the
experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
10.11 If the response for a peak exceeds
the working range of the system, prepare a
dilution of the sample with reagent water
from the aliquot in the second syringe and
reanalyze.
11. Calculations
11.1 Determine the concentration of
individual compounds in the sample.
11.1.1 If the external standard calibration
procedure is used, calculate the
concentration of the parameter being
measured from the peak response using the
calibration curve or calibration factor
determined in Section 7.3.2.
11.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.4.3 and Equation 2.
Equation 2.
Concentration (^g/L) =
(Ata)(RF)
where:
A. = Response for the parameter to be
measured.
Au = Response for the internal standard.
Cu = Concentration of the internal
standard.
11.2 Report results in >ig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
12. Method Performance
12.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Table 1 were obtained using reagent water.9
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
12.2 This method has been demonstrated
to be applicable for the concentration range
from the MDL to 1000 x MDL.' Direct
aqueous injection techniques should be used
to measure concentration levels above 1000 x
MDL
12.3 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 2.1 to 550 jig/L.9 Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
-------
44 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
\. 40 CFR Part 136. Appendix B.
2. Bellar, T.A., and Lichtenberg. J.J. Journal
American Water Works Association. 66. 739
(1974).
3. Bellar. T.A., and Lichtenberg,).). "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds." Proceedings of
Symposium on Measurement of Organic
Pollutants in Water and Wastewater.
American Society for Testing and Materials.
STP 686, C.E. Van Hall, editor. 1978.
4. "Carcinogens—Working with
Carcinogens." Department of Health,
Education, and Welfare. Public Health
Service. Center for Disease Control. National
Institute for Occupational Safety and Health.
Publication No. 77-206. August 1977.
5. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2206 (Revised,
January 1976).
6. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication. Committee on Safety, 3rd
Edition. 1979.
7. Provost, L.P.. and Elder, R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3. is two times the value 1.22 .derived in
this report.)
8."Methods 330.4 (Titrimetric. DPD-FAS)
and 330.5 (Spectrophotometric. DPD) for
Chlorine, Total Residual." Methods for
Chemical Analysis of Water and Wastes,
EPA-600/4-79-020, U.S. Environmental
Protection Agency, Office of Research and
Development, Environmental Monitoring and
Support Laboratory, Cincinnati. Ohio 45268.
March 1979.
9."EPA Method Validation Study 24,
Method 602 (Purgeable Aromatics)." Report
for EPA Contract 68-03-2856 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Pannwttr
Benign! J
TOfcMftt , _ _ _ .... ,
1 3-QicNoiQt)WMnt , - - ,--,••
Retention time (min)
Column
1
3.33
5.7$
a.2s
9.17
16.S
18.2
25.9
Column
2
2.75
4.25
(.25
(.02
18.2
15.0
19.4
Method
dctochon
Nfnrt
Oifl/U
0.2
0.2
0.2
0.2
0.3
0.4
0.4
Column 1 oonowon*: Supdcopon (100/120 mnn) coiMd with 5% SP-1200/1.75% Bwttoiw-34 pidujd in t (ft x 0.065 in. ID Uairtnt Mri column with Infant cam* gn *t 38 ml/mm
flow nta. Column wnpcratura lwU M SO 'C tor 2 min th*n progrimrMd at 6 •C/mn to 00 'C tor • frttl hold.
Column 2 oondWom: Chromowrb W-AW (60/80 mnh) OMted with 5* 1A3-Trti<2.«wo«thyoiwtoropin» ptdud in • 6 ft > 0.065 in. ID MMw MMI column w*i Mum eanWr gst at
30rM^n*»own*.C<>lurr*l«n»»riturth»ldit40X:tor2mto "^
TABLE 2.—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 602 •
15.4-24.6
16.1-23.9
13.6-26.4
14J-2S.S
13.9-26,1
116-27.4
18J-M.S
10.0-27.9
1Z7-2S.4
10.6-(7J
116-2U
11.6-28.5
10.0-26J
11J-27.7
I:
,_ ^Jd jiitojon of tour r
••AwMQ*) Moowy foe four A
idtoOCohK*
•.to
(Section 7 JUL
ign.aiemanU.4l.
«/L 0wton (£4).
K4J).
Note Th»t critorto «n» b»«d OVecfy upon ti« rmiiod perturnimc
K»»a8ui» b»tew fnM uMd to dcMtop Tabto 3.
to T«H« 3. Whtra n»cnnry, lh* fen* tor njcowy htv» bMn bioadintd tt aswra appscaMHy of «w InMi to
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 602
p*™*,
.__. J
1 HXeMomlMnnnt
1 ^ PffHffflHff'ITT «...,.. .T,. , !, , ,
TolSiw "
Aocumcy. t»
rwo«*y. X'
U4/U
0.92C+0.57
096C+002
O.UC+0.52
OJ8C-0.04
ojfjc-faw
0.94C+OJ1
0.94C+0.65
^SJUC^
"^Q/L)'
008X+OJ6
008X+OJ3
0 17X-0.04
0.15X-0.10
0.17X+0^6
0.00X+0.46
Cvtrall
pracWon, S'
(MO/U
0^1X4-086
0 ITX-fO.10
0.18H +0.06
OJOX4>0.41
O.MX+OJI
0 16f +0.71
!C-E»p»ia»d r»oov«ry tor on* or mon» r
C-Trw «Uw tor ta Conemta8gn. to (ifl/U
X>A*ma» roomy tound tor tmmunmmH* of imvtoi oontototog 8 uunuti*»tun of C, to «/L
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 45
OPTIONAL
FOAM
TRAP
•EXIT '/« IN.
0. D
— 14MM 0. D.
INLET H IN.
0. 0.
VilN.
0. D. EXIT
SAMPLE INLET
2-WAY SYRINGE VALVE
17CM 20 GAUGE SYRINGE NEEDLE
6MM. 0. 0 RUBBER SEPTUM
~10MM 0. D
-INLET
v; IN. 0.
1/16 IN. 0.0.
y STAINLESS STEa
13X MOLECULAR
SIEVE PURGE
GAS FILTER
PURGE GAS
FL0W
CONTROL
10MM GLASS FRIT
MEDIUM POROSITY
Figure 1. Purging device.
-------
46 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
PACKING PROCEDURE
GLASS
WOOL
TENAI 2:
3XOV-1
GLASS WOOL
1CM
CONSTRUCTION
COMPRESSION FITTING
NUT AND FERRULES
14FT.7A/FOOT RESISTANCE
WIRE WRAPPED SOLID
THERMOCOUPLE/
CONTROLLER
TRAP INLET
TUBNGKCM.
0.105 IN. 1.0.
0.12S IN. 0.0.
STAINLESS STEEL
Figur* 2. Trap packings and construction to include
d««orb capability.
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
47
Carrier Gas Flow Control Liquid Injection Ports
Pressure Regulator
Purge Gas
Flow Control
13X Molecular
Sieve Filter
Valve-3
Optional 4 Port Column
Selection Valve
End)
Resistance Wire
Column Oven
-. Confirmatory Column
to Detector
--Analytical Column
Velve-2
. Heeter Control
Note: All Lines Between
Trap and GC
Should be Heated
to 80°C
Figure 3. Purge and trap system • purge mode.
Carrier Gas Flow Control
Pressure Regulator
Liquid Injection Ports
Purge Gas
Flow Control \|
13X Molecular
Sieve Filer
Valve-3
Optional 4-Port Column
Selection VaKw
Trap Inlet (Tenax End)
* Resistance Wire
Column Oven
_,_ Confirmatory Column
To Detector
I "-—Analytical Column
Heater Control
Varve-2
Note: All Lines Between
Trap and GC
Should be Heated
tt>80°C
Figure 4. Purge and trap system-dry mode.
-------
48
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
Carrier Oat Flow Control Liquid Injection Ports
Preeaure Regulator
Purge OM
Flow Control \T~
13X Molecular
Sieve Filter
Valve-3
Optional 4-Port Column
Selection Valve
'rapMetfTenax
Column Own
Confirmatory Column
to Detector
Analytical Column
Heater Control
Vefeo-2
Note: All Unee Between
Trap and OC
Should be Heated
to 80°C
Figure 5. Purge and trap tystem-deaorfa moda.
Cahima: 5% SP 1200/1.75% Beateat - 34
Pregraej: M°C hx 2 •«. I«C/M la M«C
Diticun: Phemeeuiuoa, ia2 V
n
ui
v^-J
«°
02 4 • • 10 12 14 1« 10 20 22 24 26 28
Retention Tima. Min.
Figure 6. Gas chromatogram of purgeable aromatics.
MUINO OOOC MM-M-C
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 603—Acrolein and Acrylonitrile
1. Scope and Application
1.1 This method covers the determination
of acrolein and acrylonitrile. The following
parameters may be determined by this
method:
Paiameier
STORET
No.
34210
342 IS
CAS No.
107-02-8
107-13-1
1.2 This is a purge and trap gas
chromatographic (GC) method applicable to
the determination of the compounds listed
above in municipal and industrial discharges
as provided under 40 CFR 136.1. When this
method is used to analyze unfamiliar samples
for either or both of the compounds above,
compound identifications should be
supported by at least one additional
qualitative technique. This method describes
analytical conditions for a second gas
chromatographic column that can be used to
confirm measurements made with the
primary column. Method 624 provides gas
chromatograph/mass spectrometer (GC/MS)
conditions appropriate for the qualitative and
quantitative confirmation of results for the
parameters listed above, if used with the
purge and trap conditions described in this
method.
1.3 The method detection limit (MDL,
defined in Section 12.1)' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the operation of a purge and
trap system and a gas chromatograph and in
the interpretation of gas chroma tograms.
Each analyst must demonstrate the ability to
generate acceptable results with this method
using the procedure described in Section 8.2.
2. Summary of Method
2.1 An inert gas is bubbled through a 5-
mL water sample contained in a heated
purging chamber. Acrolein and acrylonitrile
are transferred from the aqueous phase to the
vapor phase. The vapor is swept through a
sorbent trap where the analytes are trapped.
After the purge is completed, the trap is
heated and backflushed with the inert gas to
desorb the compound onto a gas
chromatographic column. The gas
chromatograph is temperature programmed to
separate the analytes which are then
detected with a flame ionization detector.1 *
2.2 The method provides an optional gas
chromatographic column that may be helpful
in resolving the compounds of interest from
the interferences that may occur.
3. Interferences
3.1 Impurities in the purge gas and
organic compound outgassing from the
plumbing 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 laboratory reagent
blanks as described in Section 8.1.3. The use
of non-Teflon plastic tubing. non-Teflon
thread sealants, or flow controllers with
rubber components in the purge and trap
system 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 field reagent blank prepared
from reagent water and carried through the
sampling and handling protocol can serve as
a check on such contamination.
3.3 Contamination by carry-over can
occur whenever high level and low level
samples are sequentially analyzed. To reduce
carry-over, the purging device and sample
syringe must be rinsed between samples with
reagent water. Whenever an unusually
concentrated sample is encountered, it should
be followed by an analysis of reagent water
to check for cross contamination. For samples
containing large amounts of water-soluble
materials, suspended solids, high boiling
compounds or high analyte levels, it may be
necessary to wash the purging device with a
detergent solution, rinse it with distilled
water, and then dry it in an oven at 105 *C
between analyses. The trap and other parts
of the system are also subject to
contamination, therefore, frequent bakeout
and purging of the entire system may be
required.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this view point, 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 data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified * 6 for the information of the
analyst.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete
sampling.
5.1.1 Vial—25-mL capacity or larger,
equipped with a screw cap with a hole in the
center (Pierce #13075 or equivalent).
Detergent wash, rinse with tap and distilled
water, and dry at 105 °C before use.
5.1.2 Septum—Teflon-faced silicone
(Pierce #12722 or equivalent). Detergent
wash, rinse with tap and distilled water and
dry at 105 'C for 1 h before use.
5.2 Purge and trap system—The purge and
trap system consists of three separate pieces
of equipment: a purging device, trap, and
desorber. Several complete systems are now
commercially available.
5.2.1 The purging device must be designed
to accept 5-mL, samples with a water column
at least 3 cm deep. 1 nc gaseous heud space
between the water column and the trap must
have a total volume of less than 15 mL. The
purge gas must pass through the water
column as finely divided bubbles with u
diameter of less than 3 mm at the origin. The
purge gas must be introduced no morn tluin 5
mm from the base of the water column. The
purging device must be capable of being
heated to 85 'C within 3.0 min after transfer
of the sample to the purging device and being
held at 85 ±2 °C during the purge cycle. The
entire water column in Ihe purging device
must be heated. Design of this modification to
the standard purging device is optional.
however, use of a water bath is suggested.
5.2.1 Heating mantle—To be used to heat
water bath.
5.2.1.2 Temperature controller—Equipped
with thermocouple/sensor to accurately
control water bath temperature to ±2 "C. The
purging device illustrated in Figure 1 meets
these design criteria.
5.2.2 The trap must be at least 25 cm long
and have an inside diameter of at least 0.105
in. The trap must be packed to contain 1.0 cm
of methyl silicone coated packing (Section
6.5.2) and 23 cm of 2.6-diphenylene oxide
polymer (Section 6.5.1). The minimum
specifications for the trap are illustrated in
Figure 2.
5.2.3 The desorber must be capable of
rapidly heating the trap to 180 °C, The
desorber illustrated in Figure 2 meets these
design criteria.
5.2.4 The purge and trap system may be
assembled as a separate unit as illustrated in
Figure 3 or be coupled to a gas
chromatograph.
5.3 pH paper—Narrow pH range, about
3.5 to 5.5 (Fisher Scientific Short Range
Alkacid No. 2, #14-637-2 or equivalent).
5.4 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
columns, gases, detector, and strip-chart
recorder. A data system is recommended for
measuring peak areas.
5.4.1 Column 1—10 ft long x 2 mm ID
glass or stainless steel, packed with Porapak-
QS (80/100 mesh) or equivalent. This column
was used to develop the method performance
statements in Section 12. Guidelines for the
use of alternate column packings are
provided in Section 10.1.
5.4.2 Column 2—6 ft long x 0.1 in. ID glass
or stainless steel, packed with Chromosorb
101 (60/80 mesh) or equivalent.
5.4.3 Detector—Flame ionization detector.
This type of detector has proven effective in
the analysis of wastewaters for the
parameters listed in the scope (Section 1.1),
and was used to develop the method
performance statements in Section 12.
Guidelines for the use of alternate detectors
are provided in Section 10.1.
5.5 Syringes—5-mL, glass hypodermic
with Luerlok tip (two each).
5.6 Micro syringes—25-jiL, 0.008 in. ID
needle.
5.7 Syringe valve—2-way, with Luer ends
(three each).
-------
50 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
5.8 Bottle—15-mL screw-cap, with Teflon
cap liner.
S.9 Balance—Analytical, capable of
accurately weighing 0.0001 g.
ft Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.1.1 Reagent water can be generated by
passing tap water through a carbon filter bed
containing about 11b of activated carbon
(Filtrasorb-300, Calgon Corp., or equivalent).
6.1.2 A water purification system
(Millipore Super-Q or equivalent) may be
used to generate reagent water.
6.1.3 Regent water may also be prepared
by boiling water for 15 min. Subsequently,
while maintaining the temperature at 90 "C.
bubble a contaminant-free inert gas through
the water for 1 h. While still hot. transfer the
water to a narrow mouth screw-cap bottle
and seal with a Teflon-lined septum and cap.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL.
6.4 Hydrochloric acid (1 + 1)—Slowly, add
50 mL of concentrated HC1 (ACS) to 50 mL of
reagent water.
6.5 Trap Materials:
8.5.1 24-Diphenylene oxide polymer—
Tenax (60/80 mesh), chromatographic grade
or equivalent.
6.5.2 Methyl silicone packing—3% OV-1
on Chromotorb-W (60/80 mesh) or
equivalent.
6.6 Stock standard solutions—Stock
standard solutions may be prepared from
pure standard materials or purchased as
certified solutions. Prepare stock standard
solutions in reagent water using assayed
liquid*. Since acrolein and acrylonitrile are
lachrymators. primary dilutions of these
compounds 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.
6.6.1 Place about 9.8 mL of reagent water
into a 10-mL ground glass stoppered
volumetric flask. For acrolein standards the
reagent water must be adjusted to pH 4 to 5.
Weight the flask to the nearest 0.1 mg.
6.6.2 Using a 100-pL syringe, immediately
add two or more drops of assayed reference
material to the flask, then reweigh. Be sure
that the drops fall directly into the water
without contacting the neck of the flack.
8.6.3 Reweigh. dilute to volume, stopper.
then mix by inverting the flask several times.
Calculate the concentration in fig/fiL from
the net gain in weight. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
'standard. Optionally, stock standard
solutions may be prepared using the pure
standard material by volumetrically
measuring the appropriate amounts and
determining the weight of the material using
the density of the material. Commercially
prepared stock standards may be used at any
concentration if they are certified by the
manufactaurer or by an independent source.
6.6.4 Transfer the stock standard solution
into a Teflon-sealed screw-cap bottle. Store
at 4 'C and protect from light.
6.6.5 Prepare fresh standards daily.
6.7 Secondary dilution standards—Using
stock standard solutions, prepare secondary
dilution standards in reagent water that
contain the compounds of interest, either
singly or mixed together. The secondary
dilution standards should be prepared at
concentrations such that the aqueous
calibration standards prepared in Section
7.3.1 or 7.4.1 will bracket the working range of
the analytical system. Secondary dilution
standards should be prepared daily and
stored at 4 *C.
6.8 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Assemble a purge and trap system
that meets the specifications in Section 5.2.
Condition the trap overnight at 180 *C by
backflushing with an inert gas flow of at least
20 mL/min. Condition the trap for 10 min
once daily prior to use.
7.2 Connect the purge and trap system to
a gas chromatograph. The gas chromatograph
must be operated using temperature and flow
rate conditions equivalent to those given in
Table 1. Calibrate the purge and trap-gas
chromatographic system using either the
external standard technique (Section 7.3) or
the internal standard technique (Section 7.4).
7.3 External standard calibration
procedure:
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter by carefully adding 20.0 pL of
one or more secondary dilution standards to
100,500, or 1000 mL of reagent water. A 25-/iL
syringe with a 0.008 in. ID needle should be
used for this operation. One of the external
standards should be at a concentration near,
but above, the MDL and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector. These standards must
be prepared fresh daily.
7.3.2 Analyze each calibration standard
according to Section 10. and tabulate peak
height or area responses versus the
concentration of the standard. The results
can be used to prepare a calibration curve for
each compound. Alternatively, if the ratio of
response to concentration (calibration factor)
is a constant over the working range (< 10%
relative standard deviation. RSD), linearity
through the origin can be assumed and the
average ratio or calibration factor can be
used in place of a calibration curve.
7.4 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interference!. Became of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.4.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest as described in
Section 7.3.1.
7.4.2 Prepare a spiking solution containing
each of the internal standards using the
procedures described in Sections 6.6 and 6.7.
It is recommended that the secondary
dilution standard be prepared at a
concentration of 15 pg/mL of each internal
standard compound. The addition of 10 jiL of
this standard to 5.0 mL of sample or
calibration standard would be equivalent to
30 /ig/L.
7.4.3 Analyze each calibration standard
according to Section 10. adding 10 til of
internal standard spiking solution directly to
the syringe (Section 10.4). Tabulate peak
height or area responses against
concentration for each compound and
internal standard, and calculate response
factors (RF) for each compound using
Equation 1.
Equation 1.
RF= —
(A.HC,.)
(AJ(C,)
where:
A,=Response for the parameter to be
measured.
A*=Response for the internal standard.
0.=Concentration of the internal
standard.
C,=Concentration of the parameter to be
measured.
If the RF value over the working range is a
constant (<10* RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively.
the results can be used to plot a calibration
curve of response ratios, A./A* vs. RF.
7.5 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of a
QC check sample.
7.5.1 Prepare the QC check sample as
described in Section BA2.
7.5.2 Analyze the QC check sample
according to Section 10.
7.5.3 For each parameter, compare the
response (Q) with the corresponding
calibration acceptance criteria found in Table
2. If the responses for all parameters of
interest fall within the designated ranges.
analysis of actual samples can begin. If any
individual Q falls outside the range, a new
calibration curve, calibration factor, or RF
must be prepared for that parameter
according to Section 7.3 or 7.4.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 51
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography. the analyst is
permitted certain options (detailed in Section
10.1) to improve the separations or lower the
cost of measurements. Each time such a
modification is made to the method, the
analyst is required to repeat the procedure in
Section 8.2.
8.1.3 Each day, the analyst must analyze a
reagent water blank to demonstrate that
interferences from the analytical system are
under control.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
6.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of 25
Hg/mL in reagent water. The QC check
sample concentrate must be obtained from
the U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Prepare a QC check sample to
contain 50 /ig/L of each parameter by adding
200 ftL of QC check sample concentrate to
100 mL of reagent water.
8.2.3 Analyze four 5-mL aliquots of the
well-mixed QC check sample according to
Section 10.
8.2.4 Calculate the average recovery (X)
in jig/L, and the standard deviation of the
recovery (s) in /ig/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 3. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If either
s exceeds the precision limit or either X falls
outside the range for accuracy, the system
performance is unacceptable for that
parameter. Locate and correct the source of
the problem and repeat the lest for each
compound of interest.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If. as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 50 fig/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.2 Analyze one 5-mL sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
5-mL sample aliquot with 10 fiL of the QC
check sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 3. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.'
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
NOTE: The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 10 fiL of QC check sample concentrate
(Sections 8.2.1 or 8.3.2) to 5 mL of reagent
water. The QC check standard needs only to
contain the parameters that failed criteria in
the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is (he
true value of the standard concentration.
8.4.3 Compare the percent recovery (P.)
for each parameter with the corresponding
QC acceptance criteria found in Table 3.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sp to P+2sp. If P=90% and sD = 10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as gas chromatography with a dissimilar
column or mass spectrometer must be used.
Whenever possible, the laboratory should
analyze standard reference materials and
participate in relevant performance
evaluation studies.
ft Sample Collection. Preservation, and
Handling
9.1 All samples must be iced or
refrigerated from the time of collection until
analysis. If the sample contains free or
combined chlorine, add sodium thiosulfate
preservative (10 mg/40 mL is sufficient for up
to 5 ppm Clj) to the empty sample bottle just
prior to shipping to the sampling site. EPA
Methods 330.4 and 330.5 may be used for
measurement of residual chlorine.'Field test
kits are available for this purpose.
9.2 If acrolein is to be analyzed, collect
about 500 mL of sample in a clean glass
container. Adjust the pH of the sample to 4 to
5 using acid or base, measuring with narrow
range pH paper. Samples for acrolein
analysis receiving no pH adjustment must be
analyzed within 3 days of sampling.
9.3 Grab samples must be collected in
glass containers having a total volume of at
least 25 mL. Fill the sample bottle just to
overflowing 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. If preservative
-------
52 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
has been added, shake vigorously for 1 min.
Maintain the hermetic seul on the sample
bottle until time of analysis.
9.4 All samples must be analyzed within
14 day* of collection.1
10. Procedure
10.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
estimated retention times and MDL that can
be achieved under these conditions. An
example of the separations achieved by
Column 1 is shown in Figure 5. Other packed
columns, chromatographic conditions, or
detectors may be used if the requirements of
Section 8.2 are met.
10.2 Calibrate the system daily as
described in Section 7.
10.3 Adjust the purge gas (nitrogen or
helium) flow rate to 20 mL/min. Attach the
trap inlet to the purging device, and set the
purge and trap system to purge (Figure 3).
Open the syringe valve located on the
purging device sample introduction needle.
10.4 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 to just short of overflowing. Replace
the syringe plunger and compress the sample.
Open the syringe valve and vent any residual
air while adjusting the sample volume to 5.0
raL. 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 10.0 ul of the
internal standard spiking solution (Section
7.4.2), if applicable, through the valve bore
than close the valve.
10.5 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.
10.6 Close both valves and purge the
sample for 15.0 ± 0.1 min while heating at 85
±2'C.
10.7 After the 15-min purge time, attach
the trap to the chromatograph, adjust the
purge and trap system to the desorb mode
(Figure 4), and begin to temperature program
the gai chromatograph. Introduce the trapped
materials to the GC column by rapidly
heating the trap to 180 *C while backflushing
the trap with an inert gas between 20 and 60
mL/min for 1.5 min.
10.8 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 reagent water.
10.9 After desorbing the sample for 1.5
min. recondition the trap by returning the
purge and trap system to the purge mode.
Wail 15 s then close the syringe valve on the
purging device to begin gas flow through the
trap. The trap temperature should be
maintained at 210 *C. After approximately 7
min, turn off the trap heater and open the
syringe valve to stop the gas flow through the
trap. When the trap is cool, the next sample
can be analyzed.
10.10 Identify the parameters in the
sample by comparing the retention times of
the peaks in the sample chromalogram with
those of the peaks in standard
chromatograms. The width of the retention
time window used to make identifications
should be based upon measurements of
actual retention time variations of standards
over the course of a day. Three times the
standard deviation of a retention time for a
compound can be used to calculate a
suggested window size; however, the
experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
11. Calculations
11.1 Determine the concentration of
individual compounds in the sample.
11.1.1 If the external standard calibration
procedure is used, calculate the
concentration of the parameter being
measured from the peak response using the
calibration curve or calibration factor
determined in Section 7.3.2.
11.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.4.3 and Equation 2.
Equation 2.
Concentration (>tg/L)=
(A.KCJ
' (AJ(RF)
where:
A.=Response for the parameter to be
measured.
AH » Response for the internal standard.
Q.=Concentration of the internal
standard.
11.2 Report results in jig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
mult*.
12. Method Performance
12.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.' The MDL concentrations listed in
Table 1 were obtained using reagent water.'
The MDL actually achieved in a given
analysis will vary depending on instrument
sensitivity and matrix effects.
12.2 This method is recommended for the
concentration range from the MDL to
1.000 x MDL. Direct aqueous injection
techniques should be used to measure
concentration levels above 1,000 x MDL
12.3 In a single laboratory (Battelle-
Columbus). the average recoveries and
standard deviations presented in Table 2
were obtained.'Seven replicate samples
were analyzed at each spike level.
References
1.40 CFR Part 136, Appendix B.
2. Bellar. T.A.. and Lichtenberg. J.|.
"Determining Volatile Organic* at
Microgram-per-Litre-Levels by Gas
Chromatography," Journal American Water
Works Association. 66. 739 (1974).
3. Kerns, E.H., et al. "Determination of
Acrolein and Acrylonitrile in Water by
Heated Purge and Trap Technique," 1980,
U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268.
4. "Carcinogen*—Working With
Carcinogen*," Department of Health,
Education, and Welfare, Public Health
Service, Center for Disease Control, National
Institute for Occupational Safety and Health.
Publication No. 77-208. August 1977.
5. "OSHA Safety and Health Standards,
General Industry," (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2208 (Revised.
January 1976).
8. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication, Committee on Chemical Safety,
3rd Edition. 1979.
7. Provost, L.P., and Elder, R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory, 15, 58-63 (1983).
8. "Method* 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric. DPD) for
Chlorine. Total Residual," Methods for
Chemical Analysis of Water and Wastes.
EPA-800/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, March 1979.
9. "Evaluation of Method 603," Final report
for EPA Contract 68-03-1760 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
PorMnotor
nt*nioii
Column 1
106
12.7
mm (min)
Column Z
S.2
9.S
Moswd
dotocHuii
M|J*
07
05
Column 1 oondNon*iPorapi
OS in/100 fflMh) pasted m«10«x2mmO gtow or ilartin «Ml column wNn hotum cantor OM « 30 mL/min ton rate. Column to
uring dMorpHon), twi hMted m rapMy w pootHo to ISO "C and hold tor 20 min; column btteout«190 "C tor 10 mm.*
orb 101
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 53
TABLE 2.—SINGLE LABORATORY ACCURACY AND PRECISION—METHOD 603
Parameter
Sample
matrix
RW
RW
POTW
POTW
IW
IW
RW
RW
POTW
POTW
IW
IW
Spike
cone
(cfl'U
50
50.0
5.0
50.0
5.0
too.o
5.0
50.0
20.0
100.0
10.0
100.0
Average
recovery
0»g/L)
52
51.4
4.0
44.4
0.1
9.3
42
•51.4
20.1
101.3
9.1
104.0
Standard ,
deviation •
(lig'U '
02
0.7 i
0.2 '
0.8
01 •
1.1
02 '
1.5
0.8
1.5 :
o.e
"i
Average
percent
recovery
104
103
80
69
2
9
84
103
too
101
91
104
RW = Reagent water.
POTW = Prechlorination secondary effluent from a municipal sewage treatment plant.
IW = Industrial wastewater containing an unidentified acrolein reactant.
TABLE 3.—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 603 •
Parameter
Acn/ionit '1
ACryioni
Range for
O G»g/L)
45.9-54.1
41.2-58.6
Limit
for S
0»g/U
4.6
9.9
Range lor X
fj»g'L)
42.9-60.1
33.1-69.9
Range lor
P. P. (SI
88-118
71-135
Q= Concentration measured in QC check sample, in jig/L (Section 7.5.3).
« = Standard deviation of four recovery measurements, in jig/L (Section 8.2.4).
.. =Average recovery for four recovery measurements, in jig/L (Section 8.2.4).
P, P. = Percent recovery measured (Section 8.3.2. Section 8.4.2).
•-Criteria were calculated assuming a QC check sample concentration of 50 pg/L.
BILLING CODE 6560-50-41
-------
54
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
OPTIONAL
FOAM JW- EXIT KIN.
TRAP /^^\ 0. 0
1 — 14MM 0. D
^(lS INLET H IN.
y-^- o. D.
< I
1 1
;,' ^ SAMPLE INLET
,'.' A— 2 WAt SYRINGE VALVE
' ' — 17C1I 20 GAUGF SYRINGE NEEDLE
'/4 IN.
0 D. EXIT
6MM. 0. 0 RUBBER SEPTUM
. 0. 0
-INLET
% IN. 0. 0
1/16 IN. 00
y STAINLESS STEEL
13X MOLECULAR
SIEVE PURGE
GAS FILTER
PURGE GAS
FLOW
CONTROL
10MM GLASS FRIT
MEDIUM POROSITY
Figure 1. Purging d«vic«.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 55
PACKING PROCEDURE
GLASS suu
WOOL81"1
CONSTRUCTION
TENAI 23CU
3XOV-1
§
1CMJ
GLASS WOOL gJJ1 £
TRAP INLET
COMPRESSION FITTING
NUT AND FERRULES
14FT.7A/FOOT RESISTANCE
WIRE WRAPPED SOLID
THERMOCOUPLE/
CONTROLLER
SENSOR
ELECTRONIC
TEMPERATURE
CONTROL
AND
PYROMETER
TUBING 25CM.
.105 IN. I.D.
(1125 IN. O.D.
STAINLESS STEEL
Figure 2. Trap packing* and construction to include
desorb capability.
-------
56
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
IMMOtlCUUUI
•MVtfkVM
WATM SATM
Figure 3. Purge and trap system-purge mode.
IMMOitCtMAK
MtvtntTM
wArm MTH
Figure 4. Purge end trap system-desorb mode-.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
57
Column: Porapak QS
Program: 110°C for 15 mm. rapidly
heated to 150°C
Detector: Flame lonization
I I I I l • I l I i I
1.5 30 45 60 75 9.0 10.5 120,135 150
REftNTlON TIME. MIN.
Figure 6. Ga» chromatogram of acrolein and acrylonitriU.
BILLING CODE 6560-50-C
-------
58 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 604—Phenols
/. Scope and Application
1.1 This method covers the determination
of phenol and certain substituted phenols.
The following parameters may be determined
by this method:
Parameter
STORET
No
CAS No
4-Chkxo-3-m«tHyl(>henoi
2
Chloroptwool
2.4-OicMoroptienol
2 4.Omethytprienol
4-Onitropfwnol
2-Metry-4.6-dntroprienol
2
Nitroptwnot
4 • Nttropntnol
P«nt*chtoroptwnol
Pi
2.4.6-TncNoroprwnol
34452
34586
34601
34606
34616
34657
34591
34646
39032
34694
34621
•
59-50-7
95-57-8
120-83-2
105-67-9
51-28-5
534-52-1
88-75-5
100-02-7
87-86-5
108-95-2
88-06-2
1.2 This is a flame ionization detector gas
chromatographic (FIDCC) method applicable
to the determination of the compounds listed
above in municipal and industrial discharges
as provided under 40 CFR 136.1. When this
method is used to analyze unfamiliar samples
for any or all of the compounds above,
compound identifications should be
supported by at least one additional
qualitative technique. This method describes
analytical conditions for derivatization.
cleanup, and electron capture detector gas
chromatography (ECDGC) that can be used to
confirm measurements made by FIDGC.
Method 625 provides gas chromatograph/
mass spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
1.3 The method detection limit (MDL,
defined in Section 14.1)' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed.
depending upon the nature of interferences in
the sample matrix. The MDL listed in Table 1
for each parameter was achieved with a
flame ionization detector (FID). Comparable
results were achieved when the
derivatization cleanup and electron capture
detector (ECD) were employed (Table 2).
1.4 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L. is acidified and extracted
with methylene chloride using a separator^
funnel. The methylene chloride extract is
dried and exchanged to 2-propanol during
concentration to a volume of 10 mL or less.
The extract is separated by gas
chromatography and the phenols are then
measured with an FID.*
2.2 A preliminary sample wash under
basic conditions can be employed for
samples having high general organic and
organic base interferences.
2.3 The method also provides fur a
derivalization and column chromatography
cleanup procedure to aid in the elimination nf
interferences.13The derivatives are analyzed
by ECDGC.
— .7. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section B.I .3.
3.1.1 Glassware must be scrupulously
cleaned.4 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The
derivatization cleanup procedure in Section
12 can be used to overcome many of these
interferences, but unique samples may
require additional cleanup approaches to
achieve the MDL listed in Tables 1 and 2.
3.3 The basic sample wash (Section 10.2)
may cause significantly reduced recovery of
phenol and 2,4-dimethylphenol. The analyst
must recognize that results obtained under
these conditions are minimum
concentrations.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this mothod 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
OSIIA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling shoots should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified s~' for the information of
analyst.
4.2 Special care should be taken in
handling pcntafluorobenzyl bromide, which is
a lachrymator. and 18-crown-6-ethcr. which is
highly toxic.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 'C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol. followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (AH specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L. with Teflon
stopcock.
5.2.2 Drying column—Chromatographic
column, 400 mm long x 19 mm ID. with coarse
frit filter disc.
5.2.3 Chromatographic column—100 mm
long x 10 mm ID, with Teflon stopcock.
5.2.4 Concentrator tube. Kuderna-
Danish—10-mL, graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask, Kudema-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column. Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Snyder column, Kuderna-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
5.2.8 Vials—10 to 15-mL. amber glass,
with Teflon-lined screw cap.
5.2.9 Reaction flask—15 to 25-mL round
bottom flask, with standard tapered joint.
fitted with a water-cooled condenser and U-
shaped drying tube containing granular
calcium chloride.
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Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations 59
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2"C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighting 0.0001 g.
5.6 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
columns, gases, detector, and strip-chart
recorder. A data system is recommended for
measuring peak areas.
5.6.1 Column for underivatized phenols—
1.8 m long x 2 mm ID glass, packed with 1%
SP-1240DA on Supelcoport (80/100 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
column packings are provided in Section 11.1.
5.6.2 Column for derivatized phenols—1.8
m long x 2 mm ID glass, packed with 5% OV-
17 on Chromosorb W-AW-DMCS (80/100
mesh) or equivalent. This column has proven
effective in the analysis of wastewaters for
derivatization products of the parameters
listed in the scope (Section 1.1), and was used
to develop the method performance
statements in Section 14. Guidelines for the
use of alternate column packings are
provided in Section 11.1.
5.6.3 Detectors—Flame ionization and
electron capture detectors. The FID is used
when determining the parent phenols. The
ECD is used when determining the
derivatized phenols. Guidelines for the use of
alternatve detectors are provided in Section
11.1.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL.
6.3 Sodium hydroxide solution (1 N)—
Dissolve 4 g of NaOH (ACS) in reagent water
and dilute to 100 mL.
6.4 Sodium sulfate—(ACS) Granular.
anhydrous. Purify by heating at 400'C for 4 h
in a shallow tray.
6.5 Sodium thiosulfate—(ACS) Granular.
6.6 Sulfuric acid (1 + 1)—Slowly, add 50
mL of HjSC-4 (ACS, sp. gr. 1.84) to 50 mL of
reagent water.
6.7 Sulfuric acid (1 N)—Slowly, add 58 mL
of HiSO< (ACS, sp. gr. 1.84) to reagent water
and dilute to 1 L.
6.8 Potassium carbonate—(ACS)
Powdered.
6.9 Pentafluorobenzyl bromide (a-
Bromopentafluorotoluene)—97% minimum
purity. Note: This chemical is a lachrymator.
(See Section 4.2.)
6.10 18-crown-6-ether (1.4,7.10,13,16-
Hexaoxacyclooctadecane)—98% minimum
purity. Note: This chemical is highly toxic.
6.11 Derivatization reagent—Add 1 mL of
pentafluorobenzyl bromide and 1 g of 18-
crown-6-ether to a 50-mL volumetric flask
and dilute to volume with 2-propanol. Prepare
fresh weekly. This operation should be
carried out in a hood. Store at 4 *C and
protect from light.
6.12 Acetone, hexane. methanol.
methylene chloride. 2-propanol. toluene—
Pesticide quality or equivalent.
6.13 Silica gel—100/200 mesh. Davison.
grade-923 or equivalent. Activate at 130 'C
overnight and store in a desiccator.
6.14 Stock standard solutions (1.00 jig/
pL)—Stock standard solutions may be
prepared from pure standard materials or
purchased as certified solutions.
6.14.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in 2-propanol
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.14.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 °C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.14.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.15 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 To calibrate the FIDGC for the
anaylsis of underivatized phenols, establish
gas chromatographic operating conditions
equivalent to those given in Table 1. The gas
chromatographic system can be calibrated
using the external standard technique
(Section 7.2) or the internal standard
technique (Section 7.3).
7.2 External standard calibration
procedure for FIDGC:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
2-propanol. One of the external standards
should be at a concentration near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using injections of 2 to 5 jil. analyze
each calibration standard according to
Section 11 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (<10% relative standard
deviation, RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure for F1DCC—To use this approach,
the analyst must select one or more internal
standards thut are similar in analytical
behavior to (he compounds of interest. Thn
analyst must further demonstrate that the
measurement of the internal standard is not
affected by method or matrix interferences.
Because of these limitations, no internal
standard can be suggested that is applicable
to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with 2-propanol. One of the
standards should be at a concentration near.
but above, the MDL and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.3.2 Using injections of 2 to 5 u.L, analyze
each calibration standard according to
Section 11 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
Rr_ (A.HC..)
(Ata)(C.)
where:
A, = Response for the parameter to be
measured.
Ato = Response for the internal standard.
Cu=Concentration of the internal standard
(M8/L).
C,=Concentration of the parameter to be
measured (fig/L).
If the RF value over the working range is a
constant «10% RSD). the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios. A,/AU, vs. RF.
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%. a new
calibration curve must be prepared for that
compound.
7.5 To calibrate the ECDGC for the
analysis of phenol derivatives, establish gas
chromatographic operating conditions
equivalent to those given in Table 2.
7.5.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
2-propanol. One of the external standards
should be at a concentration near, but above,
the MDL (Table 2) and the other
concentrations should correspond to the
expected range of concentrations found in
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60 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
real samples or should define the working
range of the detector.
7.5.2 Each time samples are to be
derivatized. simultaneously treat a 1-mL
aliquot of each calibration standard as
described in Section 12.
7.5.3 After derivatization, analyze 2 to 5
jiL of each column eluate collected according
to (he method beginning in Section 12.8 and
tabulate peak height or area responses
against the calculated equivalent mass of
underivatized phenol injected. The results
can be used to prepare a calibration curve for
each compound.
7.6 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that it generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst mutt make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is establithed
a* described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography. the analyst it
permitted certain options (detailed in
Sections 10.8 and 11.1) to improve the
separations or lower the coat of
measurements. Each time such a modification
it made to the method, the analyst it required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any tamplet the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical tyttem and glassware are under
control. Each time a set of samples it
extracted or reagents are changed a reagent
water blank mutt be processed at a
safeguard against laboratory contamination.
8.1.4 The laboratory mutt, on an ongoing
basis, spike and analyze a minimum of 10% of
all tamplet to monitor and evaluate
laboratory data quality. This procedure it
described in Section 8.3.
8.1.5 The laboratory mutt on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system it in
control. This procedure it described in
Section 8.4. The frequency of the check
standard analyses it equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from tamplet (Section 8J)
meet all specified quality control criteria.
8.1.0 The laboratory mutt maintain
performance record* to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of
100 ng/mL in 2-propanol. The QC check
sample concentrate must be obtained from
the U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at a concentration of 100 ng/L by
adding 1.00 mL of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in ng/L and the standard deviation of the
recovery (t) in pg/L, for each parameter using
the four results.
8.2.5 For each parameter compare t and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 3. If t and X for all parameters
of interest meet the acceptance criteria, the
tystem performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the tyttem performance it
unacceptable for that parameter.
Note.—The large number of parameters in
Talbe 3 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameter! are
analyzed.
8.2.6 When one or more of the parameters
tested fail at leatt one of the acceptance
criteria, the analytt mutt proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.8.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.2.
8.2.6.2 Beginning with Section 8.2.2, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement tyttem. If thit occurs, locate
and correct the source of the problem and
repeat the tett for all compound* of interest
beginning with Section S.&2.
8.3 The laboratory mutt on an ongoing
basis, spike at leatt 10% of the samples from
each sample lite being monitored to assess
accuracy. For laboratories analyzing one to
ten sample* per month, at least one spiked
sample per month it required.
8.3.1 The concentration of the spike in the
sample should be determined at follow*:
8.3.1.1 If, a* in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2. whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 100 ftg/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2. whichever
concentration would be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any, or, if
none. (2) the larger of either 5 times higher
than the expected background concentration
or 100 ug/L.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) at 100(A-B)*/T, where T i* the
known true value of the spike.
84 J Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 3. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentration*, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.'If
spiking was performed at a concentration
lower than 100 jig/L, the analyst must use
either the QC acceptance criteria in Table 3,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter (1) Calculate
accuracy (X') using the equation in Table 4.
substituting the spike concentration (T) for C:
(2) calculate overall precision (S') using the
equation in Table 4. substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) ±2.44(100 S'/
T)%.»
8.3.4 If any individual P fall* ouUide the
designated range for recovery, that parameter
ha* failed the acceptance criteria. A check
standard containing each parameter the!
failed the criteria must be analyzed a*
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 84, a QC
check standard containing each parameter
that failed mutt be prepared and analyzed.
Note.—The frequency for the required
analyst* of a QC check standard will depend
upon the number of parameter* being
simultaneously tested, the complexity of the
•ample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 61
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%. where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P.)
for each parameter with the corresponding
QC acceptance criteria found in Table 3.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sp to P+2sp. If P = 90% and Sp=10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy •
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6. It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
ft Sample Collection, Preservation, and
Handling
9.1 Crab 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 *C from the time of collection
until extraction. Fill the sample bottles and, if
residual chlorine is present, add 80 mg of
sodium thiosulfate per liter of sample and
mix well. EPA Methods 330.4 and 330.5 may
be used for measurement of residual
chlorine.10 Field test kits are available for this
purpose.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.'
10. Sample Extraction
10.1 Mark the water meniscus on the side
of sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separately funnel.
10.2 For samples high in organic content.
the analyst may solvent wash the sample at
basic pH as prescribed in Sections 10.2.1 and
10.2.2 to remove potential method
interferences. Prolonged or exhaustive
contact with solvent during the wash may
result in low recovery of some of the phenols.
notably phenol and 2,4-dimethylphenol. For
relatively clean samples, the wash should be
omitted and the extraction, beginning with
Section 10.3. should be followed.
10.2.1 Adjust the pH of the sample to 12.0
or greater with sodium hydroxide solution.
10.2.2 Add 60 mL of methylene chloride to
the sample by shaking the funnel for 1 min
with periodic venting to release vapor
pressure. Discard the solvent layer. The wash
can be repeated up to two additional times if
significant color is being removed.
10.3 Adjust the sample to a pH of 1 to 2
with sulfuric acid.
10.4 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the runnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.5 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.6 Assemble a Kuderna-Oanish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.7 Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate,
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.8 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but (he chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL.
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.9 Increase the temperature of the hot
water bath to 95 to 100 "C. Remove the
Synder column and rinse the flask and its
lower joint into the concentrator tube with 1
to 2 mL of 2-propanol. A 5-mL syringe is
recommended for this operation. Attach a
two-ball micro-Snyder column to the
concentrator tube and prewet the column by
adding about 0.5 mL of 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 to 10 min. At the proper
rate of distillation the balls of the column will
actively chatter but the chambers will not
flood. When the apparent volume of liquid
reaches 2.5 mL, remove the K-D apparatus
and allow it to drain and cool for at least 10
min. Add an additional 2 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 and cool for at least 10 min.
10.10 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 refrigerated
at 4 *C if further processing will not be
performed immediately. If the extract will be
stored longer than two days, it should be
transferred to a Teflon-sealed screw-cap vial.
If the sample extract requires no further
cleanup, proceed with FIDCC analysis
(Section 11). If the sample requires further
cleanup, proceed to Section 12.
10.11 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.
11. Flame lonization Detector Cos
Chromatography
11.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. An example
of the separations achieved by this column is
shown in Figure 1. Other packed or capillary
(open-tubular) columns, chromatographic
conditions, or detectors may be used if the
requirements of Section 8.2 are met.
11.2 Calibrate the system daily as
described in Section 7.
11.3 If the internal standard calibration
procedure is used, the internal standard must
be added to the sample extract and mixed
thoroughly immediately before injection into
the gas chromatograph.
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82 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
11.4 Inject 2 to 5 pL of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique." Smaller (1.0 jiL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 uL. and the resulting peak
size in area or peak height units.
11.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound may be used
to calculate a suggested window size;
however, the experience of the analyst
should weigh heavily in the interpretation of
chromatograms.
11.6 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
11.7 If the measurement of the peak
response is prevented by the presence of
interferences, an alternative gas
chromatographic procedure is required.
Section 12 describes a derivatization and
column chromatographic procedure which
has been letted and found to be a practical
means of analyzing phenols in complex
extracts.
12. Derivatization and Electron Capture
Detector Gas Chromatography
12.1 Pipet a 1.0-mL aliquot of the 2-
propanol solution of standard or sample
extract into a glass reaction vial. Add IX) mL
of derivatizing reagent (Section 6.11). This
amount of reagent is sufficient to derivatize a
solution whose total phenolic content does
not exceed 0.3 mg/mL
1&2 Add about 3 mg of potassium
carbonate to the solution and shake gently.
12.3 Cap the mixture and heat it for 4 h at
80 'C in a hot water bath.
12.4 Remove the solution from the hot
water bath and allow it to cool.
12.5 Add 10 mL of hexane to the reaction
flask and shake vigorously for 1 min. Add 3.0
mL of distilled, deionized water to the
reaction flask and shake for 2 min. Decant a
portion of the organic layer into a
concentrator tube and cap with a glass
stopper.
12.6 Place 4.0 g of silica gel into a
chromatographic column. Tap the column to
settle the silica gel and add about 2 g of
anhydrous sodium sulfate to the top.
12.7 Preelute the column with 0 mL of
hexane. Discard the eluate and just prior to
exposure of the sodium sulfate layer to the
air, pipet onto the column 2.0 mL of the
hexane solution (Section 12.5) that contains
the derivatized sample or standard. Elute the
column with 10.0 mL of hexane and discard
the eluate. Bute the column, in order, with:
10.0 mL of 15% toluene in hexane (Fraction 1):
10.0 mL of 40* toluene in hexane (Fraction 2);
10.0 mL of 75% toluene in hexane (Fraction 3);
and 10.0 mL of 15% 2-propanol in toluene
(Fraction 4). All elution mixtures are
prepared on a volume:volume basis. Elution
patterns for the phenolic derivatives are
shown in Table 2. Fractions may be
combined as desired, depending upon the
specific phenols of interest or level of
interferences.
12.8 Analyze the fractions by ECDGC.
Table 2 summarizes the recommended
operating conditions for the gas
chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. An example
of the separations achieved by this column is
shown in Figure 2.
12.9 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
Section 7.5.
12.10 Inject 2 to 5 pL of the column
fractions into the gas chromatograph using
the solvent-flush technique. Smaller (1.0 pL)
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 or peak height units. If the peak
response exceeds the linear range of the
system, dilute the extract and reanalyze.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample analyzed
by FIDGC (without derivatization) as
indicated below.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration
where:
A=Amount of material injected (ng).
V|=Volume of extract injected (u.L).
V,—Volume of total extract (uL).
V,=Volume of water extracted (mL).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3.
Concentration (ng/L) =
Concentration (ug/L)=
(AJBJ
(AJ(RF)(V.)
where:
A,=Response for the parameter to be
measured.
A*=Response for the internal standard.
I,—Amount of internal standard added to
each extract (fig).
V.=Volume of water extracted (L).
13.2 Determine the concentration of
individual compounds in the sample analyzed
by derivatization and ECDGC according to
Equation 4.
Equation 4.
(V,)(V.HC)(E)
where:
A = Mass of underivatized phenol
represented by area of peak in sample
chromatogram, determined from
calibration curve in Section 7.5.3 (ng).
V, = Volume of eluate injected (jiL).
V,=Total volume of column eluate or
combined fractions from which V, was
taken (uL).
V,=Volume of water extracted in Section
10.10 (ml).
B=Total volume of hexane added in
Section 12.5 (mL).
C = Volume of hexane sample solution
added to cleanup column in Section 12.7
(mL).
D=Total volume of 2-propanol extract
prior to derivatization (mL).
E=Volume of 2-propanol extract carried
through derivatization in Section 12.1
(mL).
13.3 Report results in ug/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Tables 1 and 2 were obtained using reagent
water."Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked as six concentrations
over the range 12 to 450 jig/L " Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships for a flame ionization detector
are presented in Table 4.
References
1. 40 CFR Part 130, Appendix B.
2. "Determination of Phenols in Industrial
and Municipal Wastewaters," Report for EPA
Contract 68-03-2625 (In preparation).
3. Kawahara, F. K. "Microdetermination of
Derivatives of Phenols and Mercaptans by
Means of Electron Capture Gas
Chromatography," Analytical Chemistry. 40,
1009 (1968).
4. ASTM Annual Book of Standards, Part
31, D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents."
American Society for Testing and Materials,
Philadelphia.
5. "Carcinogens—Working With
Carcinogens," Department of Health.
Education, and Welfare. Public Health
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations 63
Service. Center for Disease Control. National
Institute for Occupational Safety and Health,
Publication No. 77-206. August 1977.
6. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910),
Occupational Safety and Health
Administration. OSHA 2206 (Revised,
January 1976).
7. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication, Committee on Chemical Safety.
3rd Edition, 1979.
8. Provost. L. P., and Elder. R. S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
9. ASTM Annual Book of Standards, Part
31. D3370-76. "Standard Practices for
Sampling Water." American Society for
Testing and Materials. Philadelphia.
10. "Methods 330.4 (Titrimetric. DPD-FAS)
and 330.5 (Spectrophotometric, DPD) for
Chlorine, Total Residual," Methods for
Chemical Analysis of Water and Wastes,
EPA-600/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, March 1979.
11. Burke.). A. "Cas Chromatography for
Pesticide Residue Analysis: Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists. W. 1037 (1965).
12. "Development of Detection Limits. EPA
Method 604. Phenols," Special letter report
for EPA Contract 68-03-2625, U.S.
Environmental Protection Agency.
Environmental Monitoring and Support
Laboratory. Cincinnati. Ohio 45268.
13. "EPA Method Validation Study 14.
Method 604 (Phenols)." Report for EPA
Contract 68-03-2625 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Parameter
2-Nitrophenot
Phenol
2 4-D»chlofophenol
4-tttrophenol -. -\
Retention
time (nun)
1 70
200
301
403
4 30
60S
750
1000
1024
1242
2425
Method
detection
limit (Mg/L)
0 31
045
0 14
0 32
0 39
0 64
0 36
13 0
160
7 4
26
Column conditions: Supelcoport (60/100 mesh) coated with 1% SP-12400A packed in I 1.8 m long i 2 mm ID glass column with nitrogen earner gas at 30 mL/rnn flow rate. Column
temperature was 80 'C at injection, programmed immediately at a •C/min to 150 C final temperature. MDL were determined with an FID.
TABLE 2.— SILICA GEL FRACTIONATION AND ELECTRON CAPTURE GAS CHROMATOGRAPHY OF PFBB DERIVATIVES
Parent compound
2-Nrtropnenol „ . .
Phenol
2 4-Oimethylphenol
2 4-Dichkxophenot
2 4 6-Trichkxophenol ....
4-Chkxo-3-methylphenol •
Pentachlorophenol •
4.
(percent)
99-122
38-126
44-119
24-118
30-136
12-145
43-117
13-110
36-134
23-108
53-119
»—Standard deviation of four recovery measurements, in jig/L (Section 8.2.4).
X—Average recovery for (our recovery measurements, in /ig/L (Section 8.2.4).
P. P.—Percent recovery measured (Section 8.3.2, Section 8.4.2).
NOTE.—These criteria are based directly upon the method performance data in Table 4. Where necessary, the Bmits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 4.
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64
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 4.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 604
Parameter
40*0f0.3 mrthytohral
yj^tnmntvuwl^^^^^
2 4-OMrephtno4
. f^frt^lyii
PJfrrtayfrfQfflpfE+ftfH
2 4 6-TrichtofOptMMfH
Accuracy, M
rvcovwy, X'
0»0>U
0 87C-1 97
063C-084
061C + 046
062C-1 64
084C-101
oaoc-i sa
0 81C-0 76
046C 4-018
083C+ 207
043C + 011
0660440
Stftgl«An*ly|t
prtcitwn, tV
o*'u
0 llJt-021
0 188 * 020
0 17X-002
030S-OM
0 ISX-t- 1 25
027&-1 IS
0 15^4.044
0 17X*243
0 22&-0 56
020X-OM
0 10X * 0 S3
OvwtH
pftciwjn. S
(Mfl/D
0 16X +141
02lX +0 75
0 25X + 0 46
0 19Jt + 585
0 14& + 364
0 13X + 240
X'-Enpected recovery tor one or more measurements ol a sample containing a concentration of C. in ug/L
s, -Expected single analyst standard deviation ol measurements at an average concentration found of X. in pg/l.
S *E«pec!ednterteooratory standard deviation ol measurements at an average concentration found ol X. ir -"
• True value tor the concentration, in pg/l.
'Average recovery found tor measurements of samples containing a concentration of C. in ,
•NJ
I cooe «MO-SO-*I
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
65
X 2
O . s COLUMN: IS SP-1240DA ON SUPELCOPORT
zi Q PROGRAM: 80°C AT INJECTION. IMMEDIATE fC/IWH TO 150°C
OS 4
" ~
••
DETECTOR: FLAME IONIZATION
O
_, 5
3§ | ^|
s i o o
*"§ « §2
" y P^
VJ
V.
S
3
5
8 12 16 20 24 78
RETENTION TIME. MIN.
Figure 1 Gas chromatogram of phenols-
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66
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
COLUMN 5% OV-17 ON CHROMOSORB W AW OMCS
TEMPERATURE: 200'C.
DETECTOR: aECTRON CAPTURE
A
I 12 16 20 24
RETENTION TIME. MIN.
Figure 2. Gas chromatogram of PFB derivatives of phenols.
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
67
Method 60S—Benzidines
1. Scope and Application
1.1 This method covers the determination
of certain benzidines. The following
parameters can be determined by this
method:
Parameter
S.S'-Dichkxobenjkline
Sloret No
39120
34631
CAS No.
92-87-5
91-94-1
1.2 This is a high performance liquid
chromatography (HPLC) method applicable to
the determination of the compounds listed
above in municipal and industrial discharges
as provided under 40 CFR 136.1. When this
method is used to analyze unfamiliar samples
for the compounds above, identifications
should be supported by at least one
additional qualitative technique. This method
describes electrochemical conditions at a
second potential which can be used to
confirm measurements made with this
method. Method 625 provides gas
chromatograph/mass spectrometer (CC/MS)
conditions appropriate for the qualitative and
quantitative confirmation of results for the
parameters listed above, using the extract
produced by this method.
1.3 The method detection limit (MDL
defined in Section 14.1) ' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of the
interferences in the sample matrix.
1.4 Any modification of this method.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the use of HPLC
instrumentation and in the interpretation of
liquid chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is extracted with
chloroform using liquid-liquid extractions in a
separatory funnel. The chloroform extract is
extracted with acid. The acid extract is then
neutralized and extracted with chloroform.
The final chloroform extract is exchanged to
methanol while being concentrated using a
rotary evaporator. The extract is mixed with
buffer and separated by HPLC. The benzidine
compounds are measured with an
electrochemical detector.*
2.2 The acid back-extraction acts as a
genera] purpose cleanup to aid in the
elimination of interferences.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in chromatograms. All
of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.1 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 'C
for 15 to 30 min. Some thermally stable
materials may not be eliminated by this
treatment. Solvent rinses with acetone and
pesticide quality hexane may be substituted
for the muffle furnace heating. Volumetric
ware should not be heated in a muffle
furnace. After drying and cooling, glassware
should be sealed and stored in a clean
environment to prevent any accumulation of
dust or other contaminants. Store inverted or
capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedures that are inherent in the extraction
step are used to overcome many of these
interferences, but unique samples may
require additional cleanup approaches to
achieve the MDL listed in Table 1.
3.3 Some dye plant effluents contain large
amounts of components with retention times
closed to benzidine. In these cases, it has
been found useful to reduce the electrode
potential in order to eliminate interferences
and still detect benzidine. (See Section 12.7.)
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health harzard. 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 data handling sheets should also be
made available to all personnel involved in .
the chemical analysis. Additional references
to laboratory safety are available and have
been identified *'for the information of the
analyst.
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: benzidine and 3,3'-
dichlorobenzidine. Primary standards of
these toxic compounds should be prepared in
a hood. A NIOSH/MESA approved toxic gas
respirator should be worn when the analyst
handles high concentrations of these toxic
compounds.
4.3 Exposure to chloroform should be
minimized by performing all extractions and
extract concentrations in a hood or other
well-ventiliated area.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt,
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylnne
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4°C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested):
5.2.1 Separatory funnels—2000,1000, and
250-mL. with Teflon stopcock.
5.2.2 Vials—10 to 15-mL. amber glass,
with Teflon-lined screw cap.
5.2.3 Rotary evaporator.
5.2.4 Flasks—Round bottom. 100-mL, with
24/40 joints.
5.2.5 Centrifuge tubes—Conical,
graduated, with Teflon-lined screw caps.
5.2.6 Pipettes—Pasteur, with bulbs.
5.3 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.4 High performance liquid
chromatograph (HPLC)—An analytical
system complete with column supplies, high
pressure syringes, detector, and compatible
recorder. A data system is recommended for
measuring peak areas and retention times.
5.4.1 Solvent delivery system—With pulse
damper, Altex 110A or equivalent.
5.4.2 Injection valve (optional)—Waters
U6K or equivalent.
5.4.3 Electrochemical detector—
Bioanalytical Systems LC-2A with glassy
carbon electrode, or equivalent. This detector
has proven effective in the analysis of
wastewaters for the parameters listed in the
scope (Section 1.1), and was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
detectors are provided in Section 12.1.
5.4.4 Electrode polishing kit—Princeton
Applied Research Model 9320 or equivalent.
5.4.5 Column—Lichrosorb RP-2, 5 micron
particle diameter, in a 25 cm X 4.6 mm ID
stainless steel column. This column was used
to develop the method performance
statements in Section 14. Guidelines for the
use of alternate column packings are
provided in Section 12.1.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
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Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
not observed at the MDL of the parameters of
interest.
6.2 Sodium hydroxide solution (5 N)—
Dissolve 20 g of NaOH (ACS) in reagent
water and dilute to 100 mL
8.3 Sodium hydroxide solution (1 Mi-
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 1 L.
6.4 Sodium thiosulfate—(ACS) Granular.
6.5 Sodium tribasic phosphate (0.4 M)—
Dissolve 160 g of trisodium phosphate
decahydrate (ACS) in reagent water and
dilute to 1 L.
6.6 Sulfuric acid (1 +1)—Slowly, add 50
mL of H,SO, (ACS. sp. gr. 1.84) to 50 mL of
reagent water.
6.7 Sulfuric acid (1 M)—Slowly, add 58
mL of HjSO, (ACS. sp. gr. 1.84) to reagent
water and dilute to 1 L.
6.8 Acetate buffer (0.1 M. pH 4.7)—
Dissolve 5.8 mL of glacial acetic acid (ACS)
and 13.6 g of sodium acetate trihydrate (ACS)
in reagent water which has been purified by
filtration through a RO-4 Millipore System or
equivalent and dilute to 1 L.
6.9 Acetonitrile, chloroform (preserved
with 1% ethanol). methanol—Pesticide quality
or equivalent.
6.10 Mobile phase—Place equal volumes
of filtered acetonitrile (Millipore type FH
filter or equivalent) and filtered acetate
buffer (Millipore type GS filter or equivalent)
in a narrow-mouth, glass container and mix
thoroughly. Prepare fresh weekly. Degas
daily by sonicating under vacuum, by heating
an stirring, or by purging with helium.
6.11 Stock standard solutions (1.00 pg/
jiL)—Stock standard solutions may be
prepared from pure standard materials or
purchased as certified solutions.
6.11.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in methanol
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
stand***). Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.11.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 'C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially jut prior to preparing
calibration standards from them.
6.11.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.12 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish chromatographic operating
condition* equivalent to those given in Table
1. The HPLC system can be calibrated using
the external standard technique (Section 7.2)
or the internal standard technique (Section
7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
mobile phase. One of the external standards
should be at a concentration near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using syringe injections of 5 to 25 jiL
or a constant volume injection loop, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (<10% relative standard
deviation. RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with mobile phase. One of the
standards should be at a concentration near,
but above, the MDL and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.3.2 Using syringe injections of 5 to 25 pL
or a constant volume injection loop, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF=
(AJ(CJ
where:
A.=Response for the parameter to be
measured.
A^xResponse for the internal standard.
Q.~ Concentration of the internal standard
C,—Concentration of the parameter to be
measured (ftg/L).
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively.'
the results can be used to plot a calibration
curve of response ratios. A,/A,..' vs. RF.
7.4 The working calibration curve.
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, a new
calibration curve must be prepared for that
compound. If serious loss of response occurs.
polish the electrode and recalibrate.
7.5 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.
ft Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.9,11.1. and 12.1) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8-2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing benzidine
and/or 3.3'-dichlorobenzidine at a
concentration of 50 jig/mL each in methanol.
The QC check sample concentrate must be
obtained from the U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory in Cincinnati. Ohio.
if available. If not available from that source.
the QC check sample concentrate must be
obtained from another external source. If not
available from either source above, the QC
check sample concentrate must be prepared
by the laboratory using stock standards
prepared independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at a concentration of 50 u.g/L by
adding 1.00 mL of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in /ig/L, and the standard deviation of the
recovery (s) in gg/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter. Locate and
correct the source of the problem and repeat
the test for all parameters of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess.
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2. whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 50 u.g/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any; or, if
none (2) the larger of either 5 times higher
than the expected background concentration
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T. where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 50 Mg/L. the analyst must use
either the QC acceptance criteria in Table 2,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter: (1) calculate
accuracy (X'j using the equation in Table 3,
substituting the spike concentration (T) for C;
(2) calculate overall precision (S') using the
equation in Table 3, substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X7T)±2.44(100 S'/
T)%.7
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
MOTE.— The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P,) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P.)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3.
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the nccuracy
assessment as a percent recovery interval
from P-2sptoP + 2sB. If P = 90% and sB = 10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as HPLC with a dissimilar column, gas
chromatography. or mass spectrometer must
be used. Whenever possible, the laboratory
should analyze standard reference materials
and participate in relevant performance
evaluation studies.
9. Sample Collection, Preservation, and
Handling
9.1 Grab samples must be collected in
glass containers. Conventional sampling
practices8 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 as free ss possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4'C and stored in the dark
from the time of collection until extraction.
Both benzidine and 3.3'-dichlorobenzidine are
easily oxidized. Fill the sample bottles and. if
residual chlorine is present, add 80 mg of
sodium thiosulfate per liter of sample and
mix well. EPA Methods 330.4 and 330.5 may
be used for measurement of residual
chlorine.* Field test kits are available for this
purpose. After mixing, adjust the pH of the
sample to a range of 2 to 7 with sulfuric acid.
9.3 If 1,2-diphenylhydrazine is likely to be
present, adjust the pH of the sample to 4.0±
0.2 to prevent rearrangement to benzidine.
9.4 All samples must be extracted within
7 days of collection. Extracts may be held up
to 7 days before analysis, if stored under an
inert (oxidant free) atmosphere.2 The extract
should be protected from light.
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory funnel. Check the pH of the
sample with wide-range pH paper and adjust
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70 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
to within the range of 6.5 to 7.5 with sodium
hydroxide solution or sulfuric acid.
10.2 Add 100 mL of chloroform to the
sample bottle, seal, and shake 30 s lo rinse
(he inner surface. (Caution: Handle
chloroform in a well ventilated area.)
Transfer the solvent to the scparatory funnel
and extract the sample by shaking the funnel
for 2 min with periodic venting to release
excess pressure. Allow the organic layer to
separate from the water phase for a minimum
of 10 min. If the emulsion interface between
layers is more than one-third the volume of
the solvent layer, the analyst must employ
mechanical techniques to complete the phase
separation. The optimum technique depends
upon the sample, but may include stirring,
filtration of the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the chloroform extract in a 250-mL
separator/ funnel.
10.3 Add a 50-mL volume of chloroform to
the sample bottle and repeat the extraction
procedure a second time, combining the
extracts in the separatory funnel. Perform a
third extraction in the same manner.
10.4 Separate and discard any aqueous
layer remaining in the 250-mL separatory
funnel after combining the organic extracts.
Add 25 ml of 1 M sulfuric acid and extract
the sample by shaking the funnel for 2 min.
Transfer the aqueous layer to a 250-mL
beaker. Extract with two additional 25-mL
portions of 1 M sulfuric acid and combine the
acid extracts in the beaker.
10.5 Place a stirbar in the 250-mL beaker
and stir the acid extract while carefully
adding 5 mL of 0.4 M sodium tribasic
phosphate. While monitoring with a pH
meter, neutralize the extract to a pH between
0 and 7 by dropwise addition of 5 N sodium
hydroxide solution while stirring the solution
vigorously. Approximately 25 to 30 mL of 5 N
sodium hydroxide solution will be required
and it should be added over at least a 2-min
period. Oo not allow the sample pH to exceed
8.
10.6 Transfer the neutralized extract into
a 250-mL separatory funnel. Add 30 mL of
chloroform and shake the funnel for 2 min.
Allow the phase* to-separate. amLtransfer
the organic layer to a second 250-mL
separatory funnel.
10.7 Extract the aqueous layer with two '
additional 20-mL aliquots of chloroform as
before. Combine the extracts in the 250-mL
separatory funnel.
10.8 Add 20 mL of reagent water to the
combined organic layer* and shake for 30 *.
10.9 Transfer the organic extract into a
100-mL round bottom flask. Add 20 mL of
methanol and concentrate to 5 mL with a
rotary evaporator at reduced pressure and 35
*C. An aspirator is recommended for use a*
the source of vacuum. Chill the receiver with
ice. This operation requires approximately 10
min. Other concentration technique* may be
used if the requirement* of Section 8.2 are
met.
10.10 Using a 9-in. Pasteur pipette,
transfer the extract to a 15-mL. conical,
•crew-cap centrifuge tube. Rinse the flask,
including the entire side wall, with 2-mL
portion* of methanol and combine with the
original extract.
10.11 Carefully concentrate the extract to
0.5 mL using a gentle stream of nitrogen while
heating in a 30 "C water bath. Dilute to 2 mL
with methanol. reconcentrdte to 1 mL. and
dilute to 5 mL with acetate buffer. Mix the
extract thoroughly. Cap the centrifuge tube
and store refrigerated and protected from
light if further processing will not be
performed immediately. If the extract will be
stored longer than two days, it should be
transferred to a Teflon-sealed screw-cap vial.
If the sample extract requires no further
cleanup, proceed with HPLC analysis
(Section 12). If the sample requires further
cleanup, proceed to Section 11.
10.12 Determine the original sample
volume by refilling the sample bottle to the
mark and transferring the liquid lo a 1,000-mL
graduated cylinder. Record the sample
volume to the nearest 5 mL.
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
12. High Performance Liquid
Chromatography
12.1 Table 1 summarizes the
recommended operating conditions for the
HPLC. Included in this table are retention
times, capacity factors, and MDL that can be
achieved under these conditions. An example
of the separations achieved by this HPLC
column is shown in Figure 1. Other HPLC
columns, chroma tographic conditions, or
detectors may be used if the requirements of
Section 8.2 are met. When the HPLC is idle, it
is advisable to maintain a 0.1 mL/min flow
through the column to prolong column life.
12.2 Calibrate the system daily as
described in Section 7.
12.3 If the internal standard calibration
procedure it being used, the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the instrument.
12.4 Inject 5 to 25 pL of the sample extract
or standard into the HPLC. If constant
volume injection loops are not used, record
the volume injected to the nearest 0.05 pL,
and the resulting peak size in area or peak
height units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size; however,
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.6 If the response for a peak exceeds
the working range of the system, dilute the
extract with mobile phase and reanalyze.
12.7 If the measurement of the peak
response for benzidine is prevented by the
presence of interferences, reduce the
electrode potential to +0.6 V and reanalyze.
If the ben/idine peak is still obscured by
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration (fig/L)=
where:
A = Amount of material injected (ng).
V, = Volume of extract injected (ML).
V, = Volume of total extract (/iL).
V.=Volume of water extracted (mL).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3.
Concentration (/ig/L)-
(AJ(RF)(V.)
where:
A,=Response for the parameter to be
measured.
A,, = Response for the internal standard.
I.=Amount of internal standard added to
each extract (jig).
Ve=Volume of water extracted (L).
13.2 Report results in pg/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Table 1 were obtained using reagent water.10
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
7xMDLto3000xMDL'°
14.3 This method was tested by 17
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range IX) to 70 pg/L'' Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
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Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136. Appendix B.
2. "Determination of Benzidines in
Industrial and Municipal Wastewaters."
EPA-600/4-82-022. U.S. Environmental
Protection Agency. Environmental Monitoring
and Support Laboratory. Cincinnati. Ohio
45268. May 1982.
3. ASTM Annual Book of Standards. Part
31. D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents."
American Society for Testing and Materials,
Philadelphia.
4. "Carcinogens—Working With
Carcinogens." Department of Health.
Education, and Welfare. Public Health
Service. Center for Disease Control, National
Institute for Occupational Safely and Health.
Publication No. 77-206. August 1977.
5. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910).
Occupational Safely and Health
Administration. OSHA 2200 (Revised.
January 1976).
6. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication, Committee on Chemical Safety.
3rd Edition. 1979.
7. Provost. L.P.. and Elder. R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two limes the value 1.22 derived in
this report.)
8. ASTM Annual Book of Standards. Part
31. D3370-76. "Standard Practices for
Sampling Water." American Society for
Testing and Materials. Philadelphia.
9. "Methods 330.4 (Tilrimclric. DPD-KAS)
and 330.5 (Spectrophotomclric. DPD) for
Chlorine Total Residual." Methods for
Chemical Analysis of Water and Wastes.
EPA-600/4-79-i)20. U.S. Environmental
Protection Agency. Environmental Monitoring
and Support Laboratory. Cincinnati, Ohio
45268. March 1979.
10. "Determination of Method Detection
Limit and Analytical Curve for EPA Method
605—Benzidines." Speical letter report for
EPA Contract 68-03-2624. U.S. Environmental
Protection Agency. Environmental Monitoring
and Support Laboratory. Cincinnati. Ohio
45268.
11. "EPA Method Validation Study 15.
Method 605 (Benzidines)." Report for El'A
Contract 68-03-2624 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Parameter
Retention
time
(min)
6 1
12 1
Column
capacity
factor (k1)
3 64
Method
detection
limit (ug/
L)
HPLC Column conditions: Uchrosorb RP-2. 5 micron panicle size, in a 25 cmx4.6 mm ID stainless steel column. Mobile Phase: 0.8 mL/min ot SO1* acetonitrile/50% 0.1M pH 4.7 acetate
buffer. The MDL were determined using an electrochemical detector operated at -r-0.8 V.
TABLE 2.—QC ACCEPTANCE CRITERIA—METHOD 605
Parameter
Test
cone.
(M9/U
SO
SO
Limit for
s (jig/D
187
236
Range lor
X 0»g/L)
9 1-61 0
18 7-500
Range
lor PP.
(percent)
D-140
5-128
s = Standard deviation of four recovery measurements, in pg/L (Section 8.2.4).
X = Average recovery for four recovery measurements, in jig/L (Section 8.2.4).
P. P. = Percent recovery measured (Section 8.3.2. Section 8.4.2).
0 = Detected; result must be greater than zero.
Not*.—These critena are based directly upon the method performance data in Table 3. Where necessary, the limits for recovery have been broadened to assure applicability of the limits to
concentrations below those used to develop Table 3.
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 605
Parameter
3 3'-Dichlorobenzidine
Accuracy, as
recovery.
X'Oig'L)
070C+006
0.66C + 0.23
Single analyst
precision, V
Uig'U
0.28X + 0 19
0.39X-0.05
Overall
precision. S'
(M9/L)
040X+0 18
038Xt002
X' = Expected recovery for one or more measurements of a sample containing a concentration ot C, in w)/L
s,' = Expected single analyst standard deviation ol measurements at an average concent/atipn found of XT in jig/L.
S' = Expected intertaboratory standard deviation ol measurements at an average concentration found of X, in pg/L.
C = True value lor the concentration, in |ig/L.
X = Average recovery found for measurements ol samples containing a concentration of C. in fig/L
BILLING CODE 6560-SO-M
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72
Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations
COLUMN: UCMOtOtt MP-2
MOtUfMAX: M* ACCTOMITfULE IN ACETATE SUFFER
DETECTO* aECTBOCXanCAt AT -»• 0.8 V
6 12
RETENTION TIME, MlN.
Figure 1. Liquid chromatogram
of benzidines.
MtUNO COM MM-CO-C
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 606—Phthatate Ester
/. Scope and Application
1.1 This method covers the determination
of certain phthalate esters. The following
parameters can be determined by this
method:
Parameter
8is(2.ethylrte)ryl) phthaiate
Butyl benzyl phthalate
W-tvbutyl phthalate
Diethyl phthalate
Dimethyl phthalate
0*-rH)Ctyl phlhalate
STOflET
No.
39100
34292
39110
34336
34341
34596
CAS No.
M7-61-7
85-68-7
84-74-2
84-66-2
131-11-3
117-84-0
1.2 This is a gas chromatographic (GC)
method applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes analytical conditions
for a second gas chromatographic column
that can be used to confirm measurements
made with the primary column. Method 625
provides gas chromatograph/mass
spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
1.3 The method detection limit (MDL,
defined in Section 14.1)' for each parameter
is listed in Table 1. The MDL for a specific
WLgtewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 608, 609,
. 611, and 612. Thus, a single sample may be
' extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots, as necessary, to
apply appropriate cleanup procedures. The
analyst is allowed the latitude, under Section
12, to select chromatographic conditions
appropriate for the simultaneous
measurement of combinations of these
parameters.
1.5 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is extracted with
methylene chloride using a separatory funnel.
The methylene chloride extract is dried and
exchanged to hexane during concentration to
a volume of 10 mL or less. The extract is
separated by gas chromatography and the
phthalate esters are then measured with an
electron capture detector.'
2.2 Analysis for phthalates is especially
complicated by their ubiquitous occurrence in
the environment. The method provides
Florisil and alumina column cleanup
procedures to aid in the elimination of
interferences that may be encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.9 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 'C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCS interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
dean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Phthalate esters are contaminants in
many products commonly found in the
laboratory. It is particularly important to
avoid the use of plastics because phthalates
are commonly used as plasticizers and are
easily extracted from plastic materials.
Serious phthalate contamination can result at
any time, if consistent quality control is not
practiced. Great care must be experienced to
prevent such contamination. Exhaustive
cleanup of reagents and glassware may be
required to eliminate background phthalate
contamination.41'
3.3 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedures in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
73
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified •'• for the information of the
analyst.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional]—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 "C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use. however, the compressible
tubing should be thoroughly rinsed with
methanol. followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only).
5.2.1 Separatory funnel—2-L. with Teflon
stopcock.
5.2.2 Drying column—Chromatographic
column, approximately 400 mm long X 19 mm
ID, with coarse frit filter disc.
5.2.3 Chromatographic column—300 mm
long x 10 mm ID, with Teflon stopcock and
coarse frit filter disc at bottom (Kontes K-
420540-0213 or equivalent).
5.2.4 Concentrator tube, Kuderna-
Danish—10-mL, graduated (Kontes K-
570050-1025 or equivalent). Calibration must
be checked at the volumes employed in the
test. Ground glass stopper is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask. Kudema-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column, Kudema-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Snyder column, Kudema-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
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Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
5.2.8 Vials—10 to 15-mL amber glass,
with Teflon-lined screw cap.
S.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with melhylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2 *C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 Gas chromatograph—An analytical
system complete with gas chromatograph
suitable for on-column injection and all
required accessories including syringes.
analytical columns, gaaei. detector, and (trip-
chart recorder. A data system is
recommended for measuring peak areas.
5.6.1 Column 1—1.8 m long x 4 mm ID
glass, packed with 1.5% SP-2250/1.95% SP-
2401 Supelcoport (100/120 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
column packings are provided in Section 12.1.
5.6.2 Column 2—1.8 m long X 4 mm ID
glass, packed with 3% OV-1 on Supel-coport
(100/120 mesh) or equivalent.
5.6.3 Detector—Electron capture detector.
This detector has proven effective in the
analysis of wastewaters for the parameters
listed in tht scope (Section 1.1), and was used
to develop the method performance
statement* in Section 14. Guidelines for the
use of alternate detectors are provided in
Section 12.1.
& Reagent*
6.1 Reagent water—Reagent water it
defined as • water in which an interferent is
not observed at the MDL of the parameters of
interest.
92 Acetone, hexane, isooctane,
methylene chloride, methanol—Pesticide
quality or equivalent
6J Bthyl ether—nanograde, redistilled in
glass if necessary.
6,3.1 Bthyl ether must be shown to be free
of peroxides before it is used as indicated by
EM Laboratories Quant test strips. (Available
from Scientific Products Co.. Cat. No. P1120-
8, and other suppliers.)
6.3.2 Procedures recommended for
removal of peroxides are provided with the
test strips. After cleanup. 20 mL of ethyl
alcohol preservative must be added to each
liter of ether.
6.4 Sodium sulfate—(ACS) Granular.
anhydrous. Several levels of purification may
be required in order to reduce background
phthalate levels to an acceptable level: 1)
Heat 4 h at 400 *C in a shallow tray. 2) Heat
10 h at 480 to 500 *C in a shallow tray, a)
Soxhlet extract with methylene chloride for
48 h.
04 Floristt-PR grade (80/100 mesh).
Purchase activated at 1250 *F and store in the
dark in glass containers with ground glass
stoppers or foil-lined screw caps. To prepare
for use, place 100 g of Plorisil into a 500-mL
beaker and heat for approximately 16 h at 40
*C After beating transfer to a 500-mL reagent
bottle. Tightly seal and cool to room
temperature. When cool add 3 mL of reagent
water. Mix thoroughly by shaking or rolling
for 10 min and let it stand for at least 2 h.
Keep the bottle sealed tightly.
6.6 Alumina—Neutral activity Super I,
W200 series (1CN Life Sciences Croup. No.
404583). To prepare for use, place 100 g of
alumina into a 500-mL beaker and heat for
approximately 16 h 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 reagent water. Mix
thoroughly by shaking or rolling for 10 min
and let it stand for at least 2 h. Keep the
bottle sealed tightly.
6.7 Stock standard solutions (1.00 fig/
nL)—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions.
6.7.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in isooctane
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.7.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 *C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
8.7.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.8 Quality control check sample
concentrate—See Section 8il.
7. Calibration
7.1 Establish gas chromatograph
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 72) or the
internal standard technique (Section 7.3).
72 External standard calibration
procedure:
7.2.1 Prepared calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
isooctane. One of the external standards
should be at a concentration near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
722 Using injections of 2 to 5 >tL. analyse
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over tht
working range (< 10V relative standard
deviation, RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flash. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with isooctane. One of the standards
should be at a concentraton near, but above.
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 pL. analyie
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF»
(A.HCJ
(AJ(CJ
where:
A,—Response for the parameter to be
measured.
Afc=Response for the internal standard.
Q.-Concentration of the internal standard
(W/L).
(^•Concentration of the parameter to be
measured (pg/L).
If the RF value over the working range is a
constant« 10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, AjA*, vs. RF.
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%. a new
calibration curve must be prepared for that
compound
7.5 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.
8. Quality Control
8.1 Bach laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document date quality. The laboratory must
maintain records to document the quality of
data that is generated Ongoing data quality
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
75
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4,11.1, and 12.1) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at the following
concentrations in acetone: butyl benzyl
phthalate, 10 ug/mL; bis(2-
ethylhexyljphthalate, 50 pg/mL; di-n-octyl
phthalate, 50 fig/mL any other phthlate, 25
u.g/mL. The QC check sample concentrate
must be obtained from the U.S.
Environmental Protection Agancy,
Environmental Monitoring and Support
Laboratory in Cincinnati. Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at the test concentrations shown in
Table 2 by adding 1.00 mL of QC check
sample concentrate to each of four 1-L
aliquota of reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in fig/L. and the standard deviation of the
recovery (s) in fig/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter. Locate and
correct the source of the problem and repeat
the test for all parameters of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at the test concentration in
Section 8.2.2 or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the splice concentration should be (1) the
regulatory concentration limit, if any; or, if
none (2) the larger of either 5 times higher
than the expected background concentration
or the test concentration in Section 8.2.2.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.• If
spiking was performed at a concentration
lower than the test concentration in Section
8.2.2. the analyst must use either the QC
acceptance criteria in Table 2. or optional QC
acceptance criteria calculated for the specific
spike concentration. To calculate optional
acceptance criteria for the recovery of a
parameter: (1) calculate accuracy (X') using
the equation in Table 3. substituting the spike
concentration (T) for C; (2) calculate overall
precision (S') usingjhe equation in Table 3.
substituting X' for X; (3) calculate the range
for recovery at the spike concentration as
(100 X'/T)±2.44(100 S'/T)%.»
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (8p). Express the accuracy
assessment as a percent recovery interval
from P-2s, to P+2sp. If P=90% and sp=10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the
laboratory adopt additional quality
assurance practices for use with this method.
The specific practices that ore most
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78 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
productive depend upon the need* of the
laboratory and the nature of the samples.
Field duplicates may be analyzed to assess
the precision of the environmental
measurements. When doubt exists over the
identification of a peak on the chromatogram.
confirmatory techniques such as gas
chromatography with a dissimilar column.
specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
9. Sample Collection. Preservation, and
Handling
9.1 Grab samples must be collected in
glass containers. Conventional sampling
practices10 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
mutt be as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 *C from the time of collection
until extraction.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.*
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
•ample volume. Pour the entire sample into a
2-L separately funnel.
10.2 Add 80 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the Mparatory runnel and extract the
•ample by •hiking the funnel for 2 mln. with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 mln. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
technique* to complete the phrase separation.
The optimum technique depend* upon the
sample, but may include stirring, filtration of
the emulsion through glass wool
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlennwyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the cample bottle and
repeat the extraction procedure a second
time, combining the extract* In the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentrator devices or
technique* may be used in place of the K-D
concentrator if the requirements of Section
Man met
1OS pour the combined extract through a
•ohrant-rinaed drying column containing
about 10 cm of anhydrous sodium sulfate.
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 65 'Q so that the
concentrator tube is partially immersed in the
hot water, and the entire lower rounded
surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in IS to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.7 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. Concentrate the extract
as in Section 10.6, except use hexane to
prewet the column. The elapsed time of
concentration should be 5 to 10 min.
10.8 Remove the Snyder column and rinse
the flask and its lower joint into the
concentrator tube with 1 to 2 mL of hexane. A
5-mL syringe is recommended for this
operation. Adjust the extract volume to 10
mL Stopper the concentrator tube and store
refrigerated if further processing will not be
performed immediately. If the extract will be
stored longer than two days, it should be
transferred to a Teflon-sealed screw-cap vial.
If the sample extract requires no further
cleanup, proceed with gas chromatographic
analyst* (Section 12). If the sample requires
further cleanup, proceed to Section 11.
1041 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
11. Cleanup and Separation
11. Cleanup procedure* may not be
necessary for a relatively clean sample
matrix If particular circumstances demand
the use of a cleanup procedure, the analyst
may use either procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
11.2 If the entire extract is to be cleaned
up by one of the following procedures, it must
be concentrated to 2.0 mL. To the
concentrator tube in Section 104 add a clean
boiling chip and attach a two-ball micro-
Snyder column. Prewet the column by adding
about 0.5 mL of hexane to the top. Place the
micrc-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 a* required to complete
the concentration in 5 to 10 min. At the
proper rate of distillation the ball* of the
column will actively chatter but the chambers
will not flood. When the apparent volume of
liquid reaches about M mL, remove the K-D
apparatus and allow it to drain and cool for
at least 10 min. Remove the micro-Snydar
column and rinse it* lower Joint Into the
concentrator tube with 0.2 mL of hexane.
Adjust the final volume to 2.0 mL and
proceed with one of the following cleanup
procedures.
11.3 Florisil column cleanup for phthalate
esters:
11.3.1 Place log of Florisil into a
chromatographic column. Tap the column to
settle the Florisil and add 1 cm of anhydrous
sodium sulfate to the top.
11.3.2 Preelute the column with 40 mL of
hexane. The rate for all elutions should be
about 2 mL/min. Discard the eluate and just
prior to exposure of the sodium sulfate layer
to the air, quantitatively transfer the 2-mL
sample extract onto the column using an
additional 2 mL of hexane to complete the
transfer. Just prior to exposure of the sodium
sulfate layer to the air, add 40 mL of hexane
and continue the elution of the column.
Discard this hexane eluate,
11.3.3 Next, elute the column with 100 mL
of 20% ethyl ether in hexane (V/V) into a 500-
mL K-D flask equipped with a 10-mL
concentrator tube. Concentrate the collected
fraction as in Section 10.6. No solvent
exchange is necessary. Adjust the volume of
the cleaned up extract to 10 mL in the
concentrator tube and analyze by gas
chromatography (Section 12).
11.4 Alumina column cleanup for
phthalate esters:
11.4.1 Place 10 g of alumina into a
chromatographic column. Tap the column to
settle the alumina and add 1 cm of anhydrous.
sodium sulfate to the top.
11.4.2 Preelute the column with 40 mL of
hexane. The rate for all ehitions should be
about 2 mL/min. Discard the eluate and Just
prior to exposure of the sodium sulfate layer
to the air. quantitatively transfer the 2-mL
•ample extract onto the column using an
additional 2 mL of hexane to complete the
transfer. Just prior to exposure of the sodium
sulfate layer to the air, add 35 mL of hexane
and continue the elution of the column.
Discard this hexane eluate.
11.4.3 Next elute the column with 140 mL
of 20% ethyl ether in hexane (V/V) into a 500-
mL K-D flask equipped with a 10-mL
concentrator type. Concentrate the collected
fraction as in Section 10.6. No solvent
exchange is necessary. Adjust the volume of
the cleaned up extract to 10 mL in the
concentrator tube and analyze by gas
chromatography (Section 12).
12. Gat Chromatography
12.1 Table 1 summarizes the
recommended operating condition* for the
gas chromatograph. Included in this table are
retention time* and MDL that can be
achieved under these condition*. Example* of
the separations achieved by Column i are
shown in Figure* 1 and 2. Other packed or
capillary (open-tubular) column*,
chromatographic condition*, or detectors may
be used if the requirement* of Sectional are
mat
12J Calibrate the system daily as
described in Section 7.
124 If the internal standard calibration
procedure i* being used, the internal standard
must be addad to the sample extract and
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Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations 77
mixed thoroughly immediately before
injection into the gas chromatograph.
12.4 Inject 2 to 5 fiL of the sample extract
or standard into the gas-chromatograph using
the solvent-flush technique." Smaller (1.0
jiL) volumes may be injected if automatic
devices are employed. Record the volume
injected to the nearest 0.05 fiL, and the
resulting peak size in area or peak height
units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size: however.
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.6 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.7 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration (jig/L) =
where:
A = Amount of material injected (ng).
V, = Volume of extract injected (/iL).
V, = Volume of total extract (/iL).
V.=Volume of water extracted (ml).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3-
Concentration (ng/L) =
(A5)|ls)
(A(,)(RF)(V0
where:
A, = Response for the parameter to be
measured.
Au = Response for the internal standard.
l, = Amount of internal standard added to
each extract (u,g).
V0 = Volume of water extracted (L).
13.2 Report results in ng/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Table 1 were obtained using reagent water.12
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
5 x MDL to 1000 x MDL with the following
exceptions: dimethyl and diethyl phthalate
recoveries at 1000 x MDL were low (70%);
bis-2-ethylhexyl and di-n-octyl phthalate
recoveries at 5 X MDL were low (60%).la
14.3 This method was tested by 16
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 0.7 to 106 ug/L.1* Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136. Appendix B.
2. "Determination of Phthalates in
Industrial and Municipal Wastewaters."
EPA-600/4-81-063, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory. Cincinnati. Ohio
45268. October 1981.
3. ASTM Annual Book of Standards. Part
31. D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents."
American Society for Testing and Materials.
Philadelphia.
4. Ciam. C.S., Chan. H.S.. and Nef. G.S.
"Sensitive Method for Determination of
Phthalate Ester Plasticizers in Open-Ocean
Biota Samples." Analytical Chemistry. -17.
2225 (1975).
5. Giam, C.S., and Chan. H.S. "Control of
Blanks in the Analysis of Phthalates in Air
and Ocean Biota Samples." U.S. National
Bureau of Standards, Special Publication 442.
pp. 701-708. 1976.
6. "Carcinogens—Working with
Carcinogens," Department of Health.
Education, and Welfare. Public Health
Service, Center for Disease Control. National
Institute for Occupational Safely and Health.
Publication No. 77-206. August 1977.
7. "OSHA Safety and Health Standards,
General Industry," (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2206 (Revised.
January 1976).
8. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication, Committee on Chemical Safety.
3rd Edition, 1979.
9. Provost L.P.. and Elder. R.S.
"Interpretation of Percent Recovery Data."
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
10. ASTM Annual Book of Standards, Part
31. D3370-76. "Standard Practices for
Sampling Water," American Society for
Testing and Materials. Philadelphia.
11. Burke, J.A. "Gas Chromatography for
Pesticide Residue Analysis: Some Practical
Aspects." Journal of the Association of
Official Analytical Chemists. 48.1037 (1965).
12. "Method Detection Limit and Analytical
Curve Studies, EPA Methods 606, 607, and
608," Special letter report for EPA Contract
68-03-2606. U.S. Environmental Protection
Agency, Environmental Monitoring and
Support Laboratory. Cincinnati. Ohio 45268.
13. "EPA Method Validation Study 16.
Method 606 (Phthlate Esters)," Report for
EPA Contract 68-03-2606 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Parameter
Retention lime (mm)
Column 1
2.03
2.82
8.65
a 6.94
a 8.92
a 16.2
Column 2
0.95
1.27
3.50
a 5.11
a 10.5
a 18.0
Method
detection
limit (jig/L)
0.29
0.49
0.36
0.34
2.0
3.0
Column 1 conditions: Supelcoport (100/120 mesh) coated with 1.5% SP-2250/1.95% SP-2401 packed in a 1.8 m long x 4 mm ID glass column with 5% methane/95% argon carrier gas
at 60 ml/min flow rate. Column temperature held isothermal at 180'C. except where otherwise indicated.
Column 2 conditions: Supelcoport (100/120 mesh) coated with 3% OV-1 packed in a 1.8 m long x 4 mm ID glass column with 5% methane/95% argon carrier gas at 60 ml/mm (tow
rate. Column temperature held isothermal at 200 %C. except where otherwise indicated.
• 220 *C column temperature.
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78
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 2.—QC ACCEPTANCE CRITERIA—METHOD 606
Parameter
Limit tor | Range lor
Bis<2-emylhexyl) phthalale
50 384
i 10 4.2
; 25 89
; 25 1 95
1 2-55 9
57-110
103-296
1 9-33 4
1 3-35 5
D-1S8
30-136
23-136
D-149
D-156
D-114
. , 1 I .
J- Standard deviation of four recovery measurements, m pg/L (Section 8.2 4).
• Average recovery lor four recovery measurements, in pg/L (Section 8.24)
P. P. - Percent recovery measured (Section 8.3 2. Section 64 2)
0- Detected, result must be greater than zero
Note.—These cntena are based directly upon the method performance data m Table 3. Where necessary, the limits lor recovery have been broadened to assure applicability of the limits to
wentrations below those used to develop Table 3.
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 606
Parameter
Dt-n-feutyf phtnalate
Djethyl phtnalale
DirnMhyt phtnalate .
Oi-nroctyl phthalate
Accuracy, as
recovery, X'
(MB/1-)
OS3C + 202
082C-fO 13
079C + 0 17
070C+0 13
073C+017
035C-071
Single analyst
precision, s,'
(M9/U
0 60X - 2 56
0 26X + 0 04
023X + 020
027X+005
026X+014
038X + 07t
Overall
precision. S
(Mt/t-l
0 29X * 0 06
044X4-031
0 62X -f 0 34
X= Expected recovery tor one or more measurements ot a sample containing a concentration of C, in |ia/L
s, -Expected single analyst standard deviation of measurements at an average concentration found of X. m pg/L
S -Expected Menaboratory standard deviation of measurements at an average concentration found of X. in j»g/L.
C-True value for the concentration, in pg/L
X- Average recovery found for measurements of samples containing a concentration of C. in pg/L.
O COOt
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
79
COLUMN. 1 5% SP 2250/1 95% SP-2401 ON SUPEICOPORT
TEMPERATURE: 180*C
DETECTOR; RECTRON CAPTURE
<
3
w
II
0 24 6 8 10 12
RETENTION TIME. MIN.
Figure 1. Gas chromatogram of phthalates.
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80
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
COLUMN: 15% SP-2250/1 95% SP-2401 ON SUPFI IMPORT
TEMPERATURE: 220%
DETECTOR: ELECTRON CAPTURE
8 12"
RETENTION TIME. MIN.
18
Figure 2. Gas chromatogram of phthalates
MU.HM COM t»M MM.
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Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
81
Method 607—Nilrosamines
1. Scope and Application
1.1 This method covers the determination
of certain nilrosamines. The following
parameters can be determined by this
method:
Parameter
N-Nilrosodi-n-propylamine
Store! No.
34438
34433
34428
CAS No.
62-75-9
8£-30-«
621-64-7
1.2 This is a gas chromatographic (GC)
method applicable to the determination of the
parameters listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
tht compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes analytical conditions
for a second gas chromatographic column
that can be used to confirm measurements
made with the primary column. Method 625
provides gas chromatograph/mass
spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for N-
nitrosodi-n-propylamine. In order to confirm
the presence of N-nitrosodiphenylamine. the
cleanup procedure specified in Sections 11.3
or 11.4 must be used. In order to confirm the
presence of N-nitrosodimethylamine by GC/
MS, Column 1 of this method must be
substituted for the column recommended in
Method 625. Confirmation of these
parameters using GC-high resolution mass
spectrometry or a Thermal Energy Analyzer
is also recommended. '•*
1.3 The method detection limit (MDL.
defined in Section 14.1)9 for each parameter is
listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.5 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is extracted with
methylene chloride using a separatory funnel.
The methylene chloride extract is washed
with dilute hydrochloric acid to remove free
amines, dried, and concentrated to a volume
of 10 mL or less. After the extract has been
exchanged to methanol, it is separated by gas
chromatography and the parameters are then
measured with a nitrogen-phosphorus
detector.4
2.2 The method provides Florisil and
alumina column cleanup procedures to
separate diphenylamine from the
nitrosamines and to aid in the elimination of
interferences that may he encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.9 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 *C
for 15 to 30 min. Solvent rinses with acetone
and pesticide quality hexane may be
substituted for the muffle furnace heating.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedures in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
3.3 N-Nitrosodiphenylamine is reported*'
to undergo transnitrosation reactions. Care
must be exercised in the heating or
concentrating of solutions containing this
compound in the presence of reactive amines.
3.4 The sensitive and selective Thermal
Energy Analyzer and the reductive Hall
detector may be used in place of the nitrogen-
phosphorus detector when interferences are
encountered. The Thermal Energy Analyzer
offers the highest selectivity of the non-MS
detectors.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined: however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available lo all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified m '5 for the information of tho
analyst.
4.2 These nitrosamines are known
carcinogens '*", therefore, utmost care must
be exercised in tho handling of these
materials. Nitrosamine reference standards
and standard solutions should be handled
and prepared in a ventilated glove box wilhin
a properly ventilated room.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
o, nposite sampling.
5.1.1 Grab sample bottle—1-Lorl-qt,
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 "C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flowmeter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnels—2-L and 250-
mL, with Teflon stopcock.
5.2.2 Drying column—Chromatographic
column, approximately 400 mm long X 19 mm
ID, with coarse frit filter disc.
5.2.3 Concentrator tube, Kuderna-
Danish—10-mL, graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.4 Evaporative flask, Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.5 Snyder column, Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.6 Snyder column, Kuderna-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
5.2.7 Vials—10 to 15-mJU amber glass.
with Teflon-lined screw cap.
5.2.8 Chromatographic column—
Approximately 400 mm long x 22 mm ID,
with Teflon stopcock and coarse frit filter
disc at bottom (Kontes K-420540-0234 or
equivalent), for use in Florisil column cleanup
procedure.
5.2.9 Chromatographic column—
Approximately 300 mm long X 10 mm ID,
with Teflon stopcock and coarse frit filter
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82 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
disc at bottom (Konlcs K-420540-0213 or
equivalent), for use in alumina column
cleanup procedure.
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlct
extract with methylene chloride.
5.4 Water bath—Healed, with concentric
ring cover, capable of temperature control
(±2 *C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 Gas chromatograph—An analytical
system complete with gas chromatograph
suitable for on-column injection and all
required accessories including syringes.
analytical columns, gases, detector, and strip-
chart recorder. A data system is
recommended for measuring peak areas.
5.6.1 Column 1—1.8 m long X 4 mm ID
glass, packed with 10% Carbowax 20 M/2%
KOH on Chromosorb W-AW (80/100 mesh)
or equivalent. This column was used to
develop the method performance statements
in Section 14. Guidelines for the use of
alternate column packings are provided in
Section 12.2.
5.6.2 Column 2—1.8m long x 4 mm ID
glass, packed with 10% SP-2250 on Supel-
coport (100/120 mesh) or equivalent.
5.6.3 Detector—Nitrogen-phosphorus.
reductive Hall, or Thermal Energy Analyzer
detector.'• * These detectors have proven
effective in the analysis of wastewaters for
the parameters listed in the scope (Section
1.1). A nitrogen-phosphorus detector was
used to develop the method performance
statements in Section 14. Guidelines for the
use of alternate detectors are provided in
Section 12.2.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 ml.
6.3 Sodium thiosulfate—(ACS) Granular.
6.4 Sulfuric acid (1 +1)—Slowly, add 50
ml of HtSCX (ACS, sp. gr. 1.84) to 50 mL of
reagent water.
8.5 Sodium sulfate—(ACS) Granular.
anhydrous. Purify by heating at 400 *C for 4 h
in a shallow tray.
6.6 Hydrochloric acid (1 + 9)—Add one
volume of concentrated HCI (ACS) to nine
volumes of reagent water.
6.7 Acetone, methanol, methylene
chloride, pentane—Pesticide quality or
equivalent.
6.8 Ethyl ether—Nanograde. redistilled in
glass if necessary.
6.8.1 Ethyl ether must be shown to be free
of peroxides before it is used as indicated by
EM Laboratories Quant test strips. (Available
from Scientific Products Co., Cat No. P1126-8,
and other suppliers.)
6.8.2 Procedures recommended for
removal of peroxides are provided with the
test strips. After cleanup, 20 mL of ethyl
alcohol preservative must be added to each
liter of ether.
6.9 Florisil—PR grade (60/100 mesh).
Purchase activated at 1250 *F and store in the
dark in glass containers with ground glass
stoppers or foil-lined screw caps. Before use.
activate each batch at least 16 h at 130 'C in
a foil-covered glass container and allow to
cool.
6.10 Alumina—Basic activity Super I.
W200 scries (ICN Life Sciences Group. No.
404571. or equivalent). To prepare for use.
place 100 g of alumina into a 500-ml reagent
bottle and add 2 mL of reagent water. Mix the
alumina preparation thoroughly by shaking or
rolling for 10 min and let it stand for at least 2
h. The preparation should be homogeneous
before use. Keep the bottle sealed tightly to
ensure proper activity.
6.11 Stock standard solutions (1.
u.L)—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions.
6.11.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in methanol
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.11.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 *C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.11.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.12 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish gas chromatographic
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 7.2) or the
internal standard technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
methanol. One of the external standards
should be at a concentraton near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using injections of 2 to 5 jiL, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (< 10% relative standard
deviation, RSD). linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of .1
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with methanol. One of the standards
should be at a concentration near, but above.
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 uL. analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF=
(Ato)(C.)
where:
A,=Response for the parameter to be
measured.
AH=Response for the internal standard.
Q.=Concentration of the internal standard
(M8/U-
C.=Concentration of the parameter to be
measured (M8/L)-
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A,/AU, vs. RF.
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, a new
calibration curve must be prepared for that
compound.
7.5 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
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Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
83
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in Section
10.4.11.1, and 12.2) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
.samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of 20
(ig/mL in methanol. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at a concentration of 20 pg/L by
adding 1.00 mi. of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixrd QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in f4g/L. and the standard deviation cf the
recovery (s) in fig/L. foi each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter. Locate and
correct the source of the problem and repeat
the test for all parameters of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 20 fig/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any; or. if
none (2) the larger of either 5 times higher
than the expected background concentration
or 20 jig/L
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were caluclated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.l8 If
spiking was performed at a concentration
lower than 20 ng/L, the analyst must use
either the QC acceptance criteria in Table 2.
or optional QC acceptance criteria caliirlatnd
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter (1) calculate
accuracy (X') using the equation in Table 3,
substituting the spike concentration (T) for C:
(2) calculate overall precision (S'| using the
equation in Table 3. substituting X' for X: (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) ± 2.44(100 S1/
T)%.18
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note: The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sp to P+2sp. If P=SO% and sp = 10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
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84 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
environmental measurements. When doubt
exists over the identification of a peak on the
chromalogram. confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
9. Sample Collection. Preservation, and
Handling
9.1 Crab 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 'C from the time of collection
until extraction. Fill the sample bottles and. if
residual chlorine is present, add 80 mg of
sodium thiosulfate per liter of sample and
mix well. EPA Methods 330.4 and 330.5 may
be used for measurement of residual
chlorine.10 Field test kits are available for
this purpose. If N-nitrosodiphenylamine is to
be determined, adjust the sample pH to 7 to
10 with sodium hydroxide solution or sulfuric
acid.
9.3 All samples must be extracted within
7 dayi of collection and completely analyzed
within 40 days of extraction.4
9.4 Nitrosamines are known to be light
sensitive.' Samples should be stored in
amber or foil-wrapped bottles in order to
minimize photolytic decomposition.
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separately funnel. Check the pH of the
sample with wide-range pH paper and adjust
to within the range of 5 to 9 with sodium
hydroxide solution or sulfuric acid.
10.2 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the funnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water pnase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.5 Add 10 mL of hydrochloric acid to
the combined extracts and shake for 2 min.
Allow the layers to separate. Pour the
combined extract through a solvent-rinsed
drying column containing about 10 cm of
anhydrous sodium sulfate. and collect the
extract in the K-D concentrator. Rinse the
Erlenmeyer flask and column with 20 to 30
mL of methylene chloride to complete the
quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewel the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL,
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.7 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. Stopper the
concentrator tube and store refrigerated if
further processing will not be performed
immediately. If the extract will be stored
longer than two days, it should be transferred
to a Teflon-sealed screw-cap vial. If N-
nitrosodiphenylamine is to be measured by
gas chromatography, the analyst must first
use a cleanup column to eliminate
diphenylamine interference (Section 11). If N-
nitrosodiphenylamine is of no interest, the
analyst may proceed directly with gas
chromatographic analysis (Section 12).
10.8 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.
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use either procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
Diphenylamine. if present in the original
sample extract must be separated from the
nitrosamines if N-nitrosodiphenylamine is to
be determined by this method.
11.2 If the entire extract is to be cleaned
up by one of the following procedures, it must
be concentrated to 2.0 mL To the
concentrator tube in Section 10.7, add a clean
boiling chip and attach a two-ball micro-
Snyder column. Prewet the column by adding
about 0.5 mL of methylene chloride to the top.
Place the micr-K-D apparatus on a hot waleid
bath (60 to 65 *C) so that the concentrator "
tube is partially immersed in the hot water.
Adjust (he vertical position of the apparatus
and the water temperature as required to
complete the concentration in 5 to 10 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood. When the apparent volume of
liquid reaches about 0.5 mL, remove the K-D
apparatus and allow it to drain and cool for
at least 10 min. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with 0.2 mL of methylene
chloride. Adjust the final volume to 2.0 mL
and proceed with one of the following
cleanup procedures.
11.3 Florisil column cleanup for
nitrosamines:
11.3.1 Place 22 g of activated Florisil into
a 22-mm ID chromatographic column. Tap the
column to settle the Florisil and add about 5
mm of anhydrous sodium sulfate to the top.
11.3.2 Preelute the column with 40 mL of
ethyl ether/pentane (15+85)(V/V). Discard
the eluate and just prior to exposure of the
sodium sulfate layer to the air, quantitatively
transfer the 2-mL sample extract onto the
column using an additional 2 ml of pentane
to complete the transfer.
11.3.3 Elute the column with 90 mL of
ethyl ether/pentane (15+85)(V/V) and
discard the eluate. This fraction will contain
the diphenylamine, if it is present in the
extract.
11.3.4 Next, elute the column with 100 mL
of acetone/ethyl ether (5+95)(V/V) into a
500-mL K-D flask equipped with a 10-mL
concentrator tube. This fraction will contain
all of the nitrosamines listed in the scope of
the method.
. 11.3.5 Add 15 mL of methanol to the
collected fraction and concentrate as in
Section 10.8, except use pentane to prewet
the column and set the water bath at 70 to
75 *C When the apparatus is cool, remove
the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1
to 2 mL of pentane. Analyze by gas
chromatography (Section 12).
11.4 Alumina column cleanup for
nitrosamines:
11.4.1 Place 12 g of the alumina
preparation (Section 6.10) into a 10-mm ID
chromatographic column. Tap the column to
settle the alumina and add 1 to 2 cm of
anhydrous sodium sulfate to the top.
11.4.2 Preelute the column with 10 mL of
ethyl ether/pentane (3-*-7)(V/V). Discard the
eluate (about 2 mL) and just prior to exposure
of the sodium sulfate layer to the air,
quantitatively transfer the 2 mL sample
extract onto the column using an additional 2
mL of pentane to complete the transfer.
11.4.3 Just prior to exposure of the sodium
sulfate layer to the air, add 70 mL of ethyl
ether/pentane (3+7)(V/V). Discard the first
10 mL of eluate. Collect the remainder of the
eluate in a 500-mL K-D flask equipped with a
10 mL concentrator tub*. This fraction
contains N-nitrosodiphenylamine and
probably a small amount of N-nitrosodi-n-
propylamine.
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
85
11.4.4 Next, elute the column with 60 ml
of ethyl ether/pentane (1 + 1)(V/V). collecting
the eluste in a second K-D flask equipped
with a 10-mL concentrator tube. Add 15 mL
of methanol to the K-D flask. This fraction
will contain N-nilrosodimelhylamine. most of
the N-nilrosodi-n-propylamine and any
diphenylamine that is present.
11.4.5 Concentrate both fractions as in
Section 10.6. except use pentane to prewet
the column. When the apparatus is cool.
remove the Snyder column and rinse the flask
and its lower joint into the concentrator .tube
with 1 to 2 mL of pentane. Analyze the
fractions by gas chromatography (Section 12).
12. Gas Chromatography
12.1 N-nitrosodiphenylamine completely
reacts to form diphenylamine at the normal
operating temperatures of a GC injection port
(200 to 250 *C). Thus, N-nitrosodiphenylamine
is chromatographed and detected as
diphenylamine. Accurate determination
depends on removal of diphenylamine that
may be present in the original extract prior to
GC analysis (See Section 11).
12.2 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. Examples of
the separations achieved by Column 1 are
shown in Figures 1 and 2. Other packed or
capillary (open-tubular) columns,
chromatographic conditions, or detectors may
.be used if the requirements of Section 8.2 are
met.
12.3 Calibrate the system daily as
described in Section 7.
12.4 If the extract has not been subjected
to one of the cleanup procedures in Section
11, it is necessary to exchange the solvent
from methylene chloride to methanol before
the thermionic detector can be used. To a 1 to
10-mL volume of methylene chloride extract
in a concentrator tube, add 2 mL of methanol
and a clean boiling chip. Attach a two-ball
micro-Snyder column to the concentrator
tube. Prewet the column by adding about 0.5
mL of 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
the concentration in 5 to 10 min. At the
proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood. When the apparent volume of
liquid reaches about 0.5 mL remove the K-D
apparatus and allow it to drain and cool for
at least 10 min. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with 0.2 mL of methanol.
Adjust the final volume to 2.0 mL
12.5 If the internal standard calibration
procedure is being used, the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the gas chromatograph.
12.6 Inject 2 to 5 pL of the sample extract
or standard into the gas chromatograph using
the solent-flush technique." Smaller (1.0 pL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 nL, and the resulting peak
size in area or peak height units.
12.7 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size; however,
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.8 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.9 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration (u./L] =
[VJ(VJ
where:
A=Amount of material injected (ng).
V,=Volume of extract injected (pL).
V, = Volume of total extract (/iL).
V,=Volume of water extracted (ml).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3.
Concentration (u.g/L) =
(A.K1.)
(A..HRFHV.)
where:
A,=Response for the parameter to be
measured.
A,,=Response for the internal standard.
I,=Amount of internal standard added to
each extract (jig).
V0=Volume of water extracted (L).
13.2 Report results in ftg/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.3 The MDL concentrations listed in
Table 1 were obtained using reagent water.**
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
4 x MDL to 1000 x MDL."
14.3 This method was tested by 17
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 0.8 to 55 jig/L.23 Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. Fine, D.H., Lieb, D., and Rufeh. R.
"Principle of Operation of the Thermal
Energy Analyzer for the Trace Analysis of
Volatile and Non-volatile N-nitroso
Compounds," Journal of Chromatography,
107. 351 (1975).
2. Fine, D.H., Hoffman, F., Rounbehler, D.P.,
and Belcher, N.M. "Analysis of N-nitroso
Compounds by Combined High Performance
Liquid Chromatography and Thermal Energy
Analysis," Walker, E.A., Bogovski, P. and
Griciute. L, Editors, N-nitroso Compounds—
Analysis and Formation, Lyon, International
Agency for Research on Cancer (IARC
Scientific Publications No. 14), pp. 43-50
(1976).
3. 40 CFR Part 136, Appendix B.
4. "Determination of Nitrosamines in
Industrial and Municipal Wastewaters,"
EPA-600/4-82-016, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268. May 1982.
5. ASTM Annual Book of Standards, Part
31, D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials,
Philadelphia.
6. Buglass, A.J., Challis, B.C., and Osborn,
M.R. "Transnitrosation and Decomposition of
Nitrosamines," Bogovski, P. and Walker,
E.A.. Editors, N-nitroso Compounds in the
Environment, Lyon, International Agency for
Research on Cancer (IARC Scientific
Publication No. 9), pp. 94-100 (1974).
7. Burgess, E.M., and Lavanish, f.M.
"Photochemical Decomposition of N-
nitrosamines," Tetrahedon Letters, 1221
(1964)
8. Druckrey, H., Preussmann, R., Ivankovic,
S., and Schmahl, D. "Organotrope
Carcinogene Wirkungen bei 65
Verschiedenen N-NitrosoVerbindungen an
BD-Ratten." Z. Krebsforsch., 69. 103 (1967).
9. Fiddler, W. "The Occurrence and
Determination of N-nitroso Compounds,"
Toxicol. Appl. Pharmacol., 31, 352 (1975).
10. "Carcinogens—Working With
Carcinogens," Department of Health,
-------
86
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
Education, and Welfare. Public Health
Service. Center for Disease Control, National
Institute for Occupational Safety and Health.
Publication No. 77-206. August 1977.
11. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration. OSHA 2206 (Revised.
January 1976).
12. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication. Committee on Chemical Safety.
3rd Edition. 1979.
13. Lijinsky, W. "How Nitrosamines Cause
Cancer." New Scientist. 73. 216 (1977).
14. Mirvish. S.S. "N-Nitroso compounds:
Their Chemical and in vivo Formation and
Possible Importance as Environmental
Carcinogens," /. ToxicoJ. Environ. Health. 3.
1267 (1977).
15. "Reconnaissance of Environmental
Levels of Nitrosamines in the Central United
States." EPA-330/1-77-001. National
Enforcement Investigations Center. U.S.
Environmental Protection Agency (1977).
16. "Atmospheric Nitrosamine Assessment
Report," Office of Air Quality Planning and
Standards. U.S. Environmental Protection
Agency. Research Triangle Park. North
Carolina (1976).
17. "Scientific and Technical Assessment
Report on Nitrosamines." EPA-660/6-7-001.
Office of Research and Development. U.S.
Environmental Protection Agency (1976).
16. Provost. L.P., and Elder. R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-«3 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value of 1.22 derived in
this report.)
19. ASTM Annual Book of Standards, Part
31, D3370-76. "Standard Practices for
Sampling Water," American Society for
Testing and Materials, Philadelphia.
20. "Methods 330.4 (Titrimetric. DPD-FAS)
and 330.5 (Spectrophotometric. DPD) for
Chlorine. Total Residual." Methods for
Chemical Analysis of Water and Wastes.
EPA-600/4-79-020. U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory. Cincinnati. Ohio
45268. March 1979.
21. Burke.). A. "Gas Chromatography for
Pesticide Residue Analysis: Some Practical
Aspects." Journal of the Association of
Official Analytical Chemists. 48.1037 (1965).
22. "Method Detection Limit and Analytical
Curve Studies EPA Methods 606. 607. and
608." Special letter report for EPA Contract
68-03-2606. U.S. Environmental Protection
Agency, Environmental Monitoring and
Support Laboratory, Cincinnati, Ohio 45268.
23. "EPA Method Validation Study 17,
Method 607 (Nitrosamines)," Report for EPA
Contract 68-03-2606 (In preparation).
TABLE 1.—CHROMATOORAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Parameter
Retention ttnw (nwi)
Column i
4.1
12.1
•12.8
Column Z
8.SS
42
•6.4
Method
detection
ttMbia/L)
0.1S
.46
61
Column 1 eondWorw: CNomcwxt W-AW (SO/100 m«h) cotttd with 10% Cwbowax 20 M/2% KOH pick** m • 1.8 m long > 4mm ID gton column with helium cam* gM it 40 mUmn
flow rat*. Cokm Mmptrakira h«M iaomermalat 110 *C, «opl «har« otfwrwlM indicated.
Column 2 oonMonc SupMcopoit (100/120 math) oatM wWi 10% SP-2250 packed in t 1.8 m long x 4 mm ID gfew column wtth Mum am* gtt « 40 mUmin few rate. Column
temporatm MM toothem) M 120 -C. except whan other*** Meued.
TABLE 2.—QC ACCEPTANCE CRITERIA—METHOD 607
.
•2» "C column lemperakm.
• no *C ootam Mmpmkn.
N Mtoiiiatifi n mtMMtoifrM
P. P.-P«reoni rm^ry imMurad {Sidteo 8.37,
D»t>lKHd. rwuR murt te QTM.W Mn wo.
MOM.— TVWM GriMffi tn bM«d tfrwtty upon VM
i-rtry^n*r«Hft/M hiiiraii thnM im*t In ftowsiinn TiiMil 1
PWMMf
MIIM, m M/L ««c«on 12.4).
•nit. in MO/L OtOton S.2,4).
S«e«oni4«.
Twioone.
0^/U
M
20
20
Urn* tort
(W/U
3.4
6.1
S.7
RtflgclarX
U4/U
46-20.0
2.1-24.4
11 5-26.I
lapptuMMyo
»w*
(pvctni)
19-109
O-t39
45-146
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 607
_»
N Mil ^^*«jJ TtivWM
ft NHooodl n propyloniino .
X--Ei.poc.td roooMry lor ono or mom nmmgmr* of • Mmp* .oonfcWno, • oonoi
ntraton ot C, in un/L
nMkon tound ot X. In MO/L.
Accuracy. Ǥ
'"(rt^i
0.37C+O.OO
0.64C4-O.S2
0.96C-0.07
Single analyst OiwaD
U4/I.) 04/L)
O^SJi-004 02SJ+011
0.368 1.53 046)1 047
O.ISJt+0.13 0.21X^0 IS
, .
*oe««y found tor nuMuimionli ol MH*IH containing • amo»n»»»an ol C, In «/L
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations 87
COLUMN: 10% CARBOWAX 20M / 2% KOH ON CHROMOSORB W-AW
TEMPERATURE: 110°C
DETECTOR: PHOSPHORUS/NITROGEN
ui
Z
I
X
Ui
I u>
O Z
O 5
u) 5
2 2
5 >
- &
z i
Z
-------
88
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
COLUMN: 10% CARBOWAX 20M /2% KOH ON
TEMPERATURE: 220*C
DETECTOR: PHOSPHORUS/NITROGEN
0 2 4 6 8 10 12 14 16 18
RETENTION TIME. MIN.
Figure 2. Gas chromatogram of N-nitrosodiphenylamine
as diphenylamine.
MLLMB COME •*•••*» C
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 89
Method 808—Organochlorine Pesticides and
PCBs
/. Scope and Application
1.1 This method covers the determination
of certain organochlorine pesticides and
PCBs. The following parameters can be
determined by this method:
Parameter
Aldrin
a-BHC
/3-BHC
6-BHC
y-BHC
4.4'-DDD
4.4'-DDE
4.4'-DDT
Oeldnn ... .
Endosullan II
Heptachlor
PCB-1016
PCB-1221 ... .
PCB- 1232
PCB- 1242
PCB-1248
PCB- 1254
PCB-1260
STORET No.
39330
39337
39338
34259
39340
39350
39310
39320
39300
39360
34361
34356
34351
39390
34366
39410
39420
39400
34671
39488
39492
394%
39500
39504
39508
CAS No.
309-00-2
319-84-6
•319-85-7
319-86-8
58-89-9
57-74-9
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33212-65-9
1031-07-8
72-20-8
7421-93-4
76-44-8
1024-57-3
8001-35-2
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
1.2 This is a gas chromatographic (GC)
method applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes analytical conditions
for a second gas chromatographic column
that can be used to confirm measurements
made with the primary column. Method 625
provides gas chromatograph/mass
spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
1.3 The method detection limit (MDL,
defined in Section 14.1)1 for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 606, 609,
611, and 612. Thus, a single sample may be
extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots, as necessary, to
apply appropriate cleanup procedures. The
analyst is allowed the latitude, under Section
12, to select chromatographic conditions
appropriate for the simultaneous
measurement of combinations of these
parameters.
1.5 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample.
approximately 1-L, is extracted with
methylcne chloride using a separately funnel.
The methylene chloride extract is dried and
exchanged to hexane during concentration to
a volume of 10 mL or less. The extract is
separated by gas chromatography and the
parameters are then measured with an
electron capture detector.*
2.2 The method provides a Florisil column
cleanup procedure and an elemental sulfur
removal procedure to aid in the elimination of
interferences that may be encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents,
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.3 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Interferences by phthalate esters can
pose a major problem in pesticide analysis
when using the electron capture detector.
These compounds generally appear in the
chroma togram as large late eluting peaks,
especially in the 15 and 50% fractions from
Florisil. Common flexible plastics contain
varying amounts of phthalates. These
phthalates are easily extracted or leached
from such materials during laboratory
operations. Cross contamination of clean
glassware routinely occurs when plastics are
handled during extraction steps, especially
when solvent-wetted surfaces are handled.
Interferences from phthalates can best be
minimized by avoiding the use of plastics in
(he laboratory. Exhaustive cleanup of
reagents and glassware may be required to
eliminate background phthalate
contamination.*'s The interferences from
phthalate esters can be avoided by using a
microcoulometric or electrolytic conductivity
detector.
3.3 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedures in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified "'for the information of the
analyst.
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: 4,4'-ODT. 4,4'-DDD, the BHCs,
and the PCBs. Primary standards of these
toxic compounds should be prepared in a
hood. A NIOSH/MESA approved toxic gas
respirator should be worn when the analyst
handles high concentrations of these toxic
compounds.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt,
amber glass, Titled with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4'C and protected from
light during composting. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol. followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
-------
90 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
flow meler is required lo collect flow
proportional composites.
5.2. Glassware (All specifications arc
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L. with Teflon
stopcock.
5.2.2 Drying column—Chromalographic
column, approximately 400 mm long x 19 mm
ID. with coarse frit filter disc.
5.2.3 Chromalographic column—400 mm
long x 22 mm ID. with Teflon stopcock and
coarse frit filter disc (Kontes K-42054 or
equivalent).
5.2.4 Concentrator tube. Kuderna-
Danish—10-mL, graduated (Kontes K-
570050-1025 or equivalent). Calibration must
be checked at the volumes employed in the
test. Ground glass stopper is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask, Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column, Kuderna/Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Vials—10 to 15-mL amber glass.
with Teflon-lined screw cap.
5.3. Boiing chips—Approximately 10/40
mesh. Heal to 400*C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Wajer bath—Heated, with concentric
ring cover, capable of temperature control
(±2*C). The bath should be used in a hood.
5.5. Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6. Gas chromatograph—An analytical
system complete with gas chromatograph
suitable for on-column injection and all
required accessories including syringes,
analytical columns, gases, detector, and strip-
chart recorder. A data system is
recommended for measuring peak areas.
5.6.1 Column 1—1.8 m long X 4 mm ID
glass, packed with 1.5% SP-2250/1.95% SP-
2401 on Supelcoport (100/120 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
column packings are provided in Section 12.1.
5.6.2 Column 2—1.8 m long x 4 mm ID
glass, packed with 3% OV-1 on Supelcoport
(100/120 mesh) or equivalent.
5.6.3 Detector—Electron capture detector.
This detector has proven effective in the
analysis of wastewaters for the parameters
listed in the scope (Section 1.1), and was used
to develop the method performance
statements in Section 14. Guidelines for the
use of alternate detectors are provided in
Section 12.1.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
8.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL
6.3 Sodium thiosulfate—(ACS) Granular.
6.4 Sulfuric acid (1+1)—Slowly, add 50
mL to HiSQ, (ACS. sp. gr. 1.84) to 50 mL of
reagent water.
6.5 Acetone, hexanc. isooctanc.
methylene chloride—Pesticide quality or
equivalent.
6.6 Ethyl ether—Nanograde. redistilled in
glass if necessary.
6.6.1 Ethyl ether must be shown to be free
of peroxides before it is used as indicated by
EM Laboratories Quant lest strips. (Available
from Scientific Products Co., Cat. No. P1126-
8. and other suppliers.)
6.6.2 Procedures recommended for
removal of peroxides are provided with the
test strips. After cleanup. 20 mL of ethyl
alcohol preservative must be added to each
liter of ether.
6.7 Sodium sulfate—(ACS) Granular.
anhydrous. Purify by heating at 400 "C for 4 h
in a shallow tray.
6.8 Florisil—PR grade (60/100 mesh).
Purchase activated at 1250 °F and store in the
dark in glass containers with ground glass
stoppers or foil-lined screw caps. Before use.
activate each batch at least 16 h at 130 °C in a
foil-covered glass container and allow to
cool.
6.9 Mercury—Triple distilled.
6.10 Copper powder—Activated.
6.11 Stock standard solutions (1.00 /ig/
jiL)—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions.
6.11.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in isooctane
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.11.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 *C and protect from light.
Stock standard solutions should be checked
frequently for sign* of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.11.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.12 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish gas chromatographic
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 7.2) or the
internal standard technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
isooctane. One of the external standards
should be at a concentration near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using injections of 2 to 5 >iL. analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (< 10V. relative standard
deviation. RSD). linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with isooctane. One of the standards
should be at a concentration near, but above.
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 /iL. analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF
(A.)(CJ
(AJ(C.)
where:
A,- Response for the parameter to be
measured.
A,, = Response for the internal standard.
Cu= Concentration of the internal standard
C,=Concentraton of the parameter to be
measured (
If the RF value over the working range is a
constant (< 10% RSD). the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, AjA^, vs. RF.
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, the test must
be repeated using a fresh calibration
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 91
standard. Alternatively, a new calibration
curve must be prepared for that compound.
7.5 The cleanup procedure in Section 11
utilizes Florisil column chromatography.
Florisil from different batches or sources may
vary in adsorptive capacity. To standardize
the amount of Florisil which is used, the use
of lauric acid value ' is suggested. The
referenced procedure determines the
adsorption from hexane solution of lauric
acid (mg) per g of Florisil. The amount of
Florisil to be used for each column is
calculated by dividing 110 by this ratio and
multiplying by 20 g.
7.6 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4,11.1, and 12.1) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
single-component parameter of interest at the
following concentrations in acetone: 4.4'-
DDD, 10 ng/mL: 4,4'-DDT, 10 ng/mL;
endosulfan II. 10 fig/mL; endosulfan sulfate,
10 ng/mL; endrin, 10 u,g/mL; any other single-
component pesticide, 2 fig/mL. If this method
is only to be used to analyze for PCBs,
chlordane, or toxaphene, the QC check
sample concentrate should contain the most
representative multicomponent parameter at
a concentration of 50 /ig/mL in acetone. The
QC check sample concentrate must be
obtained from the U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory in Cincinnati, Ohio,
if available. If not available from that source,
the QC check sample concentrate must be
obtained from another external source. If not
available from either source above, the QC
check sample concentrate must be prepared
by the laboratory using stock standards
prepared independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at the test concentrations shown in
Table 3 by adding 1.00 mL of QC check
sample concentrate to each of four 1-L
aliquots of reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in fig/mL; and the standard deviation of the
recovery (s) in u.g/mL, for each parameter
using the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 3. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 3 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.6 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.6.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.2.
8.2.6.2 Beginning with Section 8.2.2, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing onn lo
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at the test concentration in
Section 8.2.2 or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any: or. if
none (2) the larger of either 5 times higher
than the expected background concentration
or the test concentration in Section 8.2.2.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T. where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 3. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.10 If
spiking was performed at a concentration
lowpr than the test concentration in Section
8.2.2, the analyst must use either the QC
acceptance criteria in Table 3, or optional QC
acceptance criteria calculated for the specific
spike concentration. To calculate optional
acceptance criteria for the recovery of a
parameter: (1) Calculate accuracy (X') using
the equation in Table 4, substituting the spike
concentration (T) for C; (2) calculate overall
precision (S') using the equation in Table 4,
substituting X' for X; (3) calculate the range
for recovery at the spike concentration as
(100 X'/T)±2.44(100 S'/T)%. l°
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
-------
92
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
check aliiiulard containing each parameter
thai failed must be prepared and analyzed.
Nola.—Thr frequency for the required
amity*!* of a Qf" check standard will depend
upon the number of parameters being
sinuiltimeiMisly tested, the complexity of the
simple matrix, and the performance of the
laboratory. If the entire list of parameters in
Tublr .1 nuist be measured in the sample in
Section H.3. the probability that the analysis
of a QC check standard will be required is
hixh. In this case the QC check standard
should be routinely analyzed with the spike
sample.
H.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standards to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P.)
for each parameter with the corresponding
QC acceptance criteria found in Table 3.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. IMhe recovery of any such parameter
fulls outside the designated range, the
laboratory performance for that parameter is
fudged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewaler samples as in Section 8.3.
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (s,). Express the accuracy
assessment as a percent recovery interval
from P-2 s, to P+2 s.. If P=90% and
s,=10%. for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
ft Sample Collection, Preservation, and
Handling
9.1 Crab 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 *C from the time of collection
until extraction. If the samples will not be
extracted within 72 h of collection, the
sample should be adjusted to a pH range of
5.0 to 9.0 with sodium hydroxide solution or
sulfuric acid. Record the volume of acid or
base used. If aldrin is to be determined, add
sodium thiosulfate when residual chlorine is
present. EPA Methods 330.4 and 330.5 may be
used for measurement of residual chlorine.'2
Field test kits are available for this purpose.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.2
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separately funnel.
10.2 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the funnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optium technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Sectir.n
8.2 are met.
10.5 Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate,
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL.
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.7 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. Concentrate the extract
as in Section 10.6. except use hexane to
prewet the column. The elapsed lime of
concentration should be 5 to 10 min.
10.8 Remove the Snyder column and rinse
the flask and its lower joint into the
concentrator tube with 1 to 2 mL of hexane. A
5-mL syringe is recommended for this
operation. Stopper the concentrator tube and
store refrigerated if further processing will
not be performed immediately. If the extract
will be stored longer than two days, it should
be transferred to a Teflon-sealed screw-cap
vial. If the sample extract requires no further
cleanup, proceed with gas chromatographic
analysis (Section 12). If the sample requires
further cleanup, proceed to Section 11.
10.9 Determine the original sample
volume by refilling the sample bottle to the
mark and transferring the liquid to a 1000-mL
graduated cylinder. Record die sample
volume to the nearest S mL
11, Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use either procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
The Florisil column allows for a select
fractionatton of the compounds and will
eliminate polar interferences. Elemental
sulfur, which interferes with the electron
capture gas chroma tography of certain
pesticides, can be removed by the technique
described in Section 11.3.
11.2 Florisil column cleanup:
11.2.1 Place a weight of Florisil (nominally
20 g) predetermined by calibration (Section
7.5), into a chroma tographic column. Tap the
column to settle the Florisil and add 1 to 2 cm
of anhydrous sodium sulfate to the top.
11.2.2 Add 60 mL of hexane to wet and
rinse the sodium sulfate and Florisil. Just
prior to exposure of the sodium sulfate layer
to the air, stop the elution of the hexane by
closing the stopcock on the chromatographic
column. Discard the eluate.
11.2.3 Adjust the sample extract volume
to 10 mL with hexane and transfer it from the
K-D concentrator tube onto the column.
Rinse the tube twice with 1 to 2 mL of
hexane, adding each rinse to the column.
11.2.4 Place a 500-mL K-D flask and clean
concentrator tube under the chromatographic
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 (V/V) (Fraction 1) at a rate of
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
93
about S mL/min. Remove the K-D flask and
set it aside for later concentration. Elute the
column again, using 200 mL of 15% ethyl ether
in hexane (V/V) (Fraction 2). into a second K-
O flask. Perform the third elution using 200
mL of 50% ethyl ether in hexane (V/V)
(Fraction 3). The elution patterns for the
pesticides and PCBs are shown in Table 2.
11.2.5 Concentrate the fractions as in
Section 10.6. except use hexane to prewet the
column and set the water bath at about 85 *C.
When the apparatus is cool, remove the
Snyder column and rinse the flask and its
lower joint into the concentrator tube with
hexane. Adjust the volume of each fraction to
10 mL with hexane and analyze by gas
chromatography (Section 12).
11.3 Elemental sulfur will usually elute
entirely in Fraction 1 of the Florisil column
cleanup. 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
one to three drops of mercury and seal."
Agitate the contents of the vial for 15 to 30 s.
Prolonged shaking (2 h) may be required. If
so. this may be accomplished with a
reciprocal shaker. Alternatively, activated
copper powder may be used for sulfur
removal.14 Analyze by gas chromatography.
12, Gas Chromatography
12.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. Examples of
the separations achieved by Column 1 are
shown in Figures 1 to 10. Other packed or
capillary (open-tubular) columns,
chromatographic conditions, or detectors may
be used if the requirements of Section 8.2 are
met.
12.2 Calibrate the system daily as
described in Section 7.
12.3 If the internal standard calibration
procedure is being used, the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the gas chromatograph.
12.4 Inject 2 to 5 jiL of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique.16 Smaller (1.0 uL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 fiL, the total extract volume.
and the resulting peak size in area or peak
height units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size; however,
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.6 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.7 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
(A)(Vt)
Concentration (ug/L)= —
(VJfVJ
where:
A=Amount of material injected (ng).
V, = Volume of extract injected (>iL).
V, = Volume of total extract (>iL).
V.=Volume of water extracted (mL).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3.
Concentration (ng/L} =
(A.) (LJ
(AJfRFHV.)
where:
A.=Response for the parameter to be
measured.
Ata=Response for the internal standard.
I.=Amount of internal standard added to
each extract (ng).
V0=Volume of water extracted (L).
13.2 When it is apparent that two or more
PCB (Aroclor) mixtures are present, the
Webb and McCall procedure " may be used
to identify and quantify the Aroclors.
13.3 For muJticomponent mixtures
(chlordane, toxaphene, and PCBs) match
retention times of peaks in the standards
with peaks in the sample. Quantitate every
identifiable peak unless interference with
individual peaks persist after cleanup. Add
peak height or peak area of each identified
peak in the chromatogram. Calculate as total
response in the sample versus total response
in the standard.
13.4 Report results in /ig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.' The MDL concentrations listed in
Table 1 were obtained using reagent water.11
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
4XMDL to 1000xMDL with the following
exceptions: Chlordane recovery at 4XMDL
was low (60%); Toxaphene recovery was
demonstrated linear over the range of
10 X MDL to 1000 x MDL.1'
14.3 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations."
Concentrations used in the study ranged from
0.5 to 30 fig/L for single-component pesticides
and from 8.5 to 400 jtg/L for multicomponent
parameters. Single operator precision, overall
precision, and method accuracy were found
to be directly related to the concentration of
the parameter and essentially independent of
the sample matrix. Linear equations to
describe these relationships are presented in
Table 4.
References
1. 40 CFR Part 136, Appendix B.
2. "Determination of Pesticides and PCBs in
Industrial and Municipal Wastewaters,"
EPA-600/4-82-023. U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, June 1982.
3. ASTM Annual Book of Standards, Part
31, D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials,
Philadelphia.
4. Giam. C.S., Chan, H.S., and Nef, G.S.,
"Sensitive Method for Determination of
Phthalate Ester Plasticizers in Open-Ocean
Biota Samples." Analytical Chemistry. 47.
2225 (1975).
5. Giam, C.S., Chan, H.S. "Control of Blanks
in the Analysis of Phthalates in Air and
Ocean Biota Samples," U.S. National Bureau
of Standards, Special Publication 442, pp.
701-708,1976.
6. "Carcinogens—Working With
Carcinogens," Department of Health,
Education, and Welfare, Public Health
Service, Center for Disease Control, National
Institute for Occupational Safety and Health,
Publication No. 77-206, August 1977.
7. "OSHA Safety and Health Standards,
General Industry," (29 CFR 1910),
Occupational Safety and Health
Administration, OSHA 2206 (Revised,
January 1976).
8. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication, Committee on Chemical Safety,
3rd Edition, 1979.
9. 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).
10. Provost, L.P., and Elder, R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
-------
94
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
value 2.44 used in the equation in Section
8.3.3 it two times the value 1.22 derived in
this report.)
11. ASTM Annual Book of Standards, Part
31. 03370-76. "Standard Practices for
Sampling Water." American Society for
Testing and Materials. Philadelphia.
12. "Methods 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric. DPO) for
Chlorine. Total Residual." Methods for
Chemical Analysis of Water and Wastes.
EPA-flOO/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati. Ohio
45268. March 1979.
13. Goerlitz, O.F.. and Law, LM. Bulletin
for Environmental Contamination and
Toxicology. 6. 9 (1971).
14. "Manual of Analytical Methods for the
Analysis of Pesticides in Human and
Environmental Samples," EPA-eOO/8-80-038,
U.S. Environmental Protection Agency.
Health Effects Research Laboratory,
Research Triangle Park. North Carolina.
15. Burke. J.A. "Gas Chromatography for
Pesticide Residue Analysis; Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists, 4& 1037 (1965).
16. Webb. R.C., and McCall, A.C.
"Quantitative PCS Standards for Election
Capture Gu Chromatography," Journal of
Chromatotraph/c Science, 11,366 (1973).
17. "Method Detection Limit and Analytical
Curve Studies, EPA Methods 600.007. and
008," Special letter report for EPA Contract
00-03-2000, U.S. Environmental Protection
Agency. Environmental Monitoring and
Support Laboratory. Cincinnati. Ohio 45288.
10. "EPA Method Validation Study 18.
Method 600 (Organochlorine Pesticides and
PCBe)," Report for EPA Contract 00-03-2006
(In preparation).
TABLE 1.—CHROMATOGRAPHIC CoNomoNS
AND METHOD DETECTION LIMITS
PwnvMf
o-BHC
y-BHC
/J-BHC .
S-BHC
AUrin
EndoeuNen I
4<|'.OOE
rjlaMti
Endrin _
44'-000
EndomKann
4 4'-DDT
EndceuNan euNeJa......
CNnrdane
Tonaphene
PCS- 1016 —
PC8-1221
PC8-123I. .
Reten*
(m
Coil
1.3S
1.70
1.90
2.00
2.1S
2.40
3.50
4.50
5.13
5.46
8.56
7.13
8.00
9.40
11.62
1472
nv
mr
mr
ml
on lime
")
Cct2
182
2.13
1.97
3.35
2.20
4.10
5.00
6 JO
7.15
7.23
6.10
9.06
8J6
11.75
•JO
10.70
mr
mr
mr
mr
mr
Mettttd
d*MO-
•ret
04/L)
0.003
0.00
0.00
0.003
0.009
0.004
0.063
0.014
0.004
0.002
0.006
0.011
0.004
0.012
0.023
0.086
0.014
0.24
nd
nd
nd
TABLE 1.—CHROMATOGRAPHIC CONDITIONS
AND METHOD DETECTION LIMITS—Continued
PCB-1242
PCS- 124$
PCB-1254
PCB-1260
g/L)
0.065
nd
nd
nd
Column 1 conditions Supatooport (100/120 mesh) coeted
with 1.5% SP-2250/1.9S% SP-2401 packed in a 1.8 m long
x 4 mm ID glass column with 5% methane/95% argon
earner gas at 60 mL/mm (low rate. Column temperature held
isothermal at 200 'C. except tor PCB-1016 through PCB-
1248. should be measured at 160 'C.
Column 2 eonoHions: Supatooport (100/120 mesh) coated
with 3% OV-1 packed in a 1.6 m long x 4 mm ID glass
column with 5% melhene/95% argon earner gas at 60 ml./
mm flow rate. Column temperature held ieothermal at 200 *C
tor the pesticides; at 140 'C tor PCB-1221 and 1232; and at
170 'C tor PCB-1016 and 1242 to 1268.
mr-MuMpM peak response. See Figures 2 thru 10.
TABLE 2.—DISTRIBUTION OF CHLORINATED
PESTICIDES AND PCBs INTO FLORISIL COL-
UMN FRACTIONS *
Parameler
Aldrin.
a-BHC
/J-BHC
64HC
y-BHC. „
44'-DDD
4,4>-OOE _
4,4'-COT
OMdrtn
EndaeuMen !.._._
Endoeueen II
EndoauNaji «u»eJe...
Endrin
HaptscMor aponlfle
Toapnene
PCB-1016
PCB-1221
PC8-123*
PCS- 1242 __
PCB-1248 _.
PC8-12S4
PC8-1260
Percent
1
100
100
97
96
100
100
M
96
100
0
37
0
0
4
0
too
too
96
97
97
96
97
103
90
95
recovery by
2
100
64
7
0
9t
66
4
(radon*
3
"in
106
26
•Eluantcanportton:
FrACvon I^OT* •fnyl •fnv in nttcmc.
Fradton 2-15% ethyl ether in hexane.
Fradton 3-60% ethyl ether in hexane.
TABLE 3.—QC ACCEPTANCE CRITERIA-
METHOD 608
TABLE 3.—QC ACCEPTANCE CRITERIA-
METHOD 608—Continued
PiwiMMf
4.4--OOT
DieWnn
EndowHan 1
Endoeurfan II
Endo*uKan Suflttt
Endrin
Heptachlor
Toxaprtene.
PCB-1016. ..
PCB-1221
PCS- 1232
PCB-1242
PCS- 124*
PCS- 1254 . .
PCB-1260
Tetl
cone
(M-/
10
2.0
2.0
10
10
10
2.0
SO
50
SO
50
50
SO
50
SO
Unw
tort
(Mfl'L)
3.6
0.76
0.49
6.1
2.7
37
0.40
12.7
100
24.4
17.9
122
159
136
104
sr
(M9/L)
46-137
1.15-249
1.14-2.62
2.2-171
3.6-132
5 1-126
0.66-200
27 6-55 6
305-51 5
22 1-752
140-965
246-696
290-702
22 2-57 9
tt 7-54 9
W
PJSI
2! -160
36-146
45-153
0-202
26-144
30-147
34-111
41-126
50-114
15-176
10-215
39-150
36-158
29-131
6-127
•^Standard daniaton of tour recovery maamramaiiu. in
Mg/L (Section 8.2.4).
X« Average recov
MB/L (Secton 8J.4).
P.
.
ery tor tour recovery measurements.
..
P. P.. Percent recovery measured (Sectton 8.3.2, Section
8.4.2).
0- Detected; reauN muat be greater then zero.
Not*.— Theae criteria, are baaed aVecoY upon tie method
performance data in Table 4. Where neceaaary. the MB tar
recovery have bean broadened to aaaure appacabWy or tie
tmris to concemraliun» betow thoee ueed to develop Table
TABLE 4. METHOD ACCURACY AND PRECISION
AS FUNCTIONS OF CONCENTRATION—METH-
OD 008
4,4'-ODD
4.4 •-ODE
Teat
cone.
2.0
2.0
2.0
2.0
2.0
50
10
2.0
LMI
tora
U4/U
0.42
0.46
0.64
0.72
0.46
10.0
2.8
0.56
1.06-2.24
.96-444
0.78-4.80
1.01-&37
O.M-&32
27.6-S4.3
4.6-12.6
1.08-2.60
Accuracy, a*
fooovoiy, K
(lA/L)
OJ1C+0.04
OS4C+O.OS
OJIC-l-0.07
ojic+0.07
OJSC-0.08
OJ2C-0.04
0.64C+0.30
OJ6C+0.14
OJ9C-0.13
ojoc+0.02
0.970+0.04
OJ8C+0.34
O.WC-O97
0.86C-a04
0.68C+O.M
0.89C+0.10
0.80C-H.74
0.81C+O.SO
0.96C+0.65
0.91C + 10.T8
0.9X+070
0.97C+1.06
0.76C+2.07
0.66C+3.76
O.ieX-0.04
0.13*+0.04
.
0.16X+0.09
0.1«X+0.06
0.1JX+0.13
OJOX-0.16
0.1SS+OM
0.12X+0.19
o.iot-i-ao7
a4it-o.es
0.13X+0.33
OMt-t-OX
0.06X+0.13
0.18X-0.11
O.OBjt+3.20
013JU0.15
079X-0.76
.
0.11X+140
0.17X+041
0.15X + 1M
OvereJ
OJOX-0.01
ojaX-aoo
OJSS-OJ6
OJBX^OJM
o.iiX+o.11
O^Tt-0.14
OJeJt-0.08
OJ1S-OJ1
0.16H+0.16
0.18X+0.08
0.47X-OJO
O24X+0.36
0-J4X+0.2S
0.1«+0.06
0.2SX-O.OS
0.20X+072
0.15X+045
0.35X-0.62
0.31X+3.50
021* + 152
0.2SX-0.37
0.17X+3.82
039X-466
»E)4Mcli)d ivoowy tot on> or mora
i of •
42-122
37-134
17-147
19-140
32-127
46-119
31-141
30-146
urementa at an
9 ^ElkpMIBO
unman*) at an average
C, -True value tor •» ou
X'AvMkfli raoowy fo
oonwnino • m»o»»i.i>ion
teund eJ 1 h
O OVIMBOH Of I
found of x, ki
found for
of C. In >iO/L
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
95
COLUMN: 1.5% SP-2250/1.95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 200*C.
DETECTOR: aECTRON CAPTURE
4 8 12
RETENTION TIME. MIN .
16
Figure 1. Gas chromatogram of pesticides •
-------
96
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
COLUMN: 15% SP-2250/1 95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 2WC.
DETECTOR: ELECTRON CAPTURE
4 8 12
RETENTION TIME. MIN.
16
Figura 2. Gas chromatogram of chlordane.
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations 97
COLUMN: 1.5% SP-2250/1 95% SP 2401 ON SUPELCOPORT
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18 22 26
RETENTION TIME, MIN.
Figure 3. Gas chromatogram of toxaphene.
-------
98 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
II COLUMN: 15% SP-2250/1 95% SP-2401 ON SUPELCOPORT
II TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18 22
RETENTION TIME. MIN.
Figure 4. Gas chromatogram of PCB-1016.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
99
COLUMN: 1.5% SP 2250/1 95% SP-2401 ON SUPHCOPORT
TEMPERATURE: 160*C.
DETECTOR: aECTRON CAPTURE
u
6 10 14 18
RETENTION TIME. WIN.
22
Figure 5. Gas chromatogram of PCB-1221.
-------
100
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
COLUMN: 1.5% SP-2250/1 95% SP-2401 ON SUPaCOPORT
TBIPERATURE: 160*C.
DETECTOR: aECTRON CAPTURE
10 14 18
RETENTION TIME, MIN.
22
Figure 6. Gas chromatogram of PCB-1232.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
101
COLUMN: 1.5% SP-2250/1 95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
10 14 18
RETENTION TIME, MIN.
22
Figure 7. Gas chromatogram of PCB-1242.
-------
102
Federal Register / Vol. 49. No. 209 / Friday, October 26, 1984 / Rules and Regulations
COLUMN: 1.5% SP-2250/1 95% SP-2401 ON SUPELCOPOftT
TEMPERATURE: 160*C.
DETECTOR: ELECTRON CAPTURE
2 6 10 14 18 22
RETENTION TIME. WIIN.
Figure 8. Gas chromatogram of PCB-1248.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 103
COLUMN: 1.5% SP-225071 95% SP-2401 ON SUPELCOPORT
TEMPERATURE: 200*C
DETECTOR: ELECTRON CAPTURE
6 10 14
RETENTION TIME. MIN.
18
22
Figure 9. Gas chromatogram of PCB-1254.
-------
104
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
COLUMN: 1.5% SP 2250/1 95% SP-2401 ON SUPdCOfORT
TEMPERATURE: 200*C.
DETECTOR: ELECTRON CAPTURE
t
6
___
10 14 tt
RETENTION TIME. M|N.
Figure 10. Gas chromatogram of PCB-1260
BILLING CODE 6560-50-C
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 105
Method 609—Nitroaromalics and Isophorone
;. Scope and Application
1.1 This method covers the determination
of certain nitrouromatics and isophorone. The
following parameters may be determined by
this method:
Parameter
2 6-Oinitrotofuene
Store! No.
34611
34626
34408
34447
CasNo
121-14-2
606-20-2
78-59-1
98-95-3
1.2 This is a gas chromatographic (GC)
method applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes analytical conditions
for a second gas chromatographic column
that can be used to confirm measurements
made with the primary column. Method 625
provides gas chromatograph/mass
spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
1.3 The method detection limit (MDL,
defined in Section 14.1)' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 606, 608,
611, and 612. Thus, a single sample may be
extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots, as necessary, to
apply appropriate cleanup procedures. The
analyst is allowed the latitude, under Section
12, to select chromatographic conditions
appropriate for the simultaneous
measurement of combinations of these
parameters.
1.5 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is extracted with
methylene chloride using a separatory funnel.
The methylene chloride extract is dried and
exchanged to hexane during concentration to
a volume of 10 mL or less. Isophorone and
nitrobenzene are measured by flame
ionizalion detector gas chromatography
(FIDGC). The dinitrotoluenes are measured
by electron capture detector gas
chromatography (ECDGC).2
2.2 The method provides a Florisil column
cleanup procedure to aid in the elimination of
interferences that may be encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.3 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry. and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedure in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
* Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified *'• for the information of the
analyst.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt,
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 *C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use. however, the compressible
tubing should be thoroughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L, with Teflon
stopcock.
5.2.2 Drying column—Chromatographic
column, approximately 400 mm long x 19 mm
ID. with coarse frit filter disc.
5.2.3 Chromatographic column—100 mm
long x 10 mm ID. with Teflon stopcock.
5.2.4 Concentrator tube, Kuderna-
Danish—10-mL, graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask, Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column, Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Snyder column, Kuderna-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
5.2.3 Vials—10 to 15-mL, amber glass,
with Teflon-lined screw cap.
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control (±
2 *C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 Gas chromatograph—An analytical
system complete with gas chromatograph
suitable for on-column injection and all
required accessories including syringes,
analytical columns, gases, detector, and strip-
chart recorder. A data system is
recommended for measuring peak areas.
5.6.1 Column 1—1.2 m long x 2 or 4 mm ID
glass, packed with 1.95% QF-1/1.5S OV-17
on Gas-Chrom Q (80/100 mesh) or equivalent.
This column was used to develop the method
performance statements given in Section 14.
-------
106 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Guideline* for the ute of alternate column
packings are provided in Section 12.1.
5.6.2 Column 2—3.0 m long x 2 or 4 mm ID
glass, packed with 3% OV-101 on Gas-Chrom
Q (80/100 mesh) or equivalent.
5.0.3 Detectors—Flame ionization and
electron capture detectors. The flame
ionization detector (FID) is used when
determining isophorone and nitrobenzene.
The electron capture detector (ECD) is used
when determining the dinitrotoluenes. Both
detectors have proven effective in the
analysis of wastewaters and were used in
develop the method performance statements
in Section 14. Guidelines for the use to
alternate detectors are provided in Section
12.1.
ft Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
0.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL.
6.3 Sulfuric acid (1+1)—Slowly, add SO
mL of H,SO4 (ACS. sp. gr. 1.84) to 50 mL of
reagent water.
6.4 Acetone, hexane. methanol. methylene
chloride—Pesticide quality or equivalent.
6.5 Sodium sulfate—(ACS) Granular.
anhydrous. Purify by heating at 400 *C for 4 h
in a shallow tray.
0.0 Florisil—PR grade (60/100 mesh).
Purchase activated at 1250 *F and store in
dark in glass containers with ground glass
stoppers or foil-lined screw caps. Before use.
activate each batch at least 10 h at 200 *C in
a foil-covered glass container and allow to
cool.
6.7 Stock standard solutions (1.00 pg/
pL)—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions.
6.7.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in hexane and
dilute to volume in a 10-mL volumetric flask.
Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96* or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.7.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 *C and protect from light
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially Just prior to preparing
calibration standards from them.
6.7.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
M Quality control check sample
concentrate—See Section &2.1.
7. Calibration
7.1 Establish gas chromatographic
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 7.2) or the
internal standard technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
hexane. One of the external standards should
be at a concentration near, but above, the
MDL (Table 1) and the other concentrations
should correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.2.2 Using injections of 2 to 5 pL. analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (< 10% relative standard
deviation, RSD) linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
74.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flash. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with hexane. One of the standards
should be at a concentration near, but above,
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 pL. analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF-
(A.KCJ
(AJ(CJ
where:
A,—Response for the parameter to be
measured.
AbK Response for the internal standard.
d.-Concentration of the internal standard
(Mg/L).
C,-Concentration of the parameter to be
measured (jig/L).
If the RF value over the working range is
constant (< 10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively.
the results can be used to plot a calibration
curve of response ratios, A,/AH. vs. RF.
7.4 The working calibration curve.
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ± 15%. a new
calibration curve must be prepared for that
compound.
7.5 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements wen performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8-2.
8.1 J In recognition of advances that an
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4.11.1, and 12.1) to improve the
separations or lower the cost of
measurements. Bach time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents an changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10* of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 0^3.
8.1.5 The laboratory must on an ongoing
basis, demonstrate through the analyser of
quality control check standards that the
operation of the measurement system is in
control This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10ft of all
samples analysed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 107
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest in acetone at a
concentration of 20 n-g/mL for each
dinitrotoluene and 100 ng/mL for isophorone
and nitrobenzene. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at the test concentrations shown in
Table 2 by adding 1.00 mL of QC check
sample concentrate to each of four 1-L
aliquots of reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in fig/L. and the standard deviation of the
recovery (s) in ng/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter. Locate and
correct the source of the problem and repeat
the test for all parameters of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at the test concentration in
Section 8.2.2 or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2. whichever concentration would
be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any: or. if
none (2) the larger of either 5 times higher
than the expected background concentration
or the test concentration in Section 8.2.2.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100 (A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than the test concentration in Section
8.2.2, the analyst must use either the QC
acceptance criteria in Table 2, or optional QC
acceptance criteria calculated for the specific
spike concentration. To calculate optional
acceptance criteria for the recovery of a
parameter: (1) Calculate accuracy (X') using
the equation in Table 3, substituting the spike
concentration (T) for C; (2) calculate overall
precision (S') using the equation in Table 3,
substituting X' for XX8; (3) calculate the
range for recovery at the spike concentration
as (100 X'/T) ± 2.44 (100 S'/T)%.7
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check .
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4. If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of QC program for the
laboratory, method accuracy for waslewaler
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3.
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sB to P + 2sp. If P = 90% and SD =
10%. for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
9. Sample Collection. Preservation, and
Handling.
9.1 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 °C from the time of collection
until extraction.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.2
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory runnel. Check the pH of the
sample with wide-range pH paper and adjust
to within the range of 5 to 9 with sodium
hydroxide solution or sulfuric acid.
10.2 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory runnel and extract the
sample by shaking the runnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
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108
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
rcnthfugation, or other physical methods.
Collect the melhylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extcaction
in the same manner.
10.4 Assemble a Kuderna-Oanish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.5 Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate.
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Sections 10.7 and 10.8 describe a
procedure for exchanging the methylene
chloride solvent to hexane while
concentrating the extract volume to 1.0 mL
When it is not necessary to achieve the MDL
in Table 2, the solvent exchange may be
made by the addition of 50 mL of hexane and
concentration to 10 mL as described in
Method 608. Sections 10.7 and 10.8.
10.7 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL,
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.8 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. Add 1 to 2
mL of hexane and a clean boiling chip to the
concentrator tube and attach a two-ball
micro-Snyder column. Prewet the column by
adding about 0.5 mL of hexane to the top.
Place the micro-K-D apparatus on a hot
water bath (60 to 65 *C) so that the
concentrator tube is partially immersed in the
hot water. Adjust the vertical position of the
apparatus and the water temperature aa
required to complete the concentration in 5 to
10 min. At the proper rate of distillation the
balls of the column will actively chatter but
the chambers will not flood. When the
apparent volume of liquid reaches 0.5 mL
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.9 Remove the micro-Snyder column
and rinse its lower joint into the concentrator
tube with a minimum amount of hexane.
Adjust the extract volume to 1.0 mL. Stopper
the concentrator tube and store refrigerated if
further processing will not be performed
immediately. If the extract will be stored
longer than two days, it should be transferred
to a Teflon-sealed screw-cap vial. If the
sample extract requires no further cleanup.
proceed with gas chromatographic analysis
(Section 12). If the sample requires further
cleanup, proceed to Section 11.
10.10 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.
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use the procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
11.2 Florisil column cleanup:
11.2.1 Prepare a slurry of 10 g of activated
Florisil in methylene chloride/hexane
(1 +9)(V/V) and place the Florisil into a
chromatographic column. Tap the column to
settle the Florisil and add 1 cm of anhydrous
sodium sulfate to the top. Adjust the elution
rate to about 2 mL/min.
11.2.2 Just prior to exposure of the sodium
sulfate layer to the air, quantitatively transfer
the sample extract onto the column using an
additional 2 mL of hexane to complete the
transfer. Just prior to exposure of the sodium
sulfate layer to the air, add 30 mL of
methylene chloride/hexane (1 + 9)(V/V) and
continue the elution of the column. Discard
the eluate.
11.2.3 Next elute the column with 30 mL
of acetone/methylene chloride (1 + 9)(V/V)
into a 500-mL K-D flask equipped with a 10-
mL concentrator tube. Concentrate the
collected fraction as in Sections 10.6.10.7,
10.8, and 10.9 including the solvent exchange
to 1 mL of hexane. This fraction should
contain the nitroaromatics and isophorone.
Analyze by gas chromatography (Section 12).
12. Gas Chromatography
12.1 Isophorone and nitrobenzene are
analyzed by injection of a portion of the
extract into an FIDGC. The dinitrotoluenes
are analyzed by a separate injection into an
ECDGC. Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. Examples of
the separations achieved by Column 1 are
shown in Figures l and 2. Other packed or
capillary (open-tubular) columns,
chromatographic conditions, or detectors may
be used if the requirements of Section 8J are
met.
12.2 Calibrate the system daily as
described in Section 7.
12.3 If the internal standard calibration
procedure is being used, the internal standard
must be added to the same extract and mixed
thoroughly immediately before injection into
the gas chromalograph.
12.4 Inject 2 to 5 pL of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique.* Smaller (1.0 fiL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 pL the total extract volume.
and the resulting peak size in area or peak
height units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size: however.
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.6 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.7 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration
(VJ(V.)
where:
A=Amount of .'naterial injected (ng).
V|=Volume of extract injected (jiL).
Vt=Volume of total extract (/iL).
V,=Volume of water extracted (mL).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the samle using the response
factor (RF) determined in Section 7.3.2 and
Equation 3.
Equation 3.
Concentration (jig/L)>
(AJ(L)
(AJ(RF)(V.)
where:
A,=Response for the parameter to be
measured.
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Federal Register / Vol. 49, No. 209 / Friday, October 26. 1984 / Rules and Regulations
109
A,, = Response for the internal standard.
I, = Amount of internal standard added to
each extract (fig).
V0 = Volume of water extracted (L).
13.2 Report results in fig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.1 The MDL concentrations listed in
Table 1 were obtained using reagent water.10
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
7 x MDL to 1000 X MDL.' °
14.3 This method was tested by 18
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 1.0 to 515 u,g/L.'' Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136, Appendix B.
2. "Determination of Nitroaromatics and
Isophorone in Industrial and Municipal
Wastewaters," EPA-flOO/4-82-024, U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268, June 1982.
3. ASTM Annual Book of Standards, Part
31. D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials,
Philadelphia.
4. "Carcinogens—Working With
Carcinogens," Department of Health,
Education, and Welfare. Public Health
Service. Center for Disease Control. National
Institute for Occupational Safety and Health.
Publication No. 77-206. August 1977.
5. "OSHA Safety and Health Standards,
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration. OSHA 2206 (Revised.
January 1976).
6. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication, Committee on Chemical Safety,
3rd Edition, 1979.
7. Provost, L.P., and Elder, R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
8. ASTM Annual Book of Standards, Part
31. D3370-76. "Standard Practices for
Sampling Water," American Society for
Testing and Materials, Philadelphia.
9. Burke, J.A. "Gas Chromatography for
Pesticide Residue Analysis; Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists, 48.1037 (1965).
10. "Determination of Method Detection
Limit and Analytical Curve for EPA Method
609—Nitroaromatics and Isophorone."
Special letter report for EPA Contract 68-03-
2624, U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio 45268.
11. "EPA Method Validation Study 19,
Method 609 (Nitroaromatics and
Isophorone)," Report for EPA Contract 68-03-
2624 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS
AND METHOD DETECTION LIMITS
Parameter
Nitrobenzene
2,6-rAi'nlJOtoluone
2.4-Oinitrotoluene
Retention time
(mm)
Col. 1
3.31
3.52
4.49
5.35
Col. 2
4.31
4.75
5.72
6.54
Method detection
limfl (/ifl/L)
ECOGC
13.7
0.01
15.7
0.02
FIDGC
3.6
5.7
column temperature was held isothermal al 85 'C A 4 mm
ID column and 10% mo thane/90** argon earner gas at 44
ml/mm (low rate were used when determining the ttnitrotof-
uenes by ECDGC. The column temperature was held iso-
thermal at 145 *C.
Column 2 conditions: Gas-Chrom O (80/100 mesh) coated
with 3% OV-101 packed in a 3.0 m long \ 2 mm or 4 mm
ID glass column. A 2 mm ID column and nitrogen carrier gas
at 44 ml/mm (low rate were used when determining isophor-
one and nitrobenzene by FIDGC The column temperature
was held isothermal at 100 *C. A 4 mm ID column and 10S>
methane/90% argon earner gas at 44 ml/mm flow rate
were used when determining the dmitroto'uenes by ECDGC.
The column temperature was held isothermal at ISO *C.
TABLE 2.—QC ACCEPTANCE CRITERIA-
METHOD 609
Parameter
2.4-Dinitrotoluene
Isophorone
Test
Cone.
*L?'
20
20
100
100
Limit
lors
(jjj.
5.1
48
323
333
R_ange lor
X (M9/L)
3.6-228
3 8-23 0
8.0-1000
257-1000
for P.
P, ("*)
6-125
8-126
D-117
6-118
s = Standard deviation of tour recovery measurements, in
Mfl/L (Section 8.2.4).
X = Average recovery for (our recovery measurements, in
Mg/L (Section 6.2.4).
P. P. = Percent recovery measured (Section 8.3.2. Section
8.4.2).
D=Detected; result must be greater than zero.
Not*.—These criteria are based directly upon the method
performance data in Table 3. Where necessary, the limits lor
recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table
3.
TABLE 3.—METHOD ACCURACY AND PRECISION
AS FUNCTIONS OF CONCENTRATION—METH-
OD 609
Parameter
2,4-Oinitro-
toluene
2,6-Oinltro-
tofcwne
laophorone
Niti ober uur le
Accuracy,
as recovery,
X' Oig/U
0.65C+022
0.66C+0.20
0.49C + 2.93
0.60C+2.00
Single
analyst
precision.
V 0»0"J
0.20* + 008
0.19* +0.06
0.28X+2.77
0.25X + 2.53
Overall
precision. S'
0>g'U
0.37X 0.07
0.36X-0.00
0.46X+0.31
0.37X-0.78
Column 1 conditions: Gas-Chrom Q (80/100 mesh) coated
with 1.95% OF-1/1.5% OV-17 packed in « 1.2 m long X 2
nun or 4 mm ID glass column. A 2 mm 10 column and
nitrogen carrier gas at 44 mL/min flow rate were used when
determining isophorone and nitrobenzene by FIDGC. The
X' = Expected recovery for on* or more measurements ol a
sample containing • concentration of C, in ug/L
s,' = Expected single analyst standard deviation of meas-
urements at an average concentration found of X, in pg/L
S'=Expected interlaboratory standard deviation of meas-
urements at an average concentration found of X, in jig/L
CsTrue value tor the concentration, in pg/L
X» Average recovery found for measurements of samples
containing a concentration of C, in pg/L
BILUNQ CODE 6560-50-11
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HO Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
COIUMN: 1.5% 0V-17/1 95% QF-1 ON 8AS CMROM 0
TBNKKATIME: tS°C.
DETECTOH: FLAME IONIZATION
2 4 6 8 10 12
RETENTION TIME. MIN.
Figure 1. Gas chromatogram
of nitrobenzene
and isophorone.
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
111
COLUMN: 1.5% OV-17/1.95% QF-1 ON GAS CHROM Q
TEMPERATURE: 14S°C.
DETECTOR: &ECTRON CAPTURE
IK
- 1
| I
«* I
2468
RETENTION TIME. MIN.
Figure 2. Gas chromatogram
of dinitrotoluenes.
BILLING CODE CS60-SO-C
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112 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 610—Polynucleir Aromatic
Hydrocarbon*
/. Scope and Application
1.1 This method covers the determination
of certain polynuclear aromatic hydrocarbons
(PAH). The following parameters can be
determined by this method:
Paranwtvr
Slortt No.
CAS No
rmimrrmivi
AcanapMh»n»
Actnaphlhylww
Anthracan*
BanioUMwillvacww
0*nzo(a)pyfan«
B*fuo(b)fluorimn*n«
8«uo i*w.
63-32-9
206-96-6
120-12-7
56-55-3
50-32-6
205-99-2
191-24-2
207-06-9
216-01-9
53-70-3
206-44-0
86-73-7
193-39-5
91-20-3
65-01-6
129-00-0
1.2 This is a chromatographic method
applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyzeTjnfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
Method 625 provides gas chromatograph/
mass spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for many
of the parameters listed above, using the
extract produced by this method.
1.3 This method provides for both high
performance liquid chromatographic (HPLC)
and gas chromatographic (GC) approaches
for the determination of PAHs. The gas
chromatographic procedure does not
adequately resolve the following four pairs of
compounds: anthracene and phenanthrene:
chrysene and benzofajanthracene;
benzo(b)fluoranthene and
benzo(kjfluoranthene; and dibenzo(a.h)
anthracene and indeno (1.2,3-cd)pyrene.
Unless the purpose for the analysis can be
served by reporting the sum of an unresolved
pair, the liquid chromatographic approach
must be used for these compounds. The liquid
chromatographic method does resolve all 16
of the PAHs listed.
1.4 The method detection limit (MDL,
defined in Section 15.1) ' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.5 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 606, 608.
609, 611, and 612. Thus, a single sample may
be extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots. as necessary, to
apply appropriate cleanup procedures.
Selection of the aliquots must be made prior
to the solvent exchange steps of this method.
The analyst is allowed the latitude, under
Sections 12 and 13. to select chromatographic
conditions appropriate for the simultaneous
measurement of combinations of these
parameters.
1.6 Any modification of this method.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.7 This method is restricted to use by or
under the supervision of analysis
experienced in the use of HPLC and GC
systems and in the interpretation of liquid
and gas chroma(ograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample.
approximately 1-L. is extracted with
methylene chloride using a separately funnel.
The methylene chloride extract is dried and
concentrated to a volume of 10 mL or less.
The extract is then separated by HPLC or GC.
Ultraviolet (UV) and fluorescence detectors
are used with HPLC to identify and measure
the PAHs. A flame ionization detector is used
with GC.1
2.2 The method provides a silica gel
column cleanup procedure to aid in the
elimination of interferences that may be
encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardward that lead to discrete artifacts and/
or elevated baselines in the chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.1 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry. and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs. may not be
eliminated by this treatment. Solvent ring's
with acetone and pesticide quality hcxai,'
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedure in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achievr the MUL
listed in Table 1.
3.3 The extent of interferences that may
be encountered using liquid chromatographic
techniques has not been fully assessed.
Although the HPLC conditions described
allow for a unique resolution of the specific
PAH compounds covered by this method.
other PAH compounds may interfere.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method have 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 Pile of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified <-« for the information of the
analyst.
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian,
carcinogens: benzo(a)anthracene.
benzo(a)pyrene. and dibenzo(a.h)-
anthracene. Primary standards of these toxic
compounds should be prepared in a hood. A
NIOSH/MESA approved toxic gas respirator
should be worn when the analyst handles
high concentrations of these toxic
compounds.
5, Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 *C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
113
5.2.1 Separalory funnel—2-L. with Teflon
stopcock.
5.2.2 Drying column—Chromatographic
column, approximately 400 mm long x 19 mm
ID, with coarse frit filter disc.
5.2.3 Concentrator tube. Kuderna-
Danish—10-mL. graduated (Kontcs K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.4 Evaporative flask. Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.5 Snyder column, Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.6 Snyder column. Kuderna-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
5.2.7 Vials—10 to 15-mL, amber glass,
with Teflon-lined screw cap.
5.2.8 Chromatographic column—250 mm
long x 10 mm ID, with coarse frit filter disc at
bottom and Teflon stopcock.
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 °C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2 "C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 High performance liquid
chromatograph (HPLC)—An analytical
system complete with column supplies, high
pressure syringes, detectors, and compatible
strip-chart recorder. A data system is
recommended for measuring peak areas and
retention times.
5.6.1 Gradient pumping system—Constant
flow.
5.6.2 Reverse phase column—HC-ODS
Sil-X, 5 micron particle diameter, in a 25 cm x
2.6 mm ID stainless steel column (Perkin
Elmer No. 089-0716 or equivalent). This
column was used to develop the method
performance statements in Section 15.
Guidelines for the use of alternate column
packings are provided in Section 12.2.
5.6.3 Detectors—Fluorescence and/or UV
detectors. The fluorescence detector is used
for excitation at 280 nm and emission greater
than 389 nm cutoff (Corning 3-75 or
equivalent). Fluorometers should have
dispersive optics for excitation and can
utilize either filter or dispersive optics at the
emission detector. The UV detector is used at
254 nm and should be coupled to the
fluorescence detector. These detectors were
used to develop the method performance
statements in Section 15. Guidelines for the
use of alternate detectors are provided in
Section 12.2.
5.7 Gas chromatograph—An analytical
system complete with temperature
programmable gas chromatograph suitable
for on-column or splitless injection and all
required accessories including syringes,
analytical columns, gases, detector, and strip-
chart recorder. A data system is
recommended for measuring peak areas.
5.7.1 Column—1.8 m long x 2 mm ID glass.
packed with 3% OV-17 on Chromosorb W-
AW-DCMS (100/120 mesh) or equivalent.
This column was used to develop the
retention time data in Table 2. Guidelines for
the use of alternate column packings are
provided in Section 13.3.
5.7.2 Detector—Flame ionization detector.
This detector has proven effective in the
analysis of wastewaters for the parameters
listed in the scope (Section 1.1). excluding the
four pairs of unresolved compounds listed in
Section 1.3. Guidelines for the use of
alternate detectors are provided in Section
13.3.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an inlerferent is
not observed at the MDL of the parameters of
interest.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Cyclohexane, methanol. acetone.
methylene chloride, pentane—Pesticide
quality or equivalent.
6.4 Acetonitrile—HPLC quality, distilled
in glass.
6.5 Sodium sulfate—(ACS) Granular,
anhydrous. Purify by heating at 400 'C for 4 h
in a shallow tray.
6.6 Silica gel—100/200 mesh, desiccant.
Davison, grade-923 or equivalent. Before use.
activate for at least 16 h at 130 *C in a
shallow glass tray, loosely covered with foil.
6.7 Stock standard solutions (1.00 ug/
u,L)—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions.
6.7.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in acetonitrile
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.7.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 "C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.7.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.8 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish liquid or gas
Chromatographic operating conditions
equivalent to those given in Table 1 or 2. The
Chromatographic system can be calibrated
using the external standard technique
(Section 7.2) or the internal standard
technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
acetonitrile. One of the external standards
should be at a concentration near, but above.
the MDL (Table 1) and the other
concentrations should correspond to tho
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using injections of 5 to 25 j*L for
HPLC and 2 to 5 uL for CC. analyze each
calibration standard according to Section 12
or 13. as appropriate. Tabulate peak height or
area responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (<10% relative standard
deviation. RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with acetonitrile. One of the
standards should be at a concentration near,
but above, the MDL and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.3.2 Using injections of 5 to 25 ul for
HPLC and 2 to 5 jiL for CC, analyze each
calibration standard according to Section 12
or 13, as appropriate. Tabulate peak height or
area responses against concentration for
each compound and internal standard.
Calculate response factors (RF) for each
compound using Equation 1.
Equation 1.
RF=
(A.)(CU)
(AUJ(C.)
where:
A.=Response for the parameter to be
measured.
Au= Response for the internal standard.
Cu = Concentration of the internal standard
C. = Concentration of the parameter to be
measured (ng/L).
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A,/AU, vs. RF.
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114 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, the test must
be repealed using a fresh calibration
standard. Alternatively, a new calibration
curve must be prepared for that compound.
7.5 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 reagants.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with (hit method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4.11.1,12.2. and 13.3) to improve
the separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at the following
concentrations in acetonitrilc: 100 >ig/mL of
any of the six early-eluting PAHs
(naphthalene, acenaphthylene. acenaphthenc,
fluorene. phenanthrene, and anthracene): 5
Mg/mL of benzo(k)fluoranthene: and 10 ng/
ml of any of the other PAHs. The QC check
sample concentrate must be obtained from
the U.S. Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati. Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet. prepare QC check
samples at the test concentrations shown in
Table 3 by adding 1.00 mL of QC check
sample concentrate to each of four 1-L
aliquots of reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in fig/L. and the standard deviation of the
recovery (s) in jig/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively.
found in Table 3. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 3 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.6 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.6.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.2.
8.2.6.2 Beginning with Section 8.2.2, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2. whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at the test concentration in
Section 8.2.2 or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2. whichever concentration would
be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded).
the spike concentration should be (1) the
regulatory concentration limit, if any: or. if
none, (2) the larger of either 5 limes higher
than the expected background concentration
or the test concentration in Section 8.2.2.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100 (A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 3. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.T If
spiking was performed at a concentration
lower than the test concentration in Section
8.2.2. the analyst must use either the QC
acceptance criteria in Table 3, or optional QC
acceptance criteria calculated for the specific
spike concentration. To calculate optional
acceptance criteria for the recovery of a
parameter (1) calculate accuracy (X') using
the equation in Table 4. substituting the spike
concentration (T) for C; (2) calculate overall
precision (S') using the equation in Table 4.
substituting X' for X: (3) calculate the range
for recovery at the spike concentration as
(100 X'/T)±2.44(100 S'/T)%.7
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the critiera must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
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Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
115
laboratory. If the entire list of parameters in
Table 3 must be measured in the sample in
Section 8.3. the probability that the analysis
of a QC check standard will be required is
high. In this case the QC check standard
should be routinely analyzed wilh the spike
sample.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%. where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 3.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.S As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sp to P+2sp. If P=90% and sp=10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram, confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
ft Sample Collection. Preservation, and
Handling
9.1 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 'C from the time of collection
until extraction. PAHs are known to be light
sensitive: therefore, samples, extracts, and
standards should be stored in amber or foil-
wrapped bottles in order to minimize
photolytic decomposition. Kill the sample
bottles and. if residual chlorine is present.
add 80 mg of sodium thiosulfate per liter of
sample and mix well. EPA Methods 330.4 and
330.5 may be used for measurement of
residual chlorine.9 Field test kits are
available for this purpose.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.2
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory funnel.
10.2 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the funnel for 2 min. with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.5 Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate,
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL,
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.7 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. Stopper the
concentrator tube and store refrigerated if
further processing will not be performed
immediately. If the extract will be stored
longer than two days, it should be transferred
to a Teflon-sealed screw-cap vial and
protected from light. If the sample extract
requires no further cleanup, proceed with gas
or liquid chromatographic analysis (Section
12 or 13). If the sample requires further
cleanup, proceed to Section 11.
10.8 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.
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use the procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the methods
as revised to incorporate the cleanup
procedure.
11.2 Before the silica gel cleanup
technique can be utilized, the extract solvent
must be exchanged to cyclohexane. Add 1 to
10 mL of the sample extract (in methylene
chloride) and a boiling chip to a clean K-D
concentrator tube. Add 4 mL of cyclohexane
and attach a two-ball micro-Snyder column.
Prewet the column by adding 0.5 mL of
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 to 10 min. At the proper
rate of distillation the balls of the column will
actively chatter but the chambers will not
flood. When the apparent volume of the
liquid reaches 0.5 mL, remove the K-D
apparatus and allow it to drain and cool for
at least 10 min. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with a minimum amount of
cyclohexane. Adjust the extract volume to
about 2 mL.
11.3 Silica gel column cleanup for PAHs:
11.3.1 Prepare a slurry of 10 g of
activiated silica gel in methylene chloride
and place this into a 10-mm ID
chromatographic column. Tap the column to
settle the silica gel and elute the methylene
chloride. Add 1 to 2 cm of anhydrous sodium
sulfate to the top of the silica gel.
11.3.2 Preelute the column with 40 mL of
pentane. The rate for all elutions should be
about 2 mL/min. Discard the eluate and just
prior to exposure of the sodium sulfate layer
to the air, transfer the 2-mL cyclohexane
sample extract onto the column using an
additional 2 mL cyclohexane to complete the
transfer. Just prior to exposure of the sodium
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116 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
sulfale layer to the air, add 25 mL of penlane
and continue the elution of the column.
Discard this penlane eluate.
11.3.3 Next, elute the column with 25 mL
of mcthylene chloride/pentanc (4 + 6)(V/V)
into u 500-mL K-D flask equipped with a 10-
mL concentrator tube. Concentrate the
collected fraction to less than 10 mL as in
Section 10.6. When the apparatus is cool.
remove the Snyder column and rinse the flask
and its lower joint with pentane. Proceed
with HPLC or CC analysis.
12. High Performance Liquid
Chromatography
12.1 To the extract in the concentrator
tube, add 4 mL of acetonitrile and a new
boiling chip, then attach a two-ball micro-
Snyder column. Concentrate the solvent as in
Section 10.6, except set the water bath at 95
to 100 *C. When the apparatus is cool.
remove the micro-Snyder column and rinse
its lower joint into the concentrator tube with
about 0.2 mL of acetonitrile. Adjust the
extract volume to 1.0 mL.
12.2 Table 1 summarizes the
recommended operating conditions for the
HPLC. Included in this table are retention
times, capacity factors, and MDL that can be
achieved under these conditions. The UV
detector It recommended for the
determination of naphthalene,
acenaphthylene. acenapthene. and fluorene
and the fluorescence detector is
recommended for the remaining PAHs.
Examples of the separations achieved by this
HPLC column are shown in Figure* 1 and 2.
Other HPLC column*, chromatographic
condition!, or detectors may be used if the
requirements of Section &2 are met
12.3 Calibrate the system daily as
described in Section 7.
12.4 If the internal standard calibration
procedure is being used the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the instrument
12.5 Inject 5 to 25 pL of the sample extract
or standard into the HPLC using a high
pressure-syringe or a constant volume sample
injection loop. Record the volume injected to
the nearest 0.1 pL, and the resulting peak size
in area or peak height units. Re-equilibrate
the HPLC column at the initial gradient
conditions for at least 10 min between
injections.
12.6 Identify the parameters in the sample
by comparing the retention time of the peaks
in the sample chromatogram with those of the
peaks in standard chromatogram*. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size: however,
the experience of the analyst should weigh
heavily in the interpretation of
chroma tograms.
12.7 If the response for a peak exceeds
the working range of the system, dilute the
extract with acetonitrile and reanalyze.
12.8 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Cos Chromatography
13.1 The packed column GC procedure
will not resolve certain isomeric pairs as
indicated in Section 1.3 and Table 2. The
liquid chromalographic procedure (Section
12) must be used for these parameters.
13.2 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 the micro-K-D
apparatus on a hot water bath (60 to 65 '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 to 10 min. 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 and cool for at least 10
min. Remove the micro-Snyder column and
rinse its lower joint into the concentrator
tube with a minimum amount of methylene
chloride. Adjust the final volume to 1.0 mL
and stopper the concentrator tube.
13.3 Table 2 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times that were obtained under
these conditions. An example of the
separations achieved by this column is
shown in Figure 3. Other packed or capillary
(open-tubular) columns, chromatographic
conditions, or detectors may be used if the
requirements of Section 8.2 are met
13.4 Calibrate the gas chromatographic
system dairy as described in Section 7.
13.5 If the internal standard calibration
procedure is being used, the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the gas chromatograph.
13.0 Inject 2 to 5 ftL of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique.10 Smaller (1.0
pL) volumes may be injected if automatic
devices are employed. Record the volume
injected to the nearest 0.05 pL and the
resulting peak size in area or peak height
units.
13.7 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chroma tograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size; however,
the experience of the analyst should weigh
heavily in the interpretation of
chroma tograms.
13.8 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
13.9 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
14. Calculations
14.1 Determine the concentration of
individual compounds in the sample.
14.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration (>ig/L) =
where:
A=Amount of material injected (ng).
V, = Volume of extract injected (pL).
V,=Volume of total extract (jtL).
V.=Volume of water extracted (mL).
14.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2. and Equation 3.
Equation 3.
Concentration (jig/L)- -
(AJfRFJfVJ
where:
A.=Response for the parameter to be
measured.
Ab=Response for the internal standard.
I.=Amount of internal standard added to
each extract (jig).
V.=Volume of water extracted (L).
14.2 Report results in /ig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
15. Method Performance
15.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.' The MDL concentrations listed in
Table 1 were obtained using reagent water.''
Similar results were achieved using
representative wastewaters. MDL for the GC
approach were not determined. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
15.2 This method ha* been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
8 x MDL to 800 x MDL1 > with the following
exception: benzo(ghi)perylene recovery it 80
X and 800 X MDL were low (35* and 45%,
respectively).
15.3 This method was tested by 16
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentration*
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
117
over the range 0.1 (o 425 fig/L.1 J Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 4.
References
1. 40 CFR Part 136, Appendix B.
2. "Determination of Polynuclear Aromatic
Hydrocarbons in Industrial and Municipal
Wastewaters," EPA-600/4-82-025. U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory. Cincinnati, Ohio 45268,
September 1982.
3. ASTM Annual Book of Standards. Part
31. D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials.
Philadelphia.
4. "Carcinogens—Working With
Carcinogens," Department of Health,
Education, and Welfare, Public Health
Service. Center for Disease Control, National
Institute for Occupational Safety and Health,
Publication No. 77-206. August 1977.
5. "OSHA Safety and Health Standards,
General Industry," (29 CFR 1910),
Occupational Safety and Health
Administration, OSHA 2206 (Revised,
January 1976).
6. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication, Committee on Chemical Safety,
3rd Edition. 1979.
7. Provost, L.P., and Elder, R.S.
"Interpretation of Percent Recovery Data."
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
8. ASTM Annual Book of Standards, Part
31. D3370-76. "Standard Practices for
Sampling Water," American Society for
Testing and Materials, Philadelphia.
9. "Methods 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric. DPD) for
Chlorine, Total Residual," Methods for
Chemical Analysis of Water and Wastes,
EPA-600/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268. March 1979.
10. Burke, J.A. "Gas Chromatography for
Pesticide Residue Analysis; Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists. 48.1037 (1965).
11. Cole. T., Riggin, R., and Glaser, J.
"Evaluation of Method Detection Limits and
Analytical Curve for EPA Method 610—
PNAs." International Symposium on
Polynuclear Aromatic Hydrocarbons. 5th.
Battelle's Columbus Laboratories. Columbus.
Ohio (1980).
12. "EPA Method Validation Study 20.
Method 610 (Polynuclear Aromatic
Hydrocarbons)," Report for EPA Contract 68-
03-2624 (In preparation).
TABLE 1.—HIGH PERFORMANCE LIQUID CHRO-
MATOGRAPHY CONDITIONS AND METHOD DE-
TECTION LIMITS
TABLE 3.—QC ACCEPTANCE CRITERIA-
METHOD 610
Parameter
Naphthalene
Fluorene
Dibenzo(a.h)anthracene
Indenod 23-cd)pyrene
Reten-
tion
time
(min)
16.6
165
205
21.2
22 1
234
245
254
28.5
293
31 6
329
33.9
35.7
363
37.4
Column
capac-
ity
factor
Ik')
12.2
13.7
152
1S.S
16.6
176
185
19.1
21.6
222
240
25 1
25.9
27.4
278
28.7
Method
detec-
tion
limit
()ig'U '
1.8
23
1 8
0.21
064
066
021
027
0013
0 15
0018
0017
0.023
0.030
0076
0043
HPLC column conditions: Reverse phase HC-OOS Sil-X.
5 micron particle size, in a 25 cm x 2.6 mm ID stainless
steel column. Isocratx: elution for 5 min using acetonitrile/
water (4+6), then linear gradient elution to 100% acetonitnle
over 25 min at 0.5 ml/mm flow rate. If columns having other
internal diameters are used, the flow rate should be adjusted
to maintain a linear velocity of 2 mm/sec.
•The MDL for naphthalene, acenaphthylene, acenaph-
thene. and fluorene were determined using a UV detector. All
others were determined using a fluorescence detector.
TABLE 2.—GAS CHROMATOGRAPHIC
CONDITIONS AND RETENTION TIMES
Parameter
Naphthalene
AcenapMhyieoG
Acenflphrrume
Anthracene
Pyrene
Benzo(a)anlhracene
Banzo(b)fluoranthene
Dibenzo(a.h)anthracene
lndeno(1 2 3-cd)pyrene
Benzo(gtti)perylene
Retention
time (min)
4.5
104
108
126
15.9
159
198
206
24.7
247
280
280
294
36.2
362
38.6
Parameter
Acenaphthene
Acenaphthytene
Benzo(a)anthracer>e ....
Beruo{a)pyrene
Beruoenzo(a.h)an-
Ftooranthene
lndeno<1,2.3-
Naphthatene
Test
cone.
"ir
too
too
too
10
10
10
10
5
10
10
10
100
to
100
100
10
Limit
lor s
"tf'
40.3
45.1
267
4.0
4.0
3.1
2.3
2.S
4.2
20
30
430
30
407
377
34
Range for
* (M9'U
0-105.7
22.1-112.1
11 2-1123
3.1-11.6
0.2-11 0
1.8-13.8
0-10.7
D-7.0
0-17.5
0 3-tOO
2 7-1 1 1
D-119
1 2-100
21 5-1000
8 4-133 7
Range
lex P
P. O)
0-124
0-139
0-126
12-135
0-128
6-150
D-116
0-159
0-199
0-110
14-123
s = Standard deviation of four recovery measurements, in
(ip/L (Section 8.2.4).
X = Average recovery for lour recovery measurements, in
l»g/L (Section 8.2.4).
>. P, = Percent recovery measured (Section 8.3.2, Section
8.4.2).
0 = Delected; result must be greater than zero.
NOTE.—These criteria are based directly upon the method
performance data in Table 4. Where necessary, the limits for
recovery have been broadened to assure applicability of the
limits lo concentrations below those used to develop Table
TABLE 4.—METHOD ACCURACY AND PRECISION
AS FUNCTIONS OF CONCENTRATION—METH-
OD 610
Parameter
Acenaphtnene
Acenaphtnylene
Benzo 2 mm
ID glass column witn nitrogen carrier gas at 40 ml/mm flow
rate. Column temperature was held at 100 'C for 4 min, then
programmed at 8 'C/min to i final hold at 280 *C.
X' = Expected recovery for one or more measurements of a
sample containing a concentration of C. in ug/L
s,'=Expeded tingle analyst standard deviation of meas-
urements it an average concentration found of X, in pg/L.
S'=Expected mtertaboratory ttandard deviation of meas-
urements at an average concentration found of X, in >ig/L
C=True value for ma concentration, in )ig/L
XaAverage recovery found for measurements of samples
containing a concentration of C, in )>g/L
BILLING CODE (MO-SO-M
-------
COLUMN: HC-ODS SIL-X
MOMLE PHASE: «OK T0100% ACETONITMLE IN WATER
DETECTOR: ULTRAVIOLET AT 254nm
I
o
to
o
2
a.
a
O
o
o
a-
A
4 •12WJ024»ttM
RETENTION TIME. MIN.
Figure 1. Liquid chromatogram of polynuolear aromatic hydr<
at
a.
90
-------
COLUMN: HC-ODS SIL-I
MOBILE PHASE: 40XtO 100% ACETONITRILE
IN WATER
DETECTOR: FLUORESCENCE
3?
Q.
•73
(B
00
Z
o
a.
to
O
o
o
cr
(D
12 16 20 24
RETENTION TIME. MIN.
28
Figure 2. Liquid chromatogram of polynuclear aromatic hydrocarbons.
o
CO
0)
O.
I
o
CO
-------
COLUMN: 3X OV-170N CHROMOSOftl ff A« DCttS
PROGRAM: 100*C FOR 4 MIN. 8%/IMN TO 280«C
DETECTOR: FLAME IONIZAT10N
o
o.
eg
*<
o
"
20 34
RETENTION TIME. MIN.
32
Figure 3. Gas chromatogram of polynuclear aromatic hydrocarbons.
ye
c
s
to
Q.
§
C0
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
121
Method 811—Haloelhers
1. Scope and Application
1.1 This method covers the determination
of certain haloethers. The following
parameters can be determined by this
method:
Parameter
Bis(2-chtofoethyl) ether
Bis(2) methane
Bis(2-chlororsopropyi) ether
4-Bromophenyt phenyl ether
4-Chlorophenyl phenyl either
Suxet No.
34273
34278
34283
34636
34641
CAS No.
111-44-4
111-91-1
108-60-1
101-55-3
7005-72-3
1.2 This is a gas chromatographic (GC)
method applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes analytical conditions
for a second gas chromatographic column
that can be used to confirm measurements
made with the primary column. Method 625
provides gas chromatograph/mass
spectrometer (GC/MS) conditions
appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
W 1.3 The method detection limit (MDL,
defined in Section 14.1)' for each parameter is
listed in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 606.608,
609, and 612. Thus, a single sample may be
extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots, as necessary, to
apply appropriate cleanup procedures. The
analyst is allowed the latitude, under Section
12. to select chromatographic conditions
appropriate for the simultaneous
measurement of combinations of these
parameters.
1.5 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample.
approximately 1-U is extracted with
methylene chloride using a separatory funnel.
The methylene chloride extract is dried and
exchanged to hexane during concentration to
a volume of 10 mL or less. The extract is
separated by gas chromatography and the
parameters are then measured with a halide
specific detector.1
2.2 The method provides a Florisil column
cleanup procedure to aid in the elimination of
interferences thai may be encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents,
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.3 Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed be detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such a PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling.
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedure in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
3.3 Dichlorobenzenes 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. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of materm! data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified *"* for the information of the
analyst.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 'C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use. however, the compressible
tubing should be thoroughly rinsed with
methanol. followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L. with Teflon
stopcock.
t.2.2 Drying column—Chromatographic
column, approximately 400 mm long x 19 mm
ID, with coarse frit filter disc.
5.2.3 Chromatographic column—400 mm
long x 19 mm ID, with Teflon stopcock and
coarse frit filter disc at bottom (Konte.s K-
420540-0224 or equivalent).
5.2.4 Concentrator tube, Kuderna-
Danish—10-mL. graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask. Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column, Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Vials—10 to 15-mL. amber glass,
with Teflon-lined screw cap.
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2'C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 Gas chromatograph—An analytical
system complete with temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
-------
122
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
column*, gases, detector, and atrip-chart
recorder. A data system is recommended for
measuring peak areas.
5.6.1 Column 1—1.8 m long x 2 mm ID
glass, packed with 3% SP-1000 on
Supelcoport (100/120 mesh) or equivalent.
This column was used to develop the method
performance statements in Section 14.
Guidelines for the use of alternate column
packings are provided in Section 12.1.
5.6.2 Column 2—1.8 m long x 2 mm ID
glass, packed with 2.6-diphenylene oxide
polymer (60/80 mesh). Tenax. or equivalent.
5.6.3 Detector—Halide specific detector:
electrolytic conductivity or microcoulomelric.
These detectors have proven effective in the
analysis of wastewaters for the parameters
listed in the scope (Section 1.1). The Hall
conductivity detector was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
detectors are provided in Section 12.1.
Although less selective, an electron capture
detector is an acceptable alternative.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Acetone, hexane, methanot. methylene
chloride, petroleum ether (boiling range 30-60
•C)—Pesticide quality or equivalent.
6.4 Sodium sulfate—(ACS) Granular,
anhydrous. Purify by heating at 400 *C for 4 h
in a shallow tray.
6.5 Florisil—PR Grade (60/100 mesh).
Purchase activated at 1250 *F and store in the
dark in glass containers with ground glass
stoppers or foil-lined screw caps. Before use,
activate each batch at least 16 h at 130 *C in
a foil-covered glass container and allow to
cool.
6.6 Ethyl ether—Nanograde, redistilled in
glass if necessary.
6.6.1 Ethyl ether must be shown to be free
of peroxides before it is used •• indicated by
EM Laboratories Quant test strips. (Available
from Scientific Products Co.. Cat. No. P1126-
8, and other suppliers.)
6.6.2 Procedures recommended for
removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl
alcohol preservative must be added to each
liter of ether.
6.7 Stock standard solutions (1.00 ug/
/iL)—Stock standard solutions can be
prepared from pure standard material* or
purchased as certified solutions.
6.7.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve die material in acetone
and dilute to volume in a 10-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst When compound
purity is assayed to be 98% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source. •
6.7.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 'C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.7.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with check standards indicates a
problem.
6.8 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish gas chromatographic
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 7.2) or the
internal standard technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
hexane. One of the external standards should
be at a concentration near, but above, the
MDL (Table 1) and the other concentrations
should correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.2.2 Using injections of 2 to 5 pL, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (<10% relative standard
deviation, RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with hexane. One of the standards
should be at a concentration near, but above.
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 pL. analyse
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF =
(A.HCJ
(AJ(C.)
where:
A, = Response for the parameter to be
measured.
A,. = Response for the internal standard.
Ctt=Concentration of the internal standard
(Mg/L).
C. = Concentration of the parameter to be
measured (pg/L).
If the RF value over the working range is a
constant (< 10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios. A./AU, vs. RF.
7.4 The working calibration curve.
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, a new
calibration curve must be prepared for that
compound.
7.5 The cleanup procedure in Section 11
utilizes Florisil column chromatography.
Florisil from different batches or sources may
vary in adsorptive capacity. To standardize
the amount of Florisil which is used, the use
of lauric acid value* is suggested. The
referenced procedure determines the
adsorption from hexane solution of lauric
acid (mg) per g of Florisil. The amount of
Florisil to be used for each column is
calculated by dividing 110 by this ratio and
multiplying by 20 g.
7.6. 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements wen performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section Bi
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4, ll.l, and 12.1) to improve the
separations or lower the cost of
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 123
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of
100 Mg/mL in acetone. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati, Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at a concentration of 100 jig/L by
adding 1.00 ml of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in /ig/L, and the standard deviation of the
recovery (a) in pg/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter. Locate and
correct the source of the problem and repeat
the test for all parameters of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spiked
sample per month is required.
8.3.1. The concentration of the spike in
the sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 100 fig/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any: or, if
none (2) the larger of either 5 times higher
than the expected background concentration
or 100 /ig/L.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 ml of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 100 fig/L, the analyst must use
either the QC acceptance criteria in Table 2,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter (1) calculate
accuracy (X') using the equation in Table 3,
substituting the spike concentration (T) for C;
(2) calculate overall precision (S') using the
equation in Table 3, substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) ±2.44(100 S'/
T)%.»
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3. a QC
check standard containing each parameter
that failed must be prepared and analyzed.
NOTE.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 m/L of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2Sp to P+2sp. If P=90% and sp = 10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.6 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. When doubt
exists over the identification of a peak on the
chromatogram. confirmatory techniques such
as gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
9. Sample Collection, Preservation, and
Handling
9.1 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 as free as possible of Tygon tubing
and other potential sources of contamination.
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124 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
9.2 All samples must be iced or
refrigerated at 4'C from the time of collection
until extraction. Fill the sample bottles and. if
residual chlorine is present, add 80 mg of
sodium thiosulfate per liter of sample and
mix well. EPA Methods 330.4 and 330.5 may
be used for measurement of residual
chlorine.10 Field test kits are available for this
purpose.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.1
10. Sample Extraction
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory funnel.
10.2 Add 60 mL methylene chloride to the
sample bottle, seal, and shake 30 s to rinse
the inner surface. Transfer the solvent to the
separatory funnel and extract the sample by
shaking the funnel for 2 min with periodic
venting to release excess pressure. Allow the
organic layer to separate from the water
phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation. or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 00-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extract! in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kudema-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a SOO-mL evaporative
flask. Other concentration device* or
techniques may be used in place of the K-D
concentrator if the requirements of Section
&2 are met
1O5 Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate,
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
th* top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL,
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
Note.—Some of the haloethers are very
volatile and significant losses will 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 to 2 mL before
removing the K-D apparatus from the hot
water bath.
10.7 Momentarily remove the Snyder
column, add 50 mL of hexane and a new
boiling chip, and rcattach the Snyder column.
Raise the temperature of the water bath to 85
to 90 *C. Concentrate the extract as in
Section 10.6, except use hexane to prewet the
column. The elapsed time of concentration
should be 5 to 10 min.
10.8 Remove the Snyder column and rinse
the flask and its lower joint into the
concentrator tube with 1 to 2 mL of hexane. A
5-mL syringe is recommended for this
operation. Stopper the concentrator tube and
store refrigerated if further processing will
not be performed immediately. If the extract
will be stored longer than two days, it should
be transferred to a Teflon-sealed screw-cap
vial. If the sample extract requires no further
cleanup, proceed with gas chromatographic
analysis (Section 12). If the sample requires
further cleanup, proceed to Section 11.
10.9 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
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use the procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section &2 can be met using the method as
revised to incorporate the cleanup procedure.
11.2 Florisil column cleanup for
haloethers:
11.2.1 Adjust the sample extract volume
tolOmL
11.2J Place a weight of Florisil (nominally
20 g) predetermined by calibration (Section
7.5), into a chromatographic column. Tap the
column to settle the Florisil and add 1 to 2 cm
of anhydrous sodium sulfate to the top.
11.2.3 Preelute the column with 50 to 60
mL of petroleum ether. Discard the eluate and
just prior to exposure of the sodium sulfate
layer to the air, quantitatively transfer the
sample extract onto 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
ethyl ether/petroleum ether (6+94) (V/V).
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.
11.2.4 Concentrate the fraction as in
Section 10.6, except use hexane to prewet the
column. When the apparatus is cool, remove
the Snyder column and rinse the flask and its
lower joint into the concentrator tube with
hexane. Adjust the volume of the cleaned up
extract to 10 mL with hexane and analyze by
gas chromatography (Section 12).
12. Gas Chromatography
12.1 Table 1 summarizes (he
recommended operating conditions for the
giis chromatograph. Included in this la.ble are
retention times and MDL (hat can be
achieved under these conditions. Examples of
the separations achieved by Columns 1 and 2
are shown in Figures 1 and 2, respectively.
Other packed or capillary (open-tubular)
columns, chromatographic conditions, or
detectors may be used if the requirements of
Section 8.2 are met.
12.2 Calibrate the system daily as
described in Section 7.
12.3 If the internal standard calibration
procedure is being used, the internal standard
must be added to the sample extract and
mixed thoroughly immediately before
injection into the gas chromatrograph.
12.4 Inject 2 to 5 fit of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique." Smaller (1.0 fiL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 uL. the total extract volume.
and the resulting peak size in area or peak
height units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromatograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over the course of a
day. Three times the standard deviation of a
retention time for a compound can be used to
calculate a suggested window size; however,
the experience of the analyst should weight
heavily in the interpretation of
chromatograms.
124 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.7 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculation*
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration, Oig/L)«
(A)(VJ
(VJ(VJ
where:
A—Amount of material injected (ng).
V,- Volume of extract injected fjtL).
V,- Volume of total extract (jiL).
V.- Volume of water extracted (mL).
13.17 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
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Federal Register / Vol. 49, No, 209 / Friday. October 26. 1984 / Rules and Regulations
125
Equation 3.
Concentration (u,g/L)=-
(Ate)(RF)(V0)
where:
A. = Response for the parameter to be
measured.
Alf = Response for the internal standard.
I. = Amount of internal standard added to
each extract (ng).
V0 = Volume of water extracted (L).
13.2 Report results in ng/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.'The MDL concentrations listed in
Table 1 were obtained using reagent water.12
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
4 X MDL to 1000 X MDL.12
14.3 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 1.0 to 626 fi/L.11 Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1.40 CFR Part 136, Appendix B.
2. "Determination of Haloethers in
Industrial and Municipal Wastewaters,"
EPA-600/4-81-062, U.S. Environmental
Protection Agency, Environmental
Monitoring and Support Laboratory.
Cincinnati. Ohio 45268, December 1981.
3. ASTM Annual Book of Standards, Part
31, D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constitutents,"
American Society for Testing and Materials, -.
Philadelphia.
4. "Carcinogens—Working Carcinogens, "
Department of Health, Education, and
Welfare, Public Health Services, Center for
Disease Control. National Institute for
Occupational Safety and Health. Publication
No. 77-206, August 1977.
5. "OSHA Safety and Health Standards.
General Industry," (29 CFR 1910).
Occupational Safety and Health
Administration. OSHA 2200 (Revised,
January 1976).
6. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication. Committee on Chemical Safety,
3rd Edition. 1979.
7. 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. Provost. L.P., and Elder. R.S.
"Interpretation of Percent Recovery Data."
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
9. ASTM Annual Book of Standards, Part
31, D3370-76. "Standard Practices for
Sampling Water." American Society for
Testing and Materials. Philadelphia.
10. "Methods 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric, DPD) for
Chlorine, Total Residual," Methods for
Chemical Analysis of Water and Wastes,
EPA-600/4-79-020. U.S. Environmental
Protection Agency. Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, March 1979.
11. Burke, J.A. "Gas Chromatography for
Pesticide Residue Analysis; Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists. 48,1037 (1965).
12. "EPA Method Validation Study 21,
Method 611 (Haloethers)," Report for EPA
Contract 68-03-2633 (In preparation).
TABLE 1.—CHROMATOGRAPHIC CONDITIONS
AND METHODS DETECTION LIMITS
Parameters
Bis(2-chloroisopropyl) ether
Bis(2-criloroetriyi) ether ,
Bis(2-chkxoethoxy) methane
4-Cnlorophenyl ether
4-Bromophenyl phenyl ether
Retention time
(min)
Col-
umn t
8.4
9.3
13.1
19.4
21.2
Col-
umn 2
9.7
9.1
10.0
15.0
16.2
Column 1 conditions: Supelcocort (100/120 mesh
with 3% SP-1000 packed in a 1.8 m long > 2 mm
Meth-
od
detec-
tion
limit
0.6
0.3
O.S
3.9
2.3
coated
D glass
column with helium carrier gas at 40 mL/min flow rate.
Column temperature held at 60 'C for 2 mm after injection
then programmed at 8 ' C/min to 230 'C and nek) for 4 min.
Under these conditions the retention time for Aktrin is 22.6
min.
Column 2 conditions: Tenax-GC (60/80 mesh) packed in
a 1.8 m long x 2mm ID glass column with helium carrier gas
at 40 mL/mV flow rate. Column temperature held at 150 'C
tor 4 min after injection then programmed at 18 •C/min to
310 'C Under these conditions the retention time lor Aldnn
is 16.4 mm.
TABLE 2.—QC ACCEPTANCE CRITERIA-
METHOD 611
Parameter
Bis (2
chloroethyl)ether....
Bis (2-
chloroethoxylmethe
Bra (2-
chlorasopropyl)etr>
4-Bromophenyl
phenyl ether
4-Chlorophenyl
phenyl ether
Test
cone.
W
too
•w.100
f. 100
too
100
Limit
lors
-------
126
Federal Register / Vol. 49. No. 209 / Friday, October 26.1984 / Rules and Regulations
COLUMN: W1M8W OH MPKCOPOtT
PM6MM 60
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Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
127
COLUMN: TEN AX GC
PROGRAM: 150°C FOR 4 MIN. 16«C/MIN TO 310°C
DETECTOR: HALL ELECTROLYTIC CONDUCTIVITY
0 48 12 16 20
RETENTION TIME..MIN.
Figure 2. Gas chromatogram of haloethers.
BtLUNQ CODE 6MO-60-C
24
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128
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
Method 612—Chlorinated Hydrocarbon!
I. Scope and Application
1.1 Thi* method covers the determination
of certain chlorinated hydrocarbon*. The
following parameters can be determined by
this method:
U-OcNarabtnnnr..
l.d^OptMorotMnztn*]...
l.4-Ocmonib«ni«nr
1,2.4-TricMorebw
STOHET
No.
34SS1
34536
34571
39700
34391
343S6
34396
34551
CAS No
91-56-7
95-50-1
541-73-1
106-46-7
118-74-1
87-68-3
77-47-4
87-72-1
120-62-1
1.2 This is a gas chromatographic (CC)
method applicable to the determination of the
compounds listed above in municipal and
industrial discharges as provided under 40
CFR 136.1. When this method is used to
analyze unfamiliar samples for any or all of
the compounds above, compound
identifications should be supported by at
least one additional qualitative technique.
This method describes a second gas
chromatographic column that can be used to
confirm measurements made with the
primary column. Method 625 provides gas
chromatograph/mass spectrometer (GC/MS)
condition* appropriate for the qualitative and
quantitative confirmation of results for all of
the parameters listed above, using the extract
produced by this method.
14 The method detection limit (MDL,
defined in Section 14.1)' for each parameter is
lifted in Table 1. The MDL for a specific
wastewater may differ from those listed,
depending upon the nature of interferences in
the sample matrix.
1.4 The sample extraction and
concentration steps in this method are
essentially the same as in Methods 606,60S,
609, and 611. Thus, a single sample may be
extracted to measure the parameters
included in the scope of each of these
methods. When cleanup is required, the
concentration levels must be high enough to
permit selecting aliquots. as necessary, to
apply appropriate cleanup procedures. The
analyst is allowed the latitude, under Section
12, to select chromatographic conditions
appropriate for the simultaneous
measurement of combinations of these
parameters.
1.5 Any modification of this method.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
14 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph and in the interpretation of
gas chromatograms. Each analyst must
demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section &2.
2 Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is extracted with
methylene chloride using a separatory runnel.
The methylene chloride extract is dried and
exchanged to hexane during concentration to
a volume of 10 mL or less. The extract is
separated by gas chromatography and the
parameters are then measured with an
electron capture detector.'
2.2 The method provides a Florisil column
cleanup procedure to aid in the elimination of
interferences that may be encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents,
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in gas chromatograms.
All of these materials must be routinely
demonstrated to be free from interferences
under the conditions of the analysis by
running laboratory reagent blanks as
described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.'Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry, and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. The cleanup
procedure in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches to achieve the MDL
listed in Table 1.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining • current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified **for the information of the
analyst.
5. Apparatus and Material*
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Crab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 *C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use. however, the compressible
tubing should be thoroughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L. with Teflon
stopcock.
5.12 Drying column—Chromatographic
column, approximately 400 mm long x 19 mm
ID, with coarse frit filter disc.
SA3 Chromatographic column—300 long
x 10 mm 0). with Teflon stopcock and coarse
frit filter disc at bottom.
5A4 Concentrator tube. Kuderna-
Danish—10-rnL, graduated (Kontes K-5700SO-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test
Ground glass stopper Is used to prevent
evaporation of extracts.
5.2.5 Evaporative flask, Kudema-
Danish—500-mL (Kontes K-670001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.6 Snyder column, Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2.7 Vials—10 to 15-mL, amber glass,
with Teflon-lined screw cap.
54 Boiling chip*—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control (±
2'C). The bath should be used in a hood.
54 Balance—Analytical, capable of
accurately weighing 00001 g.
5.6 Gas chromatograph—An analytical
system complete with gaa chromatograph
suitable for on-column infection and all
required accessories including syringes,
analytical columns, gases, detector, and strip-
chart recorder. A data system is
recommended for measuring peak anas.
5.6.1 Column 1—14 m long x 2 mm ID
glass, packed with 1% SP-1000 on
Supelcoport (100/120 mesh) or equivalent.
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 129
Guidelines for the use of alternate column
packings are provide in Section 12.1.
5.6.2 Column 2—1.8 m long x 2 mm ID
glass, packed with 1.5% OV-1/2.4% OV-225
on Supelcoport (80/100 mesh) or equivalent.
This column was used to develop the method
performance statements in Section 14.
5.8.3 Detector— Electron capture detector.
This detector has proven effective in the
analysis of wastewaters for the parameters
listed in the scope (Section 1.1), and was used
to develop the method performance
statements in Section 14. Guidelines for the
use of alternate detectors are provided in
Section 12.1.
6. Reagents
6.1 Reagent water— Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.2 Acetone, hexane, isooctane, methanol,
methylene chloride, petroleum ether (boiling
range 30 to 60 *C)—Pesticide quality or
equivalent.
6.3 Sodium sulfate—(ACS) Granular,
anhydrous. Purify heating at 400 'C for 4 h in
a shallow tray.
6.4 Florisil—PR grade (60/100 mesh).
Purchase activated at 1250 *F and store in the
dark in glass containers with ground glass
stoppers or foil-lined screw caps. Before use,
activate each batch at least 16 h at 130 * C in
a foil-covered glass container and allow to
cool.
6.5 Stock standard solution (1.00 /ig/fiL}—
Stock standard solutions can be prepared
from pure standard materials or purchased as
certified solutions.
6.5.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in isooctane
and dilute to volume in a 120-mL volumetric
flask. Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight can be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards can be used at any concentration if
they are certified by the manufacturer or by
an independent source.
6.5.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 'C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
6.5.3 Stock standard solutions must be
replaced after six months, or sooner if
comparision with check standards indicates a
problem.
6.6 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish gas chromatographic
operating conditions equivalent to those
given in Table 1. The gas chromatographic
system can be calibrated using the external
standard technique (Section 7.2) or the
internal standard technique (Section 7.3).
7.2 External standard calibration
procedure:
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask and diluting to volume with
isooctane. One of the external standards
should be at a concentration near, but above,
the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the detector.
7.2.2 Using injections of 2 to 5 fiL, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against the mass injected. The
results can be used to prepare a calibration
curve for each compound. Alternatively, if
the ratio of response to amount injected
(calibration factor) is a constant over the
working range (<10% relative standard
deviation, RSD), linearity through the origin
can be assumed and the average ratio or
calibration factor can be used in place of a
calibration curve.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select one or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Because of
these limitations, no internal standard can be
suggested that is applicable to all samples.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding volumes
of one or more stock standards to a
volumetric flask. To each calibration
standard, add a known constant amount of
one or more internal standards, and dilute to
volume with isooctane. One of the standards
should be at a concentration near, but above,
the MDL and the other concentrations should
correspond to the expected range of
concentrations found in real samples or
should define the working range of the
detector.
7.3.2 Using injections of 2 to 5 jiL, analyze
each calibration standard according to
Section 12 and tabulate peak height or area
responses against concentration for each
compound and internal standard. Calculate
response factors (RF) for each compound
using Equation 1.
Equation 1.
RF=
(AU)(C.)
where:
A,=Response for the parameter to be
measured.
A|,=Response for the internal standard.
Cu=Concentration of the internal standard
(W/U-
C,=Concentration of the parameter to be
measured (fig/L).
If the RF value over the working range is a
constant (<10% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A./At,, vs. RF.
7.4 The working calibration curve,
calibration factor, or RF must be verified on
each working day by the measurement of one
or more calibration standards. If the response
for any parameter varies from the predicted
response by more than ±15%, a new
calibration curve must be prepared for thai
compound.
7.5 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
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When the
results of sample spikes indicate atypical
method performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.4,11.1, and 12.1) to improve the
separations or lower the cost of
measurements. Each time such modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 10% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 10% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
465-028 O - 85 - 5
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at the following
concentrations in acetone: Hexachloro-
aubstiluted parameters. 10 pg/mL: any other
chlorinated hydrocarbon. 100 ug/mL. The QC
check sample concentrate must be obtained
from the U.S. Environmental Protection
Agency. Environmental Monitoring and
Support Laboratory in Cincinnati. Ohio, if
available. If not available from that source.
the QC check sample concentrate must be
obtained from another external source. If not
available from either source above, the QC
check sample concentrate must be prepared
by the laboratory using stock standards
prepared independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at the test concentrations shown in
Table 2 by adding 1.00 mL of QC check
sample concentrate to each of four l-L
aliquots of reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in (ig/L. and the standard deviation of the
recovery (s) in pg/L. for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively.
found in Table 2. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual i exceed* the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 2 portent a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analysed.
8.2.8 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.8.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.2.
8.2.6.2 Beginning with Section 8.2.2, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2^.
8.3 The laboratory must on an ongoing
basis, spike at least 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten samples per month, at least one spike
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined a* follows:
8.3.1.1 If, aa in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at the test concentration in
Section 8.2.2 or 1 to S times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any: or, if
none by (2) the larger of either 5 times higher
than the expected background concentration
or the test concentration in Section 8.2.2.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. In necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100 (A-B)%/T. where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than the test concentration in Section
8.2.2, the analyst must use either the QC
acceptance criteria in Table 2, or optional QC
acceptance criteria calculated for the specific
spike concentration. To calculate optional
acceptance criteria for the recovery of a
parameter. (1) calculate accuracy (X') using
the equation in Table 3, substituting the spike
concentration (T) for C; (2) calculate overall
precision (S') using the equation in Table 3,
substituting X* for X: (3) calculate the range
for recovery at the spike concentration as
(100 X'/T) ± 2.44 (100 S'/T)%.7
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4. If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix and the performance of the
laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (P.) as 100 (A/T)%. where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 2.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for thai parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (s,). Express the accuracy
assessment as a percent recovery interval
from P-2s, to P-l-2s,. If P-90% and s,~10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter on a regular
basis (e.g. after each five to ten new accuracy
measurement*).
8.6 It is recommended that the laboratory
adopt additional quality assurance practice*
for use with this method The specific
practices that are most productive depend
upon the need* of the laboratory and the
nature of the sample*. Field duplicate* may
be analyzed to assess the precision of the
environmental measurement*. When doubt
exist* over the identification of a peak on the
chromatogram, confirmatory technique* such
a* gas chromatography with a dissimilar
column, specific element detector, or mass
spectrometer must be used. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevent performance evaluation studies.
ft Sample Collection, Preservation, and
Handling
9.1 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 container*
in accordance with the requirement* of the
program. Automatic sampling equipment
must be as free a* possible of Tygon tubing
and other potential sources of contamination.
9.2 All sample* muit be iced or
refrigerated at 4 *C from the time of collection
until extraction.
9.3 All samples must be extracted within
7 days of collection and completely analyzed
within 40 day* of extraction.1
10. Sample Extraction
10.1 Mark the water meniscus on the tidm
of the sample bottle for later determination oj
•ample volume. Pour the entire sample into a
2-L separatory funnel
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 131
10.2 Add 60 mL of melhylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separately funnel and extract the
sample by shaking the funnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool.
centrifugation. or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.3 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.4 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.S Pour the combined extract through a
solvent-rinsed drying column containing
about 10 cm of anhydrous sodium sulfate,
and collect the extract in the K-D
concentrator. Rinse the Erlenmeyer flask and
column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.6 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 65 'Q so that the
concentrator tube is partially immersed in the
hot water, and the entire lower rounded
surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 to 2
mL, remove the K-D apparatus and allow it
to drain and cool for at least 10 min.
Note.—The dichloribenzenes 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 to 2 mL before removing the K-D
apparatus from the hot water bath.
10.7 Momentarily remove the Snyder
column, add 50 mL of hexane and a new
boiling chip, and reattach the Snyder column.
Raise the tempeature of the water bath to 85
to 90 *C. Concentrate the extract as in
Section 10.6, except use hexane to prewet the
column. The elapsed time of concentration
should be 5 to 10 min.
i 10.8 Remove the Snyder column and rinse
fche flask and its lower joint into the
concentrator tube with 1 to 2 mL of hexane. A
5-mL syringe is recommended for this
operation. Stopper the concentrator tube and
store refrigerated if further processing will
not be performed immediately. If the extract
will be stored longer than two days, it should
be transferred to a Teflon-sealed screw-cap
vial. If Ihe sample extract requires no further
cleanup, proceed with gas chrumatographic
analysis (Section 12). If the sample requires
further cleanup, proceed to Section 11.
10.9 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.
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix. If particular circumstances demand
the use of a cleanup procedure, the analyst
may use the procedure below or any other
appropriate procedure. However, the analyst
first must demonstrate that the requirements
of Section 8.2 can be met using the method as
revised to incorporate the cleanup procedure.
11.2 Florisil column cleanup for
chlorinated hydrocarbons:
11.2.1 Adjust the sample extract to 10 mL
with hexane.
11.2.2 Place 12 g of Florisil into a
chromatographic column. Tap the column to
settle the Florisil and add 1 to 2 cm of
anhydrous sodium sulfate to the top.
11.2.3 Preelute the column with 100 mL of
petroleum ether. Discard the eluate and just
prior to exposure of the sodium sulfate layer
to the air, quantitatively transfer the sample
extract onto 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 200 mL of 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.
11.2.4 Concentrate the fraction as in
Section 10.6, except use hexane to prewet the
column. When the apparatus is cool, remove
the Snyder column and rinse the flask and its
lower joint into the concentrator tube with
hexane. Analyze by gas chromatography
(Section 12).
12. Gas Chromatography
12.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. Examples of
the separations achieved by Column 2 are
shown in Figures 1 and 2. Other packed or
capillary (open-tubular) columns,
chromatographic conditions, or detectors may
be used if the requirements of Section 8.2 are
met.
12.2 Calibrate the system daily as
described in Section 7.
12.3 If the internal standard calibration
procedure is being used, the internal standard
must be added to the sample extract and
mixed throughly immediately before injection
into the gas chromatograph.
12.4 Inject 2 to 5 \iL of the sample extract
or standard into the gas chromatograph using
the solvent-flush technique.9 Smaller (1.0 /iL)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 jiL. Ihe total extract volume.
and the resulting peak size in area or peak
height units.
12.5 Identify the parameters in the sample
by comparing the retention times of the peaks
in the sample chromatogram with those of the
peaks in standard chromalograms. The width
of the retention time window used to make
identifications should be based upon
measurements of actual retention time
variations of standards over Ihe course of a
day. Three times Ihe standard deviation of a
retention time for a compound can be used to
calculate a suggested window size: however.
the experience of the analyst should weigh
heavily in the interpretation of
chromatograms.
12.6 If the response for a peak exceeds
the working range of the system, dilute the
extract and reanalyze.
12.7 If the measurement of the peak
response is prevented by the presence of
interferences, further cleanup is required.
13. Calculations
13.1 Determine the concentration of
individual compounds in the sample.
13.1.1 If the external standard calibration
procedure is used, calculate the amount of
material injected from the peak response
using the calibration curve or calibration
factor determined in Section 7.2.2. The
concentration in the sample can be
calculated from Equation 2.
Equation 2.
Concentration (fig/L) =
(VJ(VJ
where:
. A=Amount of material injected (ng).
V, = Volume of extract injected (fiL).
V,=Volume of total extract (fiL).
V.=Volume of water extracted (mL).
13.1.2 If the internal standard calibration
procedure is used, calculate the
concentration in the sample using the
response factor (RF) determined in Section
7.3.2 and Equation 3.
Equation 3.
Concentration (jig/L)= -
(AJ(RF)(VJ
where:
A,=Response for the parameter to be
measured.
A,,=Response for the internal standard.
I, —Amount of internal standard added to
each extract (fig).
V0=Volume of water extracted (L).
13.2 Report results in fig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
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132
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.' The MDL concentrations listed in
Table 1 were obtained using reagent water.10
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method has been tested for
linearity of spike recovery from reagent
water and has been demonstrated to be
applicable over the concentration range from
4 X MDL to 1000 X MDL."
14.3 This method was tested by 20
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 1.0 to 356 pg/L " Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136. Appendix B.
2. "Determination of Chlorinated
Hydrocarbons In Industrial and Municipal
Wastewaters." Report for EPA Contract 68-
03-2625 (In preparation).
3. ASTM Annual Book of Standards, Part
31. D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials,
Philadelphia.
4. "Carcinogens—Working With
Carcinogens." Department of Health.
Education, and Welfare. Public Health
Service. Center for Disease Control. National
Institute for Occupational Safety and Health,
Publication No. 77-206. August 1977.
5. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration. OSHA 2206 (Revised.
January 1976).
6. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication. Committee on Chemical Safety,
3rd Edition, 1979.
7. Provost. L.P.. and Elder, R.S.
"Interpretation of Percent Recovery
Date."American Laboratory. 15. 58-63 (1983).
(The value 2.44 used in the equation in
Section 8.3.3 is two times the value 1.22
derived in this report.)
8. ASTM Annual Book of Standards. Part
31. D3370-76. "Standard Practices for
Sampling Water." American Society for
Testing and Materials. Philadelphia.
9. Burke, ).A. "Gas Chromatography for
Pesticide Residue Analysis; Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists. 48.1037 (1965).
10. "Development of Detection Limits. EPA
Method 612. Chlorinated Hydrocarbons,"
Special letter report for EPA Contract 68-03-
2625, U.S. Environmental Protection Agency.
Environmental Monitoring and Support
Laboratory, Cincinnati. Ohio 45268.
11. "EPA Method Validation Study 22.
Method 612 (Chlorinated Hydrocarbons),"
Report for EPA Contract 68-03-2625 (In
preparation).
12. "Method Performance for
Hexachlorocyclopentadiene by Method 612,"
Memorandum from R. Slater. U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory. Cincinnati. Ohio 45268.
December 7,1983.
TABLE 1.—CHROMATOORAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Column 1 oondMonc Supatooport (100/120 rnaah) OHM wMi 1% SP-1000 psotod in • 1 J m « 2 mm 10 ghas column «Mn 8% matim/M% argon cantor ga> « 25 mL/mki Sew rato.
Column Mmparakm Md texharmal * 68 •& ampt wliara otwrwto* tadtoatod.
CakmnTeontWon: Supatooport (80/100 me*) oc«M w«i 1.8% OV-1/2.4% OV-22S psetod to s 1 * m > 2 mm e gtas oakmn •» 5% tMltan^l6« Mgon cwlv ass « 2S mL/
mm How mis. Column Mmpmhm KM Kotwmri « 75 "C. ampt «m*n> oftanrtM krtastod.
nd-NMc
•180 "Ce
-
< 100 -C column I
TABLE 2.— QC ACCEPTANCE CRITERIA— METHOD 612
Taat
cone.
04/L)
LMttor
Rang* tor it
(MO/U
Rang*
(ptrcint'l
U-Otchtorobara
1,3-Otohtorobara
100
100
100
100
10
10
10
10
100
37.3
28.3
28.4
20.8
2.4
U
it
3J
31.8
28.S-128.8
23.8-146.1
7.2-138.8
22.7-128.8
2.8-14.8
0-12.7
0-10.4
2.4-1U
20J-13J.7
8-148
8-180
0-180
13-137
15-158
0-138
0-111
8-138
8-14*
(SacSon 8*4).
(Sacson
NOTE.-T1MM eriMa an oaaad
12.4):
i dMai hi Tcbto 3. Whet1*) McnMry, stit Into tor racovwy hew bMn brotdvnvd to i
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 012
oaaad ctaoty upon ft* rnadod pwtaman
uaad IB
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations 133
TABLE 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 612—Continued
Parameter
! Acccuracy. as
recovery, X'
iMS/D
072Ct280
. 087C-002
061C + 003
047C
0 74C 002
0 76C + 0 98
Sngte analyst
precision. V
(MS/L) .
0 16X -0 48
0 t4$ f 007
0 1 eX +008
0 24X
0 23)t f 0 07
0 23X - 0 44
Overall
precision, S'
IMS/LI
0 351X -0 57
0 36X - 0 19
0 36X 000
0 40X- t 37
X* = Expected recovery for one or more measurements of a sample containing a concentration of C. in na/L.
V = Expected single analyst standard deviation of measurements at an average concentration found of X, in jig/L.
S1 •* Expected interlaboratory standard deviation of measurements at an average concentration found of X, in jig/L.
C = True value for the concentration, in pg/l.
X= Average recovery found lor measurements of samples containing a concentration of C. in jig/L.
• Estimates based upon the performance in a single laboratory."
BILLING CODE 8560-50-M
-------
134 Federal Register / Vol. 49, No. 209 / Friday. October 26,1984 / Rules and Regulations
COLUMN: 1.5% (MM/14% OV-22S ON SUffLCOPM?
TBJKMnMC; Tit
omen* afcnm CAPTURE
0 4 I 12 II 20 24
RETENTION TIME. WIN.
Figure 1. Gas chromatogram of chlorinated hydrocarbons.
-------
Federal Register / Vol. 49. No. 209 / Friday, October 28, 1984 / Rules and Regulations
135
COLUMN: 1.6% 0V-1/2.4% OV-225 ON SUPELCOPORT
TEMPERATURE: 16S*C
DETECTOR: ELECTRON CAPTURE
£
(M
§
2
g
s
0 4 8 12
RETENTION TIME, MIN.
Figure 2. Gas chromatogram of chlorinated hydrocarbons.
MUJNQ COOt M*0-(0-C
-------
136
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 613—2 ,3.7,8-Tetrachlorodibeiuo-p-
Oioxin
1. Scope and Application
1.1 This method covers the determination
of 2.3,7,8-telrachlorodibenzo-p-dioxin (2,3,7,8-
TCDO). The following parameter may be
determined by this method:
PirwncMr
j 3 7 S-TCOO
STORET
No.
3487S
GAS No
1746-01 -6
1.2 This is a ga* chromatographic/mass
spectrometer (GC/MS) method applicable to
the determination of 2,3,7,8-TCDD in
municipal and industrial discharges as
provided under 40 CFR 136.1. Method 625
may be used to screen samples for 2,3,7.6-
TCDD. When the screening test is positive.
the final qualitative confirmation and
quantification mlust be made using Method
613.
1.3 The method detection limit (MDL.
defined in Section 14.1)' for 2,3,7,8-TCDD is
listed in Table 1. The MDL for a specific
wastewater may be different from that listed.
depending upon the nature of interferences in
the sample matrix.
1.4 Because of the extreme toxicity of this
compound, the analyst must prevent
exposure to himself, of to others, by materials
knows or believed to contain 2,3,7,8-TCDD.
Section 4 of this method contains guidelines
and protocols that serve as minimum safe-
handling standards in a limited-access
laboratory.
14 Any modification of this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph/mass spectrometer and in the
interpretation of mass spectra. Each analyst
must demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is spiked with an internal
standard of labeled 2.3.7.8-TCDD and
extracted with methylene chloride using a
separatory funnel. The methylene chloride
extract is exchanged to hexane during
concentration to a volume of 1.0 mL or less.
The extract is then analyzed by capillary
column GC/MS to separate and measure
2A7A-TCDD."
L2 The method provides selected column
chromatographic cleanup proceudres to aid in
the elimination of interferences that may be
encountered.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents,
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated backgrounds at the masses (m/z)
monitored. All of these materials must be
routinely demonstrated to be free from
interferences under the conditions of the
analysis by running laboratory reagent
blanks as described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.' Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry. and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by the treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to mininmize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are coextracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled. 2,3,7,8-TCDD is
often associated with other interfering
chlorinated compounds which are at
concentrations several magnitudes higher
than that of 2.3,7,8-TCDD. The cleanup
producers in Section 11 can be used to
overcome many of these interferences, but
unique samples may require additional
cleanup approaches '••''to eliminate false
positives and achieve the MDL listed in Table
1.
3 J The primary column, SP-2330 or
equivalent, resolves 2,3,7.8-TCDD from the
other 21TCDD insomers. Positive results
using any other gas chromatographic column
must be confirmed using the primary column.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a currant awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified •»for the information of die
analyst Benzene and 2,3,7,8-TCDD have been
identified as suspected human or mammalian
carcinogens.
4.2 Each laboratory must develop a strict
safety program for handling 2,3,7,8-TCDD.
The following laboratory practices are
recommended:
4.2.1 Contamination of the laboratory will
be minimized by conducting all
manipulations in a hood.
4.2.2 The effluents of sample splitters for
the gas chromatograph and roughing pumps
on the GC/MS should pass through either a
column of activated charcoal or be bubbled
through a trap containing oil or high-boiling
alcohols.
4.2.3 Liquid waste should be dissolved in
methanol or ethanol and irradiated with
ultraviolet light with a wavelength greater
than 290 nm for several days. (Use F 40 BL
lamps or equivalent). Analyze liquid wastes
and dispose of the solutions when 2.3.7,8-
TCDD can no longer be detected.
4.3 Dow Chemical U.S.A. has issued the
following precautions (revised November
1978) for safe handling of 2,3,7,8-TCDD in the
laboratory.
4.3.1 The following statements on safe
handling are as complete as possible on the
basis of available lexicological information.
The precautions for safe handling and use are
necessarily general in nature since detailed,
specific recommendations can be made only
for the particular exposure and circumstances
of each individual use. Inquiries about
specific operations or uses may be addressed
to the Dow Chemical Company. Assistance in
evaluating the health hazards of particular
plant conditions may be obtained from
certain consulting laboratories and from
State Departments of Health or of Labor,
many of which have an industrial health
service. 2,3,7,8-TCDD is extremely toxic to
laboratory animals. However, it has been
handled for yean without injury in analytical
and biological laboratories. Techniques used
in handling radioactive and infectious
materials an applicable to 2,3,7,8,-TCDD.
4.3.1.1 Protective equipment—Throw-
away plastic gloves, apron or lab coat, safety
glasses, and a lab hood adequate for
radioactive work.
4.3.1.2 Training—Workers must be
trained in the proper method of removing
contaminated gloves and clothing without
contacting the exterior surfaces.
4.3.1.3 Personal hygiene—Thorough
washing of hands and forearms after each
manipulation and before breaks (coffee,
lunch, and shift).
4.3.1.4 Confinement—Isolated work area,
posted with signs, segregated glassware and
tools, plastic-backed absorbent paper on
benchtops.
4.3.1 J Waste—Good technique includes
minimizing contaminated waste. Plastic bag
liners should be used in waste cans. Janitors
must be trained in the safe handling of waste.
4.3.1.6 Disposal of wastes—2A7.8-TCDD
decomposes above 600 *C Low-level waste
such as absorbent paper, tissues, animal
remains, and plastic gloves may be burned in
a good incinerator. Gross quantities
(milligrams) should be packaged securely and
disposed through commercial or
<
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 137
4.3.1.7 Decontamination—For personal
decontamination, use any mild soap with
plenty of scrubbing action. For
decontamination of glassware, tools, and
surfaces, Chlorothene NU Solvent
(Trademark of the Dow Chemical Company)
is the least toxic solvent shown to be
effective. Satisfactory cleaning may be
accomplished by rinsing with Chlorothene,
then washing with any detergent and water.
Dishwater may be disposed to the sewer. It is
prudent to minimize solvent wastes because
they may require special disposal through
commercial sources which are expensive.
4.3.1.8 Laundry'—Clothing known to be
contaminated should be disposed with the
precautions described under Section 4.3.1.6.
Lab coats or other clothing worn in 2,3,7,8-
TCDD work areas may be laundered.
Clothing should be collected in plastic
bags. Persons who convey the bags and
launder the clothing should be advised of the
hazard and trained in proper handling. The
clothing may be put into a washer without
contact if the launderer knows the problem.
The washer should be run through a cycle
before being used again for other clothing.
4.3.1.9 Wipe tests—A useful method of
determining cleanliness of work surfaces and
tools is to wipe the surface with a piece of
filter paper. Extraction and analysis by gas
chromatography can achieve a limit of
sensitivity of 0.1 fig per wipe. Less than 1 fig
of 2,3.7.8-TCDD per sample indicates
acceptable cleanliness; anything higher
warrants further cleaning. More than 10 fig
on a wipe sample constitutes an acute hazard
and requires prompt cleaning before further
use of the equipment or work space. A high
(>10 fig) 2,3,7,8-TCDD level indicates that
unacceptable work practices have been
employed in the past.
4.3.1.10 Inhalation—Any procedure that
may produce airborne contamination must be
done with good ventilation. Gross losses to a
ventilation system must not be allowed.
Handling of the dilute solutions normally
used in analytical and animal work presents
no inhalation hazards except in the case of
an accident.
4.3.1.11 Accidents—Remove
contaminated clothing immediately, taking
precautions not to contaminate skin or other
articles. Wash exposed skin vigorously and
repeatedly until medical attention is
obtained.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composite sampling.
5.1.1 Grab sample bottle—1-L or 1-qt.
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 *C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used. Before use, however, the compressible
tubing should be thoroughly rinsed with
methanol. followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.1.3 Clearly label all samples as
"POISON" and ship according to U.S.
Department of Transportation regulations.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnels—2-L and 125-mL,
with Teflon stopcock.
5.2.2 Concentrator tube, Kuderna-
Danish—lOmL, graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
5.2.3 Evaporative flask, Kuderna-
Danish—500-mL (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with
springs.
5.2.4 Snyder column. Kuderna-Danish—
Three-ball macro (Kontes K-503000-0121 or
equivalent).
5.2,5 Snyder column, Kuderna-Danish—
Two-ball micro (Kontes K-569001-0219 or
equivalent).
5.2.6 Vials—10 to 15-mL, amber glass.
with Teflon-lined screw cap.
5.2.7 Chromatographic column—300 mm
long X 10 mm ID, with Teflon stopcock and
coarse frit filter disc at bottom.
5.2.8 Chromatographic column—400 mm
long x 11 mm ID, with Teflon stopcock and
coarse frit filter disc at bottom.
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min or Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2 *C). The bath should be used in a hood.
5.5 GC/MS system:
5.5.1 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph and all
required accessories including syringes,
analytical columns, and gases. The injection
port must be designed for capillary columns.
Either split, splitless, or on-column injection
techniques may be employed, as long as the
requirements of Section 7.1.1 are achieved.
5.5.2 Column—60 m long X 0.25 mm ID
glass or fused silica, coated with SP-2330 (or
equivalent) with a film thickness of 0.2 fim.
Any equivalent column must resolve 2, 3, 7,
B-TCDD from the other 21 TCDD isomers."
5.5.3 Mass spectrometer—Either a low
resolution mass spectrometer (LRMS) or a
high resolution mass spectrometer (HRMS)
may be used. The mass spectrometer must be
equipped with a 70 V (nominal) ion source
and be capable of aquiring m/z abundance
data in real time selected ion monitoring
(SIM) for groups of four or more masses.
5.5.4 GC/MS interface—Any GC to MS
interface can be used that achieves the
requirements of Section 7.1.1. GC to MS
interfaces constructed of all glass or glass-
lined materials are recommended. Glass
surfaces can be deactivated by silanizing
with dichlorodimethylsilane. To achieve
maximum sensitivity, the exit end of the
capillary column should be placed in the ion
source. A short piece of fused silica capillary
can be used as the interface to overcome
problems associated with straightening the
exit end of glass capillary columns.
5.5.5 The SIM data acquired during the
Chromatographic program is defined as the
Selected Ion Current Profile (SICP). The SICP
can be acquired under computer control or as
a real time analog output. If computer control
is used, there must be software available to
plot the SICP and report peak height or area
data for any m/z in the SICP between
specified time or scan number limits.
5.6 Balance—Analytical, capable of
accurately weighing 0.0001 g.
ft Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of 2, 3, 7, 8-TCDD.
6.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL. Wash the
solution with methylene chloride and hexane
before use.
6.3 Sodium thiosulfate—(ACS) Granular.
6.4 Sulfuric acid—Concentrated (ACS, sp.
gr. 1.84).
6.5 Acetone, methylene chloride, hexane.
benzene, ortho-xylene, tetradecane—
Pesticide quality or equivalent.
6.6 Sodium sulfate—(ACS) Granular,
anhydrous. Purify by heating at 400 *C for 4 h
in a shallow tray.
6.7 Alumina—Neutral, 80/200 mesh
(Fisher Scientific Co., No. A-540 or
equivalent). Before use, activate for 24 h at
130 'C in a foil-covered glass container.
6.8 Silica gel—High purity grade, 100/120
mesh (Fisher Scientific Co., No. S-679 or
equivalent).
6.9 Stock standard solutions (1.00 fig/
fiL]—Stock standard solutions can be
prepared from pure standard materials or
purchased as certified solutions. Acetone
should be used as the solvent for spiking
solutions; ortho-xylene is recommended for
calibration standards for split injectors; and
tetradecane is recommended for splitless or
on-colum injectors. Analyze stock internal
standards to verify the absence of native
2,3,7,8-TCDD.
6.9.1 Prepare stock standard solutions of
2,3.7,8-TCDD (mol wt 320) and either "C14
2,3,7,8-TCDD (mol wt 328) or "Clu 2,3,7,8-
TCDD (mol wt 332) in an isolated area by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in pesticide
quality solvent and dilute to volume in a 10-
mL volumetric flask. When compound purity
is assayed to be 96% or greater, the weight
can be used without correction to calculate
the concentration of the stock standard.
Commercially prepared stock standards can
be used at any concentration if they are
certified by the manufacturer or by an
independent source.
6.9.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store in an isolated refrigerator
protected from light. Stock standard solutions
should be checked frequently for signs of
degradation or evaporation, especially just
-------
138
Federal Roaster / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
prior to preparing calibrnllon standards or
spiking solutions from them.
6.9.3 Stock standard solutions must be
replaced after six monlhn. or sooner if
comparison with check ulnmliirds indicates a
problem.
6.10 Internal standunl »pikin« solution (25
ng/mL)—Using stock standard solution,
prepare a spiking solution in ucotone of
either"Cl,, or "Cl4 2.:i,7.H-TCl)U H! a
concentration of 25 ng/ml.. (Sue Section 10.2)
6.11 Quality control <:hi!i:k sample
concentrate—See Section H.2.1.
7. Calibration
7.1 Establish gas chromiilograhic
operating conditions equivalent to those
given in Table 1 and SIM conditions for the
mass spectrometer as described in Section
12.2 The GC/MS system must be calibrated
using the internal standard technique.
7.1.1 Using slock standards, prepare
calibration standards that will allow
measurement of relative response factors of
at least three concentration ratios of 2,3.7,8-
TCDD to internal standard. Each calibration
standard must be prepared to contain the
internal standard at a concentration of 25 ng/
mL If any Interferences are contributed by
the internal standard at m/z 320 and 322, its
concentration may be reduced In the
calibration standard* and In the Internal
standard spiking solution (Section 6.10). One
of the calibration standards should contain
2,3,7,8-TCDD at a concentration near, but
above, the MDL and the other 2,3.7.8-TCDD
concentration* should correspond to the
expected range of concentration* found In
real samples or should define the working
range of the GC/MS system.
7.1.2 Using injections of 2 to 5 pU analyse
each calibration standard according to
Section 12 and tabulate peak height or ana
response against the concentration of 2.3,7,8-
TCDD and internal standard. Calculate
response factor* fRF) for 2.3,7.8-TCDD using
Equation 1.
Equation 1.
RF=
(A.) (CJ
(AJ (C.)
where: ,
A.=SIM response for 2.3.7.8-TCDD m/z
320.
A»-SIM response for the internal
standard. m/z 332 for »Cw 2A7>TCDD
m/z 328 for "a 2.3.7 ,8-TCDD.
(^.-Concentration of the internal standard
.
(^-Concentration of 2,3,7,8-TCDD fjig/L).
If the RF value over the working range is a
constant « 10% relative standard deviation,
RSD), the RF can be assumed to be invariant
and the average RF can be used for
calculation*. Alternatively, the reiult* can be
used to plot a calibration curve of response
ratios, AjAfc. v*. RF.
7.1.3 The working calibration curve or RF
must be verified on each working day by the
measurement of one or more 2A73-TCDD
calibration rtandard*. If the re*pon*e for
2.3.7.8-TCDD varies from the predicted
response by more than ±15%, the te*t mu*t
be repeated using a fresh calibration
standard. Alternatively, a new calibration
curve must be prepared.
7.2 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.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability ia established
as described In Section 8\2.
8.1.2 In recognition of advance* that an
occurring in chromatography, the analyst is
permitted certain option* (detailed in
Section* 10.5,11.1. and 12.1) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8\2
8.1.3 Before processing any sample*, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware an under
control. Each time a set of sample* is
extracted or reagents an changed, a reagent
water blank must be processed a* a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basil, spike and analyze a minimum of 10% of
all samples with native 2,3,7,8-TCDD to
monitor and evaluate laboratory data quality.
This procedure is described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control This procedure I* described in
Section 8.4. The frequency of the check
standard analyse* i* equivalent to 10% of all
•ample* analyzed but may be reduced if
•pike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance record* to document the quality
of data that I* generated. This procedure I*
described in Section 8.5.
84 To establish the ability to generate
acceptable accuracy and prediion, the
analyst must perform the following
operation*.
8.2.1 A quality control (QQ check sample
concentrate I* required containing 2,3,7,8-
TCDD at a concentration of 0.100 pg/mL in
acetone. The QC check sample concentrate
must be obtained from the U.S.
Environmental Protection Agency.
Environmental Monitoring and Support
Laboratory in Cincinnati. Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Using a pipet. prepare QC check
samples at a concentration of 0.100 pg/L (100
ng/L) by adding 1.00 mL of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10.
8.2.4 Calculate the average recovery (X)
in ng/L, and the standard deviation of the
recovery (s) in jig/L, for 2,3,7,8-TCDD using
the four results.
8.2.5 Compare s and (X) with the
corresponding acceptance criteria for
precision and accuracy, respectively, found in
Table 2. If s and X meet the acceptance
criteria, the system performance I*
acceptable and analysis of actual samples
can begin. If • exceed* the precision limit or
X fall* outside the range for accuracy, the
system performance I* unacceptable for
23,7,8-TCDD. Locate and correct the source
of the problem and repeat the test beginning
with Section 8JJ,
8~3 The laboratory must, on an ongoing
basis, spike at leact 10% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing one to
ten sample* per month, at least one spiked
sample per month i* required.
8J.1 The concentration of the spike in the
sample should be determined as follows:
8J.1.1 If, as in compliance monitoring, the
concentration of 2A7.8-TCDD in the sample
i* being checked against a regulatory
concentration limit the spike should be at
that limit or 1 to 5 time* higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of 2.3,7,8-
TCDD in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 0.100 pg/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8J.1J If it U impractical to determine
background level* before spiking (e.g..
maximum holding time* will be exceeded).
the *pike concentration should be (1) the
regulatory concentration limit if any; or, if
none (2) the larger of either S time* higher
than the expected background concentration
or 0.100 pg/L.
8J.2 Analyze one sample aliquot to
determine the background concentration (B)
of 2.3,7,8-TCDD. If nece**ary. prepare a new
QC check (ample concentrate (Section oil)
appropriate for the background concentration
in the sample. Spike a second sample aliquot
with 1.0 mL of the QC check sample
concentrate and analyze it to determine the
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 139
concentration after spiking (A) of 2.3.7.8-
TCDD. Calculate percent recovery (P) as
100(A-B)%T, where T is the known true value
of the spike.
8.3.3 Compare the percent recovery (P) for
2,3.7,8-TCDD with the corresponding QC
acceptance criteria found in Table 2. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1." If
spiking was performed at a concentration
lower than 0.100 ng/L, the analyst must use
either the QC acceptance criteria in Table 2,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of 2.3,7,8-TCDD: (1) calculate
accuracy (X') using the equation in Table 3,
substituting the spike concentration (T) for C:
(2) calculate overall precision (S') using the
equation in Table 3. substituting X' for X: (3)
calculate the range for recovery at the spike
concentration as (100 X'/T)±2.44{100 S'/
T)%. "
8.3.4 If the recovery of 2,3,7,8-TCDD falls
outside the designated range for recovery, a
check standard must be analyzed as
described in Section 8.4.
8.4 If the recovery of 2,3,7,8-TCDD fails
the acceptance criteria for recovery in
Section 8.3, a QC check standard must be
prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the complexity of the sample matrix
and the performance of the laboratory.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Section 8.2.1 or 8.3.2] to 1 L of
reagent water.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
2,3,7,8-TCDD. Calculate the percent recovery
(P.) as 100 (A/T)%. where T is the true value
of the standard concentration.
8.4.3 Compare the percent recovery (P.)
with the corresponding QC acceptance
criteria found in Table 2. If the recovery of
2,3.7.8-TCDD falls outside the designated
range, the laboratory performance is judged
to be out of control, and the problem must be
immediately identified and corrected. The
analytical result for 2,3.7,8-TCDD in the
unspiked sample is suspect and may not be
reported for regulatory compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the spandard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P-2sp to P+2sp. If P=90% and sp=10%,
for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment on a regular basis (e.g. after each
five to ten new accuracy measurements).
8.6 It is recommended that the
laborataory adopt additional quality
assurance practices for use with this method.
The specific practices that are most
productive depend upon the needs of the
laboratory and the nature of the samples.
Field duplicates may be analyzed to assess
the precision of the environmental
measurements. Whenever possible, the
laboratory should analyze standard reference
materials and participate in relevant
performance evaluation studies.
ft Sample Collection, Preservation, and
Handling
9.1 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All samples must be iced or
refrigerated at 4 *C and protected from light
from the time of collection until extraction.
Fill the sample bottles and. if residual
chlorine is present, add 80 mg of sodium
thiosulfate per liter of sample and mix well.
EPA Methods 330.4 and 330.5 may be used for
measurement of residual chlorine.13 Field test
kits are available for this purpose.
9.3 Label all samples and containers
"POISON" and ship according to applicable
U.S. Department of Transportation
regulations.
9.4 All samples must be extracted within
7 days of collection and completely analyzed
within 40 days of extraction.*
10. Sample Extraction
Caution: When using this method to
analyze for 2,3,7,8-TCDD. all of the following
operations must be performed in a limited-
access laboratory with the analyst wearing
full protective covering for all exposed skin
surfaces. See Section 4.2.
10.1 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory funnel.
10.2 Add 1.00 mL of internal standard
spiking solution to the sample in the
separatory runnel. If the final extract will be
concentrated to a fixed volume below 1.00
mL (Section 12.3), only that volume of spiking
solution should be added to the sample so
that the final extract will contain 25 ng/mL of
internal standard at the time of analysis.
10.3 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the funnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layers is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugetion. or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask.
10.4 Add a second 60-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner.
10.5 Assemble a Kuderna-Danish (K-D)
concentrator by attaching a 10-mL
concentrator tube to a 500-mL evaporative
flask. Other concentration devices or
techniques may be used in place of the K-D
concentrator if the requirements of Section
8.2 are met.
10.6 Pour the combined extract into the
K-D concentrator. Rinse the Erlenmeyer flask
with 20 to 30 mL of methylene chloride to
complete the quantitative transfer.
10.7 Add one or two clean boiling chips to
the evaporative flask and attach a three-ball
Snyder column. Prewet the Snyder column by
adding about 1 mL of methylene chloride to
the top. Place the K-D apparatus on a hot
water bath (60 to 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 with hot vapor.
Adjust the vertical position of the apparatus
and the water temperature as required to
complete the concentration in 15 to 20 min. At
the proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent. When
the apparent volume of liquid reaches 1 mL,
remove the K-D apparatus and allow it to
drain and cool for at least 10 min.
10.8 Momentarily remove the Snyder
column, add 50 mL of hexane and a new
boiling chip, and reattach the Snyder column.
Raise the temperature of the water bath to 85
to 90*C. Concentrate the extract as in Section
10.7, 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 to 2 mL of hexane. A 5-mL syringe
is recommended for this operation. Set aside
the K-D glassware for reuse in Section 10.14.
10.9 Pour the hexane extract from the
concentrator tube into a 125-mL separatory
funnel. Rinse the concentrator tube four times
with 10-mL aliquots of hexane. Combine all
rinses in the 125-mL separatory funnel.
10.10 Add 50 mL of sodium hydroxide
solution to the funnel and shake for 30 to 60 s.
Discard the aqueous phase.
10.11 Perform a second wash of the
organic layer with 50 mL of reagent water.
Discard the aqueous phase.
10.12 Wash the hexane layer with a least
two 50-mL aliquots of concentrated sulfuric
acid. Continue washing the hexane layer with
50-mL aliquots of concentrated sulfuric acid
until the acid layer remains colorless. Discard
all acid fractions.
10.13 Wash the hexane layer with two 50-
mL aliquots of reagent water. Discard the
aqueous phases.
10.14 Transfer the hexane extract into a
125-mL Erlenmeyer flask containing 1 to 2 g
of anhydrous sodium sulfate. Swirl the flask
for 30 s and decant the hexane extract into
the reassembled K-D apparatus. Complete
the quantitative transfer with two 10-mL
hexane rinses of the Erlenmeyer flask.
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
10.15 Replace the one or two clean boiling
chips and concentrate the extract to 6 to 10
mL a* in Section 10.8.
10.10 Add a clean boiling chip to the
concentrator tube and attach a two-ball
micro-Snyder column. Prewet the column by
ddding about 1 mL of hexane 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
the concentration in 5 to 10 min. At the
proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood. When the apparent volume of
liquid reaches about 0.5 mL remove the K-D
apparatus and allow it to drain and cool for
at least 10 min. Remove the micro-Snyder
column and rinse its lower joint into the
concentrator tube with 0.2 mL of hexane.
Adjust the extract volume to 1.0 mL with
hexane. Stopper the concentrator tube and
store refrigerated and protected from light if
further processing will not be performed
immediately. If the extract will be stored
longer than two days, it should be transferred
to a Teflon-sealed screw-cap vial. If the
sample extract requires no further cleanup,
proceed with GC/MS analysis (Section 12). If
the sample requires further cleanup, proceed
to Section 11.
10.17 Determine the original sample
volume by refilling the sample bottle to the
mark and transferring the liquid to a lOOO-mL
graduated cylinder. Record the sample
volume to the nearest 5 mL
11. Cleanup and Separation
11.1 Cleanup procedures may not be
necessary for a relatively clean sample
matrix If particular circumstances demand
the use of a cleanup procedure, the analyst
may use either procedure below or any other
appropriate procedure.1-*'However, the
analyst first must demonstrate that the
requirements of Section 8.2 can be met using
the method as revised to incorporate the
cleanup procedure. Two cleanup column
options are offered to the analyst in this
section. The alumina column should be used
first to overcome interferences. If background
problems are still encountered, the silica gel
column may be helpful.
11.2 Alumina column cleanup for 2,3,7,8-
TCDD:
11.2.1 Fill a 300 mm long x 10 mm ID
chromatographic column with activated
alumina to the ISO mm level. Tap the column
gently to settle the alumina and add 10 mm of
anhydrous sodium sulfate to the top.
11.2.2 Preelute the column with SO ml of
hexane. Adjust the elution rate to 1 mL/min.
Discard the eluate and just prior to exposure
of the sodium sulfate layer to the air,
quantitatively transfer the 1.0-mL sample
extract onto the column using two 2-mL
portions of hexane to complete the transfer.
11.2.3 Just prior to exposure of the sodium
sulfate layer to the air, add 50 mL of 3%
methylene chloride/97% hexane (V/V) and
continue the elution of the column. Discard
the eluate.
11.2.4 Next, elute the column with 50 mL
of 20% methylene chloride/80% hexane (V/V)
into a 500-mL K-D flask equipped with a 10-
mL concentrator tube. Concentrate the
collected fraction to 1.0 mL as in Section
10.18 and analyze by GC/MS (Section 12).
11.3 Silica gel column cleanup for 2,3.7,8-
TCDD:
11.3.1 Fill a 400 mm long x 11 mm ID
chromatographic column with silica gel to the
300 mm level. Tap the column gently to settle
the silica gel and add 10 mm of anhydrous
sodium sulfate to the top.
11.3.2 Preelute the column with 50 mL of
20% benzene/80% hexane (V/V). Adjust the
elution rate to 1 mL/min. Discard the eluate
and just prior to exposure of the sodium
sulfate layer to the air, quantitatively transfer
the 1.0-mL sample extract onto the column
using two 2-mL portions of 20% benzene/80%
hexane to complete the transfer.
11.3.3 Just prior to exposure of the sodium
sulfate layer to the air, add 40 mL of 20%
benzene/80% hexane to the column. Collect
the eluate in a clean 500-mL K-D flask
equipped with a 10-mL concentrator tube.
Concentrate the collected fraction to 1.0 mL
as in Section 10.18 and analyze by GC/MS.
12. GC/MS Analysis
12.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. Other
capillary columns or chromatographic
conditions may be used if the requirement* of
Sections 5.5.2 and 8.2 are met.
12.2 Analyze standards and samples with
the mass spectrometer operating in the
selected ion monitoring (SIM) mode using a
dwell time to give at least seven points per
peak. For LRMS, use masses at m/z 320,322,
and 257 for 2,3,7,8-TCDD and either m/z 328
for "CU 2,3,7,8-TCDD or m/z 332 for "Cu
2,3,7.8-TCDD. For HRMS, use masses at m/z
319.8865 and 321.8936 for 2,3.7,8-TCDD and
either m/z 327.8847 for "Cl 2,3,7,8-TCDD or
m/z 331.9367 for **Cu 2,3,7,8-TCDD.
12.3 If lower detection limits are required,
the extract may be carefully evaporated to
dryness under a gentle stream of nitrogen
with the concentrator tube in a water bath at
about 40 'C. Conduct this operation
immediately before GC/MS analysis.
Redissolve the extract in the desired final
volume of ortho-xylene or tetradecane.
12.4 Calibrate the system daily as
described in Section 7.
12.5 Inject 2 to 5 j*L of the sample extract
into the gas chromatograph. The volume of
calibration standard injected must be
measured, or be the same as all sample
injection volumes.
12.6 The presence of 2,3,7,8-TCDD is
qualitatively confirmed if all of the following
criteria are achieved:
12.6.1 The gas chromatographic column
must resolve 2,3,7,8-TCDD from the other 21
TCDD isomers.
12A2 The masses for native 2,3,7,8-TCDD
(LRMS-m/z 320, 322, and 257 and HRMS-m/z
320 and 322) and labeled 2,3,7,8-TCDD (m/z
328 or 332) must exhibit a simultaneous
maximum at a retention time that matches
that of native 2.3,7,8-TCDD in the calibration
standard, with the performance specifications
of the analytical system.
12.6.3 The chlorine isotope ratio at m/z
320 and m/z 322 must agree to within ±10% of
that in the calibration standard.
12.8.4 The signal of all peaks must be
greater than 2.5 times the noise level.
12.7 For quantitation. measure the
response of the m/z 320 peak for 2.3.7.8-
TCDD and the m/z 332 peak for "Ct, 2.3,7,8-
TCDD or the m/z 328 peak for "CU 2,3.7.8-
TCDD.
12.8 Co-eluting impurities are suspected if
all criteria are achieved except those in
Section 12.6.3. In this case, another SIM
analysis using masses at m/z 257, 259. 320
and either m/a 328 or m/z 322 can be
performed. The masses at m/z 257 and m/z
259 are indicative of the loss of one chlorine
and one carbonyl group from 2.3.7,8-TCDD. If
masses m/z 257 and m/z 259 give a chlorine
isotope ratio that agrees to within ±10% of
the same cluster in the calibration standards,
then the presence of TCDD can be confirmed.
Co-eluting DDD, DDE, and PCB residues can
be confirmed, but will require another
injection using the appropriate SIM masses or
full repetitive mass scans. If the response for
"CU 2,3.7,8-TCDD at m/z 328 is too large.
PCB contamination is suspected and can be
confirmed by examining the response at both
m/z 326 and m/z 328. The 'K& 24,7,8-TCDD
internal standard gives negligible response at
m/z 326. These pesticide residues can be
removed using the alumina column cleanup
procedure.
12.9 If broad background interference
restrict* the sensitivity of the GC/MS
analysis, the analyst should employ
additional cleanup procedures and reanalyze
by GC/MS.
12.10 In those circumstances where these
procedures do not yield a definitive
conclusion, the use of high resolution mass
spectrometry is suggested.*
13. Calculations
13.1 Calculate the concentration of 2.3,7,8-
TCDD in the sample using the response factor
(RF) determined in Section 7.1.2 and Equation
2.
Equation 2:
Concentration (jig/L)=
(A,.)(RF)(V0)
where:
A.-SIM response for 2,3,7,8-TCDD at m/z
320.
Ab—SIM response for the internal
standard at m/z 328 or 332.
I. a Amount of internal standard added to
each extract (ug).
V.- Volume of water extracted (L).
13.2 For each sample, calculate the
percent recovery of the internal standard by
comparing the ana of the m/z peak
measured in the sample to the area of the
same peak in the calibration standard. If the
recovery is below 50%, the analyst should
review all aspects of his analytical technique.
13.3 Report results in pg/L without
correction for recovery data. All QC data
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Federal Register / Vol. 49. No. 209 / Friday, October 26, 1984 / Rules and Regulations
141
obtained should be reported with (he sample
results.
14. Method Performance
14.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.' The MDL concentration listed in Table
1 was obtained using reagent water."The
MDL actually achieved in a given analysis
will vary depending on instrument sensitivity
and matrix effects.
14.2 This method was tested by 11
laboratories using reagent water, drinking
water, surface water, and three industrial
wastewaters spiked at six concentrations
over the range 0.02 to 0.20 jig/L.15 Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 3.
References
1. 40 CFR Part 136, Appendix B.
2. "Determination of 2.3,7,8-TCDD in
Industrial and Municipal Wastewaters."
EPA-600/4-82-028. U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, June 1982.
3. Buser. H.R., and Rappe, C. "High
Resolution Gas Chromatography of the 22
Tetrachlorodibenzo-p-dioxinlsomers,"
Analytical Chemistry. 52. 2257 (1980).
4. ASTM Annual Book of Standards, Part
31, D3694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials,
Philadelphia.
5. Harless, R. L, Oswald, E. O., and
Wilkinson, M. K. "Sample Preparation and
Gas Chromatography/Mass Spectrometry
Determination of 2,3.7,8-Tutrachlorodibenzo-
p-dioxin," Analytical Chemistry. 52. 1239
(1980).
6. Lamparski, L. L.. and Nestrick. T. J.
"Determination of Tetra-, Hepta-, and
Octachlorodibenzo-p-dioxin Isomers in
Particulate Samples at Parts per Trillion
Levels." Analytical Chemistry. 52. 2045
(1980).
7. Longhorst, M, L., and Shadoff. L. A.
"Determination of Parts-per-Trillion
Concentrations of Tetra-, Hexa-, and
Octachlorodibenzo-p-dioxins in Human
Milk," Analytical Chemistry. 52. 2037 (1980).
8. "Carcinogens—Working with
Carcinogens." Department of Health,
Education, and Welfare. Public Health
Service. Center for Disease Control. National
Institute for Occupational Safety and Health,
Publication No. 77-206, August 1977.
9. "OSHA Safety and Health Standards,
General Industry," (29 CFR 1910).
Occuptional Safety and Health
Administration. OSHA 2206 (Revised,
January 1976).
10. "Safety in Academic Chemistry
Laboratories." American Chemical Society
Publication. Committee on Chemical Safety,
3rd Edition, 1979.
11. Provost, L. P., and Elder, R. S.,
"Interpretation of Percent Recovery Data,"
American Laboratory. 15, 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
12. ASTM Annual Book of Standards, Part
31. D3370-78, "Standard Practices for
Sampling Water," American Society for
Testing and Materials, Philadelphia.
13. "Methods, 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric DPD) for
Chlorine. Total Residual," Methods for
Chemical Analysis of Water and Wastes,
EPA-600/4-79-020, U.S. Environmental
Protection Agency. Environmental Monitoring
and Support Laboratory. Cincinnati, Ohio
45268, March 1979.
14. Wong, A.S. et al. "The Determination of
2,3.7.8-TCDD in Industrial and Municipal
Wastewaters, Method 613. Part 1—
Development and Detection Limits." G.
Choudhay, L. Keith, and C. Ruppe. ed..
Butterworlh Inc.. (1983).
15. "EPA Method Validation Study 23.
Method 613 (2.3.7,8-Tetrachlorodiben7o-p-
dioxin)," Report for EPA Contract 68-03-2863
(In preparation).
TABLE 1.—Chromatographic Conditions and
Method Detection Limit
Parameter
2.3.7.8,-TCOO
Retention
time
(min)
13 1
Method
Detection
limit (|ig/
U
Column conditions: SP-2330 coated on a GO m long x
0 25 mm ID glass column with hydrogen carrier gas at 40
cm/sec linear velocity, splitless injection using letradecane.
Column temperature held isothermal at 200'C for 1 min. then
programmed at 8'C/nwi to 250 'C and held. Use of helium
earner gas will approximately double the retention time.
TABLE 2.—QC Acceptance Criteria—Method
613
Parameter
2.3.7.6-TCOO
Test
cone.
"if
0.100
Unit
tors
Ojc/
0.0276
Range lor X
Oig/L)
0.0523-0.1226
Range
lor IT
P. 1*1
45-128
5 = Standard deviation of four recovery measurements, in
M8/L (Section 8.2.4).
X=Average recovery for four recovery measurements, in
Mfl/L (Section 8.2.4).
P. P.=Percent reco
8.4.2).
«.-v
I recovery measured (Section 8.3.2. Section
Not*.—These criteria are based directly upon the method
performance data in Table 3. Where necessary, the limits for
recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table
TABLE. 3.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 613
Parameter
2.3 7 8-TCOO
Accuracy, as
recovery. X'
04/1)
086C+000145
Single analyst
precision, t, '
(M/L)
Overall
precision, S '
Oi/g/U
X'-Expected recovery for one or more measurements, of a sample containing a concentration of C, in ug/L
s,'=Expected single analyst standard deviation of measurements at an average concentration found of X, in jig/L
S' = Expected intertaboratory standard deviation of measurements at an average concentration found of X. in
C = True value for the concentration, in jig/L.
X=Average recovery found for measurements of samples containing a concentration of C, in jjg/L
Method 624—Purgeables
1. Scope and Application
1.1 This method covers the determination
of a number of purgeable organics. The
following parameters may be determined by
this method:
Parameter
Bramotorm
Carton tetracftforxfe
ChJoroethane
2-Chtoroethytviny* ether
STORET
No.
34030
32101
32104
34413
32102
34301
34311
34576
32106
CAS No.
71-43-2
75-27-4
75-25-2
74-63-9
56-23-5
108-90-7
75-00-3
110-75-8
67-66-3
Parameter
1.3-Oichlorobenzene
1 4-Oichlorobenzene
1 1-Oicnloroetnane..
1 ,2-Dichloroelhane
1 1-Dlchlcroetnane
trans- 1.2-Dichloroethane • ••<
trans- 1 3-Oichlorooropane
Ethyl benzene
Mothytene chloride
1 1 2,2-Tetrachkxoethane
Toluene
1.1.2-Trichloroethane
STORET
No.
34418
34536
34566
34571
34496
34531
34501
34546
34541
34704
34699
34371
34423
34516
34475
34010
34511
CAS No.
74-67-3
95-50-1
541-73-1
106-46-7
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
10061-01-5
10061-02-6
100-41-4
75-09-2
79-34-5
127-16-4
106-66-3
79-00-5
Parameter
VmylcNoricto
STORET
No.
39180
39175
CAS No.
79-01-6
75-01-4
1.2 The method may be extended to
screen samples for acrolein (STORET No.
34210, CAS No. 107-02-8) and acrylonitrile
(STORET No. 34215. CAS No. 107-13-1),
however, the preferred method for these two
compounds in Method 603.
1.3 This is a purge and trap gas
chromatographic/mass spectrometer (GC/
MS) method applicable to the determination
of the compounds listed above in municipal
-------
142
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
•nd industrial discharges as provided under
40 CFR 136.1.
1.4 The method detection limit (MDL,
defined in Section 14.1)' for each parameter
is listed in Table 1. The MDL for a specific
wastewater may differ from those listed.
depending upon the nature of interferences in
the sample matrix.
1.5 Any modification to this method,
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
Depending upon the nature of the
modification and the extent of intended use,
the applicant may be required to demonstrate
that the modifications will produce
equivalent results when applied to relevant
wastewaters.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the operation of a purge and
trap system and a gas chromatograph/mass
spectrometer and in the interpretation of
mast spectra. Each analyst must demonstrate
the ability to generate acceptable results with
this method using the procedure described in
Section 8.2.
2. Summary of Method
2.1 An inert gas is bubbled through a 5-
mL water sample contained in a specially-
designed purging chamber at ambient
temperature. The purgeables are efficiently
transferred from the aqueous phase to the
vapor phase. The vapor is swept through a
sorbent trap where the purgeables are
trapped. After purging is completed, the trap
is heated and backflushed with the inert gas
to desorb the purgeables onto a gas
chromatographic column. The gas
chromatograph is temperature programmed to
separate the purgeables which are then
detected with a mass spectrometer.11
3. Interferences
3.1 Impurities in the purge gas. organic
compounds outgassing from the plumbing
ahead of the trap, and solvent vapors in the
laboratory account for the majority of
contamination problems. The analytical
system must be demonstated to be free from
contamination under the conditions of the
analysis by running laboratory reagent
blanks as described in Section 8.1.3. The use
of non-Teflon plastic tubing, non-Teflon
thread sealants, or flow controllers with
rubber components in the purge and trap
system should be avoided.
3.2 Samples can be contaminated by
diffusion of volatile organics (particularly
fluorocarbons and methylene chloride)
through the septum seal into the sample
during shipment and storage. A field reagent
blank prepared from reagent water and
carried through the sampling and handling
protocol can serve as a check on such
contamination.
3.3 Contamination by carry-over can
occur whenever high level and low level
samples are sequentially analyzed. To reduce
carry-over, the purging device and sample
syringe must be rinsed with reagent water
between sample analyses. Whenever an
unusually concentrated sample is
encountered, it should be followed by an
analysis of reagent water to check for cross
contamination. For samples containing large
amounts of water-soluble materials.
suspended solids, high boiling compounds or
high purgeable levels, it may be necessary to
wash the purging device with a detergent
solution, rinse it with distilled water, and
then dry it in a 105 ' C oven between
analyses. The trap and other parts of the
system are also subject to contamination:
therefore, frequent bakeout and purging of
the entire system may be required.
4. Safety
4.1 The toxicity or carcinogenicity of each
reagent used in this method has not been
precisely defined: however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint, exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is responsible for
maintaining a current awareness file of
OSHA regulations regarding the safe
handling of the chemicals specified in this
method. A reference file of material data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified4'' for the information of the
analyst.
4.2. The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: benzene, carbon tetrachloride,
chloroform, 1,4-dichlorobenzene, and vinyl
chloride. Primary standards of these toxic
compounds should be prepared in a hood. A
NIOSH/MESA approved toxic gas respirator
should be worn when the analyst handles
high concentrations of these toxic
compounds.
5. Apparatus and Materials
S.I Sampling equipment, for discrete
sampling.
5.1.1 Vial—25-mL capacity or larger,
equipped with a screw cap with a hole in the
center (Pierce #13075 or equivalent).
Detergent wash, rinse with tap and distilled
water, and dry at 105 *C before use.
5.1.2 Septum—Teflon-faced silicane
(Pierce =12722 or equivalent). Detergent
wash, rinse with tap and distilled water, and
dry at 105 'C for 1 h before use.
5.2 Purge and trap system—The purge and
trap system consists of three separate pieces
of equipment: a purging device, trap, and
desorber. Several complete systems are now
commercially available.
5.2.1 The purging device must be designed
to accept 5-mL samples with a water column
at least 3 cm deep. The gaseous head space
between the water column and the trap must
have a total volume of less than 15 mL. The
purge gas must pass though the water column
as finely divided bubbles with a diameter of
lets than 3 mm at the origin. The purge gas
must be introduced no more than 5 mm from
the base of the water column. The purging
device illustrated in Figure 1 meets these
design criteria.
5.2.2 The trap must be at least 25 cm long
and have an inside diameter of at least 0.105
in. The trap must be packed to contain the
following minimum lengths of adsorbents: 1.0
cm of methyl silicone coaled packing (Section
6.3.2). 15 cm of 2.8-dyphenylene oxide
polymer (Section 6.3*1). and 8 cm of silica gel
(Section 6.3.3). The minimum specifications
for the trap are illustrated in Figure 2.
5.2.3 The desorber should be capable of
rapidly heating the trap to 180 'C. The
polymer section of the trap should not be
heated higher than 180 'C and the remaining
sections should not exceed 200 'C. The
desorber illustrated in Figure 2 meets these
design criteria.
5.2.4 The purge and trap system may be
assembled as a separate unit or be coupled to
a gas chromatograph as illustrated in Figures
3 and 4.
5.3 CC/MS system:
5.3.1 Gas chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph suitable
for on-column injection and all required
accessories including syringes, analytical
columns, and gases.
5.3.2 Column—6 ft long x 0.1 in ID
stainless steel or glass, packed with 1% SP-
1000 on Carbopack B (60/80 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 14. Guidelines for the use of alternate
column packings are provided in Section 11.1.
5.3.3 Mass spectrometer—Capable of
scanning from 20 to 200 amu every 7 s or leas,
utilizing 70 V (nominal) electron energy in the
electron impact ionization mode, and
producing a mass spectrum which meets all
the criteria in Table 2 when 50 ng of 4-
bromofluorobenzene (BFB) is injected through
the GC inlet.
5.3.4 GC/MS interface—Any GC to MS
interface that gives acceptable calibration
points at 50 ng or less per injection for each
of the parameters of interest and achieves all
acceptable performance criteria (Section 10)
may be used. GC to MS interfaces
constructed of all glass or glass-lined
materials are recommended. Glass can be
deactivated by silanizing with
dichlorodimethylsilane.
5.3.5 Data system—A computer system
must be interfaced to the mass spectrometer
that allows the continuous acquisition and
storage on machine-readable media of all
mass spectra obtained throughout the
duration of the chromatographic program.
The computer must have software that allows
searching any GC/MS data file for specific
m/z (masses) and plotting such m/z
abundances versus time or scan number. This
type of plot is defined as an Extracted Ion
Current Profile (E1CP). Software must also be
available that allows integrating the
abundance in any EICP between specified
time or scan number limits.
5.4 Syringes—5-mL. glass hypodermic
with Luerlok tip (two each), if applicable to
the purging device.
5.5 Micro syringes—25-pL, 0.008 in. ID
needle.
5.6 Syringe valve—2-way. with Luer ends
(three each).
5.7 Syringe—5-mL. gas-tight with shut-off
valve.
5.8 Bottle—15-mL, screw-cap, with Teflon
cap liner.
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 143
5.9 Balance—Analytical, capable of
accurately weighing 0.0001 g.
ft Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.1.1 Reagent water can be generated by
passing tap water through a carbon filter bed
containing about 1 Ib of activated carbon
(Filtrasorb-300. Calgon Corp.. or equivalent).
6.1.2 A water purification system
(Millipore Super-Q or equivalent) may be
used to generate reagent water.
6.1.3 Reagent water may also be prepared
by boiling water for 15 min. Subsequently,
while maintaining the temperature at 90 'C.
bubble a contaminant-free inert gas through
the water for 1 h. While still hot, transfer the
water to a narrow mouth screw-cap bottle
and seal with a Teflon-lined septum and cap.
6.2 Sodium thiosulfate—(ACS) Granular.
6.3 Trap materials:
6.3.1 2,6-Diphenylene oxide polymer—
Tenax. (60/80 mesh), chromatographic grade
or equivalent.
6.3.2 Methyl silicone packing—3% OV-1
on Chromosorb-W (60/80 mesh) or
equivalent.
6.3.3 Silica gel—35/60 mesh, Davison.
grade-15 or equivalent.
6.4 Methanol—Pesticide quality or
equivalent.
6.5 Stock standard solutions—Stock
standard solutions may be prepared from
pure standard materials or purchased as
certified solutions. Prepare stock standard
solutions in methanol using assayed liquids
or gases as appropriate. Because of the
toxicity of some of the compounds, 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.
6.5.1 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 min or until all alcohol wetted
surfaces have dried. Weigh the flask to the
nearest 0.1 mg.
6.5.2 Add the assayed reference material:
6.5.2.1 Liquids—Using a 100-jtL syringe,
immediately add two or more 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.
6.5.2.2 Gases—To prepare standards for
any of the four halocarbons that boil below
30 *C (bromomethane. chloroethane.
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 methanol
meniscus. Slowly introduce the reference
standard above the surface of the liquid (the
heavy gas will rapidly dissolve in the
methanol).
6.5.3 Reweigh. dilute to volume, stopper,
then mix by inverting the flask several times.
Calculate the concentration in pg/fiL from
the net gain in weight. When compound
purity is assayed to be 96% or greater, the
weight may be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards may be used at any concentration
if they are certified by the manufacturer or by
an independent source.
6.5.4 Transfer the stock standard solution
into a Teflon-sealed screw-cap bottle. Store.
with minimal headspace. at -10 to -20 "C
and protect from light.
6.5.5 Prepare fresh standards weekly for
the four gases and 2-chloroethylvinyl ether.
All other standards must be replaced after
one month, or sooner if comparison with
check standards indicates a problem.
6.6 Secondary dilution standards—Using
stock solutions, prepare secondary dilution
standards in methanol that contain the
compounds of interest, either singly or mixed
together. The secondary dilution standards
should be prepared at concentrations such
that the aqueous calibration standards
prepared in Section 7.3 will bracket the
working range of the analytical system.
Secondary dilution standards should be
stored with minimal headspace and should
be checked frequently for signs of
degradation or evaporation, especially just
prior to preparing calibration standards from
them.
6.7 Surrogate standard spiking solution-
Select a minimum of three surrogate
compounds from Table 3. Prepare stock
standard solutions for each surrogate
standard in methanol as described in Section
6.5. Prepare a surrogate standard spiking
solution from these stock standards at a
concentration of 15 pg/mL in water. Store the
solutions at 4 *C in Teflon-sealed glass
containers with a minimum of headspace.
The solutions should be checked frequently
for stability. The addition of 10 /iL of this
solution of 5 mL of sample or standard is
equivalent to a concentration of 30 ftg/L of
each surrogate standard.
6.8 BFB Standard—Prepare a 25 ng/mL
solution of BFB in methanol.
6.9 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Assemble a purge and trap system
that meets the specifications in Section 5.2.
Condition the trap overnight at 180 *C by
backflushing with an inert gas flow of at least
20 mL/min. Condition the trap for 10 min
once daily prior to use.
7.2 Connect the purge and trap system to
a gas chromatograph. The gas chromatograph
must be operated using temperature and flow
rate conditions equivalent to those given in
Table 1.
7.3 Internal standard calibration
procedure—To use this approach, the analyst
must select three or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standard is not affected by
method or matrix interferences. Some
recommended internal standards are listed in
Table 3.
7.3.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter by carefully adding 20.0 \iL of
one or more secondary dilution standards to
50, 250, or 500 mL of reagent water. A 25-/iL
syringe with a 0.006 in. ID needle should be
used for this operation. One of the calibration
standards should be at a concentration near,
but above, the MDL (Table 1) and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the GC/MS system. These aqueous
standards can be stored up to 24 h. if held in
scaled vials with zero headspace as
described in Section 9.2. If not so stored, they
must be discarded after 1 h.
7.3.2 Prepare a spiking solution containing
each of the internal standards using the
procedures described in Sections 6.5 and 6.6.
It is recommended that the secondary
dilution standard be prepared at a
concentration of 15 /ig/mL of each internal
standard compound. The addition of 10 ^L of
this standard to 5.0 mL of sample or
calibration standard would be equivalent to
7.3.3 Analyze each calibration standard
according to Section 11, adding 10 p.L of
internal standard spiking solution directly to
the syringe (Section 11.4). Tabulate the area
response of the characteristic m/z against
concentration for each compound and
internal standard, and calculate response
factors (RF) for each compound using
Equation 1.
Equation 1.
RF =
(A.)(CU)
(Au)(C.)
where:
A,=Area of the characteristic m/z for the
parameter to be measured.
Ato=Area of the characteristic m/z for the
inernal standard.
Cu=Concentration of the internal
standard.
C,=Concentration of the parameter to be
measured.
If the RF value over the working range is a
constant (<35% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A./AU, vs. RF.
7.4 The working calibration curve or RF
must be verified on each working day by the
measurement of a QC check sample.
7.4.1 Prepare the QC check sample as
described in Section 8.2.2.
7.4.2 Analyze the QC check sample
according to the method beginning in Section
10.
7.4.3 For each parameter, compare the
response (Q) with the corresponding
calibration acceptance criteria found in Table
5. If the responses for all parameters of
interest fall within the designated ranges,
analysis of actual samples can begin. If any
individual Q falls outside the range, proceed
according to Section 7.4.4.
Note.—The large number of parameters in
Table 5 present a substantial probability that
one or more will not meet the calibration
-------
144 Federal Register / Vol. 49. No. 209 / Friday. October 26, 1964 / Rules and Regulations
acceptance criteria when all parameters are
analyzed.
7.4.4 Repeat the test only for those
parameters that failed to meet the calibration
acceptance criteria. If the response for a
parameter does not fall within the range in
this second test, a new calibration curve or
RF must be prepared for that parameter
according to Section 7.3.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial.
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.1 In recognition of advances that are
occurring in chromatography, the analyst is
permitted certain options (detailed in Section
11.1) to improve the separations or lower the
cost of measurements. Bach time such a
modification is made to the method, the
analyst is required to repeat the procedure in
Section 8.2.
8.1.3 Each day, the analyst must analyze a
reagent water blank to demonstrate that
interferences from the analytical system are
under control.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 5% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section 8.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 5* of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3)
meet all specified quality control criteria.
8.1.8 The laboratory must spike all
samples with surrogate standards to monitor
continuing laboratory performance. This
procedure is described in Section 8.5.
8,1.7 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.6.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
12.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of 10
pg/mL in methanol. The QC check sample
concentrate must be obtained from the U.S.
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory in Cincinnati. Ohio, if available. If
not available from that source, the QC check
sample concentrate must be obtained from
another external source. If not available from
either source above, the QC check sample
concentrate must be prepared by the
laboratory using stock standards prepared
independently from those used for
calibration.
8.2.2 Prepare a QC check sample to
contain 20 jig/L of each parameter by adding
200 pL of QC check sample concentrate to
100 mL of reagent water.
8.2.3 Analyze four 5-mL aliquots of the
well-mixed QC check sample according to
the method beginning in Section 10.
8.2.4 Calculate the average recovery (X)
in fig/L, and the standard deviation of the
recovery (s) in pg/L for each parameter of
interest using the four results.
8.2.S For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively.
found in Table 5. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 5 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.6 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.8.1 or 8.2.6.2.
8.2.6.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8^.3.
8.2.6.2 Beginning with Section 6.2.3, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2.3.
8.3 The laboratory must, on an ongoing
basis, spike at least 5% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing 1 to 20
samples per month, at least one spiked
sample per month is required.
8.3.1 The concentration of the spike in the
sample should be determined as follows:
8.3.1.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 20 pg/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2. whichever
concentration would be larger.
8.3.2 Analyze one 5-mL sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
5-mL sample aliquot with 10 fit of the QC
check sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T. where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 5. These
acceptance criteria wer calculated to include
an allowance for error in measurement of
both the background and spike
concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 20 ug/L, the analyst must use
either the QC acceptance criteria in Table 5,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recoveryof a parameter: (1) calculate
accuracy (X') using the equation in Table 6,
substituting the spike concentration (T) for G
(2) calculate overall precision (S') usingthe
equation in Table 6, substituting X1 for X; (3)
calculate the range for recovery at the spike
concentration as (100 X'/T) (±2.44(100 S'/
T)*.'
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
anlaysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory. If the entire list of parameters in
Table 5 must be measured in the sample in
Section 8.3. the probability that the analysis
of a QC check standard will be required is
high. In this case the QC check standard
should be routinely analyzed with the spiked
sample.
8.4.1 Prepare the QC check standard by
adding 10 pL of QC check sample concentrate
(Sections 8.2.1 or 8.3.2) to 5 mL of reagent
water. The QC check standard needs only to
contain the parameters that failed criteria in
the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (PJ as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (PJ
for each parameter with the corresponding
QC acceptance criteria found in Table S.
-------
Federal Register / Vol. 49, No. 209 / Friday. October 28, 1984 / Rules and Regulations
145
.Only parameters that failed the test in
•Section 8.3 need to be compared with these
PEriteria. If the recovery of any such parameter
falls outside the designated range, the
laboratory performance for that parameter is
judged to be out of control, and the problem
must be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As a quality control check, the
laboratory must spike all samples with the
surrogate standard spiking solutions as
described in Section 11.4, and calculate the
percent recovery of each surrogate
compound.
8.6 As part of the QC program for the
laboratory, method accuracy for wastewater
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section 8.3,
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (sp). Express the accuracy
assessment as a percent recovery interval
from P—2sp to P + 2s0. If P=90% and
Sp=10%, for example, the accuracy interval is
expressed as 70-110%. Update the accuracy
assessment for each parameter a regular
basis (e.g. after each five to ten new accuracy
measurements).
8.7 If is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that are most productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analyzed to assess the precision of the
environmental measurements. Whenever
possible, the laboratory should analyze
standard reference materials and participate
in relevant performance evaluation studies.
ft Sample Collection, Preservation, and
Handling
9.1 All samples must be iced or
refrigerated from the time of collection until
analysis. If the sample contains residual
chlorine, add sodium thiosulfate preservative
(10 mg/40 mL is sufficient for up to 5 ppm Cli)
to the empty sample bottle just prior to
shipping to the sampling site. EPA Methods
330.4 and 330.5 may be used for measurement
of residual chlorine.8 Field test kits are
available for this purpose.
9.2 Crab samples must be collected in
glass containers having a total volume of at
least 25 mL. Fill the sample bottle just to
overflowing 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. If preservative
has been added, shake vigorously for 1 min.
Maintain the hermetic seal on the sample
bottle until time of analysis.
9.3 Experimental evidence indicates that
some aromatic compounds, notably benzene,
toluene, and ethyl benzene are susceptible to
rapid biological degradation under certain
environmental conditions.9 Refrigeration
alone may not be adequate to preserve these
compounds in wastewaters for more than
seven days. For this reason, a separate
sample should be collected, acidified, and
analyzed when these aromatics are to be
determined. Collect about 500 mL of sample
in a clean container. Adjust the pH of the
sample to about 2 by adding 1+1 HC1 while
stirring vigorously, Check pH with narrow
range (1.4 to 2.8) pH paper. Fill a sample
container as described in Section 9.2.
9.4 All samples must be analyzed within
14 days of collection.9
10. Daily CC/MS Performance Tests
10.1 At the beginning of each day that
analyses are to be performed, the GC/MS
system must be checked to see if acceptable
performance criteria are achieved for BFB.9
The performance test must be passed before
any samples, blanks, or standards are
analyzed, unless the instrument has met the
DFTPP test described in Method 625 earlier in
the day.10
10.2 These performance tests require the
following instrumental parameters:
Electron Energy: 70 V (nominal)
Mass Range: 20 to 260 amu
Scan Time: To give at least 5 scans per
peak but not to exceed 7 s per scan.
10.3 At the beginning of each day, inject 2
fiL of BFB solution directly on the column.
Alternatively, add 2 ui. of BFB solution to 5.0
mL of reagent water or standard solution and
analyze the solution according to section 11.
Obtain a background-corrected mass
spectrum of BFB and confirm that all the key
m/z criteria in Table 2 are achieved. If all the
criteria are not achieved, the analyst must
retune the mass spectrometer and repeat the
test until all criteria are achieved.
11. Sample Purging and Gas Chromatography
11.1 Table 1 summarizes the
recommended operating conditions for the
gas chromatograph. Included in this table are
retention times and MDL that can be
achieved under these conditions. An example
of the separations achieved by this column is
shown in Figure 5. Other packed columns or
chromatographic conditions may be used if
the requirements of Section 8.2 are met.
11.2 After achieving the key m/z
abundance criteria in Section 10, calibrate
the system daiy as described in Section 7.
11.3 Adjust the purge gas (helium) flow
rate to 40 mL/min. Attach the trap inlet to the
purging device, and set the purge and trap
system to purge (Figure 3). Open the syringe
valve located on the purging device sample
introduction needle.
11.4 Allow the sample to come to ambient
temperature prior to introducing it into the
syringe. 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 to just short of overflowing. Replace
the syringe plunger and compress the sample.
Open the syringe valve and vent any residual
air while adjusting the sample volume to 5.0
mL. 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 10.0 pL of the
surrogate spiking solution (Section 8.7) and
10.0 pL of the internal standard spiking
solution (Section 7.3.2) through the valve
bore, then close the valve. The surrogate and
internal standards may be mixed and added
as a single spiking solution.
11.5 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.
11.6 Close both valves and purge the
sample for 11.0±0.1 min at ambient
temperature.
11.7 After the 11-min purge time, attach
the trap to the chromatograph. adjust the
purge and trap system to the desorb mode
(Figure 4), and begin to temperature program
the gas chromatograph. Introduce the trapped
materials to the GC column by rapidly
heating the trap to 180 'C while backflushing
the trap with an inert gas between 20 and 60
mL/min for 4 min. If rapid heating of the trap
cannot be achieved, the GC cloumn must be
used as a secondary trap by cooling it to 30
*C (subambient temperature, if problems
persist) instead of the initial program
temperature of 45 *C.
11.8 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 reagent water.
11.9 After desorbing the sample for 4 min,
recondition the trap by returning the purge
and trap system to the purge mode. Wait 15 s
then close the syringe valve on the purging
device to begin gas flow through the trap. The
trap temperature should be maintained at
180 "C. After approximately 7 min, turn off
the trap heater and open the syringe valve to
stop the gas flow through the trap. When the
trap is cool, the next sample can be analyzed.
11.10 If the response for any m/z exceeds
the working range of the system, prepare a
dilution of the sample with reagent water
from the aliquot in the second syringe and
reanalyze.
12. Qualitative Identification
12.1 Obtain EICPs for the primary m/z
(Table 4) and at least two secondary masses
for each parameter of interest. The following
criteria must be met to make a qualitative
identification:
12.1.1 The characteristic masses of each
parameter of interest must maximize in the
same or within one scan of each other.
12.1.2 The retention time must fall within
±30 s of the retention time of the authentic
compound.
12.1.3 The relative peak heights of the
three characteristic masses in the EICPs must
fall within ± 20% of the relative intensities of
these masses in a reference mass spectrum.
The reference mass spectrum can be obtained
from a standard analyzed in the GC/MS
system or from a reference library.
12.2 Structural isomers that have very
similar mass spectra and less than 30 s
difference in retention time, can be explicitly
identified only if the resolution between
authentic isomers in a standard mix is
acceptable. Acceptable resolution is achieved
if the baseline to valley height between the
isomers is less than 25% of the sum of the two
peak heights. Otherwise, structural isomers
are identified as isomeric pairs.
13. Calculations
13.1 When a parameter has been
identified, the quantitation of that parameter
should be based on the integrated abundance
-------
146
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
from the EICP of the primary characteristic
m/i given in Table 4. If the (ample produces
an interference for the primary m/z. use a
secondary characteristic m/s to quantitate.
Calculate the concentration in the sample
using the response factor (RF) determined in
Section 7.3.3 and Equation 2.
Equation 2.
Concentration (fig/L) =
(A.MCJ
(AJ(RF)
where:
A,=Area of the characteristic m/z for the
parameter or surrogate standard to be
measured.
A*=Area of the characteristic m/z for the
internal standard.
C»=Concentration of the internal
standard.
13.2 Report results in pg/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
14. Method Performance
14.1 The method detection limit (MDL) it
defined as the rntnimiim concentration of a
substance that can be measured and reported
with 98% confidence that the value it above
zero.1 The MDL concentrations listed in Table
1 were obtained using reagent water."
Similar results were achieved using
representative wastewaters. The MDL
actually achieved in a given analysis will
vary depending on instrument sensitivity and
matrix effects.
14.2 This method was tested by 15
laboratories using reagent water, drinking
water, surface water, and industrial
wastewaters spiked at six concentrations
over the range 5-000 pg/L " Single operator
precision, overall precision, and method
accuracy were found to be directly related to
the concentration of the parameter and
essentially independent of the sample matrix.
Linear equations to describe these
relationships are presented in Table 5.
References
1. 40 CFR Part 136, Appendix B.
2. Bellar. T.A.. and Lichtenberg, J.J.
"Determining Volatile Organics at
Microgram-per-Utre Levels by Gas
QuomttQfftphy."/oumai American Water
Works Astociation. 06,739 (1974).
3. Bellar. T.A., and Lichtenberg. J.J. "Semi-
Automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds." Measurement
of Organic Pollutants in Water and
Wastewater. CE. Van Hall, editor, American
Society for Testing and Materials,
Philadelphia, PA. Special Technical
Publication 066,1978.
4. "Carcinogens—Working With
Carcinogens." Department of Health.
Education, and Welfare, Public Health
Service. Center for Disease Control, National
Institute for Occupational Safety and Health.
Publication No. 77-208, August 1977.
5. "OSHA Safety and Health Standards.
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration. OSHA 2208 (Revised.
January 1978).
8. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication. Committee on Chemical Safety,
3rd Edition. 1979.
7. Provost. L.P.. and Elder. R.S.
"Interpretation of Percent Recovery Data."
American Laboratory. 15, 58-83 (1983). (The
value 2.44 used in the equation in Section
8.2.3 is two times the value 1.22 derived in
this report.)
8. "Methods 330.4 (Titrimetric, DPD-FAS)
and 330.5 (Spectrophotometric, DPD) for
Chlorine. Total Residual." Methods for
Chemical Analysis of Water and Wastes.
EPA-600/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory. Cincinnati, Ohio
45268, March 1979.
9. Budde. W.L.. and Eichelberger. J.W.
"Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass
Spectrometry Equipment and Laboratories."
EPA-600/4-80-025, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio
45268, April 1980.
10. Eichelberger. J.W., Harris, LE.. and
Budde. WJ. "Reference Compound to
Calibrate Ion Abundance Measurement in
Gas Chromatography—Mass Spectrometry
Systems," Analytical Chemiftry. 47,996-1000
(1975).
11. "Method Detection Limit for Methods
624 and 625." Olynyk, P.. Budde. W.L, and
Eichelberger. J.W. Unpublished report,
October I960.
12. "Interiaboratory Method Study for EPA
Method 624—Purgeables." Final Report for
EPA Contract 86-03-3102.
13."Method Performance Data for Method
624," Memorandum from R. Slater and T.
Pressley, U.S. Environmental Protection
Agency, Environmental Monitoring and
Support Laboratory, Cincinnati, Ohio 45286,
January 17,1964.
TABLE 1.—CHROMATOGRAPHIC CONDITIONS
AND METHOD DETECTION LIMITS
Chkxonwtfwns)..
Bmmmnnthans)..
U^StM^IM^ «4^M6J4SI
aaOTnyiw waw•!•..,
TricNoroRuoroimM
1.1
l.l-CtohtonMeian*
M»1>OtoMamMiMni....
CNorotorm
1>OtoMoroMhsM.~
l.l.1.Tnohtoro»ew».
Ctfbon tRksKtiiOnBi)«
U-OtcMoropro»»...-
cto-I^OeMoioeropMi
TitohtoroitMi«.
DtvonncntorornMhira..
1.1Z2-T««HNORM«N
MUM (min)
2.3
3.1
3.6
4.6
6.4
6.3
6.0
10.1
10.8
11.4
12.1
13.4
13.7
14.3
16.7
16.8
17.0
17.1
17J
IM
19J
22.1
nd
nd
nd
nd
2.8
nd
2.8
4.7
1.6
1.6
2.8
t8
U
6.0
5.0
1.8
4.4
3.1
5.0
nd
nd
4.7
6.8
4.1
TABLE 1.—CHROMATOORAPHIC CONDITIONS
AND METHOD DETECTION LIMITS—Continued
_
TokiMW
Effiyl b«ni«oi
1 3-OerHorob«ni«o«
i 2*Dicrtlorobffnnnt
1 .4-Ochtorobwmni
RdcnHon
MM (mm)
23.5
246
26.4
33.9
35.0
35.4
»F
6.0
6.0
1.2
nd
nd
nd
Column condNtont Cirboplk B (60/60 mMh) eottod MV)
1% SP-1000 ptdud in • 6 n by 0.1 in. 10 aton column M»
iMtum own* get M 30 mL/min (tow nto. Column tomptra-
turt hdd « 45*0 tor 3 mn. «Mn programmtd it 6'C/rnn to
nd.
TABLE 2.—BFB KEY M/Z ABUNDANCE CRITERIA
so
7$
•S
96
173
174
17S
176
177._ .„. ... _.
, »L -. -~ •
15 to 40% Ol IMM 66.
30 to SO* otmtMK
BMt PMk, 100% flilMui
Abundmo*.
5 toS% of rnntM
<-j% nl mm 174
>SO% of mm 96
S to *% ol m*M 174.
> B0% but < 101 % Of fltoM
174.
S to 1% Of MM 17*.
TABLE 3.-Si
INTERNAL STANDARDS
ATE AND
Compound
Eti»6j«mn» d-10...
FkorobMsm.
17.0
26J
111
1S.S
26.4
26.4
1S.4
23.5
SJ
1U
m/i
S4
96
lOt
114
111
166
126
n
H
•For i
dWoM. IM TahW 1.
TABLE 4.—CHARACTERISTIC MASSES FOR
PUROEABLE ORQAMCS
Vinyl cntoMt,
TrichtoroSuoroinieiano..
1.2-acNORMtli
1.1.1
CMonM
•Mono*.
>OtoMoroerap<
TitohtoioMhira
I.U-TricNarostll
okvl^Otontonpn
50
64
62
64
64
101
67
117
127
112
78
130
76
127
67
76
106
Steondvy
52.
66.
64.
66.
46,51. and 66.
103.
61 end 66.
66. A 66. 66.
•ndlOO.
61 and 66.
66.
6t 64, and 100.
66,117. and
116.
116 and 121.
63.66, Vd 126.
63,66, and 114.
77.
06, 67. and 132.
126. 206. Md
906.
63.66,66.132
and 134.
77.
63 and 66.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 147
TABLE 4.—CHARACTERISTIC MASSES FOR
PURGEABLE ORGANICS—Continued
TABLE 4.—CHARACTERISTIC MASSES FOR
PURGEABLE ORGANICS—Continued
TABLE 4.—CHARACTERISTIC MASSES FOR
PURGEABLE ORGANICS—Continued
Parameter
1,1,2.2-Tetrachloroethane
Pri-
mary
173
168
Secondary
171 175.250
252. 254. and
256.
83. 85. 131. 133.
and 166.
Parameter
Toluene
Chkxobenzene
Ethvl benzene
Pn-
mary
164
92
112
106
Secondary
129 131 and
166.
91.
114.
91.
Parameter
t ,3-Ochloroben2ene
1 .2-Ochlorooeruene
1 ,4-Oichkyobenzene
Pn-
mary
146
146
146
Secondary
148 and 1 13
1 48 and 1 1 3
148 and 1 13
TABLE 5.—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 624"
Parameter
Totuene • •
Range lor O (pi
g/U
128-27.2
13 1-269
14.2-25.8
2.8-37.2
146-254
132-268
7 6 - 32 4
0-44 8
135-265
0-408
135-265
126-27 4
146-25.4
126-274
145-255
136-26 4
101-299
139-261
68-332
4.6-35.2
100-300
11 8-282
121 27 9
12.1-27.9
14.7-25.3
14.9 25.1
150 250
14.2-25.8
13.3-26.7
9.6-30.4
0.8-39.2
Limit lor
*(M/g/
L)
69
64
5.4
179
52
63
11 4
25 9
6 1
198
61
7 1
55
7 1
51
60
91
57
138
158
104
75
74
74
50
48
46
5.5
6.6
100
200
Range tor X (M/
g/U
152 260
10 1 -260
11.4-31.1
0-41.2
172-235
164 274
64 40 4
O 50 4
137 242
D-459
138 266
11 8-34 7
170 288
11 8-347
14 2-28 5
14 3-27 4
37 423
136-28 5
38 362
10 390
76 324
174 267
D 41 0
135-272
170 266
166 26 7
13 7-30 1
14.3 27 1
18.6 27 6
89-31 5
0 435
Range lor P.
P. (S)
37 - 151
35 155
45 169
0 242
70 140
37 160
14 230
D 305
51 138
D 273
53- 149
59 156
18-190
59 155
49 - 1 55
0-234
54- 156
0-210
D-227
17 1S3
37 162
D 221
46-157
64- 148
47-150
52 162
52-150
71 157
17- 181
D 251
= Concentration measur
red in QC check sample, in jig/L (Section 7.5.3).
s= Standard deviation of lour recovery measurements, in pg/L (Section 8.2.4).
X = Average recovery of lour recovery measurements, in ng/L (Section 8.2.4).
,
P P. = Percent recovery measured, (Section 8.3.2, Section 8.4.2).
D= Detected; result must be greater than zero.
•Criteria were calculated assuming a QC cheek sample concentration ol 20 jig/L.
NOTE.— These criteria are based directly upon the method performance data in Table 6. Where necessary, the limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 6.
TABLE 6.—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 624
Parameter
wlothylono chloride
Pi l^i-Tet/achloroethane -
Tetnchloroethene
Accuracy, as
recovery. X
0*9/U
0.93C+2.00
1.03C-1.58
1.18C-2.35
1.00C
1. IOC- 1.68
0.98C+2.28
1.18C+0.81
1.00C
0.93C+0.33
1.03C-1.81
1 .010-0.03
0.94C+4.47
1.06C+1.68
0.94C+4.47
1.05C+0.36
1.020+0.45
1.12C+0.6I
1.05C+0.03
1.00C
1 OOC
1.00C
0.98C+2.4S
0.87C+1.88
0.93C+1.76
1.08C+0.60
Single analyst
precision, s,'
G*9/L)
0.26X-1 74
0.15X + 0.59
0.123+0.34
0.43X
0.12X+0.25
0.16X-0.09
0 14X + 278
0.62X
0.16X + 0.22
0.37X+2.14
0.17X-0.18
0.22X-1.45
0.14X-0.48
0.22X-1.45
0.13X-0.05
0.17X-0.32
0.17X+1.06
0.14X+O.OB
033X
038X
0.25X
0.14X+1.00
0.15X+1.07
0.18X + 0.69
0.13X-0.18
Overall
precision. S
(H9'M
0 25X - 1 33
0.20X+1.13
0.17X+1.38
0.58X
0.11X+0.37
026X-1 92
029X+1 75
0.84X
0.18X + 0.16
0.58X +0.43
0.17X + 0.49
0.30X-1.20
0.18X-0.82
0.30X-1.20
0.16X+0.47
0.21 X- 0.38
0.43X-0.22
0.19X+0.17
045X
052X
0.34X
0.26X-1.72
0.32X + 4.00
0.20X + 0.41
0.16X-0.4S
-------
148
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 6.— METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION— METHOD 624— Continued
Accuracy, u
Parameter recovery. X
(MQ'1.1
Toluene 098CU203
Tnchtoroethene 104C + 227
TnchtofOftouromethene ,. ; 099C + 0.39
X - Expected recovery for one or more measurements of • sample containing a concentration of C, in ug/L.
S -• Eipected mtflabofilofy ilandwd devotion of measurement! at an average concentration found olx. in (ig/L
C = True value lor the concentration, in »i9/L
• Eitimjie* btMd upon ttw performanct m a tmgta laboratory. >*
» Oua lo cnrornatoyapntc nMdution probl«ms. parformanca sUtemants for tha«a nomars are based upon the sum* of their concentrations.
Single analyst
precision, a,
(M«'U
01SX-071
012X-015
0 MX +002
013X4-036
033X 148
04«X
Overall
preoaton. S
(Mg'D
022X-1 71
0 21X 03S
018X^000
012X+059
034X^039
085X
mCOOCCMO-MMI
-------
149 Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations
OPTIONAL
FOAM J\
TRAP X
M?
If
i >
1 1
i'
i»
•i
H IN. _n
0. D. EXIT. V;—
-I
I
q
0
I
3
?
I
/
«*•••• «** * 4»A ^Mft«
—EXIT V. IN.
>|
0. D.
— 14MM 0. D.
S INLET '/4 IN.
•—
0. D.
^-SAMPLE INLET
| ||-«-2.»AY SYRINGE
f
)
'
\
L
uu
-*l
VALVE
— 17CM. 20 GAUGE SYRINGE NEEDLE
V^6MM. 0. D. RUBBER SEPTUM
/
I
I
\
/
» I V
^~10MM. 0. D. 1/18 IN. O.D.
^
^
tf
*r
V--INLET
% IN. 0.
— j y" STAINLESS STttL
D.
/fel13l MOLECULAR
^/ SIEVE PURGE
^ GAS FILTER
I
*Wm PURGE GAS
T CONTROL
MEDIUM POROSITY
Figure 1. Purging titvicc.
-------
150
Federal Register / Vol. 49. No. 209 / Friday, October 26,1984 / Rules and Regulations
PACKING PROCEDURE
GLASS
WOOL
GRADE 15
SILICA
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26.1984 / Rules and Regulations
CARRIER GAS FLO* CONTROL
PRESSURE REGULATOR
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SllVt FILTER
LIOUIQ INJECTION PORT*
^ COLUMN OVEN
r- CONFIRMATORY COLUMN
TO OETf CTOR
-- ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SELECTION VALVI
TRAP INLET
/ RESISTANCE WIRE
e' CHEATER "CONTROL
PURGING
DEVICE
NoU ALL LINES BETWEEN
TRAP AND OC
SHOULD K HEATED
TOtQX
Figure 3. Purge and trap system • purge mode.
CARRIER GAS
I CONTROL
PRESSURE
REGULATOR
LIQUID INJECTION PORTS
.COLUMN OVEN
PURGE GAS
FLOW CONTROL^
13X MOLEQULAR
SIEVE FILTER
CONFIRMATORY COLUMN
Jinnf-l" ^-ANALYTICAL COLUMN
OPTIONAL 4-PORT COLUMN
SELECTION VALVE
6-PORT TRAP INLET
VALVE J RESISTANCE WIRE HEATEB
*~ r"" CONTROL
TRAP
T?0°C
PURGING
DEVICE
Note:
ALL LINES BETWEEN
TRAP AND GC
SHOULD BE HEATED
TO 95°C.
Figure 4. Purge and trap system - desorb mode.
-------
152
Fudcr.il KKgiklut / Vul \> No. 209 / Fruluy Uctolx'r 2tt. IKtH / Rule* and
COLUMN: ?% SP-100U ON CARBOPACK B
PROGRAM 45
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
153
Method 825—Base/Neutrals and Adds
7. Scope and Application
1.1 This method covers the determination
of a number of organic compounds that are
partitioned into an organic solvent and are
amenable to gas chromatography. The
parameters listed in Tables 1 and 2 may be
qualitatively and quantitatively determined
using this method.
1.2 The method may be extended to
include the parameters listed in Table 3.
Benzidine can be subject to oxidative losses
during solvent concentration. Under the
alkaline conditions of the extraction step, a-
BHC. y-BHC, endosulfan I and II. and endrin
are subject to decomposition.
Hexachlorocyclopentadiene is subject to
thermal decomposition in the inlet of the gas
chromatograph. chemical reaction in acetone
solution, and photochemical decomposition.
N-nitrosodimethylamine is difficult to
separate from the solvent under the
chromatographic conditions described. N-
nitrosodiphenylamine decomposes in the gas
chromatographic inlet and cannot be
separated from diphenylamine. The preferred
method for each of these parameters is listed
in Table 3.
1.3 This is a gas chromatographic/mass
spectrometry (GC/MS) method applicable to
the determination of the compounds listed in
Tables 1, 2, and 3 in municipal and industrial
discharges as provided under 40 CFR 136.1.
1.4 The method detection limit (MDL,
defined in Section 16.1)' for each parameter
is listed in Tables 4 and 5. The MDL for a
specific wastewater may differ from those
listed, depending upon the nature of
interferences in the sample matrix.
1.5 Any modification to this method.
beyond those expressly permitted, shall be
considered as a major modification subject to
application and approval of alternate test
procedures under 40 CFR 136.4 and 136.5.
Depending upon the nature of the
modification and the extent of intended use,
the applicant may be required to demonstrate
that the modifications will produce
equivalent results when applied to relevant
wastewaters.
1.6 This method is restricted to use by or
under the supervision of analysts
experienced in the use of a gas
chromatograph/mass spectrometer and in the
interpretation of mass spectra. Each analyst
must demonstrate the ability to generate
acceptable results with this method using the
procedure described in Section 8.2.
2. Summary of Method
2.1 A measured volume of sample,
approximately 1-L, is serially extracted with
methylene chloride at a pH greater than 11
and again at a pH less than 2 using a
separatory funnel or a continuous extractor.
The methylene chloride extract is dried,
concentrated to a volume of 1 mL, and
analyzed by GC/MS. Qualitative
identification of the parameters in the extract
is performed using the retention time and the
relative abundance of three characteristic
masses (m/z). Quantitative analysis is
performed using either external or internal
standard techniques with a single
characteristic m/z.
3. Interferences
3.1 Method interferences may be caused
by contaminants in solvents, reagents.
glassware, and other sample processing
hardware that lead to discrete artifacts and/
or elevated baselines in the total ion current
profiles. All of these materials must be
routinely demonstrated to be free from
interferences under the conditions of the
analysis by running laboratory reagent
blanks as described in Section 8.1.3.
3.1.1 Glassware must be scrupulously
cleaned.'Clean all glassware as soon as
possible after use by rinsing with the last
solvent used in it. Solvent rinsing should be
followed by detergent washing with hot
water, and rinses with tap water and distilled
water. The glassware should then be drained
dry. and heated in a muffle furnace at 400 *C
for 15 to 30 min. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment. Solvent rinses
with acetone and pesticide quality hexane
may be substituted for the muffle furnace
heating. Thorough rinsing with such solvents
usually eliminates PCB interference.
Volumetric ware should not be heated in a
muffle furnace. After drying and cooling,
glassware should be sealed and stored in a
clean environment to prevent any
accumulation of dust or other contaminants.
Store inverted or capped with aluminum foil.
3.1.2 The use of high purity reagents and
solvents helps to minimize interference
problems. Purification of solvents by
distillation in all-glass systems may be
required.
3.2 Matrix interferences may be caused
by contaminants that are co-extracted from
the sample. The extent of matrix
interferences will vary considerably from
source to source, depending upon the nature
and diversity of the industrial complex or
municipality being sampled.
3.3 The base-neutral extraction may
cause significantly reduced recovery of
phenol. 2-methylphenol, and 2.4-
dimethylphenol. The analyst must recognize
that results obtained under these conditions
are minimum concentrations.
3.4 The packed gas chromatographic
columns recommended for the basic fraction
may not exhibit sufficient resolution for
certain isomeric pairs including the following:
anthracene and phenanthrene; chrysene and
benzo(a)anthracene; and
benzo(b)fluoranthene and
benzo(k)fluoranthene. The gas
chromatographic retention time and mass
spectra for these pairs of compounds are not
sufficiently different to make an
unambiguous identification. Alternative
techniques should be used to identify and
quantify these specific compounds, such as
Method 610.
3.5 In samples that contain an inordinate
number of interferences, the use of chemical
ionization (CI) mass spectrometry may make
identification easier. Tables 6 and 7 give
characteristic Cl ions for most of the
compounds covered by this method. The use
of CI mass spectrometry to support electron
ionization (El) mass spectrometry is
encouraged but not required.
4. Safety.
4.1 The toxicity or carcinogenicity of each
reagent used in this method have 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 data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified *6 for the information of the
analyst.
4.2 The following parameters covered by
this method have been tentatively classified
as known or suspected, human or mammalian
carcinogens: benzo(a)anthracene, benzidine.
3,3'-dichlorobenzidine, benzo(a)pyrene. a-
BHC, 0-BHC, 6-BHC, y-BHC,
dibenzo(a,h)anthracene, N-
nitrosodimethylamine. 4,4'-DDT, and
polychlorinated biphenyls (PCBs). Primary
standards of these toxic compounds should
be prepared in a hood. A N1OSH/MESA
approved toxic gas respirator should be worn
when the analyst handles high concentrations
of these toxic compounds.
5. Apparatus and Materials
5.1 Sampling equipment, for discrete or
composit sampling.
5.1.1 Grab sample bottle—1-L or 1-gt,
amber glass, fitted with a screw cap lined
with Teflon. Foil may be substituted for
Teflon if the sample is not corrosive. If amber
bottles are not available, protect samples
from light. The bottle and cap liner must be
washed, rinsed with acetone or methylene
chloride, and dried before use to minimize
contamination.
5.1.2 Automatic sampler (optional)—The
sampler must incorporate glass sample
containers for the collection of a minimum of
250 mL of sample. Sample containers must be
kept refrigerated at 4 *C and protected from
light during compositing. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used, before use, however, the compressible
tubing should be throughly rinsed with
methanol, followed by repeated rinsings with
distilled water to minimize the potential for
contamination of the sample. An integrating
flow meter is required to collect flow
proportional composites.
5.2 Glassware (All specifications are
suggested. Catalog numbers are included for
illustration only.):
5.2.1 Separatory funnel—2-L, with Teflon
stopcock.
5.2.2 Drying column—Chromatographic
column, 19 mm ID, with coarse frit filter disc.
5.2.3 Concentrator tube, Kuderna-
Danish—10-mL, graduated (Kontes K-570050-
1025 or equivalent). Calibration must be
checked at the volumes employed in the test.
Ground glass stopper is used to prevent
evaporation of extracts.
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154 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
5.2.4 Evaporative flask. Kuderna-
Danish—500-mL (Kontes K-57001-0500 or
equivalent). Attach to concentrator tube with
spring*.
5.2.5 Snyder column, Kuderna-Danish—
Three all macro (Kontes K-503000-0121 or
equivalent).
5.2.6 Snyder column. Kuderna-Danish—
Two-ball macro (Kontes K-569001-0219 or
equivalent).
5.2.7. Vials—10 to 15-mL, amber glass.
with Teflon-lined screw cap.
5.2.8 Continuous liquid—liquid
extractor—Equipped with Teflon or glass
connecting joints and stopcocks requiring no
lubrication. (Hershberg-Wolf Extractor. Ace
Glass Company. Vineland. N.J., P/N 6841-10
or equivalent.)
5.3 Boiling chips—Approximately 10/40
mesh. Heat to 400 *C for 30 min of Soxhlet
extract with methylene chloride.
5.4 Water bath—Heated, with concentric
ring cover, capable of temperature control
(±2*C). The bath should be used in a hood.
5.5 Balance—Analytical, capable of
accurately weighing 0.0001 g.
5.6 GC/MS system:
5.6.1 Cat Chromatograph—An analytical
system complete with a temperature
programmable gas chromatograph and all
required accessores including syringes,
analytical column*, and gases. The injection
port mutt be designed for on-column injection
when using packed columns and for spUtless
injection when using capillary columns.
5.0.2 Column for base/neutrals—1.8 m
long x 2 mm ID glass, packed with 3% SP-
2250 on Supelcoport (100/120 mesh) or
equivalent. This column was used to develop
the method performance statements in
Section 16. Guidelines for the use of alternate
column packings are provided in Section 13.1.
5.6.3 Column for acids—13 m long x 2 mm
ID glass, packed with 1* SP-1240DA on
Supelcoport (100/120 mesh) or equivalent.
This column was used to develop the method
performance statements in Section 16.
Guidelines for the use of alternate column
packings are given in Section 13.1.
5.8.4 Mass spectrometer—Capable of
scanning from 35 to 450 amu every 7 s or less.
utilizing a 70 V (nominal) electron energy in
the electron impact ionization mode, and
producing a mass spectrum which meets all
the criteria in Table 9 when 50 ng of
decafluorotriphenyl phosphine (DFTPP;
bis(perfluorophenyl) phenyl phosphine) is
injected through the GC inlet.
5.0.5 GC/MS interface—Any GC to MS
interface that gives acceptable calibration
points at 50 ng per injection for each of the
parameters of interest and achieves all
acceptable performance criteria (Section 12)
may be used. GC to MS interfaces
constructed of all glass or glass-lined
materials are recommended. Glass can be
deactivated by silanizing with
dichlorodimethylsilane.
5.6.6 Data system—A computer system
must be interfaced to the mass spectrometer
that allows the continuous acquisition and
storage on machine-readable media of all
mass spectra obtained throughout the
duration of the chromatographic program.
The computer must have software that allows
searching any GC/MS data file for specific
m/z and plotting such m/z abundances
versus time or scan number. This type of plot
is defined as an Extracted Ion Current Profile
(EICP). Software must also be available that
allows integrating the abundance in any EICP
between specified time or scan number
limits.
6. Reagents
6.1 Reagent water—Reagent water is
defined as a water in which an interferent is
not observed at the MDL of the parameters of
interest.
6.2 Sodium hydroxide solution (10 N)—
Dissolve 40 g of NaOH (ACS) in reagent
water and dilute to 100 mL.
6.3 Sodium thiosulfate—(ACS) Granular.
6.4 Sulfuric acid (1 +1)—Slowly. add 50
mL of HtSC-4 (ACS, sp. gr. 1.84) to 50 mL of
reagent water.
6.5 Acetone, methanol, methlylene
chloride—Pesticide quality or equivalent.
6.0 Sodium sulfate—(ACS) Granular,
anhydrous. Purify by heating at 400 *C for 4 h
in a shallow tray.
6.7 Stock standard solutions (1.00 fig/
pL)—standard solutions can be prepared
from pure standard materials or purchased as
certified solutions.
6.7.1 Prepare stock standard solutions by
accurately weighing about 0.0100 g of pure
material. Dissolve the material in pesticide
quality acetone or other suitable solvent and
dilute to volume in a 10-mL volumetric flask.
Larger volumes can be used at the
convenience of the analyst. When compound
purity is assayed to be 96% or greater, the
weight may be used without correction to
calculate the concentration of the stock
standard. Commercially prepared stock
standards may be used at any concentration
if they are certified by the manufacturer or by
an independent source.
0.7.2 Transfer the stock standard
solutions into Teflon-sealed screw-cap
bottles. Store at 4 *C and protect from light.
Stock standard solutions should be checked
frequently for signs of degradation or
evaporation, especially just prior to preparing
calibration standards from them.
0.7.3 Stock standard solutions must be
replaced after six months, or sooner if
comparison with quality control check
samples indicate a probelm.
0.6 Surrogate standard spiking solution—
Select a minimum of three surrogate
compounds from Table 8. Prepare a surrogate
standard spiking solution containing each
selected surrogate compound at a
concentration of 100 pg/mL in acetone.
Addition of 1.00 mL of this solution to 1000
mL of sample is equivalent to a concentration
of 100 WJ/L of each surrogate standard. Store
the spiking solution at 4 *C in Teflon-sealed
glass container. The solution should be
checked frequently for stability. The solution
must be replaced after six months, or sooner
if comparison with quality control check
standards indicates a problem.
OA DFTPP standard—Prepare a 25 pg/mL
solution of DFTPP in acetone.
6.10 Quality control check sample
concentrate—See Section 8.2.1.
7. Calibration
7.1 Establish gas chromatographic
operating parameters equivalent to those
indicated in Tables 4 or 5.
7.2 Internal standard calibration
procedure—To use this approach, the analyst
must select three or more internal standards
that are similar in analytical behavior to the
compounds of interest. The analyst must
further demonstrate that the measurement of
the internal standards is not affected by
method or matrix interferences. Some
recommended internal standards are listed in
Table 8. Use the base peak m/z as the
primary m/z for quantification of the
standards. If interferences are noted, use one
of the next two most intense masses for
quantification.
7.2.1 Prepare calibration standards at a
minimum of three concentration levels for
each parameter of interest by adding
appropriate volumes of one or more stock
standards to a volumetric flask. To each
calibration standard or standard mixture, add
a known constant amount of one or more
internal standards, and and dilute to volume
with acetone. One of the calibration
standards should be at a concentration near,
but above, the MDL and the other
concentrations should correspond to the
expected range of concentrations found in
real samples or should define the working
range of the GC/MS system.
7.2J Using injections of 2 to 5 pL, analyze
each calibration standard according to
Section 13 and tabulate the area of the
primary characteristic m/z (Tables 4 and 5)
against concentration for each compound and
internal standard. Calculate response factors
(RF) for each compound using Equation 1.
Equation 1.
(A.HQ.)
RF-
where:
A.=Area of the characteristic m/z for the
parameter to be measured.
A,,=Area of the characteristic m/z for the
internal standard.
Cu=Concentration of the internal standard
(M8/L).
C,=Concentration of the parameter to be
measured (pg/L).
If the RF value over the working range is a
constant (<35% RSD), the RF can be
assumed to be invariant and the average RF
can be used for calculations. Alternatively,
the results can be used to plot a calibration
curve of response ratios, A./A^ vs. RF.
7.3 The working calibration curve or RF
must be verified on each working day by the
measurement of one or more calibration
standards. If the response for any parameter
varies from the predicted response by more
than ±20%, the test must be repeated uning a
fresh calibration standard. Alternatively, a
new calibration curve must be prepared for
that compound.
8. Quality Control
8.1 Each laboratory that uses this method
is required to operate a formal quality control
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 155
program. The minimum requirements of this
program consist of an initial demonstration of
laboratory capability and an ongoing
analysis of spiked samples to evaluate and
document data quality. The laboratory must
maintain records to document the quality of
data that is generated. Ongoing data quality
checks are compared with established
performance criteria to determine if the
results of analyses meet the performance
characteristics of the method. When results
of sample spikes indicate atypical method
performance, a quality control check
standard must be analyzed to confirm that
the measurements were performed in an in-
control mode of operation.
8.1.1 The analyst must make an initial,
one-time, demonstration of the ability to
generate acceptable accuracy and precision
with this method. This ability is established
as described in Section 8.2.
8.1.2 In recognition of advances that are
occuring in chromatography, the analyst is
permitted certain options (detailed in
Sections 10.6 and 13.1) to improve the
separations or lower the cost of
measurements. Each time such a modification
is made to the method, the analyst is required
to repeat the procedure in Section 8.2.
8.1.3 Before processing any samples, the
analyst must analyze a reagent water blank
to demonstrate that interferences from the
analytical system and glassware are under
control. Each time a set of samples is
extracted or reagents are changed, a reagent
water blank must be processed as a
safeguard against laboratory contamination.
8.1.4 The laboratory must, on an ongoing
basis, spike and analyze a minimum of 5% of
all samples to monitor and evaluate
laboratory data quality. This procedure is
described in Section B.3.
8.1.5 The laboratory must, on an ongoing
basis, demonstrate through the analyses of
quality control check standards that the
operation of the measurement system is in
control. This procedure is described in
Section 8.4. The frequency of the check
standard analyses is equivalent to 5% of all
samples analyzed but may be reduced if
spike recoveries from samples (Section 8.3}
meet all specified quality control criteria.
8.1.6 The laboratory must maintain
performance records to document the quality
of data that is generated. This procedure is
described in Section 8.5.
8.2 To establish the ability to generate
acceptable accuracy and precision, the
analyst must perform the following
operations.
8.2.1 A quality control (QC) check sample
concentrate is required containing each
parameter of interest at a concentration of
100 /ig/mL in acetone. Multiple solutions may
be required. PCBs and multicomponent
pesticides may be omitted from this test. The
QC check sample concentrate must be
obtained from the U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory in Cincinnati, Ohio,
if available. If not available from that source,
the QC check sample concentrate must be
obtained from another external source. If not
available from either source above, the QC
check sample concentrate must be prepared
by the laboratory using stock standards
prepared independently from those used for
calibration.
8.2.2 Using a pipet, prepare QC check
samples at a concentration of 100 (ig/L by
adding 1.00 mL of QC check sample
concentrate to each of four 1-L aliquots of
reagent water.
8.2.3 Analyze the well-mixed QC check
samples according to the method beginning in
Section 10 or 11.
8.2.4 Calculate the average recovery (X)
in fig/U and the standard deviation of the
recovery (s) in ng/L, for each parameter using
the four results.
8.2.5 For each parameter compare s and X
with the corresponding acceptance criteria
for precision and accuracy, respectively,
found in Table 6. If s and X for all parameters
of interest meet the acceptance criteria, the
system performance is acceptable and
analysis of actual samples can begin. If any
individual s exceeds the precision limit or
any individual X falls outside the range for
accuracy, the system performance is
unacceptable for that parameter.
Note.—The large number of parameters in
Table 6 present a substantial probability that
one or more will fail at least one of the
acceptance criteria when all parameters are
analyzed.
8.2.6 When one or more of the parameters
tested fail at least one of the acceptance
criteria, the analyst must proceed according
to Section 8.2.6.1 or 8.2.6.2.
8.2.6.1 Locate and correct the source of
the problem and repeat the test for all
parameters of interest beginning with Section
8.2.2.
8.2.6.2 Beginning with Section 8.2.2, repeat
the test only for those parameters that failed
to meet criteria. Repeated failure, however,
will confirm a general problem with the
measurement system. If this occurs, locate
and correct the source of the problem and
repeat the test for all compounds of interest
beginning with Section 8.2.2.
8.3 The laboratory must, on an ongoing
basis, spike at least 5% of the samples from
each sample site being monitored to assess
accuracy. For laboratories analyzing 1 to 20
samples per month, at least one spiked
sample per month is required.
8.3.1. The concentration of the spike in
the sample should be determined as follows:
8.3.1 If, as in compliance monitoring, the
concentration of a specific parameter in the
sample is being checked against a regulatory
concentration limit, the spike should be at
that limit or 1 to 5 times higher than the
background concentration determined in
Section 8.3.2, whichever concentration would
be larger.
8.3.1.2 If the concentration of a specific
parameter in the sample is not being checked
against a limit specific to that parameter, the
spike should be at 100 /xg/L or 1 to 5 times
higher than the background concentration
determined in Section 8.3.2, whichever
concentration would be larger.
8.3.1.3 If it is impractical to determine
background levels before spiking (e.g.,
maximum holding times will be exceeded),
the spike concentration should be (1) the
regulatory concentration limit, if any; or, if
none (2) the larger of either 5 times higher
than the expected background concentration
or 100 pg/L.
8.3.2 Analyze one sample aliquot to
determine the background concentration (B)
of each parameter. If necessary, prepare a
new QC check sample concentrate (Section
8.2.1) appropriate for the background
concentrations in the sample. Spike a second
sample aliquot with 1.0 mL of the QC check
sample concentrate and analyze it to
determine the concentration after spiking (A)
of each parameter. Calculate each percent
recovery (P) as 100(A-B)%/T, where T is the
known true value of the spike.
8.3.3 Compare the percent recovery (P) for
each parameter with the corresponding QC
acceptance criteria found in Table 6. These
acceptance criteria were calculated to
include an allowance for error in
measurement of both the background and
spike concentrations, assuming a spike to
background ratio of 5:1. This error will be
accounted for to the extent that the analyst's
spike to background ratio approaches 5:1.' If
spiking was performed at a concentration
lower than 100 fig/L. the analyst must use
either the QC acceptance criteria in Table 6,
or optional QC acceptance criteria calculated
for the specific spike concentration. To
calculate optional acceptance criteria for the
recovery of a parameter (1) calculate
accuracy (X') using the equation in Table 7.
substituting the spike concentration (T) for C;
(2) calculate overall precision (S'} using the
equation in Table 7, substituting X' for X; (3)
calculate the range for recovery at the spike
concentration as (100 X7T)±2.44(100 S'/T)%'
8.3.4 If any individual P falls outside the
designated range for recovery, that parameter
has failed the acceptance criteria. A check
standard containing each parameter that
failed the criteria must be analyzed as
described in Section 8.4.
8.4 If any parameter fails the acceptance
criteria for recovery in Section 8.3, a QC
check standard containing each parameter
that failed must be prepared and analyzed.
Note.—The frequency for the required
analysis of a QC check standard will depend
upon the number of parameters being
simultaneously tested, the complexity of the
sample matrix, and the performance of the
laboratory. If the entire list of single-
component parameters in Table 6 must be
measured in the sample in Section 8.3. the
probability that the analysis of a QC check
standard will be required is high. In this case
the QC check standard should be routinely
analyzed with the spike sample.
8.4.1 Prepare the QC check standard by
adding 1.0 mL of QC check sample
concentrate (Sections 8.2.1 or 8.3.2) to 1 L of
reagent water. The QC check standard needs
only to contain the parameters that failed
criteria in the test in Section 8.3.
8.4.2 Analyze the QC check standard to
determine the concentration measured (A) of
each parameter. Calculate each percent
recovery (Pa) as 100 (A/T)%, where T is the
true value of the standard concentration.
8.4.3 Compare the percent recovery (P,)
for each parameter with the corresponding
QC acceptance criteria found in Table 6.
Only parameters that failed the test in
Section 8.3 need to be compared with these
criteria. If the recovery of any such parameter
falls outside the designated range, the
-------
156
Federal RegUter / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulation!
laboratory performance for that parameter it
fudged to be out of control, and the problem
mu»l be immediately identified and
corrected. The analytical result for that
parameter in the unspiked sample is suspect
and may not be reported for regulatory
compliance purposes.
8.5 As part of the QC program for the
laboratory, method accuracy for waste water
samples must be assessed and records must
be maintained. After the analysis of five
spiked wastewater samples as in Section S.3.
calculate the average percent recovery (P)
and the standard deviation of the percent
recovery (s,). Express the accuracy
assessment as a percent interval from P-2s,
to P+2sr If P=90% and s,=10%, for
example, the accuracy interval is expressed
as 70-110%. Update the accuracy
assessment for each parameter on a regular
basil (e.g. after each five to ten new accuracy
measurements).
8.6 As a quality control check, the
laboratory must spike all samples with the
surrogate standard spiking solution as
described in Section 10.2. and calculate the
percent recovery of each surrogate
compound.
8,7 It is recommended that the laboratory
adopt additional quality assurance practices
for use with this method. The specific
practices that an moat productive depend
upon the needs of the laboratory and the
nature of the samples. Field duplicates may
be analysed to assess the precision of the
environmental measurements. Whenever
possible, the laboratory should analyse
standard reference materials and participate
in relevant performance evaluation studies.
9. Sample Collection, Preservation, and
Handling
9.1 Crab 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 as free as possible of Tygon tubing
and other potential sources of contamination.
9.2 All sampling must be iced or
refrigerated at 4 'C from the time of collection
until extraction. Fill the sample bottles and, if
residual chlorine is present, add 80 mg of
sodium thiosulfate per liter of sample and
mix well. EPA Methods 330.4 and 330.5 may
be used for measurement of residual
chlorine.* Field test kits are available for this
purpose.
9.3 All samples must be extracted within
7 days of collection and completely analysed
within 40 days of extraction.
10. Separately Funnel Extraction
10.1 Samples are usually extracted using
separately funnel techniques. If emulsions
will prevent achieving acceptable solvent
recovery with separatory funnel extractions,
continuous extraction (Section 11} may be
used. The separatory funnel extraction
scheme described below assumes a sample
volume of 1L When sample volumes of 2 L
are to be extracted, use 250.100, and 100-mL
volumes of methylene chloride for the serial
extraction of the base/neutrals and 200,100,
and 100-mL volumes of methylene chloride
for the acids.
10.2 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Pour the entire sample into a
2-L separatory funnel. Pipet 1.00 mL of the
surrogate standard spiking solution into the
separatory funnel and mix well. Check the
pH of the sample with wide-range pH paper
and adjust to pH>ll with sodium hydroxide
solution.
10.3 Add 60 mL of methylene chloride to
the sample bottle, seal, and shake for 30 s to
rinse the inner surface. Transfer the solvent
to the separatory funnel and extract the
sample by shaking the funnel for 2 min with
periodic venting to release excess pressure.
Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the
emulsion interface between layer* is more
than one-third the volume of the solvent
layer, the analyst must employ mechanical
techniques to complete the phase separation.
The optimum technique depends upon the
sample, but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Collect the methylene chloride extract in a
250-mL Erlenmeyer flask. If the emulsion
cannot be broken (recovery of leas than 80ft
of the methylene chloride, corrected for the
water solubility of methylene chloride),
transfer the sample, solvent and emulsion
into the extraction chamber of a continuous
extractor and proceed as described in Section
11.3.
10.4 Add a second 80-mL volume of
methylene chloride to the sample bottle and
repeat the extraction procedure a second
time, combining the extracts in the
Erlenmeyer flask. Perform a third extraction
in the same manner. Label the combined
extract as the base/neutral fraction.
10.5 Adjust the pH of the aqueous phase
to less than 2 using sulfuric add. Serially
extract the acidified aqueous phase three
times with 80-mL aliquots of methylene
chloride. Collect and combine the extracts in
a 250-mL Erlenmeyer flask and label the
combined extracts as the acid fraction.
10.6 For each fraction, assemble a
Kudema-Danish (K-D) concentrator by
attaching a 10-mL concentrator tube to a 500-
mL evaporative flask. Other concentration
devices or techniques may be used in place of
the K-D concentrator if the requirements of
Section 8.2 are met.
10.7 For each fraction, pour the combined
extract through a solvent-rinsed drying
column containing about 10 cm of anhydrous
sodium sulfate, and collect the extract in the
K-D concentrator. Rinse the Erlenmeyer flask
and column with 20 to 30 mL of methylene
chloride to complete the quantitative transfer.
10.8 Add one or two clean boiling chips
and attach a three-ball Snyder column to the
evaporative flask for each fraction. Prewet
each Snyder column by adding about 1 mL of
methylene chloride to the top. Place the K-D
apparatus on a hot water bath (60 to 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
with hot vapor. Adjust the vertical position of
the apparatus and the water temperature as
required to complete the concentration in 15
to 20 min. At the proper rate of distillation the
balls of the column will actively chatter but
the chambers will not flood with condensed
solvent. When the apparent volume of liquid
reaches 1 mL remove the K-D apparatus
from the water bath and allow it to drain and
cool for at least 10 min. 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 S-mL syringe is
recommended for this operation.
10.9 Add another one or two clean boiling
chips to the concentrator tube for each
fraction and attach a two-ball micro-Snyder
column. Prewet the Snyder column by adding
about 0.5 mL of methylene chloride to the top.
Place the K-D apparatus on a hot water bath
(80 to 85 *C) so that the concentrator tube is
partially immersed in hot water. Adjust the
vertical position of the apparatus and the
water temperature as required to complete
the concentration in 5 to 10 min. At the
proper rate of distillation the balls of the
column will actively chatter but the chambers
will not flood with condensed solvent When
the apparent volume of liquid reaches about
0.5 mL remove the K-D apparatus from the
water bath and allow it to drain and cool for
at least 10 min. Remove the Snyder column
and rinse the flask and its lower joint Into the
concentrator tuba with approximately O2 mL
of acetone or methylene chloride. Adjust the
final volume to 1.0 mL with the solvent
Stopper the concentrator tuba and store
refrigerated if further processing will not be
performed immediately. If the extracts will be
stored longer than two days, they should be
transferred to Teflon-sealed screw-cap vials
and labeled base/neutral or add fraction as
appropriate.
10.10 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
11. Continuant Extraction
11.1 When experience with a sample from
a given source indicates that a serious
emulsion problem will result or an emulsion
is encountered using a separatory funnel in
Section 10.3, a continuous extractor should be
used.
11.2 Mark the water meniscus on the side
of the sample bottle for later determination of
sample volume. Check the pH of the sample
with wide-range pH paper and adjust to pH
>11 with sodium hydroxide solution.
Transfer the sample to the continuous
extractor and using a pipet add 1.00 mL of
surrogate standard spiking solution and mix
well. Add 80 mL of methylene chloride to the
sample bottle, seal, and shake for 30 s to
rinse the inner surface. Transfer the solvent
to the extractor.
11.3 Repeat the sample bottle rinse with
an additional 50 to 100-mL portion of
methylene chloride and add the rinse to the
extractor.
11.4 Add 200 to 500 mL of methylene
chloride to the distillinf flask, add suffldent
reagent water to ensure proper operation.
and extract for 24 h. Allow to cool then
detach the distilling flask. Dry, concentrate.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 157
and seal the extract as in Sections 10.6
through 10.9.
11.5 Charge a clean distilling flask with
500 mL of methylene chloride and attach it to
the continuous extractor. Carefully, while
stirring, adjust the pH of the aqueous phase
to less than 2 using sulfuric acid. Extract for
24 h. Dry. concentrate, and seal the extract as
in Sections 10.6 through 10.9.
12. Daily GC/MS Performance Tests
12.1 At the beginning of each day that
analyses are to be performed, the GC/MS
system must be checked to see if acceptable
performance criteria are achieved for
DFTPP.10Each day that benzidine is to be
determined, the tailing factor criterion
described in Section 12.4 must be achieved.
Each day that the acids are to be determined,
the tailing factor criterion in Section 12.5
must be achieved.
12.2 These performance tests require the
following instrumental parameters:
Electron Energy: 70 V (nominal)
Mass Range: 35 to 450 amu
Scan Time: To give at least 5 scans per
peak but not to exceed 7 s per scan.
12.3 DFTPP performance test—At the
beginning of each day, inject 2 jiiL (50 ng) of
DFTPP standard solution. Obtain a
background-corrected mass spectra of DFTPP
and confirm that all the key m/z criteria in
Table 9 are achieved. If all the criteria are not
achieved, the analyst must retime the mass
spectrometer and repeat the test until all
criteria are achieved. The performance
criteria must be achieved before any samples,
blanks, or standards are analyzed. The tailing
factor tests in Sections 12.4 and 12.5 may be
performed simultaneously with the DFTPP
test.
12.4 Column performance test for base/
neutrals—At the beginning of each day that
the base/neutral fraction is to be analyzed
for benzidine, the benzidine tailing factor
must be calculated. Inject 100 ng of benzidine
either separately or as a part of a standard
mixture that may contain DFTPP and
calculate the tailing factor. The benzidine
tailing factor must be less than 3.0.
Calculation of the tailing factor is illustrated
in Figure 13." Replace the column packing if
the tailing factor criterion cannot be
achieved.
12.5 Column performance test for acids—
At the beginning of each day that the acids
are to be determined, inject 50 ng of
pentachlorophenol either separately or as a
part of a standard mix that may contain
DFTPP. The tailing factor for
pentachlorophenol must be less than 5.
Calculation of the tailing factor is illustrated
in Figure 13." Replace the column packing if
the tailing factor criterion cannot be
achieved.
13. Gas Chromatography/Mass Spectrometry
13.1 Table 4 summarizes the
recommended gas chromatographic operating
conditions for the base/neutral fraction.
Table 5 summarizes the recommended gas
chromatographic operating conditions for the
acid fraction. Included in these tables are
retention times and MDL that can be
achieved under these conditions. Examples of
the separations achieved by these columns
are shown in Figures 1 through 12. Other
packed or capillary (open-tubular) columns or
chromatographic conditions may be used if
the requirements of Section 8.2 are met.
13.2 After conducting the GC/MS
performance tests in Section 12, calibrate the
system daily as described in Section 7.
13.3 If the internal standard calibration
procedure is being used, the internal standard
must be added to sample extract and mixed
thoroughly immediately before injection into
the instrument. This procedure minimizes
losses due to adsorption, chemical reaction or
evaporation.
13.4 Inject 2 to 5 /*L of the sample extract
or standard into the GC/MS system using the
solvent-flush technique." Smaller (1.0 fit)
volumes may be injected if automatic devices
are employed. Record the volume injected to
the nearest 0.05 fiL
13.5 If the response for any m/z exceeds
the working range of the GC/MS system,
dilute the extract and reanalyze.
13.6 Perform all qualitative and
quantitative measurements as described in
Sections 14 and 15. When the extracts are not
being used for analyses, store them
refrigerated at 4*C, protected from light in
screw-cap vials equipped with unpierced
Teflon-lined septa.
14. Qualitative Identification
14.1 Obtain EICPs for the primary m/z
and the two other masses listed in Tables 4
and 5. See Section 7.3 for masses to be used
with internal and surrogate standards. The
following criteria must be met to make a
qualitative identification:
14.1.1 The characteristic masses of each
parameter of interest must maximize in the
same or within one scan of each other.
14.1.2 The retention time must fall within
±30 s of the retention time of the authentic
compound.
14.1.3 The relative peak heights of the
three characteristic masses in the EICPs must
fall within ±20% of the relative intensities of
these masses in a reference mass spectrum.
The reference mass spectrum can be obtained
from a standard analyzed in the GC/MS
system or from a reference library.
14.2 Structural isomers that have very
similar mass spectra and less than 30 s
difference in retention time, can be explicitly
identified only if the resolution between
authentic isomers in a standard mix is
acceptable. Acceptable resolution is achieved
if the baseline to valley height between the
isomers is less than 25% of the sum of the two
peak heights. Otherwise, structural isomers
are identified as isomeric pairs.
15. Calculations
15.1 When a parameter has been
identified, the quantitation of that parameter
will be based on the integrated abundance
from the EICP of the primary characteristic
m/z in Tables 4 and 5. Use the base peak m/z
for internal and surrogate standards. If the
sample produces an interference for the
primary m/z, use a secondary characteristic
m/z to quantitate.
Calculate the concentration in the sample
using the response factor (RF) determined in
Section 7.2.2 and Equation 3.
Equation 3.
Concentration (w!/L) =
(A.KU
(AU)(RF)(V0)
where:
A, = Area of the characteristic m/z for the
parameter or surrogate standard to be
measured.
Ato=Area of the characteristic m/z for the
internal standard.
I, = Amount of internal standard added to
each extract (jig).
V0=Volume of water extracted (L).
15.2 Report results in jig/L without
correction for recovery data. All QC data
obtained should be reported with the sample
results.
16. Method Performance
16.1 The method detection limit (MDL) is
defined as the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the value is above
zero.'The MDL concentrations listed in
Tables 4 and 5 were obtained using reagent
water."The MDL actually achieved in a
given analysis will vary depending on
instrument sensitivity and matrix effects.
16.2 This method was tested by 15
laboratories using reagent water, drinking
water, surface water, and industrial
wastewaters spiked at six concentrations
over the range 5 to 1300 pg/L." Single
operator precision, overall precision, and
method accuracy were found to be directly
related to the concentration of the parameter
and essentially independent of the sample
matrix. Linear equations to describe these
relationships are presented in Table 7.
17. Screening Procedure for 2.3.7,8-
Tetrachlorodibenzo-p-dioxin (2.3,7,8- TCDD)
17.1 If the sample must be screened for
the presence of 2,3,7,8-TCDD, it is
recommended that the reference material not
be handled in the laboratory unless extensive
safety precautions are employed. It is
sufficient to analyze the base/neutral extract
by selected ion monitoring (SIM) GC/MS
techniques, as follows:
17.1.1 Concentrate the base/neutral
extract to a final volume of 0.2 ml.
17.1.2 Adjust the temperature of the base/
neutral column (Section 5.6.2) to 220 *C.
17.1.3 Operate the mass spectrometer to
acquire data in the SIM mode using the ions
at m/z 257, 320 and 322 and a dwell time no
greater than 333 milliseconds per mass.
17.1.4 Inject 5 to 7 jiL of the base/neutral
extract. Collect SIM data for a total of 10 min.
17.1.5 The possible presence of 2,3,7,8-
TCDD is indicated if all three masses exhibit
simultaneous peaks at any point in the
selected ion current profiles.
17.1.6 For each occurrence where the
possible presence of 2,3,7,8-TCDD is
indicated, calculate and retain the relative
abundances of each of the three masses.
17.2 False positives to this test may be
caused by the presence of single or coeluting
combinations of compounds whose mass
spectra contain all of these masses.
17.3 Conclusive results of the presence
and concentration level of 2,3,7,8-TCDD can
465-028 n -
-------
158
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rulet and Regulation*
b« obtained only from a properly equipped
laboratory through the use of EPA Method
013 or other approved alternate teit
procedures.
References
1.40 CFR Part 136. Appendix B.
2. "Sampling and Analysis Procedures for
Screening of Industrial Effluents for Priority
Pollutants," U.S. Environmental Protection
Agency. Environmental Monitoring and
Support Laboratory. Cincinnati, Ohio 45268,
March 1977. Revised April 1977. Available
from Effluent Guidelines Diviiion,
Washington, DC 20460.
3. ASTM Annual Book of Standards. Part
31.03694-78. "Standard Practices for
Preparation of Sample Containers and for
Preservation of Organic Constituents,"
American Society for Testing and Materials.
Philadelphia.
4. "Carcinogens—Working With
Carcinogens," Department of Health,
Education, and Welfare, Public Health
Service, Center for Disease Control, National
Institute for Occupational Safety and Health.
Publication No. 77-206, August 1977.
5. "OSHA Safety and Health Standards,
General Industry." (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2208 (Revised.
January 1976).
6. "Safety in Academic Chemistry
Laboratories."American Chemical Society
Publication. Committee on Chemical Safety.
3rd Edition. 1979.
7. Provost, L.P., and Elder. R.S.
"Interpretation of Percent Recovery Data,"
American Laboratory. 15. 58-63 (1983). (The
value 2.44 used in the equation in Section
8.3.3 is two times the value 1.22 derived in
this report.)
8. ASTM Annual Book of Standards. Part
31. D3370-76. "Standard Practice! for
Sampling Water," American Society for
Testing and Materials. Philadelphia.
9. "Methods 330.4 (Titrimetric. DPD-FAS)
and 330.5 (Spectrophotometric, DPD) for
Chlorine. Total Residual." Methods for
Chemical Analysis of Water and Wastes.
EPA-eoo/4-79-020, U.S. Environmental
Protection Agency, Environmental Monitoring
and Support Laboratory. Cincinnati, Ohio
45268. March 1979.
10. Eichelberger. |.W.. Harris. L.E., and
Budde. W.L. "Reference Compound to
Calibrate Ion Abundance Measurement in
Cas Chromatography-Mass Spectometry,"
Analytical Chemistry. 47,995 (1975).
11. McNair. N.M. and Bonelli. E.J. "Basic
Chromatography," Consolidated Printing,
Berkeley, California, p. 52,1969.
12. Burke. J.A. "Gas Chromatography for
Pesticide Residue Analysis: Some Practical
Aspects," Journal of the Association of
Official Analytical Chemists. 48.1037 (1965).
13. Olynyk. P.. Budde. W.L, and
Eichelberger. J.W. "Method Detection Limit
for Methods 624 and 625." Unpublished
report. October 1980.
14. "Interlaboratory Method Study for EPA
Method 625—Base/Neutrals, Acids, and
Pesticides," Final report for EPA Contract 68-
03-3102 (In preparation).
TABLE 1.—BASE/NEUTRAL EXTRACTABLES
AompMMM..
Attkv
Bfell-cNoraMprapiiQttMr
4*omoph«i»< phw
CMari
DlMnyi pfNnNiw ••
EndOMMnwSMt..
No.
94206
94200
94110
34616
94242
34247
94611
34261
94266
94273
34278
38100
94263
34698
34681
94641
34320
38910
38320
98110
94671
94691
34396
34341
34611
34626
34351
CAS No.
89-32-8
206-86-6
120-12-7
20746-6
60-91-6
181-24-2
66-66-7
916-66-7
111-81-1
117-81-7
131-11-9
121-14-2
606-20-2
117-64-0
1031-07-8
TABLE 1.—BASE/NEUTRAL EXTRACTABLES-
ConthHMd
N-Nttrotodi n propyttvnntx..
PC8-1018
PC8-1221
PCfl-1232
PCB-1242
PCB-1246
PCB-1264 —
pctj-i2eo
1.2,4-TricNMafei
STORE!
No.
3*376
343S1
39410
38420
38700
34381
3438S
34403
34447
34428
34871
38482
34481
as
00
•1
CAS No.
7421-83-4
208-444
88-73-7
78-44-8
1024-57-3
118-74-1
87-48-3
87-72-1
183-38-5
78-58-1
81-20-3
88-85-3
821-64-7
12874-11-2
11104-28-2
11141-18-5
S34S8->1-8
12872-28-4
11087-88-1
11088-82-5
85-01-8
128-00-0
8001-35-t
110-81-1
TABLE 2.—ACID EXTRACTABLEB
2.44MM0WI01..
24SMPM
101-86-3
57-74-8
81-68-7
7005-72-3
218-01-8
72-64-8
72-86-8
50-28-3
69-70-3
84-74-2
541-73-1
86-80-1 MHC_
108-46-7
81-84-1
80-87-1
2.4»ThoMoru|ih»iiel-.-
No.
34801
94616
94667
34881
38032
34684
34621
CAS No.
85-87-8
166-67-6
51-68-4
534-61-1
88-75-6
100-01-7
S7-(
TABLE 3.—ADDITIONAL EXTRACTABLE
PARAMETERS•
BmUn
MHC-
EndomMn «....
Endrin
No.
36110
14961
94491
CAS NO.
82-87-6
31844-6
39119-68-6
72-20-6
77-47-4
82-75-6
612
607
807
•SMSK*on1.2.
TABLE 4.—CHROMATOORAPHIC CONDITIONS, METHOD DETECTION LIMITS. AND CHARACTERISTIC MASSES FOR BASE/NEUTRAL EXTRACTABLES
161
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 159
TABLE 4.—CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LIMITS. AND CHARACTERISTIC MASSES FOR BASE/NEUTRAL EXTRACTABLES—
Continued
Parameter
Diethylphthalate
0-BHC*
6-BHC*
A.RHC
Heptacttior
(.BMC
Aldrin
Endosulfan I"
Dieldrtn
4 4''OOE • • •
*
Endrin*
Endosulfan II*
4 4'g/u
1.9
1.6
1.9
1.9
4.2
5.7
1.9
1.9
1.9
1.9
5.4
1.9
4.2
1.9
3.1
1.9
2.5
2.2
2.2
2.5
5.6
1.9
2.8
44
4.7
5.6
2.5
2.5
2.5
7.8
16.5
2.5
4.8
2.5
2.5
3.7
2.5
4.1
30
36
Characteristic masses
Electron impact
Primary
154
163
165
166
204
165
149
169
2B4
183
248
183
178
178
181
100
183
66
149
353
237
202
79
246
202
81
237
235
184
235
272
67
149
149
228
228
252
149
252
252
252
276
278
276
42
373
159
224
190
190
224
294
294
330
Second-
ary
153
194
89
165
206
63
177
168
142
181
250
161
179
179
163
272
109
263
150
355
338
101
263
248
101
263
339
237
92
237
387
345
91
167
226
229
254
253
253
253
138
139
138
74
375
231
260
224
224
260
330
330
362
Second-
ary
152
164
121
167
141
182
150
167
249
109
141
109
176
176
109
274
181
220
104
351
341
100
279
176
100
82
341
165
185
165
422
250
206
279
229
226
126
125
125
125
277
279
277
44
377
233
294
260
260
294
262
362
394
Chemical iontzation
Meth-
ane
1S4
151
183
166
183
177
169
264
249
178
178
149
203
203
185
149
149
228
228
252
252
252
276
278
278
Meth-
ane
155
163
211
167
211
223
170
286
251
179
179
205
231
231
213
299
229
229
253
253
253
277
279
277
'
Meth-
ane
183
164
223
195
223
251
198
268
277
207
207
279
243
243
225
327
257
257
281
281
281
305
307
305
•See Section 1.2.
* These compounds are mixtures of various isomers. (See figures 2 thru 12.)
Column conditions: Supecoport (100/120 mesh) coated with 3% SP-2250 packed in a 1.8 m long x 2mm ID glass column with helium carrier gas at 30 mL/min now rate. Column
temperature held isothermal at 50 'C for 4 min. then programmed at 8 'C/rrun to 270 "C and held for 30 min.
TABLE 5.—CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LIMITS, AND CHARACTERISTIC MASSES FOR ACID EXTRACTABLES
Parameter
Phenol •
2 4-Otehtorophenol
2.4 6*Trichlorophenol
4'Nltropheool • •'
Reten-
tion time
(min)
5.9
6.5
8.0
9.4
9.8
11.8
13.2
15.9
16.2
17.5
20.3
Method
detec-
tion Birin
(wi'L)
3.3
3.6
1.5
2.7
2.7
2.7
3.0
42
24
3.6
2.4
Characteristic masses
Electron Impact
Primary
128
139
94
122
162
196
142
184
198
266
65
Second-
ary
64
65
65
107
164
198
107
63
182
264
139
Second-
ary
130
109
66
121
98
200
144
154
77
268
109
Chemical ionization
Meth-
ane
129
140
95
123
163
197
143
185
199
267
140
Meth-
ane
131
168
123
151
165
199
171
213
227
265
168
Meth-
ane
157
122
135
163
167
201
183
225
239
269
122
Column conditions: Supelcoporl (100/120 mesh) coated Witt) 1% SP-1240DA packed in a 1.8 m long x 2mm ID glass column with helium carrier gas at 30 mL/min How rate. Column
temperature held isothermal at 70 'C for 2 min then programmed at 8 'C/min to 200 'C.
-------
160
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1964 / Rules and Regulations
TABLE 6.—OC ACCEPTANCE CRITERIA—METHOD 625
PwvmMr
Ac*mtpMh*n*
Akttn .... ., . , ,
ftanmlilanthractnt
ltoninftiHliytfmtfv«a
ptfqo(ft)fluoraniht*>t
nmnmtitAtarvtuM
•V8HC
B>.BHC ..
Pttff cMufUflhyJ)t|ihoicy)m<ODO
44*-OOt
4 4'-OOT •
l^tmnmnim KlftntfVBIMnS
Hi n iMtftd (iMftMBBta
1 J'PJCMOtflfrtfttfrT .. -- -..--• ""--• ------ .-----... -r,— ,„„—-,.,- ,,, n, — -.., ,,, ...
3-l*~OtA~M^^BJU^feM
OMhyl pMlvlcto
"** «•— •» frfrit^i^ita
j^l OrtHijmmna
1.6 Prt>olGfcMn»
Ot*H)cM)Mh0ftAi
Endrin *Jtfttiyd> - ,„-...,--,--....„,-, --„... ,,....,„„ „
RunwNhm
nMwmm
Httilvttar
1 Tipiir'ltnT tpcwUt .
"^'^ifrnntiiiiMiaiM
lB>MM IlkHll^lMfcl
tni>ttTCf1.2,3'
ttoghonjne >
••WtMhl^iVM
iS|]i|gn¥>ni|
tl NH muul n propyltirtnt .,
PCS- 1 260
pyf«O* . « «
1.2,4 Trtchtorabcrmn* , , ,,
2*d^Di^tfMiWL.
2.4-OrtMphMOl » -» »—
3 Mfcoptunol
4 NUmtunijI
PinlKMirtXitfunnl
Ptt«no4 ~
P. P.-MnMt neotwv nwMMd (Section 8.3A 8«*onT«it"" ~ '
T«M condmion
(M8/L)
100
100
100
100
100
100
100
too
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Umtt tor • (MS/
27 6
402
390
32.0
276
3tt
323
390
5(9
234
31 S
216
SSO
345
46.3
41.1
230
130
334
46.3
310
320
61.6
700
16.7
309
41 7
32.1
714
307
26.5
23£
216
29.6
314
16.7
32.5
32.6
20.7
37.2
547
24.9
26.3
245
446
63.3
30 1
993
554
54.2
206
2U
2S.1
37,2
26.7
26.4
26.1
4*6
93.2
354
472
46.9
22.6
31 7
Rang* toe X(|ig/
60 1-1323
535-1260
72-1522
43 4-1 16 0
41 6-1330
420-1404
252-145 7
31 7-1460
0-1950
0-1399
41 5-1306
0-1000
429-1260
49 2-164 7
62.6-136.6
26.9-136.6
64 9-1144
64 5-1135
441-1399
0-1345
197-1197
D-1706
0-1967
64-111 0
46 6-1 12.0
16.7-1S3.6
37 3-106 7
6.2*312.5
443-119.3
D-1000
0-100.0
47 5-126.6
66.1-136.7
16.6-131 6
D-103.5
0-16M
42.9-121 J
71 6-106.4
D-17U
709-1094
76-141 5
376-1024
554-1000
0-1509
46.6-1604
35.6-1196
543-1576
13 6-197.9
163-1210
654-106.7
696-100.0
57 3-1294
406-1279
364-120.4
52.5-121.7
416-1060
0-172.9
530-1000
45.0-166.7
13.0-106.5
36.1-151.6
16.6-1000
524-1292
Rang* tor f. P.
(P«c*nt)
33.145
0-166
27-133
33-143
24-159
17-163
D-219
EM 52
0-110
12-156
35-166
6-156
60-116
17-166
0-146
4*136
0-909
O427
1-116
IVM9
16-136
0-112
36-136
50-156
4-146
0-107
0-206
16-137
56-121
0*161
26-155
O-ISf i
24*i td
40»1iy
0-171
21*166
35-160
D-230
O»164
54*120
52*115
44-142
23-114
36-136
31*116
D-191
D-161
29-162
0-132
14-176
5-112
37-144
D
HatK TtaM oritarH «• biMd dMc6y iiwn
cuiororKtom tavtow «WM uMd to dw*top TtM
I dtf* In THito 7. Wlwra niOJMHY. 9» InMt tor raeoMiy Km* DMA brmtonMl to MM* DWioMty ol th* mM to
TABLE 7. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 625
"wmv,
Skigto
tncSan. «,'
Bmyl Dm* pMMM.....
MHC—
mt»-«HanMHA»t»
0.96C4O16
0.66C4O74
0.70C+1.69
040C4O66
0.99C-0.60
0.93C-1.60
047C-1J6
0.90C-0.13
0.96C-OJ6
0.66C-1.66
047C-0.64
04K-1.06
OJ6C-1.54
0.15X-0.12
044X-1.06
047X-146
041X-042
O.ISX+0.93
042X+0.43
0.19X+1.03
049X+2.40
0.16X+0.94
040X-0.56
0.34X+0.98
040X-0.66
041X-0.67
0.26X-OM
O43X41.13
047X-044
048X-046
046X4066
OJ6X4O40
042X4144
0.53X+
0.30X-
0.63X-O1
045X40.10
i
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations 161
TABLE 7. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 625—Continued
I Accuracy, as
! recovery. X' (jjg
i U
Single analyst
precision. V (Mg/
U
• Overall precision.
S' (
Bis<2-chloroethoxy)metnane I 1 12C -5.04
Bis(2-chtoroisopropyl)ether 1 03C--2.31
Bis<2-ethylhexyl)phthalate 0.84C -1.18
4-Bromophenyl phanyl ether 0.91C -1.34
2-Chloronaphthalene 089C + 0.01
4-Chlorophenyl pnenyl ether 0.91C-1-0.53
Chrysene 0.93C-1.00
4.4'-DOO 0.56C-0.40
4.4'-DDE 0.70C-0.54
4,4'-DOT 0.79C-3.28
DibenK>(a,h)anthracene < 0.68C + 4.72
Oi-n-butyl phlhalate I 0.59C + 0.71
1.2-Oichlorobenrene 080C + 0.28
1.3-Dichlorobenzene 0.86C-0.70
1,4-Dichlorobenjene 073C-1.47
3.3'-Dicnlorobenzidine 1.23C-12.65
Dieldrin O.B2C-0.16
Diethyl phthalate 0.43C + 1.00
Oimelhyl phthalate 0.20C + 1.03
2.4-Oinitrotoluene 0.92C-4.81
2.6-Oin«rotoluene 1.06C-3.60
Di-n-oetylphthalate 0.76C-0.79
Endosurfan aultate 0.39C+0.41
Endrin aldehyde 0.76C-3.86
Fluoranthene 0.81C + 1.10
Ruorene 0.90C-0.00
Heptachtor 0.87C-2.97
Heptachlor epoxkte 0.92C-1.87
Hexachlorobenzene 0.74C+0.66
Hexachlorobutadiene 0.71C-1.01
Hextchloroethane 0.73C-0.83
lndeno<1.2,3-cd)pyrene 0.78C-3.10
Isophorone 1.12C-M.41
Naphthalene 0.76C+1.58
Nitrobenzene 1.09C-3.05
N-Nitrosodi-n-propyfamine 1.12C-6.22
PCB-1260 0.81C-10.86
Phenanthrene 0.87C-0.06
Pyrene 0.84C-0.16
1.2,4-Trichloroben2ene 0.94C-0.79
4-Chkxo-3-methy1phenot 0.84C+0.35
2-Chlorophenol 0.78C + 0.29
2.4-Dichlorophenol 0.87C + 0.13
2.4-Dimethylphenol 0.71C+4.41
2.4-Dinitrophenol 0.81C-18.04
2-Methyl-4.6-dinitrophenol 1.04C-28.04
2-Nitrophenol 1.07C-1.15
4-Nitrophenol 0.61C-1.22
Pemachlorophenot 0.93C+1.99
Phenol 0.43C+1.26
2,4.6-Trichlorophenol 0.91C-0.18
0.18*+ 134
0.24X40.28
0.26X + 0.73
0.13X + 0.66
0.07X + 0.52
0.20X-0.94
0.28X + 0.13
0.29X-0.32
0.26X-1.17
0.42X^0.19
0.30X + 851
0.13X + 1.16
0.20X + 0.47
0.25X+068
0.24X + 0.23
0.28X-I-7.33
0.20X-0.16
0.28X+1 44
0.54X+0.19
0.12X+1.06
0.14X+1.26
0.21X + 1.19
0.12X+2.47
0.18X+3.91
0.22X-0.73
0.12X+0.26
0.24X-O.S6
0.33X-0.46
0.18X-0.10
0.19X+0.92
0.17X+0.67
0.29X+1.48
0.27X+0.77
0.21 X-0.41
0.19X+0.92
0.27X + 0.68
035X+3.61
0.12X + 0.57
0.16X+006
0.15X + 0.05
0.23X + 0.75
0.18*+1.46
0.15JJ+1.25
0.16X+1.21
0.38X + 2.36
0.1 OX+42.29
0.16X + 1.94
0.38X+2.57
0.24X + 3.03
0.26X + 0.73
0.16X + 2.J2
0.26Xi2.0l
0.25X *. 1 04
036X^06-
0.13X+0.34
0.30X-046
0.33^-009
0.66X-096
0.39X-1.04
0.65X-0.58
0.59X + 0.25
0.39X+0.6C
0.24X i 0.39
0.41^+0.11
0.29X+0.36
0.47)< + 3.45
0.26X-0.07
0.52^ + 0.22
1.05X-0.92
0.21 X+ 1.50
0.19X+0.35
0.37X + 1.19
0.63X-1.03
0.73X-0.62
0.28X-060
0.13X + 0.61
O.SOit-0.23
0.28^ + 0.64
0.43X-0.52
0.26X+0.49
0.17X + 0.80
0.50X + 0.44
0.33X + 0.26
0.30X-0.68
0.27X+0.21
0.44X + 0.47
0.43X+1.82
0.15X + 0.25
O.tSX + 0.31
0.21 X + 0.39
0.29X + 1.31
0.28X + 0.97
0.21 X+ 1.28
0.22X+1.31
0.42X + 26.29
0.26X + 23.10
0.27 j( + 2.60
0.44X + 3.24
0.30* + 4.33
035X + 0.58
0.22X+181
X' = Expected recovery for one or more measurements of a sample containing a concentration of C. in ua/L
s/=Expected single analyst standard deviation of measurements at an average concentration found of X, in jig/L
S= Expected intertaboratory standard deviation of measurements at an average concentration found of X. in fig/L.
C = True value for the concentration, in ug/L.
x=Average recovery found for measurements of samples containing a concentration of C, i
TABLE 8.—SUGGESTED INTERNAL AND
SURROGATE STANDARDS
Base/neutral fraction
Aniline-d,
Anthracene-dio
Benzo(a)anthracene-dn
4,4'-Dtbromobiphenyl
4,4'-
Dibromooctafluorobiphenyl.
Decafluorobiphenyl
2.2 '-Difluorobiphenyl
4-Fhjoroaniline
1 -Fluoronaphthylene
2-Fluoronaphthylene
Naphthatone-rJ,
Nitrobenzene-di
2.3.4.5,6-Pentatluorobipnenyl.
Phenanthrene-dio
Pyridine-d.
Acid fraction
2-Fluorophenol.
Pentaftuorophenol.
PhenoMk
2-Parfluoromethyt phenol.
TABLE 9.—DFTPP KEY MASSES AND
ABUNDANCE CRITERIA
Mass
51
68
70
127
197
198
199
275
365
441
442
443
m/2 Abundance criteria
30-60 percent of mass 198.
Less than 2 percent of mass 69.
Less than 2 percent of mass 69.
40-60 percent of mass 198.
Less than 1 percent of mass 198.
Base peak. 100 percent relative abundance.
5-9 percent of mass 198.
10-30 percent of mass 198.
Greater than 1 percent of mass 198.
Present but less than mass 443.
Greater than 40 percent of mass 198.
17-23 percent of mass 442.
BILLING COOT 6560-SO-U
-------
COLUMN: 3% SP-2260 OH SUKLCOPORT
PUKKAH: Mt FOR 4 MUL Iff• Mil TO 21TC
DETECTOR: MASS SPECTROMETER
fi
2.4-OINITMOTOLUiNE^ N-NITKOSO OIPHENVLAMINE
o
o
51
10
15
20 25 30
RETENTION TIME. MIN.
35
40
•45
Figure 1. Gas chromatogram of base/neutral fraction.
90
55"
u
01
a.
O
CO
-------
COLUMN: 1% SP-1240DA ON SUPaCOPOftT
PROGRAM: 70«C FOR 2 MW 8*C/M1N TO 200*C
DETECTOi: MASS SPECTROMETER.
8 10 12 14
RETENTION TIME. MIN .
16
18
20
Figure 2. Gas chromatogram of acid fraction .
JO
CD
o
a.
50
CD
o
CO
-------
COUJMN: 3% SP-22SO ON SUPELCOFORT
PROGRAM: 50«C FOR 4 MM. 8«C/MW TO 270«C
DETECTOR: MASS SPECTROMETER
£
o
g
9
d
5
o
9
1
CO
o
a.
o>
o
o
o
er
CD
10
IS
20
25
30
RETENTION TIME. MIN.
Figure 3. G«s chromatogram of pasticida fraction.
50
re"
ca
0)
a.
o
CO
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
165
COLUIM: 3ft 8P-2250 ON SUPELCOPORT
PfiOGEAM: SO*C FOR 4 Ml* 8*C/MIN TO 270*C
DETECTOR: MASS SPECTROMETER
Hi/z-35 TO 450
m/z-377
m/z-375
m/z-373
18
202224»2«303234
RETENTION TIME. MIN.
*
Figure 4. Gas chromatogram of chlordane.
-------
166
Federal Register / Vol. 49. No. 209 / Friday, October 28,1984 / Rules and Regulations
COLUMN: 3% SP-2250 ON SUP&COPORT
PROGRAM: SO*C FOR 4 Ml* 8*C/MM TO 270«C
DETECTOR: MASS SPECTROMETER
2224X3303294 MM
RETENTION TIME. MIN.
Figure 5. Gas chromatogram of toxaphene.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
167
COLUMN: 3* SP-2250 ON SUPELCOPORT
PROGRAM: 50°C FOR 4 MIN. 8°C/MIN TO 270*C
DETECTOR: MASS SPECTROMETER
30 32
18 20 22 24 26 28
RETENTION TIME. MIN.
Figure 6. Gas chromatograrh of PCB-1016.
-------
168
Federal Register / Vol. 49, No. 209 / Friday. October 26,1984 / Rules and Regulations
COLUMN: 3X SP-22SI ON SUPELCOKMT
PROGRAM: 50*C FOR 4Mm.lt/MrX TO 270*C
DETECTOR: MASS SPECTROMETER
10 20 22 24 20 2f 30 32
RETENTION TIME. MIN.
Figure 7. Gas chromatogram of PCB-1221
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26, 1984 / Rules and Regulations
169
COLUMN: 3% SP-2250 ON SUPELCOPORT
PROGRAM: 50°C FOR 4 MIN, 8°C/MIN TO 270°C
DETECTOR: MASS SPECTROMETER
n/z*35 TO 450
is 20
32
22 24 26 28 30
RETENTION TIME, MIN.
Figure 8. Gas chromatogram of PCB-1232.
-------
170
Federal Register / Vol. 49. No. 209 / Friday, October 28. 1984 / Rules and Regulations
COLUMN: 3% SP-2210 ON SUPaCOPORT
PROGRAM: 50*C FOR 4 Ml* 8«C/MIN TO 270«C
DETECTOR: MASS SPECTROMETER
m/z-35 TO 450
22 24 a a 30
RETENTION TIME. MIN
Figure 9. Gas chromatogVam of PCB-1242.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
171
COLUMN: 3X SP-2250 ON SUPELCOPORT
PROGRAM: 50*C FOR 4 MIN. 8°C/MIN TO 270°C
DETECTOR: MASS SPECTROMETER
*
RETENTION TIME.
Figure 10. Gas chromatogram of PCB-1248.
-------
172
Federal Register / Vol. 49, No. 209 / Friday, October 26,1984 / Rules and Regulations
COLUMN; » SM2M ON SUPaCOPOftT
NIQ8RAM: 60«C FOR 4 MM. I«C/MIN TO 270-C
DETECTOR: MASS SKCTMOMETI
24 M M 36 32
RETENTION TIME. MIN.
Figure 11. Ga« chromatogram of ^CB-1264.
-------
Federal Register / Vol. 49, No. 209 / Friday, October 28,1984 / Rules and Regulations
173
COLUMN: 3% SP-2250 ON SUPELCOPORT
PROGRAM: 50°C FOR 4 MIN. 8°C/MIN TO 270°C
DETECTOR: MASS SPECTROMETER
m/z=35 TO 450
m/z-354
m/i-362
m/z-330
18
20 22
24
32 -34
RETENTION TtME, MtN.
Figure 12. Gas chromatogram of PCB-1260.
-------
174
Federal Register / Vol. 49. No. 209 / Friday, October 26.1984 / Rules and Regulations
TAILING FACfOM =
calculatieo: Peak ttoifM « BE m^Mmm
WX Peak H^fM-IO-ttwui
Peak W*h M MX PMk rHifkt mAC* 23 1
Faclw « —
Figur* 13. Tailing factor calculation.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 175
Method 1624 Revision B—Volatile Organic
Compounds by Isotope Dilution GC/MS
1 Scope and application
1.1 This method is designed to determine
the volatile toxic organic pollutants
associated with the 1976 Consent Decree and
additional compounds amenable to purge and
trap gas chromatography-mass spectrometry
(GC/MS).
1.2 The chemical compounds listed in
table 1 may be determined in municipal and
industrial discharges by this method. The
method is designed to meet the survey
requirements of Effluent Guidelines Division
(BCD) and the National Pollutants Discharge
Elimination System (NPDES) under 40 CFR
136.1 and 136.5. Any modifications 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 136.4
and 136.5.
1.3 The detection limit of this method is
usually dependent on the level of
interferences rather than instrumental
limitations. The limits in table 2 represent the
minimum quantity that can be detected with
no interferences present.
1.4 The GC/MS portions of this method
are for use only by analysts experienced with
GC/MS or under the close supervision of
such qualified persons. Laboratories
unfamiliar with the analyses of
environmental samples by GC/MS should run
the performance tests in reference 1 before
beginning.
2 Summary of method
2.1 Stable isotopically labeled analogs of
the compounds of interest are added to a 5
mL water sample. The sample is purged at
20-25 *C with an inert gas in a specially
designed chamber. The volatile organic
compounds are transferred from the aqueous
phase into the gaseous phase where they are
passed into a sorbent column and trapped.
After purging is completed, the trap is
backflushed and heated rapidly to desorb the
compounds into a gas chromatograph (GC).
The compounds are separated by the GC and
detected by a mass spectrometer (MS)
(references 2 and 3). The labeled compounds
serve to correct the variability of the
analytical technique.
2.2 Identification of a compound
(qualitative analysis) is performed by
comparing the GC retention time and the
background corrected characteristic spectral
masses with those of authentic standards.
2.3 Quantitative analysis is performed by
GC/MS using extracted ion current profile
(EICP) areas. Isotope dilution is used when
labeled compounds are available; otherwise,
an internal or external standard method is
used.
2.4 Quality is assured through
reproducible calibration and testing of the
purge and trap and GC/MS systems.
3 Contamination and interferences
3.1 Impurities in the purge gas, organic
compounds out-gassing from the plumbing
upstream of the trap, and solvent vapors in
the laboratory account for the majority of
contamination problems. The analytical
system is demonstrated to be free from
interferences under conditions of the analysis
by analyzing blanks initially and with each
sample lot (samples analyzed on the same 8
hr shift), as described in section 8.5.
3.2 Samples can be contaminated by
diffusion of volatile organic compounds
(particularly methylene chloride) through the
bottle seal during shipment and storage. A
field blank prepared from reagent water and
carried through the sampling and handling
protocol serves as a check on such
contamination.
3.3 Contamination by carry-over can
occur when high level and low level samples
are analyzed sequentially. To reduce carry-
over, the purging device and sample syringe
are rinsed between samples with reagent
water. When an unusually concentrated
sample is encountered, it is followed by
analysis of a reagent water blank to check for
carry-over. For samples containing large
amounts of water soluble materials.
suspended solids, high boiling compounds, or
high levels or purgeable compounds, the
purge device is washed with soap solution,
rinsed with tap and distilled water, and dried
in an oven at 100-125 *C. The trap and other
parts of the system are also subject to
contamination: therefore, frequent bakeout
and purging of the entire system may be
required.
3.4 Interferences resulting from samples
will vary considerably from source to source,
depending on the diversity of the industrial
complex or municipality being sampled.
4 Safety
4.1 The toxicity or carcinogenicity of each
compound or reagent used in this method has
not been precisely determined; however, each
chemical compound should be treated as a
potential health hazard. Exposure to these
compounds should be reduced to the lowest
possible level. 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 data handling
sheets should also be made available to all
personnel involved in these analyses.
Additional information on laboratory safety
can be found in references 4-6.
4.2 The following compounds covered by
this method have been tentatively classified
as known or suspected human or mammalian
carcinogens: benzene, carbon tetrachloride,
chloroform, and vinyl chloride. Primary
standards of these toxic compounds should
be prepared in a hood, and a NIOSH/MESA
approved toxic gas respirator should be worn
when high concentrations are handled.
5 Apparatus and materials
5.1 Sample bottles for discrete sampling
5.1.1 Bottle—25 to 40 mL with screw cap
(Pierce 13075, or equivalent). Detergent wash,
rinse with tap and distilled water, and dry at
>105 *C for one hr minimum before use.
5.1.2 Septum—Teflon-faced silicone
(Pierce 12722, or equivalent), cleaned as
above and baked at 100-200 *C, for one hour
minimum.
5.2 Purge and trap device—consists of
purging device, trap, and desorber. Complete
devices are commercially available.
5.2.1 Purging device—designed to accept 5
mL samples with water column at least 3 cm
deep. The volume of the gaseous head space
between the water and trap shall be less than
15 mL. The purge gas shall be introduced less
than 5 mm from the base of the water column
and shall pass through the water as bubbles
with a diameter less than 3 mm. The purging
device shown in figure 1 meets these criteria.
5.2.2 Trap—25 to 30 cm x 2.5 mm i.d.
minimum, containing the following:
5.2.2.1 Methyl silicone packing—one ±
0.2 cm. 3 percent OV-1 on 60/80 mesh
Chromosorb W, or equivalent.
5.2.2.2 Porous polymer—15 ± 1.0 cm.
Tenax GC (2,6-diphenylene oxide polymer),
60/80 mesh, chromatographic grade, or
equivalent.
5.2.2.3 Silica gel—8 ± 1.0 cm. Davison
Chemical, 35/60 mesh, grade 15. or
equivalent. The trap shown in figure 2 meets
these specifications.
5.2.3 Desorber—shall heat the trap to 175
± 5 'C in 45 seconds or less. The polymer
section of the trap shall not exceed 180 'C,
and the remaining sections shall not exceed
220 "C. The desorber shown in figure 2 meets
these specifications.
5.2.4 The purge and trap device may be a
separate unit or coupled to a GC as shown in
figures 3 and 4.
5.3 Gas chromatograph—shall be linearly
temperature programmable with initial and
final holds, shall contain a glass jet separator
as the MS interface, and shall produce results
which meet the calibration (section 7), quality
assurance (section 8), and performance tests
(section 11) of this method.
5.3.1 Column—2.8 ± 0.4 m x 2 ± 0.5 mm i.
d. glass, packekd with one percent SP-1000
on Carbopak B, 60/80 mesh, or equivalent.
5.4 Mass spectrometer—70 eV electron
impact ionization; shall repetitively scan from
20 to 250 amu every 2-3 seconds, and produce
a unit resolution (valleys between m/z 174-
176 less than 10 percent of the height of the
m/z 175 peak), background corrected mass
spectrum from 50 ng 4-bromo-fluorobenzene
(BFB) injected into the GC. The BFB spectrum
shall meet the mass-intensity criteria in table
3. All portions of the GC column, transfer
lines, and separator which connect the GC
column to the ion source shall remain at or
above the column temperature during
analysis to preclude condensation of less
volatile compounds.
5.5 Data system—shall collect and record
MS data, store mass intensity data in spectral
libraries, process GC/MS data and generate
reports, and shall calculate and record
response factors.
5.5.1 Data acquisition—mass spectra shall
be collected continuously throughout the
analysis and stored on a mass storage device.
5.5.2 Mass spectral libraries—user
created libraries containing mass spectra
obtained from analysis of authentic
standards shall be employed to reverse
search GC/MS runs for the compounds of
interest (section 7.2).
5.5.3 Data processing—the data system
shall be used to search, locate, identify, and
quantify the compounds of interest in each
GC/MS analysis. Software routines shall be
employed to compute retention times and
EICP areas. Displays of spectra, mass
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178 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
chromatograma. and library comparisons are
required (o verify results.
5.5.4 Response factors and multipoint
calibrations—the data system shall be used
to record and maintain lists of response
factors (response ratios for isotope dilution)
and generate multi-point calibration curves
(section 7). Computations of relative standard
deviation (coefficient of variation) are useful
for testing calibration linearity. Statistics on
initial and on-going performance shall be
maintained (sections a and 11).
5.6 Syringes—5 mL glass hypodermic,
with Luer-lok tips.
5.7 Micro syringes—10, 25. and 100 uL
5.8 Syringe valves—2-way, with Luer
ends (Telfon or Kel-F).
5.9 Syringe—5 mL gas-tight, with shut-off
valve.
5.10 Bottles—15 mL. screw-cap with
Telfon liner.
5.11 Balance—analytical, capable of
weighing 0.1 mg.
6 Reagents and standards
6.1 Reagent water—water in which the
compounds of interest and interfering
compounds are not detected by this method
(section 11.7). It may be generated by any of
the following methods:
6.1.1 Activated carbon—pass tap water
through a carbon bed (Calgon Filtrasorb-300,
or equivalent).
6.1.2 Water purifier—pass tap water
through a purifier (Millipore Super Q, or
equivalent).
6.1.3 Boil and purge—heat tap water to
90-100 *C and bubble contaminant free inert
gas through it for approx one hour. While still
hot. transfer the water to screw-cap bottles
and seal with a Teflon-lined cap.
&2 Sodium tniosulfate—ACS granular.
64 Methanol—pesticide quality or
equivalent
6.4 Standard solutions—purchased as
solution or mixtures with certification to their
purity, concentration, and authenticity, or
prepared from materials of known purity and
composition. If compound purity is 96 percent
or greater, the weight may be used without
correction to calculate the concentration of
the standard.
6.5 Preparation of stock solutions—
prepare in methanol using liquid or gaseous
standards per the steps below. Observe the
safety precautions given in section 4.
6.5.1 Place approx 9.8 mL of methanol in a
10 mL ground glass stoppered volumetric
flask. Allow the flask to stand unstoppered
for approximately 10 minutes or until all
methanol wetted surfaces have dried In each
case, weigh the flask, immediately add the
compound, then immediately reweigh to
prevent evaporation losses from affecting the
measurement.
6.5.1.1 Liquids—using a 100 pL syringe,
permit 2 drops of liquid to fall into the
methanol without contacting the neck of the
flask. Alternatively, inject a known volume of
the compound into the methanol in the flask
using a micro-syringe.
64.1.2 Cases (chloromethane,
bromomethane. chloroethane. vinyl
chloride)—fill a valved 5 mL gas-tight syringe
with the compound. Lower the needle to
approx 5 mm above the methanol meniscus.
Slowly introduce the compound above the
surface of the meniscus. The gas will dissolve
rapidly in the methanol.
6.5.2 Fill the flask to volume, stopper, then
mix by inverting several times. Calculate the
concentration in mg/mL (jig/jtL ) from the
weight gain (or density if a known volume
was injected).
6.5.3 Transfer the stock solution to a
Teflon sealed screw-cap-bottle. Store, with
minimal headspace. in the dark at -10 to
-20'C.
6.5.4 Prepare fresh standards weekly for
the gases and 2-chloroethylvinyl ether. All
other standards are replaced after one month,
or sooner if comparison with check standards
indicate a change in concentration. Quality
control check standards that can be used to
determine the accuracy of calibration
standards are available from the US
Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio.
6.6 Labeled compound spiking solution—
from stock standard solutions prepared as
above, or from mixtures, prepare the spiking
solution to contain a concentration such that
a 5-10 pL spike into each 5 mL sample, blank.
or aqueous standard analyzed will result in a
concentration of 20 pg/L of each labeled
compound. For the gases and for the water
soluble compounds (acrolein, acrylonitrile.
acetone, diethyl ether, and MEK), a
concentration of 100 pg/L may be used.
Include the internal standards (section 7.5) in
this solution so that a concentration of 20 fig/
L in each sample, blank, or aqueous standard
will be produced.
6.7 Secondary standards—using stock
solutions, prepare a secondary standard in
methanol to contain each pollutant at a
concentration of 800 pg/mL For the gases and
water soluble compounds (section 6.6), a
concentration of 2.5 mg/mL may be used.
6.7.1 Aqueous calibration standards-
using a 25 pL syringe, add 20 jiL of the
secondary standard (section 6.7) to 50.100,
200.500, and 1000 mL of reagent water to
produce concentrations of 200,100,50,20,
and 10 /ig/L respectively. If the higher
concentration standard for the gases and
water soluble compounds was chosen
(section 6.6), these compounds will be at
cocentrations of 1000,500.250, loo, and 50
Mg/L in the aqueous calibration standards.
6.7.2 Aqueous performance standard—an
aqueous standard containing all pollutants,
internal standards, labeled compounds, and
BFB is prepared daily, and analyzed each
shift to demonstrate performance (section 11).
This standard shall contain either 20 or 100
ftg/L of the labeled and pollutant gases and
water soluble compounds. 10 pg/L BFB, and
20 jtg/L of all other pollutants, labeled
compounds, and internal standards. It may be
the nominal 20 ftg/L aqueous calibration
standard (section 6.7.1).
6.7.3 A methanolic standard containing
all pollutants and internal standards is
prepared to demonstrate recovery of these
compounds when syringe injection and purge
and trap analyses an compared. This
standard shall contain either 100 Mg/mL or
500 |ig/mL of the gases and water soluble
compounds, and 100 Mg/mL of the remaining
pollutants and internal standards (consistent
with the amounts in the aqueous performance
standard in 6.7.2).
6.7.4 Othe standards which may be
needed are those for test of BFB performance
(section 7.1) and for collection of mass
spectra for storage in spectral libraries
(section 7.2).
7 Calibration
7.1 Assemble the gas chromatographic
apparatus and establish operating conditions
given in table 2. By injecting standards into
the GC. demonstrate that the analytical
system meets the detection limits in table 2
and the mass-intensity criteria in table 3 for
SO ng BFB.
7.2 Mass spectral libraries—detection and
identification of the compound of interest are
dependent upon the spectra stored in user
created libraries.
7.2.1 Obtain a mass spectrumof each
pollutant and labeled compound and each
internal standard by analyzing an authentic
standard either singly or as part of a mixture
in which there is no interference between
closely eluted components. That only a single
compound is present is determined by
examination of the spectrum. Fragments not
attributable to the compound under study
indicate the presence of an interfering
compound. Adjust the analytical conditions
and scan rate (for this test only) to produce
an undistorted spectrum at the GC peak
maximum. An undistorted spectrum will
usually be obtained if five complete spectra
are collected across the upper half of the GC
peak. Software algorithms designed to
"enhance" the spectrum may eliminate
distortion, but may also eliminate authentic
m/z's or introduce other distortion.
7.2.3 The authentic reference spectrum is
obtained under BFB tuning conditions
(section 7.1 and table 3) to normalize it to
spectra from other instruments.
7.2.4 The spectrum to edited by saving the
5 most intense mass spectral peaks and all
other mass spectral peaks greater than 10
percent of the base peak. lUs spectrum to
stored for reverse search and for compound
confirmation.
7.3 Assemble the purge and trap device.
Pack the trap as shown in figure 2 and
condition overnight at 170-180 *C by
backflushing with an inert gas at a flow rate
of 20-30 mL/min. Condition traps daily for a
minimum of 10 minutes prior to use.
7.3.1 Analyze the aqueous performance
standard (section 6.7.2) according to the
purge and trap procedure in section 10.
Compute the ana at the primary m/i (table
4) for each compound. Compare these areas
to those obtained by Injecting one pL of the
methanolic standard (section 6.7.3) to
determine compound recovery. The recovery
shall be greater than 20 percent for the water
soluble compound*, and 60-110 percent for
all other compounds. This recovery to
demonstrated initially for each purge and
trap GC/MS system. The test to repeated only
if the purge and trap or GC/MS systems are
modified in any way that might result in a
change in recovery.
7.3.2 Demonstrate that 100 ng toluene (or
toluene-d8) produces an area at m/z 91 (or
99] approx one-tenth that required to exceed
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 177
the linear range of the system. The exact
value must be determined by experience for
each instrument. It is used to match the
calibration range of the instrument to the
analytical range and detection limits
required.
7.4 . Calibration by isotope dilution— the
isotope dilution approach is used for the
purgeable organic compounds when
appropriate labeled compounds are available
and when interferences do not preclude the
analysis. If labeled compounds are not
available, or interferences are present,
internal or external standard methods
(section 7.5 or 7.6) are used. A calibration
curve encompassing the concentration range
of interest is prepared for each compound
determined. The relative response (RR) vs
concentration (fig/L) is plotted or computed
using a linear regression. An example of a
calibration curve for toluene using toluene-d8
is given in figure 5. Also shown are the ±10
percent error limits (dotted lines). Relative
response is determined according to the
procedures described below. A minimun of
five data points are required for calibration
(section 7.4.4).
7.4.1 The relative response (RR) of
pollutant to labeled compound is determined
from isotope ratio values calculated from
acquired data. Three isotope ratios are used
in this process:
R,=the isotope ratio measured in the pure
pollutant (figure 6A).
R,=the isotope ratio of pure labeled
compound (figure 6B).
Rm=the isotope ratio measured in the
analytical mixture of the pollutant and
labeled compounds (figure 6C).
The correct way to calculate RR is:
RR=(R,-R.,) (R,+l)/fRn-RJ(R,+l) If R=,
is not between 2R, and 0.5R,, the method
does not apply and the sample is analyzed by
internal or external standard methods
(section 7.5 or 7.6).
7.4.2 In most cases, the retention times of
the pollutant and labeled compound are the
same and isotope ratios (R's) can be
calculated from the EICP areas, where:
R=(area at mi/z)/(area at nh/z) If either of
the areas is zero, it is assigned a value of one
in the calculations; that is, if: area of m>/
2=50721. and area of m»/z=0, then
R= 50721 /I =50720. The m/z's are always
selected such that R,>R,. When there is a
difference in retention times (RT) between
the pollutant and labeled compounds, special
precautions are required to determine the
isotope ratios.
RI, RT, and Rm are defined as follows:
R,=l/[area nh/z (at RTi)]
Rm=[area mi/z (at RTi)]/[area nh/z (at
RT,)]
7.4.3 An example of the above
calculations can be taken from the data
plotted in figure 5 for toluene and toluene-dB.
For these data. R,= 168920/1 =168900. R,=l/
60960=0.00001040, and R_=98868/
82508=1.174. The RR for the above data is
then calculated using the equation given in
section 7.4.1. For the example, RR= 1.174.
Note: Not all labeled compounds elute before
their pollutant analogs.
7.4.4 To calibrate the analytical system by
isotope dilution, analyze a 5 mL aliquot of
each of the aqueous calibration standards
(section 6.7.1) spiked with an appropriate
constant amount of the labeled compound
spiking solution (section 6.6). using the purge
and trap procedure in section 10. Compute
the RR at each concentration.
7.4.5 Linearity—if the ratio of relative
response to concentration for any compound
is constant (less than 20 percent coefficient of
variation) over the 5 point calibration range,
an averaged relative response/concentration
ratio may be used for that compound:
otherwise, the complete calibration curve for
that compound shall be used over the 5 point
calibration range.
7.5 Calibration by internal standard—
used when criteria for isotope dilution
(section 7.4) cannot be met. The method is
applied to pollutants having no labeled
analog and to the labeled compounds. The
internal standards used for volatiles analyses
are bromochloromethane, 2-bromo-l-
chloropropane, and 1.4-dichlorobutane.
Concentrations of the labeled compounds
and pollutants without labeled analogs are
computed relative to the nearest eluted
internal standard, as shown in table 2.
7.5.1 Response factors—calibration
requires the determination of response
factors (RF) which are defined by the
following equation: RF=(A.xC,.)/(AtoxC,),
where A, is the EICP area at the
characteristic m/z for the compound in the
daily standard. Ata is the EICP area at the
characteristic m/z for the internal standard.
Cu is the concentration (ug/L) of the
internal standard
C, is the concentration of the pollutant in
the daily standard.
7.5.2 The response factor is determined at
10, 20. 50,100, and 200 ug/L for the pollutants
(optionally at five times these concentrations
for gases and water soluble pollutants—see
section 6.7), in a way analogous to that for
calibration by isotope dilution (section 7.4.4).
The RF is plotted against concentration for
each compound in the standard (C,) to
produce a calibration curve.
7.5.3 Linearity—if the response factor (RF)
for any compound is constant (less than 35
percent coefficient of variation) over the 5
point calibration range, an averaged response
factor may be used for that compound;
otherwise, the complete calibration curve for
that compound shall be used over the 5 point
range.
7.8 Combined calibration—by adding the
isotopically labeled compounds and internal
standards (section 6.6) to the aqueous
calibration standards (section 8.7.1), a single
set of analyses can be used to produce
calibration curves for the isotope dilution and
internal standard methods. These curves are
verified each shift (section 11.5) by purging
the aqueous performance standard (section
6.7.2). Recalibration is required only if
calibration and on-going performance
(section 11.5) criteria cannot be met.
8 Quality assurance/quality control
8.1 Each laboratory that uses this method
is required to operate a formal quality
assurance program. The minimum
requirements of this program consist of an
initial demonstration of laboratory capability,
analysis of samples spiked with labeled
compounds to evaluate and document data
quality, and analysis of standards and blanks
as tests of continued performance.
Laboratory performance is compared to
established performance criteria to determine
if the results of analyses meet the
performance characteristics of the method.
8.1.1 The analyst shall make an initial
demonstration of the ability to generate
acceptable accuracy and precision with this
method. This ability is established as
described in section 8.2.
8.1.2 The analyst is permitted to modify
this method to improve separations or lower
the costs of measurements, provided all
performance specifications are met. Each
time a modification is made to the method,
the analyst is required to repeat the
procedure in section 8.2 to demonstrate
method performance.
8.1.3 Analyses of blanks are required to
demonstrate freedom from contamination
and that the compounds of interest and
interfering compounds have not been carried
over from a previous analysis (section 3). The
procedures and criteria for analysis of a
blank are described in sections 8.5 and 11.7.
8.1.4 The laboratory shall spike all
samples with labeled compounds to monitor
method performance. This test is described in
section 8.3. When results of these spikes
indicate atypical method performance for
samples, the samples are diluted to bring
method performance within acceptable limits
(section 14.2).
8.1.5 The laboratory shall, on an on-going
basis, demonstrate through the analysis of
the aqueous performance standard (section
6.7.2) that the analysis system is in control.
This procedure is described in sections 11.1
and 11.5.
8.1.6 The laboratory shall maintain
records to define the quality of data that is
generated. Development of accuracy
statements is described in sections 8.4 and
11.5.2.
8.2 Initial precision and accuracy—to
establish the ability to generate acceptable
precision and accuracy, the analyst shall
perform the following operations:
8.2.1 Analyze two sets of four 5-mL
aliquots (8 aliquots total) of the aqueous
performance standard (section 6.7.2)
according to the method beginning in section
10.
8.2.2 Using results of the first set of four
analyses in section 8.2.1, compute the average
recovery (X) in ug/L and the standard
deviation of the recovery (s) in ug/L for each
compound, by isotope dilution for polluitants
with a labeled analog, and by internal
standard for labeled compounds and
pollutants with no labeled analog.
8.2.3 For each compound, compare s and
X with the corresponding limits for initial
precision and accuracy found in table 5. If s
and X for all compounds meet the acceptance
criteria, system performance is acceptable
and analysis of blanks and samples may
begin. If individual X falls outside the range
for accuracy, system performance is
unacceptable for that compound. NOTE: The
large number of compounds in table 5 present
a substantial probability that one or more
will fail one of the acceptance criteria when
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Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
all compounds are analyzed. To determine if
the analytical system is out of control, or if
the failure can be attributed to probability.
proceed as follows:
8.2.4 Using the results of the second set of
four analyses, compute s and X for only those
compounds which failed the test of the first
set of four analyses (section 8.2.3). If these
compounds now pass, system performance is
acceptable for all compounds and analysis of
blanks and samples may begin. If, however,
any of the same compounds fail again, the
analysis system is not performing properly
for the compound(s) in question. In this event,
correct the problem and repeat the entire test
(section 8.2.1).
8.3 The laboratory shall spike all samples
with labeled compounds to assess method
performance on the sample matrix.
8.3.1 Spike and analyze each sample
according to the method beginning in section
10.
8.3.2 Compute the percent recovery (P) of
the labeled compounds using the internal
standard method (section 7.5).
8.3.3 Compare the percent recovery for
each compound with the corresponding
labeled compound recovery limit in table 5. If
the recovery of any compound falls outside
its warning limit, method performance is
unacceptable for that compound in that
sample. Therefore, the sample matrix is
complex and the sample is to be diluted and
reanalyzed, per section 14.2.
8.4 As part of the QA program for the
laboratory, method accuracy for wastewater
sample* shall be assessed and records shall
be maintained. After the analysis of five
wastewater samples for which the labeled
compounds pass the tests in section 8.3.3,
compute the average percent recovery (P) and
the standard deviation of the percent
recovery (s,) for the labeled compounds only.
Express the accuracy assessment as a
percent recovery interval from P—2s, to
P+2Sp. For example, if P-90% and s,
the accuracy interval is expressed as 70-
110%. Update the accuracy assessment for
each compound on a regular basis (e.g. after
each 5-10 new accuracy measurements).
8.5 Blanks—reagent water blanks are
analyzed to demonstrate freedom from carry-
over (section 3) and contamination.
8.5.1 The level at which the purge and
trap system will carry greater than 5 ug/L of
a pollutant of interest (table 1) into a
succeeding blank shall be determined by
analyzing successively larger concentrations
of these compounds. When a sample contains
this concentration or more, a blank shall be
analyzed immediately following this sample
to demonstrate no carry-over at the 5 pg/L
level.
M.2 With each sample lot (samples
analysed on the same 8 hr shift), a blank
shall be analyzed immediately after analysis
of the aqueous performance standard (section
11.1) to demonstrate freedom from
contamination. If any of the compounds of
interest (table 1) or any potentially interfering
compound i* found in a blank at greater than
10 pf/L (assuming a response factor of 1
relative to the nearest eluted internal
standard for compounds not listed in table 1),
analysis of samples 1* halted until the source
of contamination is eliminated and a blank
shows no evidence of contamination at this
level.
8.8 The specifications contained in this
method can be met if the apparatus used is
calibrated properly, then maintained in a
calibrated state.
The standards used for calibration (section
7), calibration verification (section 11.5) and
for initial (section 8.2) and on-going (section
11.5) precision and accuracy should be
identical, so that the most precise results will
be obtained. The GC/MS instrument in
particular will provide the most reproducible
results if dedicated to the settings and
conditions required for the analyses of
volatile* by this method.
8.7 Depending on specific program
requirements, field replicates may be
collected to determine the precision of the
sampling technique, and spiked samples may
be required to determine the accuracy of the
analysis when internal or external standard
methods are used.
9 Sample collection, preservation, and
handling
9.1 Grab samples are collected in glass
containers having a total volume greater than
20 mL. Fill sample bottles so that no air
bubbles pass through the sample as the bottle
is filled. Seal each bottle so that no air
bubbles are entrapped. Maintain the hermetic
seal on the sample bottle until time of
analysis.
9.2 Samples are maintained at 0-4 *C
from the time of collection until analysis. If
the sample contains residual chlorine, add
sodium thiosulfate preservative (10 mg/40
mL) to the empty sample bottles just prior to
shipment to the sample site. EPA Methods
330.4 and 330.5 may be used for measurement
of residual chlorine (reference 8). If
preservative has been added, shake bottle
vigorously for one minute Immediately after
filling.
9.3 Experimental evidence indicates that
some aromatic compounds, notably benzene,
toluene, and ethyl benzene are susceptible to
rapid biological degradation under certain
environmental conditions. Refrigeration alone
may not be adequate to preserve these
compounds in wastewaters for more than
seven days. For this reason, a separate
sample should be collected, acidified, and
analyzed when these aromatics are to be
determined. Collect about 500 mL of sample
in a clean containers.
Adjust the pH of the sample to about 2 by
adding HO (1+1) while stirring. Check pH
with narrow range (1.4 to 2.8) pH paper. Fill a
sample container as described in section 9.1.
If residual chlorine is present, add sodium
thiosulfate to a separate sample container
and fill as in section 9.1.
9.4 All samples shall be analyzed within
14 days of collection.
10 Purge, trap, and GC/MS analysis
10.1 Remove standards and samples from
cold storage and bring to 20-25 '.
10.2 Adjust the purge gas flow rate to 40
±4 mL/min. Attach the trap inlet to the
purging device and set the valve to the purge
mode (figure 3). Open the syringe valve
located on the purging device sample
introduction needle (figure 1).
10.3 Remove the plunger from a 5-mL
syringe and attach a closed syringe valve.
Open the sample bottle and carefully pour
the sample into the syringe barrel until it
overflows. Replace the plunger and compress
the sample. Open the syringe valve and vent
any residual air while adjusting the sample
volume to 5.0 mL Because this process of
taking an aliquot destroys the validity of the
sample for future analysis, fill a second
syringe at this time to protect against
possible loss of data. Add an appropriate
amount of the labeled compound spiking
solution (section 8.8) through the valve bore,
then close the valve.
10.4 Attach the syringe valve assembly to
the syringe valve on the purging device. Open
both syringe valves and inject the sample
into the purging chamber.
10.5 Close both valves and purge the
sample for 11.0 ± 0.1 minutes at 20-25 *C.
10.6 After the 11 minute purge time,
attach the trap to the chromatograph and set
the purge and trap apparatus to the desorb
mode (figure 4). Desorb the trapped
compounds into the GC column by heating
the trap to 170-180 *C while backflushing
with carrier gas at 20-60 mL/min for four
minutes. Start MS data acquisition upon start
of the desorb cycle, and start the GC column
temperature program 3 minutes later. Table 1
summarizes the recommended operating
conditions for the gas chromatograph.
Included in this table are retention times and
detection limits that were achieved under
these conditions. An example of the
separations achieved by the column listed is
shown in figure 5. Other columns may be
used provided the requirements in section 8
can be met If the priority pollutant gases
produce GC peak* so broad that the precision
and recovery specifications (section 8£)
cannot be met, the column may be cooled to
ambient or sub-ambient temperatures to
sharpen these peaks.
10.7 While analysis of the desorbed
compounds proceeds, empty the purging
chamber using the sample introduction
syringe. Wash the chamber with two 5-mL
portions of reagent water. After the purging
device has been emptied, allow the purge gas
to vent through the chamber until the frit is
dry, so that it is ready for the next sample.
10.8 After desorbing the sample for four
minutes, recondition the trap by returning 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 170-180 *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 i* ready for the next sample.
11 System performance
11.1 At the beginning of each 8 hr shift
during which analyses are performed, system
calibration and performance shall be verified
for all pollutants and labeled compounds. For
these testa, analysis of the aqueous
performance standard (section 6.7.2) shall be
used to verify aU performance criteria.
Adjustment and/or ^calibration (per section
7) shall be performed until all performance
criteria are met. Only after all performance
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 179
criteria are met may blanks and samples be
analyzed.
11.2 BFB spectrum validity—the criteria in
table 3 shall be met.
11.3 Retention times—the absolute
retention times of all compounds shall
approximate those given in table 2.
11.4 GC resolution—the valley height
between toluene and to!uene-d8 (at m/z 91
and 99 plotted on the same graph) shall be
less than 10 percent of the taller of the two
peaks.
11.5 Calibration verification and on-going
precision and accuracy—compute the
concentration of each polutant (table 1) by
isotope dilution (section 7.4) for those
compounds which have labeled analogs.
Compute the concentration of each pollutant
(table 1) which has no labeled analog by the
internal standard method (section 7.5).
Compute the concentration of the labeled
compounds by the internal standard method.
These concentrations are computed based on
the calibration data determined in section 7.
11.5.1 For each pollutant and labeled
compound, compare the concentration with
the corresponding limit for on-going accuracy
in table 5. If all compounds meet the
acceptance criteria, system performance is
acceptable and analysis of blanks and
samples may continue. If any individual
value falls outside the range given, system
performance is unacceptable for that
compound. NOTE: The large number of
compounds in table 5 present a substantial
probability that one or more will fail the
acceptance criteria when all compounds are
analyzed. To determine if the analytical
system is out of control, or if the failure may
be attributed to probability, proceed as
follows:
11.5.1.1 Analyze a second aliquot of the
aqueous performance standard (section 6.7.2).
11.5.1.2 Compute the concentration for
only those compounds which failed the first
test (section 11.5.1). If these compounds now
pass, system performance is acceptable for
all compounds and analyses of blanks and
samples may proceed. If, however, any of the
compounds fail again, the measurement
system is not performing properly for these
compounds. In this event, locate and correct
the problem or recalibrate the system
(section 7), and repeat the entire test (section
11.1) for all compounds.
11.5.2 Add results which pass the
specification in 11.5.1.2 to initial (section 8.2)
and previous on-going data. Update QC
charts to form a graphic representation of
laboratory performance (Figure 7). Develop a
statement of accuracy for each pollutant and
labeled compound by calculating the average
percentage recovery (R) and the standard
deviation of percent recovery (sr). Express
the accuracy as a recovery interval from
R-2sr to R+2sn For example, if R=95% and
s,=5%, the accuracy is 85-105 percent.
12 Qualitative determination—
accomplished by comparison of data from
analysis of a sample or blank with data from
analysis of the shift standard (section 11.1).
Identification is confirmed when spectra and
retention times agree per the criteria below.
12.1 Labeled compounds and pollutants
having no labeled analog:
12.1.1 The signals for all characteristic
masses stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
12.1.2 Either (1) the background corrected
EICP areas, or (2) the corrected relative
intensities of the mass spectral peaks at the
CC peak maximum shall agree within a
factor of two (0.5 to 2 times) for all masses
stored in the library.
12.1.3 The retention time relative to the
nearest eluted internal standard shall be
within ±7 scans or ±20 seconds, whichever
is greater of this difference in the shift
standard (section 11.1).
12.2 Pollutants having a labeled analog:
12.2.1 The signals for all characteristic
masses stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
12.2.2 Either (1) the background corrected
EICP areas, or (2) the corrected relative
intensities of the mass spectral peaks at the
GC peak maximum shall agree within a
factor of two for all masses stored in the
spectral library.
12.2.3 The retention time difference
between the pollutant and its labeled analog
shall agree within ±2 scans or ±6 seconds
(whichever is greater) of this difference in the
shift standard (section 11.1).
12.3 Masses present in the experimental
mass spectrum that are not present in the
reference mass spectrum shall be accounted
for by contaminant or background ions. If the
experimental mass spectrum is contaminated,
an experienced spectrometrist (section 1.4) is
to determine the presence or absence of the
compound.
13 Quantitative determination
13.1 Isotope dilution—by adding a known
amount of a labeled compound to every
sample prior to purging, correction for
recovery of the pollutant can be made
because the pollutant and its labeled analog
exhibit the same effects upon purging,
desorption, and gas chromatography. Relative
response (RR) values for sample mixtures are
used in conjunction with calibration curves
described in section 7.4 to determine
concentrations directly, so long as labeled
compound spiking levels are constant. For the
toluene example given in figure 6 (section
7.4.3), RR would be equal to 1.174. For this RR
value, the toluene calibration curve given in
figure 5 indicates a concentration of 31.8 fig/
L
13.2 Internal standard—calculate the
concentration using the response factor
determined from calibration data (section 7.5)
and the following equation:
Concentration =(A, x Ci,)/(Au x RF)
where the terms are as defined in section
7.5.1.
13.3 If the EICP area at the quantitation
mass for any compound exceeds the
calibration range of the system, the sample is
diluted by successive factors of 10 and these
dilutions are analyzed until the area is within
the calibration range.
13.4 Report results for all pollutants and
labeled compounds (table 1) found in all
standards, blanks, and samples, in u.g/L to
three significant figures. Results for samples
which have been diluted are reported at the
least dilute level at which the area at the
quantitation mass is within the calibration
range (section 13.3) and the labeled
compound recovery is within the normal
range for the Method (section 14.2).
14 Analysis of complex samples
14.1 Untreated effluents and other
samples frequently contain high levels
(> 1000 ftg/L) of the compounds of interest
and of interfering compounds. Some samples
will foam excessively when purged: others
will overload the trap/or GC column.
14.2 Dilute 0.5 mL of sample with 4.5 mL
of reagent water and analyze this diluted
sample when labeled compound recovery is
outside the range given in table 5. If the
recovery remains outside of the range for this
diluted sample, the aqueous performance
standard shall be analyzed (section 11) and
calibration verified (section 11.5). If the
recovery for the labeled compound in the
aqueous performance standard is outside the
range given in table 5, the analytical system
is out of control. In this case, the instrument
shall be repaired, the performance
specifications in section 11 shall be met, and
the analysis of the undiluted sample shall be
repeated. If the recovery for the aqueous
performance standard is within the range
given in table 5, the method does not work on
the sample being analyzed and the result may
not be reported for regulatory compliance
purposes.
14.3 Reverse search computer programs
can misinterpret the spectrum of
chromatographically unresolved pollutant
and labeled compound pairs with overlapping
spectra when a high level of the pollutant is
present. Examine each chromatogram for
peaks greater than the height of the internal
standard peaks. These peaks can obscure the
compounds of interest.
15 Method performance
15.1 The specifications for this method
were taken from the inter-laboratory
validation of EPA Method 624 (reference 9).
Method 1624 has been shown to yield slightly
better performance on treated effluents than
Method 624. Additional method performance
data can be found in Reference 10.
References
1. "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass
Spectrometry Equipment and Laboratories,"
USEPA, EMSL/Cincinnati, OH 45268, EPA-
600/4-80-025 (April 1980).
2. Bellar, T.A. and Lichtenberg, ].}., "Journal
American Water Works Association." 66. 739
(1974).
3. Bellar, T.A. and Lichtenberg, J.J., "Semi-
automated Headspace Analysis of Drinking
Waters and Industrial Waters for Purgeable
Volatile Organic Compounds." in
Measurement of Organic Pollutants Water
and Wastewater. C.E. VanHall. ed..
American Society for Testing Materials,
Philadelphia, PA. Special Technical
Publication 688, (1978).
4. "Working with Carcinogens," DHEW,
PHS, NIOSH. Publication 77-206 (1977).
5. "OSHA Safety and Health Standards.
General Industry," 29 CFR 1910. OSHA 2206.
(1976).
-------
180 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
6. "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publication. Committee on Chemical Safety
(1979).
7. "Handbook of Analytical Quality Control
in Water and Wastewater Laboratories."
USEPA. EMSL/Cincinnati, OH 45268. EPA-4-
79-019 (March 1979).
8. "Methods 330.4 and 330.5 for Total
Residual Chlorine." USEPA. EMSL/
Cincinnati. OH 45268. EPA-4-79-020 (March
1979).
9. 'Test Method: Purgeables—Method 624,"
USEPA. EMSL/Cincinnati. OH 45268.
10. "Colby. B.N., Beimer, R.G.. Rushneck,
D.R.. and Telliard. W.A.. "Isotope Dilution
Gas Chromatography-Mass Spectrometry for
the Determination of Priority Pollutants in
Industrial Effluents," USEPA, Effluent
Guidelines Division, Washington, DC 20460
(1880).
TABLE 1 .—VOLATILE ORGANIC COMPOUNDS
ANALYZED BY ISOTOPE DILUTION Gc/MS
Compound
Gabon
2
c«l lyiteff
EGO
No.
ID
161
245
345
246
344
268
366
2"
316
244
716
002
229
329
213
313
615
71S
230
330
614
714
223
204
304
251
351
214
314
019
182
247
347
216
315
286
366
143
244
386
207
307
238
338
166
Compound
Biuiiiuchlorom^run*(IS) ....
ChlorornMhin*-d3
BTomom*twt*....
Vlnjw cNonov43..
Vinyl cntorid*
ChtofOMYMno^S..
M*lnyten* eNorid*-d2
Anton*...
AaytarMr*MO
Adyta***
1.1-dfcNoro*m»n*-d2..
I.UfcMoroofun*
1.HtcMororth«n*-(13
l.lxflcMorosilhsVW
OiMhyl •ttttr-dlO
Olvtnyt tvMr
Tnn*-l ,2-dtehtaoMh*n»d2..
Tnnt-U-dkMoroMh*n*
MMiyl omyl kMon»d3
1.2-dkNoi
Tnr*>1>
Trara-14-
TrlcMonM«Mn-t3C1
TitchloroOten*
CMorodbromonwVwn* ..
1-1X1.
1.U-«oMoro**WM
•ramofomviXl
1.1AW
us.....
1.1Z2-MkKMofO*«lirM
TMneMoro*«MM-1X2
TokvMonMlhMi*
1 »4*4jicMofQouisyw (ini (to).....
Tob*r»d6
Tokmw
Chtarob*ni*n*-d5
BromoSuorobmon*..
EGO
No.
161
181
245
161
246
161
268
161
216
181
244
181
616
181
161
203
181
229
191
213
181
615
181
230
181
614
181
223
181
210
181
211
181
182
206
162
248
162
232
162
233
162
267
182
204
182
251
162
214
162
162
162
247
163
215
163
286
183
183
266
163
207
163
238
163
SO p*re*M of mn* 95.
96 to 100 p*rc*nt ol mm 174.
5 to 9 PMO*M ol mwa 176,
TABLE 4.—VOLATILE ORGANIC COMPOUND
CHARACTERISTIC MASSES
LfltMltd compound
Aerate*
Tokwn*
1,1,1-MeMoroMhtn*
1,U-McMoro*tt*n*
TrichloroMhiM
Vinyl cntoridi
Amtog
dS
IX
IX
d3
IX
dS
dS
d7
IX
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Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rule3 and Regulations
181
TABLE 5.—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
Acceptance criteria tt 20 ng/L
Compound
Initial precision and accuracy Labeled
section 8.2.3 compound
recovery
tec 8.3 and
14.2
P (percent)
i (ng/LI 8 (jig/L)
On-going
accuracy
uc 11.5
Acetone
Acrotein
Acrytonrtnto
Benzene
Bromodicfiloromeihane
Brornolorm
Bromomethane
Carbon tetrachlorid*
CMorobenzene
CrUoroethane
2-chloroethyrvinyl ether
Chloroform
Chloromerhane
Obromochlorometharie
1.1-dtehloroetriarw
1.2-dfchloroethane
1,1-dichloroelhen*
Tran*- t ,2-dKfilorQethon6
1,2-dtehloropropane
Ci»-1.3-oTchloropropen*
Trara-1.3-dichkxopropen»...
Oietrryt ether
P-dioxane
Ethyl benzene
Methylene chloride
Methyl ethyl ketone
1,1,2^-tetrachkxoethan*
TetracMoroethen*
Totmrw
1,1,1-ttiehloroethane
t, 1,2-tncNoroethane
Trichloroethene
Vinyl eWorld*
9.0
8.2
7.0
25.0
6.9
8.2
M.8
36.0
7.9
26.0
7.9
8.7
7.7
11.7
7.4
19.2
221
14.5
9.8
9.8
6.6
6.3
5.9
7.1
8.9
27.1
note t
note 2
note 2
13.0-28.2
6.5-31.5
7.4-35.1
Ct-543
15.9-24.8
14.2-29.6
2.1-46.7
d-«9.8
11.8-26.3
d-55.5
11.2-29.1
11.4-31.4
11.6-30.1
d-49.8
10.5-31.5
d-46.8
d-51.0
ct-402
notal
not* 1
15.6-28.5
d-49.8
not»1
10.7-30.0
15.1-28.5
14.5-28.7
10.5-33.4
11.8-29.7
16.6-29.5
d-56.5
ru-196
na-199
n*-214
ns-414
42-165
na-205
ns-308
ns-554
18-172
na-410
16-185
23-191
12-192
n»-315
15-195
r»-343
ns-381
nt-2S4
nt-203
nt-318
5-199
31-181
4-193
12-200
21-184
35-196
na-4SZ
4-33
4-34
6-36
d-61
12-30
4-35
d-51
d-79
6-30
d-64
8-32
9-33
9-03
d-52
8-34
d-51
0-56
d-44
5-35
0-50
7-34
11-32
6-33
8-35
9-32
12-34
d-65
d'detected: resutt must be greater (Man zero.
n»=no specification; limrl would be below detection tor*.
Note 1: Specification* not available, tor these compound* at lima of rateaa* of (hi* method.
Note t Specification* not developed tor these compound*; use method 603.
HLUNO CODE (SaO-SO-M
-------
182
Federal Register / Vol. 49, No. 209 / Friday. October 26.1984 / Rules and Regulations
IVMOKCUUMMt
•UtWOMMItH
hOMJCQMVMQl
:» MDtlCIAM
MVf fU
FIGURE 1 Purging Device.
FIGURES Schematic of Purge afMl Trap
Device—Purge Mode.
. 9 MM OlA»'M*
I
FIUUHC 2 Tiap Packings and Construction to
Include Oeaorb CapaoMlty
FIGURE 4 Schematic ol Purge and Tiap
Device-Daeer* Mede.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
133
10 H
g 10-
Ot-
10 20 SO 100 200
CONCENTRATION (ug/U
5
,j "•»
*
K inonn -
TOLUENE Oj
. * «
I ^ ™
»3»
- • -1*
1214S67I910
ANALYSIS NUMBER
u§
Ul u)
!l
j
0 90 .
llllllllf
TOLUENE
-. - r^~- ,- t ^
e/i
6/1 on w2 «i w e/«
DATE ANALYZED
FIGURE 5 Relative Response Calibration Curve
for Toluene. The Dotted Lines Enclose a x 10
Percent Error Window.
FIGURE 7 Quality Control Charts Showing Area
(top graph) and Relative Response of Toluene to
Toluene-d, (lower graph) Plotted as a Function of
Time or Analysis Number.
— AfitA
• M 299
•MZ92
(Bl
AREA MI960
• M f*t
• M.Z92
(Cl
M /d.;' M>H6tt
M/2 99 62506
•M/Z M
• M/Z92
FIGURE 6 Extracted Ion Current Profiles for
(A) Toluene. (B) Toluene d., and a Mixture of
Toluene and Toluene-d,.
MUJMQCOOCl
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184
Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
Method 1625 Revision B—SemivolatUe
Organic Compound* by Isotope Dilution GC/
MS
1 Scope and application
1.1 This method is designed to determine
the »emivolatile toxic organic pollutants
associated with the 1976 Consent Decree and
additional compounds amenable to
extraction and analysis by capillary column
gas chromatography-mass spectrometry (GO/
MS).
1.2 The chemical compounds listed in
tables 1 and 2 may be determined in
municipal and industrial discharges by this
method. The method is designed to meet the
survey requirements of Effluent Guidelines
Division (ECD) and the National Pollutants
Discharge Elimination System (NPDES) under
40 CFR 136.1. Any modifications 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 136.4
and 136.5.
1.3 The detection limit of this method is
usually dependent on the level of
interferences rather than instrumental
limitations. The limits listed in tables 3 and 4
represent the minimum quantity that can be
detected with no interferences present.
1.4 The GC/MS portions of this method
are for use only by analysts experienced with
GC/MS or under the close supervision of
such qualified persons. Laboratories
unfamiliar with analyses of environmental
samples by GC/MS should run the
performance tests in reference 1 before
beginning.
2 Summary of method
2.1 Stable isotopically labeled analogs of
the compounds of interest are added to a one
liter wastewater sample. The sample is
extracted at pH 12-13, then at pH <2 with
methylene chloride using continuous
extraction techniques. The extract is dried
over sodium sulfate and concentrated to a
volume of one mL An internal standard is
added to the extract, and the extract is
injected into the gas chromatograph (GC).
The compounds are separated by GC and
detected by a mass spectrometer (MS). The
labeled compounds serve to correct the
variability of the analytical technique.
2.2 Identification of a compound
(qualitative analysis) is performed by
comparing the GC retention time and
background corrected characteristic spectral
masses with those of authentic standards.
2.3 Quantitative analysis is performed by
GC/MS using extracted ion current profile
(EICP) areas. Isotope dilution is used when
labeled compounds are available; otherwise,
an internal or external standard method is
used.
2.4 Quality is assured through
reproducible calibration and testing of the
extraction and GC/MS systems.
3 Contamination and interferences
3.1 Solvents, reagents, glassware, and
other sample processing hardware may yield
artifacts and/or elevated baselines causing
misinterpretation of chromatograms and
spectra. All materials shall be demonstrated
to be free from interferences under the
conditions of analysis by running method
blanks initially and with each sample lot
(samples started through the extraction
process on a given 8 hr shift, to a maximum of
20). Specific selection of reagents and
purification of solvents by distillation in all-
glass systems may be required. Glassware
and, where possible, reagents are cleaned by
solvent rinse and baking at 450 'C for one
hour minimum.
3.2 Interferences coextracted from
samples will vary considerably from source
to source, depending on the diversity of the
industrial complex or municipality being
samples.
4 Safety
4.1 The toxicity or carcinogenicity of each
compound or reagent used in this method has
not been precisely determined: however, each
chemical compound should be treated as a
potential health hazard. Exposure to these
compounds should be reduced to the lowest
possible level. 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 data handling
sheets should also be made available to all
personnel involved in these analyses.
Additional information on laboratory safety
can be found in references 2-4.
4.2 The following compounds covered by
this method have been tentatively classified
as known or suspected human or mammalian
carcinogens: benzo(a)anthracene, 3,3'-
dichlorobenzidine, benzo(a)pyrene,
dibenzo(a,h)anthracene, N-
nitrosodimethylamine, and B-naphthylamine.
Primary standards of these compounds shall
be prepared in a hood, and a NIOSH/MESA
approved toxic gas respirator should be worn
when high concentrations are handled.
5 Appartaus and materials
5.1 Sampling equipment for discrete or
composite sampling.
5.1.1 Sample bottle, amber glass, 1.1 liters
minimum. If amber bottles are not available,
samples shall be protected from light. Bottles
are detergent water washed, then solvent
rinsed or baked at 450 *C for one hour
minimum before use.
5.1.2 Bottle caps—threaded to fit sample
bottles. Caps are lined with Teflon.
Aluminum foil may be substituted if the
sample is not corrosive. Liners are detergent
water washed, then reagent water (section
6.5) and solvent rinsed, and baked at
approximately 200 'C for one hour minimum
before use.
5.1.3 Compositing equipment—automatic
or manual compositing system incorporating
glass containers for collection of a minimum
1.1 liters. Sample containers are kept at 0 to 4
*C during sampling. Glass or Teflon tubing
only shall be used. If the sampler uses a
peristaltic pump, a minimum length of
compressible silicone rubber tubing may be
used in the pump only. Before use, the tubing
is thoroughly rinsed with methanol, followed
by repeated rinsings with reagent water
(section 6.5) to minimize sample
contamination. An integrating flow meter is
used to collect proportional composite
samples.
5.2 Continuous liquid-liquid extractor—
Teflon or glass conncecting joints and
stopcocks without lubrication (Hershberg-
Wolf Extractor) one liter capacity. Ace Glass
6841-10. or equivalent.
5.3 Drying column—15 to 20 mm i.d. Pyrex
chromatographic column equipped with
coarse glass frit or glass wool plug.
5.4 Kuderna-Danish (K-D) apparatus
5.4.1 Concentrator tube—lOmL graduated
(Kontes K-570050-1025. or equivalent) with
calibration verified. Ground glass stopper
(size 19/22 joint) is used to prevent
evaporation of extracts.
5.4.2 Evaporation flask—500 mL (Kontes
K-570001-0500, or equivalent), attached to
concentrator tube with springs (Kontes K-
662750-0012).
5.4.3 Snyder column—three ball macro
(Kontes K-503000-0232, or equivalent).
5.4.4 Snyder column—two ball micro
(Kontes K-469002-0219, or equivalent).
5.4.5 Boiling chips—approx 10/40 mesh.
extracted with methylene chloride and baked
at 450 *C for one hr minimum.
5.5 Water bath—heated, with concentric
ring cover, capable of temperature control (q
2 *C), installed in a fume hood.
5.6 Sample vials—amber glass, 2-5 mL
with Teflon-lined screw cap.
5.7 Analytical balance—capable of
weighing 0.1 mg.
5.8 Gas chromatograph—shall have
splitless or on-column injection port for
capillary column, temperature program with
30 "C hold, and shall meet all of the
performance specifications in section 12.
5.8.1 Column—30±5mx0.25±0.02 mm
i.d. 5% phenyl, 94% methyl, 1% vinyl silicone
bonded phase fused silica capillary column Q
ft W DB-5, or equivalent).
5.9 Mass spectrometer—70 eV electron
impact ionization, shall repetitively scan from
35 to 450 amu in 0.95 to 1.00 second, and shall
produce a unit resolution (valleys between
ra/x 441-442 less than 10 percent of the height
of the 441 peak), backgound corrected mass
spectrum from 50 ng
decafluorotriphenylphosphine (DFTPP)
introduced through the GC inlet. The
spectrum shall meet the mass-intensity
criteria in table 5 (reference 5). The mass
spectrometer shall be interfaced to the GC
such that the end of the capillary column
terminates within one centimeter of the ion
source but does not intercept the electron or
ion beams. All portions of the column which
connect the GC to the ion source shall remain
at or above the column temperature during
analysis to preclude condensation of less
volatile compounds.
5.10 Data system—shall collect and
record MS data, store mass-intensity data in
spectral libraries, process GC/MS data,
generate reports, and shall compute and
record response factors.
5.10.1 Data acquisition—mass spectra
shall be collected continuously throughout
the analysis and stored on a mass storage
device.
5.10.2 Mass spectral libraries—user
created libraries containing mass spectra
obtained from analysis of authentic
standards shall be employed to reverse
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185
search CC/MS runs for the compounds of
interest (section 7.2).
5.10.3 Data processing—the data system
shall be used to search, locate, identify, and
quantify the compounds of interest in each
GC/MS analysis. Software routines shall be
employed to compute retention times and
peak areas. Displays of spectra, mass
chromatograms, and library comparisons are
required to verify results.
5.10.4 Response factors and multipoint
calibrations—the data system shall be used
to record and maintain lists of response
factors (response ratios for isotope dilution)
and multipoint calibration curves (section 7).
Computations of relative standard deviation
(coefficient of variation) are useful for testing
calibration linearity. Statistics on initial
(section 8.2) and on-going (section 12.7)
performance shall be computed and
maintained.
6 Reagents and standards
8.1 Sodium hydroxide—reagent grade, 6N
in reagent water.
6.2 Sulfuric acid—reagent grade. 8N in
reagent water.
6.3 Sodium sulfate—reagent grade.
granular anhydrous, rinsed with methylene
chloride (20 mL/g) and conditioned at 450 *C
for one hour minimum.
6.4 Methylene chloride—distilled in glass
(Burdick and Jackson, or equivalent).
6.5 Reagent water—water in which the
compounds of interest and interfering
compounds are not detected by this method.
6.6 Standard solutions—purchased as
solutions or mixtures with certification to
their purity, concentration, and authenticity,
or prepared from materials of known purity
and composition. If compound purity is 96
percent or greater, the weight may be used
without correction to compute the
concentration of the standard. When not
being used, standards are stored in the dark
at -20 to -10 "C in screw-capped vials with
Teflon-lined lids. A mark is placed on the vial
at the level of the solution so that solvent
evaporation loss can be detected. The vials
are brought to room temperature prior to use.
Any precipitate is redissolved and solvent is
added if solvent loss has occurred.
6.7 Preparation of stock solutions—
prepare in methylene chloride, benzene, p-
dioxane. or a mixture of these solvents per
the steps below. Observe the safety
precautions in section 4. The large number of
labeled and unlabeled acid, base/neutral,
and Appendix C compounds used for
combined calibration (section 7) and
calibration verification (12.5) require high
concentrations (approx 40 mg/mL) when
individual stock solutions are prepared, so
that dilutions of mixtures will permit
calibration with all compounds in a single set
of solutions. The working range for most
compounds is 10-200 ug/mL. Compounds
with a reduced MS response may be prepared
at higher concentrations.
6.7.1 Dissolve an appropriate amount of
assayed reference material in a suitable
solvent. For example, weigh 400 mg
naphthalene in a 10 mL ground glass
stoppered volumetric flask and fill to the
mark with benzene. After the naphthalene is
completely dissolved, transfer the solution to
a 15 mL vial with Teflon-lined cap.
6.7.2 Stock standard solutions should be
checked for signs of degradation prior to the
preparation of calibration or performance test
standards. Quality control check samples
that can be used to determine the accuracy of
calibration standards are available from the
US Environmental Protection Agency,
Environmental Monitoring and Support
Laboratory. Cincinnati, Ohio 45268.
6.7.3 Stock standard solutions shall be
replaced after six months, or sooner if
comparison with quality control check
samples indicates a change in concentration.
6.8 Labeled compound spiking solution—
from stock standard solutions prepared as
above, or from mixtures, prepare the spiking
solution at a concentration of 200 /ig/mL, or
at a concentration appropriate to the MS
response of each compound.
6.9 Secondary standard—using stock
solutions (section 6.7). prepare a secondary
standard containing all of the compounds in
tables 1 and 2 at a concentration of 400 fig/
mL, or higher concentration appropriate to
the MS response of the compound.
6.10 Internal standard solution—prepare
2,2'-difluorobiphenyl (DFB) at a concentration
of 10 mg/mL in benzene.
6.11 DFTPP solution—prepare at 50 fig/
mL in acetone.
6.12 Solutions for obtaining authentic
mass spectra (section 7.2)—prepare mixtures
of compounds at concentrations which will
assure authentic spectra are obtained for
storage in libraries.
6.13 Calibration solutions—combine 0.5
mL of the solution in section 6.8 with 25, 50,
125. 250, and 500 fiL of the solution in section
6.9 and bring to 1.00 mL total volume each.
This will produce calibration solutions of
nominal 10, 20, 50,100, and 200 fig/mL of the
pollutants and a constant nominal 100 fig/mL
of the labeled compounds. Spike each
solution with 10 uL of the internal standard
solution (section 6.10). These solutions permit
the relative response (labeled to unlabeled)
to be measured as a function of concentration
(section 7.4).
6.14 Precision and recovery standard—
used for determination of initial (section 8.2)
and on-going (section 12.7) precision and
recovery. This solution shall contain the
pollutants and labeled compounds at a
nominal concentration of 100 fig/mL.
6.15 Stability of solutions—all standard
solutions (sections 6.8-6.14) shall be analyzed
within 48 hours of preparation and on a
monthly basis thereafter for signs of
degradation. Standards will remain
acceptable if the peak area at the
quantitation mass relative to the DFB internal
standard remains within ±15 percent of the
area obtained in the initial analysis of the
standard.
7 Calibration
7.1 Assemble the GC/MS and establish
the operating conditions in table 3. Analyze
standards per the procedure in section 11 to
demonstrate that the analytical system meets
the detection limits in tables 3 and 4, and the
mass-intensity criteria in table 5 for 50 ng
DFTPP.
7.2 Mass spectral libraries—detection and
identification of compounds of interest are
dependent upon spectra stored in user
created libraries.
7.2.1 Obtain a mass spectrum of each
pollutant, labeled compound, and the internal
standard by analyzing an authentic standard
either singly or as part of a mixture in which
there is no interference between closely
eluted components. That only a single
compound is present is determined by
examination of the spectrum. Fragments not
attributable to the compound under study
indicate the presence of an interfering
compound.
7.2.2 Adjust the analytical conditions and
scan rate (for this test only) to produce an
undistorted spectrum at the GC peak
maximum. An undistorted spectrum will
usually be obtained if five complete spectra
are collected across the upper half of the GC
peak. Software algorithms designed to
"enhance" the spectrum may eliminate
distortion, but may also eliminate authentic
masses or introduce other distortion.
7.2.3 The authentic reference spectrum is
obtained under DFTPP tuning conditions
(section 7.1 and table 5) to normalize it to
spectra from other instruments.
7.2.4 The spectrum is edited by saving the
5 most intense mass spectral peaks and all
other mass spectral peaks greater than 10
percent of the base peak. This edited
spectrum is stored for reverse search and for
compound confirmation.
7.3 Analytical range—demonstrate that 20
ng anthracene or phenanthrene produces an
area at m/z 178 approx one-tenth that
required to exceed the linear range of the
system. The exact value must be determined
by experience for each instrument. It is used
to match the calibration range of the
instrument to the analytical range and
detection limits required, and to diagnose
instrument sensitivity problems (section 15.4).
The 20 ug/mL calibration standard (section
6.13) can be used to demonstrate this
performance.
7.3.1 Polar compound detection—
demonstrate that unlabeled
pentachlorophenol and benzidine are
detectable at the 50 ug/mL level (per all
criteria in section 13). The 50 ug/mL
calibration standard (section 6.13) can be
used to demonstrate this performance.
7.4 Calibration with Isotope dilution—
isotope dilution is used when 1) labeled
compounds are available, 2) interferences do
not preclude its use, and 3) the quantitation
mass extracted ion current profile (E1CP) area
for the compound is in the calibration range.
If any of these conditions preclude isotope
dilution, internal or external standard
methods (section 7.5 or 7.6) are used.
7.4.1 A calibration curve encompassing
the concentration range is prepared for each
compound to be determined. The relative
response (pollutant to labeled) vs
concentration in standard solutions is plotted
or computed using a linear regression. The
example in Figure 1 shows a calibration
curve for phenol using phenol-dS as the
isotopic diluent Also shown are the ± 10
percent error limits (dotted lines). Relative
Reponse (RR) is determined according to the
procedures described below. A minimum of
five data points are employed for calibration.
7.4.2 The relative response of a pollutant
to its labeled analog is determined from
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186 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
isotope ratio values computed from acquired
data. Three isotope ratios are used in this
process:
R, = the isotope ratio measured for the
pure pollutant.
R, = the isotope ratio measured for the
labeled compound.
R. = the isotope ratio of an analytical
mixture of pollutant and labeled compounds.
The m/z's are selected such that R,>Rr. If
R» is not between 2R, and 0.5R., the method
does not apply and the sample is analyzed by
internal or external standard methods.
7.4.3 Capillary columns usually separate
the pollutant-labeled pair, with the labeled
compound eluted first (figure 2). For this case.
R, « [area mi/z]/l, at the retention time of
the pollutant (RTi). R, = l/[area im/z. at the
retention time of the labeled compound RTi).
R. - (area at m,/z (at RT,)]/[area at RT,)),
as measured in the mixture of the pollutant
and labeled compounds (figure 2), and RR =
R.
7.4.4 Special precautions are taken when
the pollutant-labeled pair is not separated, or
when another labeled compound with
interfering spectral masses overlaps the
pollutant (a case which can occur with
isomeric compounds). In this case, it is
necessary to determine the respective
contributions of the pollutant and labeled
compounds to the respective EICP areas. If
the peaks are separated well enough to
permit the data system or operator to remove
the contributions of the compounds to each
other, the equations in section 7.4.3 apply.
This usually occurs when the height of the
valley between the two GC peaks at the same
m/z is less than 10 percent of the height of
the shorter of the two peaks. If significant GC
and spectral overlap occur, RR is computed
using the following equation: RR = (R, - RJ
(R. + D/(R.-R.)(R, + l). where R. ia
measured as shown in figure 3A, R, is
measured as shown in figure 3B, and R. is
measured as shown in figure 3C. For
example, R, - 48100/4780 = 9.844, R, -
2850/43800 - 0.0808, R. - 49200/48300 =
1.019. amd RR - 1.114.
7.4.5 To calibrate the analytical system by
isotope dilution, analyze a 1.0 pL aliquot of
each of the calibration standards (section
8.13) using the procedure in section 11.
Compute the RR at each concentration.
7.4.8 Linearity—if the ratio of relative
response to concentration for any compound
is constant (less than 20 percent coefficient of
variation) over the 5 point calibration range,
and averaged relative response/
concentration ratio may be used for that
compound: otherwise, the complete
calibration curve for that compound shall be
used over the 5 point calibration range.
7.5 Calibration by internal standard-
used when criteria for istope dilution (section
7.4) cannot be met. The internal standard to
be used for both acid and base/neutral
analyses is 2.2'-difluorobiphenyl. The
internal standard method is also applied to
determination of compounds having no
labeled analog, and to measurement of
labeled compounds for intra-laboratory
statistics (sections 8.4 and 12.7.4).
7.5.1 Response factors—calibration
requires the determination of response
factors (RF) which are defined by the
following equation:
RF = (A. X CJ/IA,. X C,), where
A. is the area of the characteristic mass for
the compound in the daily standard
AH is the area of the characteristic mass for
the internal standard
Cu is the concentration of the internal
standard (pg/mL)
C, is the concentration of the compound in
the daily standard (pg/mL)
7.5.1.1 The response factor is determined
for at least five concentrations appropriate to
the response of each compound (section 8.13);
nominally, 10,20.50,100, and 200 pg/mL The
amount of internal standard added to each
extract is the same (100 pg/mL) so that Q.
remains constant. The RF is plotted vs
concentration for each compound in the
standard (C.) to produce a calibration curve.
7.5.1.2 Linearity—if the response factor
(RF) for any compound is constant (less than
35 percent coefficient of variation) over the 5
point calibration range, an averaged response
factor may be used for that compound;
otherwise, the complete calibration curve for
that compound shall be used over the 5 point
range.
7.8 Combined calibration—by using
calibration solutions (section 8.13) containing
the pollutants, labeled compounds, and the
internal standard, a single set of analyses can
be used to produce calibration curves for the
isotope dilution and internal standard
methods. These curves are verified each shift
(section 12.5) by analyzing the 100 0g/mL
calibration standard (section 8.13).
Recalibration is required only if calibration
verification (section 12.5) criteria cannot be
met.
8 Quality assurance/quality control
B.1 Each laboratory that uses this method
is required to operate a formal quality
assurance program. The minimum
requirements of this program consist of an
initial demonstration of laboratory capability,
analysis of samples spiked with labeled
compounds to evaluate and document data
quality, and analysis of standards and blanks
as tests of continued performance.
Laboratory performance is compared to
established performance criteria to determine
if the results of analyses meet the
performance characteristics of the method.
8.1.1 The analyst shall make an initial
demonstration of the ability to generate
acceptable accuracy and precision with this
method. This ability is established as
described in section 8.2.
8.1.2 The analyst is permitted to modify
this method to improve separations or lower
the costs of measurements, provided all
performance specifications are met. Each
time a modification is made to the method,
the analyst is required to repeat the
procedure in section 8.2 to demonstrate
method performance.
8.1.3 Analyses of blanks are required to
demonstrate freedom from contamination.
The procedures and criteria for analysis of a
blank are described in section 8.5.
8.1.4 The laboratory shall spike all
samples with labeled compounds to monitor
method performance. This test is described in
section 8.3. When results of these spikes
indicate atypical method performance for
samples, the samples are diluted to bring
method performance within acceptable limits
(section 15).
8.1.5 The laboratory shall, on an on-going
basis, demonstrate through calibration
verification and the analysis of the precision
and recovery standard (section 8.14) that the
analysis system is in control. These
procedures are described in sections 12.1.
12.5, and 12.7.
8.1.8 The laboratory shall maintain
records to define the quality of data that is
generated. Development of accuracy
statements is described in section 8.4.
8.2 Initial precision and accuracy—to
establish the ability to generate acceptable
precision and accuracy, the analyst shall
perform the following operations:
8.2.1 Extract, concentrate, and analyze
two sets of four one-liter aliquots (8 aliquots
total) of the precision and recovery standard
(section 6.14) according to the procedure in
section 10.
8.2.2 Using results of the first set of four
analyses, compute the average recovery (X)
in fig/mL and the standard deviation of the
recovery (s) in 0g/mL for each compound, by
isotope dilution for pollutants with a labeled
analog, and by internal standard for labeled
compounds and pollutants with no labeled
analog.
8.2.3 For each compound, compare s and
X with the corresponding limits for initial
precision and accuracy in table 8. If s and X
for all compounds meet the acceptance
criteria, system performance is acceptable
and analysis of blanks and samples may
begin. If, however, any individual s exceeds
the precision limit or any individual X falls
outside the range for accuracy, system
performance is unacceptable for that
compound. Note: The large number of
compounds in table 8 present a substantial
probability that one or more will fail the
acceptance criteria when all compounds are
analyzed. To determine if the analytical
system is out of control, or if the failure can
be attributed to probability, proceed as
follows:
8.2.4 Using the results of the second set of
four analyses, compute s and X for only those
compounds which failed the test of the first
set of four analyses (section 8^.3). If these
compounds now pass, system performance is
acceptable for all compounds and analysis of
blanks and samples may begin. If. however,
any of the same compounds fail again, the
analysis system is not performing properly
for these compounds. In this event, correct
the problem and repeat the entire test
(section 8.2.1).
8.3 The laboratory shall spike all samples
with labeled compounds to assess method
performance on the sample matrix.
8.3.1 Analyze each sample according to
the method in section 10.
8.3.2 Compute the percent recovery (P) of
the labeled compounds using the internal
standard method (section 7.5).
8.3.3 Compare the labeled compound
recovery for each compound with the
corresponding limits in table 8. If the
recovery of any compounds falls outside its
warning limit, method performance is
unacceptable for that compound in that
sample. Therefore, the sample is complex and
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Federal Register / Vol. 49, No. 209 / Friday. October 26. 1984 / Rules and Regulations
187
is to be diluted and reanalyzed per section
1S.4.
8.4 As part of the QA program for the
laboratory, method accuracy for wastewater
samples shall be assessed and records shall
be maintained. After the analysis of five
wastewater samples for which the labeled
compounds pass the tests in section 8.3,
compute the average percent recovery (P) and
the standard deviation of the percent
recovery (sp) for the labeled compounds only.
Express the accuracy assessment as a
percent recovery interval from P—2 ,„ to
P + 2,p. For example, if P=90% and sp = 10%.
the accuracy interval is expressed as 70-
100%. Update the accuracy assessment for
each compound on a regular basis (e.g. after
each 5-10 new accuracy measurements).
8.S Blanks—reagent water blanks are
analyzed to demonstrate freedom from
contamination.
8.5.1 Extract and concentrate a blank
with each sample lot (samples started
through the extraction process on the same 8
hr shift, to a maximum of 20 samples).
Analyze the blank immediately after analysis
of the precision and recovery standard
(section 6.14) to demonstrate freedom from
contamination.
6.5.2 If any of the compounds of interest
(tables 1 and 2) or any potentially interfering
compound is found in a blank at greater than
10 ng/L (assuming a response factor of 1
relative to the internal standard for
compounds not listed in tables 1 and 2),
analysis of samples is halted until the source
of contamination is eliminated and a blank
shows no evidence of contamination at this
level.
8.6 The specifications contained in this
method can be met if the apparatus used is
calibrated properly, then maintained in a
calibrated state. The standards used for
calibration (section 7), calibration
verification (section 12.5), and for initial
(section 8.2) and on-going (section 12.7)
precision and recovery should be identical, so
that the most precise results will be obtained.
The GC/MS instrument in particular will
provide the most reproducible results if
dedicated to the settings and conditions
required for the analysis of semi-volatiles by
this method.
8.7 Depending on specific program
requirements. Held replicates may be
collected to determine the precision of the
sampling technique, and spiked samples may
be required to determine the accuracy of the
analysis when internal or external standard
methods are used.
9 Sample collection, preservation, and
handling
9.1 Collect samples in glass containers
following conventional sampling practices
(reference 7). Composite samples are
collected in refrigerated glass containers
(section 5.1.3) in accordance with the
requirements of the sampling program.
9.2 Maintain samples at 0-4 'C from the
time collection until extraction. If residual
chlorine is present, add 60 mg sodium
thiosulfate per liter of water. EPA methods
330.4 and 330.5 may be used to measure
residual chlorine (references 8).
9.3 Begin sample extraction within seven
days of collection, and analyze all extracts
within 40 days of extraction.
10 Sample extraction and concentration
(See figure 1)
10.1 Labeled compound spiking—measure
1.00 ± 0.01 liter of sample into a glass
container. For untreated effluents, and
samples which are expected to be difficult to
extract and/or concentrate, measure an
additional 10.0 ± 0.1 mL and dilute to a final
volume of 1.00 ± 0.01 liter with reagent water
in a glass container.
10.1.1 For each sample or sample lot (to a
maximum of 20) to be extracted at the same
time, place three 1.00 ± 0.10 liter aliquots of
reagent water in glass containers.
10.1.2 Spike 0.5 ml of the labeled
compound spiking solution (section 6.8) into
all samples and one reagant water aliquot.
10.1.3 Spike 1.0 mL of the precision and
recovery standard (section 6.14) into the two
remaining reagent water aliquots.
10.1.4 Stir and equilibrate all solutions for
1-2 hr.
10.2 Base/neutral extraction—place 100-
150 mL methylene chloride in each
continuous extractor and 200-300 in each
distilling flask.
10.2.1 Pour the sample(s), blank, and
standard aliquots into the extractors. Rinse
the glass containers with 50-100 mL
methylene chloride and add to the respective
extractor.
10.2.2 Adjust the pH of the waters in the
extractors to 12-13 with 6N NaOH while
monitoring with a pH meter. Begin the
extraction by heating the flask until the
methylene chloride is boiling. When properly
adjusted, 1-2 drops of methylene chloride per
second will fall from the condenser tip into
the water. After 1-2 hours of extraction, test
the pH and readjust to 12-13 if required.
Extract for 18-24 hours.
10.2.3 Remove the distilling flask.
estimate and record the volume of extract (to
the nearest 100 mL), and pour the contents
through a drying column containing 7 to 10
cm anhydrous sodium sulfate. Rinse the
distilling flask with 30-50 mL of methylene
chloride and pour through the drying column.
Collect the solution in a 500 mL K-D
evaporator flask equipped with a 10 mL
concentrator tube. Seal, label as the base/
neutral fraction, and concentrate per sections
10.4 to 10.5.
10.3 Acid extraction—adjust the pH of the
waters in the extractors to 2 or less using 8N
sulfuric acid. Charge clean distilling flasks
with 300-400 mL of methylene chloride. Test
and adjust the pH of the waters after the first
1-2 hr of extraction. Extract for 18-24 hours.
10.3.1 Repeat section 10.2.3, except label
as the acid fraction.
10.4 Concentration—concentrate the
extracts in separate 500 mL K-D flasks
equipped with 10 mL concentrator tubes.
10.4.1 Add 1 to 2 clean boiling chips to the
flask and attach a three-ball macro Snyder
column. Prewet the column by adding
approximately one mL of methylene chloride
through the top. Place the K-D apparatus in a
hot water bath so that the entire lower
rounded surface of the flask is bathed with
steam. 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 will
actively chatter but the chambers will not
flood. When the liquid has reached an
apparent volume of 1 mL. remove the K-D
apparatus from the bath and allow the
solvent to drain and cool for at least 10
minutes. 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.
10.4.2 For performance standards
(sections 8.2 and 12.7) and for blanks (section
8.5). combine the acid and base/neutral
extracts for each at this point. Do not
combine the acid and base/neutral extracts
for samples.
10.5 Add a clean boiling chip and attach a
two ball micro Snyder column to the
concentrator tube. Prewet the column by
adding approx 0.5 mL methylene chloride
through the top. Place the apparatus in the
hot water bath. Adjust the vertical position
and the water temperature as required to
complete the concentration in 5-10 minutes.
At the proper rate of distillation, the balls of
the column will actively chatter but the
chambers will not flood. When the liquid
reaches an apparent volume of approx 0.5
mL, remove the apparatus from the water
bath and allow to drain and cool for at least
10 minutes. Remove the micro Snyder column
and rinse its lower joint into the concentrator
tube with approx 0.2 mL of methylene
chloride. Adjust the final volume to 1.0 mL
10.6 Transfer the concentrated extract to
a clean screw-cap vial. Seal the vial with a
Teflon-lined lid, and mark the level on the
vial. Label with the sample number and
fraction, and store in the dark at —20 to
— 10 *C until ready for analysis.
11 GC/MS analysis
11.1 Establish the operating conditions
given in tables 3 or 4 for analysis of the base/
neutral or acid extracts, respectively. For
analysis of combined extracts (section 10.4.2),
use the operating conditions in table 3.
11.2 Bring the concentrated extract
(section 10.6) or standard (sections 6.13-6.14)
to room temperature and verify that any
precipitate has redissolved. Verify the level
on the extract (sections 6.6 and 10.6) and
bring to the mark with solvent if required.
11.3 Add the internal standard solution
(section 6.10) to the extract (use 1.0 uL of
solution per 0.1 mL of extract) immediately
prior to injection to minimize the possibility
of loss by evaporation, adsorption, or
reaction. Mix thoroughly.
11.4 Inject a volume of the standard
solution or extract such that 100 ng of the
internal standard will be injected, using on-
column or splitless injection. For 1 mL
extracts, this volume will be 1.0 uL Start the
GC column initial isothermal hold upon
injection. Start MS data collection after the
solvent peak elutes. Stop data collection after
the benzo (ghi) perylene or
pentachlorophenol peak elutes for the base/
neutral or acid fraction, respectively. Return
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188 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
the column to the initial temperature for
analysis of the next cample.
12 System and laboratory performance
12.1 At the beginning of each 8 hr shift
during which analyses are performed. GC/
MS system performance and calibration are
verified for all pollutants and labeled
compounds. For these test, analysis of the 100
jig/mL calibration standard (section 6.13)
shall be used to verify all performance
criteria. Adjustment and/or recalibration (per
section 7) shall be performed until all
performance criteria are met. Only after all
performance criteria are met may samples,
blanks, and precision and recovery standards
be analyzed.
12.2 DFTPP spectrum validity—inject 1 pL
of the DFTPP solution (section 8.11) either
separately or within a few seconds of
injection of the standard (section 12.1)
analyzed at the beginning of each shift. The
criteria in table 5 shall be met.
12.3 Retention times—the absolute
retention time of 2,2'-difluorobiphenyl shall
be within the range of 1078 to 1248 seconds
and the relative retention times of all
pollutants and labeled compounds shall fall
within the limits given in tables 3 and 4.
12.4 GC resolution—the valley height
between anthracene and phenanthrene at
m/x 178 (or the analogs at m/z 188) shall not
exceed 10 percent of the taller of the two
peaks.
12.5 Calibration verification—compute
the concentration of each pollutant (tables 1
and 2) by isotope dilution (section 7.4) for
those compounds which have labeled
analogs. Compute the concentration of each
pollutant which has no labeled analog by the
internal standard method (section 7.5).
Compute the concentration of the labeled
compounds by the internal standard method.
These concentrations are computed based on
the calibration data determined in section 7.
12.5.1 For each pollutant and labeled
compound being tested, compare the
concentration with the calibration
verification limit in table 8. If all compounds
meet the acceptance criteria, calibration has
been verified and analysis of blanks.
samples, and precision and recovery
standards may proceed. If. however, any
compound fails, the measurement system is
not performing properly for that compound.
In this event, prepare a fresh calibration
standard or correct the problem causing the
failure and repeat the test (section 12.1). or
recalibrate (section 7).
12.8 Multiple peaks—each compound
injected shall give a single, distinct GC peak.
12.7 On-going precision and accuracy.
12.7.1 Analyze the extract of one of the
pair of precision and recovery standards
(section 10.1.3) prior to analysis of samples
from the same lot.
12.7.2 Compute the concentration of each
pollutant (tables 1 and 2) by isotope dilution
(section 7.4) for those compounds which have
labeled analogs. Compute the concentration
of each pollutant which has no labeled
analog by the internal standard method
(section 7.5). Compute the concentration of
the labeled compounds by the internal
standard method.
12.7.3 For each pollutant and labeled
compound, compare the concentration with
the limits for on-going accuracy in table 8. If
all compounds meet the acceptance criteria,
system performance is acceptable and
analysis of blanks and samples may proceed.
If. however, any individual concentration
falls outside of the range given, system
performance is unacceptable for that
compound. NOTE: The large number of
compounds in table 8 present a substantial
probability that one or more will fail when all
compounds are analyzed. To determine if the
extraction/concentration system is out of
control or if the failure is caused by
probability, proceed as follows:
12.7.3.1 Analyze the second aliquot of the
pair of precision and recovery standard
(section 10.1.3).
12.7.3.2 Compute the concentration of
only those pollutants or labeled compounds
that failed the previous test (section 12.7.3). If
these compounds now pass, the extraction/
concentration processes are in control and
analysis of blanks and samples may proceed.
If, however, any of the same compounds fail
again, the extraction/concentration processes
are not being performed properly for these
compounds, hi this event, correct the
problem, re-extract the sample lot (section 10)
and repeat the on-going precision and
recovery test (section 12.7).
12.7.4 Add results which pass the
specifications in section 12.7.2 to initial and
previous on-going data. Update QC charts to
perform a graphic representation of
continued laboratory performance (Figure 5).
Develop a statement of laboratory accuracy
for each pollutant and labeled compound by
calculating the average percent recovery (R)
and the standard deviation of percent
recovery (sr). Express the accuracy as a
recovery interval from R—2», to R+2*,. For
example, if R=95% and »,=5%. the accuracy
is 85 -105%.
13 Qualitative determination
13.1 Qualitative determination is
accomplished by comparison of data from
analysis of a sample or blank with data from
analysis of the shift standard (section 12.1)
and with data stored in the spectral libraries
(section 7.2.4). Identification is confirmed
when spectra and retention times agree per
the criteria below.
13.2 Labeled compounds and pollutants
having no labeled analog:
13.2.1 The signals for all characteristic
masses stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
13.2.2 Either (1) the background corrected
EICP areas, or (2) the corrected relative
intensities of the mass spectral peaks at the
GC peak maximum shall agree within a
factor of two (0.5 to 2 times) for all masses
stored in the library.
13.2.3 The retention time relative to the
nearest eluted internal standard shall be
within ±15 scans or ±15 seconds, whichever
is greater.
13.3 Pollutants having a tabled analog:
13.3.1 The signals for all characteristic
masses stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
13.3.2. Either (1) the background corrected
EICP areas, or (2) the corrected relative
intensities of the mass spectral peaks at the
GC peak maximum shall agree within a
factor of two for all masses stored in the
spectral library.
13.3.3. The retention time difference
between the pollutant and its labeled analog
shall agree within ± 8 scans or ± 8 seconds
(whichever is greater) of this difference in the
shift standard (section 12.1).
13.4 Masses present in the experimental
mass spectrum that are not present in the
reference mass spectrum shall be accounted
for by contaminant or background ions. If the
experimental mass spectrum is contaminated.
an experienced spectrometrist (section 1.4) is
to determine the presence or absence of the
compound.
14 Quantitative determination
14.1 Isotope dilution—by adding a known
amount of a labeled compound to every
sample prior to extraction, correction for
recovery of the pollutant can be made
because the pollutant and its labeled analog
exhibit the same effects upon extraction,
concentration, and gas chroma lography.
Relative response (RR) values for mixtures
are used in conjunction with calibration
curves described in section 7.4 to determine
concentrations directly, so long as labeled
compound spiking levels are constant For the
phenol example given in figure 1 (section
7.4.1). RR would be equal to 1.114. For this RR
value, the phenol calibration curve given in
figure 1 indicates a concentration of 27 pg/
mL in the sample extract (Cra).
14.2 Internal standard—compute the
concentration in the extract using the
response factor determined from calibration
data (section 7.5) and the following equation:
CM(Mg/mL)=(A. x CJ(A» x RF) where C« is
the concentration of the compound in the
extract, and the other terms are as defined in
section 7.5.1.
14.3 The concentration of the pollutant in
water is computed using the volumes of the
original water sample (section 10.1) and the
final extract volume (section 10.5), as follows:
Concentration in water (jig/Ll-fC^, x Vn)/
V. where Vn is the extract volume in mL, and
V, is the sample volume in liters.
14.4 If the EICP area at the quantitiation
mass for any compound exceeds the
calibration range of the system, the extract of
the dilute aliquot (section 10.1) is analyzed by
isotope dilution; otherwise, the extract is
diluted by a factor of 10,9 pL of internal
standard solution (section 8.10) are added to
a 1.0 mL aliquot, and this diluted extract is
analyzed by the internal standard method
(section 14.2). Quantify each compound at the
highest concentration level within the
calibration range.
14.5 Report results for all pollutants and
labeled compounds (tables 1 and 2) found in
all standards, blanks, and samples in pg/L to
three significant figures. Results for samples
which have been diluted are reported at the
least dilute level at which the area at the
quantitation mass is within the calibration
range (section 14.4) and the labeled
compound recovery is within the normal
range for the method (section 15.4).
-------
Federal Register / Vol. 49, No. 209 / Friday. October 26, 1984 / Rules and Regulations
189
15 Analysis of complex samples
15.1 Untreated effluents and other
samples frequently contain high levels
(> 1000 fig/L) of the compounds of interest,
interfering compounds, and/or polymeric
materials. Some samples will not concentrate
to one mL (section 10.5): others will overload
the CC column and/or mass spectrometer.
15.2 Analyze the dilute aliquot (section
10.1) when the sample will not concentrate to
1.0 mL. If a dilute aliquot was not extracted.
and the sample holding time (section 9.3) has
not been exceeded, dilute an aliquot of the
sample with reagent water and re-extract
(section 10.1); otherwise, dilute the extract
(section 14.4) and analyze by the internal
standard method (section 14.2).
15.3 Recovery of internal standard— the
E1CP area of the internal standard should be
within a factor of two of the area in the shift
standard (section 12.1). If the absolute areas
of the labeled compounds are within a factor
of two of the respective areas in the shift
standard, and the internal standard area is
less than one-half of its respective area, then
internal standard loss in the extract has
occurred. In this case, use one of the labeled
compounds (perferably a polynuclear
aromatic hydrocarbon) to compute the
concentration of a pollutant with no labeled
analog.
15.4 Recovery of labeled compounds— in
most samples, labeled compound recoveries
will be similar to those from reagent water
(section 12.7). If the labeled compound
recovery is outside the limits given in table 8,
the dilute extract (section 10.1) is analyzed as
in section 14.4. If the recoveries of all labeled
compounds and the internal standard are low
(per the criteria above), then a loss in
instrument sensitivity is the most likely
cause. In this case, the 100 Mg/mL calibration
standard (section 12.1) shall be analyzed and
calibration verified (section 12.5). If a loss in
sensitivity has occurred, the instrument shall
be repaired, the performance specifications in
section 12 shall be met, and the extract
reanalyzed. If a loss in instrument sensitivity
has not occurred, the method does not work
on the sample being analyzed and the result
may not be reported for regulatory
compliance purposes.
76 Method performance
16.1 Intel-laboratory performance for this
method is detailed in references 9 and 10.
16.2 A chromatogram of the 100 fig/ml
acid/base/neutral calibration standard
(section 6.13) is shown in figure 6.
References
1. "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass
Spectrometry Equipment and Laboratories"
USEPA, EMSL/Cincinnati, OH 45268. EPA-
600/4-80-025 (April 1980).
2. "Working with Carcinogens," DHEW,
PHS, CDC, N1OSH, Publication 77-208. (Aug
1977).
TABLE 1 .—Base/Neutral Extractable Compounds
3. "OSHA Safety and Health Standards.
General Industry" OSHA 2208. 29 CFR 1910
()an 1976).
4. "Safety in Academic Chemistry
Laboratories. " ACS Committee on Chemical
Safety (1979).
5. "Reference Compound to Calibrate Ion
Abundance Measurement in Gas
Chromatography-Mass Spectrometry
Systems." J.W. Eichelberger, L.E. Harris, and
W.L. Budde, Anal. Chem., 47. 955 (1975).
6. "Handbook of Analytical Quality Control
in Water and Wastewater Laboratories,"
USEPA, EMSL/Cincinnati, OH 45268. EPA-
600/4-79-019 (March 1979).
7. "Standard Practice for Sampling Water."
ASTM Annual Book of Standards. ASTM,
Philadelphia. PA. 76 (1980).
8. "Methods 330.4 and 330.5 for Total
Residual Chlorine," USEPA, EMSL/
Cincinnati, OH 45268. EPA 600/4-70-020
(March 1979).
9. Colby, B.N., Beimer. R.G., Rushneck,
D.R.. and Telliard. W.A.. "Isotope Dilution
Gas Chromatography-Mass Spectrometry for
the determination of Priority Pollutants in
Industrial Effluents." USEPA, Effluent
Guidelines Division, Washington. DC 20460
(1980).
10. "Inter-laboratory Validation of US
Environmental Protection Agency Method
1625." USEPA. Effluent Guidelines Division.
Washington. DC 20480 (June 15.1984).
Compound
Acenaphtnytene
Anthracene . ...
p+naoinq
0 enrnfajanthracfl*^
BenzoWfluorantnene
Benzo(a)pyTono
Benzo(ahi)porytonfl
Btohenyl (Appendix C)
8ia(g-chtoroetnyO ether
B^f? . <«,., ,,
FHa(? rWorniaopfopyO ether
Biafg-etfiyfhftrryl) pMNrtat* , ,,
Butyl benzyl phthalato
R-C12 {Appendbt Q
n-Cl4 (Appendbt C)
n-Cl6 (Appendix C)
n-C10 (Appendbt C) .
rvC20 (Appendbt C)
n-C22 (Appendbt O .
n-C24 (Appendbt C)
n-C20 (Appendbt C)
n-C30 (Appendix C) ...
Cartoaznte (4c)
Gnrytane
OMvlxityf phthaJatt -
1 3-dtohlorobenzene ,
1 4 dteMoroberoene
3 3* dfcNoJUtiemitine
nttttri nhthalata
2.4 ilrttroloiuene
2.6-dnftrotokjene
U-n-ocM Dhtnaiate
Store!
34205
34200
34220
39120
34526
34230
34242
34247
34521
81513
34273
34278
34283
38100
34636
34292
77427
77588
77691
77757
77804
77830
77859
77888
77901
78118
78117
77571
34581
34641
34320
77356
34556
61302
39110
34536
34566
34571
34631
34336
34606
34341
34611
34626
34596
CASregntty
83-32-9
206-96-8
120-12-7
92-87-5
56-55-3
205-99-2
207-06-9
50-32-6
191-24-2
92-52-4
111-44-4
111-91-1
108-60-1
117-61-7
101-55-3
85-68-7
124-16-5
112-40-2
629-59-4
544-76-3
593-45-3
112-95-8
629-97-0
646-31-1
630-01-3
630-02-4
636-6B-6
86-74-6
91-56-7
7005-72-3
216-01-9
99-87-6
53-70-3
132-64-8
132-65-0
64-74-2
85-50-1
541-73-1
106-46-7
91-94-1
64-66-2
105-67-8
131-11-3
121-14-2
606-20-2
117-64-0
EPA-EGD
001 B
077 B
078 B
005 B
072 B
074 B
075 B
073 B
079 B
512 B
018 B
0438
042 B
066 B
041 B
087 B
517 B
506 B
518 B
519 B
520 B
521 B
5226
523 B
524 B
525 B
526 B
528 B
020 B
040 B
076 B
513 B
062B
5056
5048
068B
025 B
026B
027B
028 B
070 B
034 A
071 B
0358
036 B
069B
NPOES
001 B
002 B
003 B
004 B
005 6
007 B
009 B
006 6
008 6
011 6
0108
012 B
013 B
014 B
015 B
016 B
017 6
0188
019 B
0266
020 6
021 B
0226
0236
024 B
003 A
0256
0276
0286
0286
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 1.—Base/Neutral Extractabto Compounds-Continued
Compound
Dtohtnyl ilfr" CAiiumdli C) • ,,.,,,
« 3 iMMMtiui>Mrt» .
Fluororthont
FhiofWW
1 inMMnrnhuMtono
... " j-Tjjtma
I Inartfr m«" ir|nn wfectan*)
(nriffftofl i S-orOpyrv**
NnMHtoni
H i^itflhylMiiim (Apptndbt C) .,..—,
MtobonMnt . >
H nfrotodMvp>opv- <
fipKf ploofcn dT • •
•bftt ptooftw
eftn6MJSl Hit
n r uniMM dH ..
-• nLrfl
pht^noi ~
lilip i^mtijiQ •tfwdO
bit(«*fhlonwlhyl) Mhor
II OOLtlM Illl •
n-dsjcan* ,. -
1 !9-(jtet^orobs)nx*nsvd4
tVjtVM. itiinTinsi rU
1J-iW6kimb4iiHn«
idiH jliLmifrisiiiiiimn i>Mf dll > ..,,
b^H^itoraiMpropyO otMr
M lUJLIJllM
BBfBT^BfWfVMrfM
2 4-Jtiyi4tfiutei1>nol
nsirkt^tB^BV^MM
nartitfiiltni
n rfcttron>tlM
n-ftodMCW i< -
Mm (we)
1163
365
417
426
546
546
742
755
666
700
(66
704
666
720
722
724
737
740
756
760
766
766
616
(23
630
645
648
661
668
(21
(24
•36
(65
tf»
(63
(67
(73
(75
(53
(61
RMwwon ttiv
EOOB*
164
164
164
603
164
610
164
613
164
265
164
218
164
617
164
226
164
227
164
225
164
242
164
212
164
164
256
164
254
164
234
164
164
206
164
256
164
606
164
606
•
Rtfttv*
t 000-1000
OM6-0363
1.006-1.028
0460-0466
1.002-1.008
0.624-0.662
1008-1023
0.564-0.613
0.966-1 010
0.564-0.607
1 007-1.016
0.565-0.615
1.022-1.036
OHM 0 630
0(66-1006
0 601-0.666
0.667-1.008
0632-0667
O.M5-1.006
0664-0.681
1.010-1.016
0660-0717
0.666-1.001
0.706-0.727
1002-1007
0.747-0.767
O.N6-1.017
0781-0603
0.886-1.003
0.813-0.630
1.000-1.005
0.616-0.636
1.001-1.006
0.626-0.644
0.9*6-1.006
0.730-0906
0.666-1.051
Pturton
MHGift/L)
10
^0
50
59
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
10
10
10
10
10
10
10
10
10
1CU
1?
ll
10
10
10
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Rules and Regulations 191
TABLE 3.—GAS CHROMATOGRAPHY or BASE/NEUTRAL EXTRACT ABLE COMPOUNDS—Continued
EGO
No.1
Compound
Retention time
Mean (tec)
EGO Ret
Relative
Detection
529 1.2.3-trichloroDetuene
252 hexachtorooutadieoe-13C4
352 hexechtoroDutadiene
253 heucMorocyctoperw-13C4..
353 henachlorocycJopentadiene
220 2-chloronapr>thalene-d7
320 2-ehkxonapnthelene
518 n-tetredecane
612 Biphem>d10
712 Biphenyl
60S Diphenyt ether-d10
708 Oiphenyl ether
277 Acenaphthylene-dB
377 Acenapnthylene
271 Dimethyl phthalate-d4
371 Dimethyl phlhalate
236 2,6-dWtrotoluene-d3
338 2.6-dinitFOtoluene
201 Acenaphthene-dlO
301 Acenapnthene
605 Oioenzofuran-dS
705 Dfcenzofuran
602 Beta-riaj>rithylamine-d7
702 Beta-naphthytamine
280 Fluoreoe-d10
380 FHwrene
240 4-chkxophenyl phenyl ether-dS
340 4-chkxophenyl phenyl ether
270 Diethyl phthalate-d4
370 Diethyl phthalate
619 n-hexadecane-d34
719 n-hexadecane
235 2.4-d«trotoluene-d3
335 2,4-dWtrotoluene
237 1,2-dipheny1hydraiine-d8
337 1,2-diphenylriydrazine (*)
807 Diphenylaroine-dlO
707 Diphenylamine
262 N-nHro90diphenylamlne-d6
362 N-nitn»odiphenylamine I4)
041 4-tromophenyl phenyl ether
209 Hexachloro6anzene-13Ce
309 HexacMorooenzene
281 Phenanthrene-010
520 n-octadecane
361 Pttonfifithrono
278 Anthr»C8ne-d10
378 Anthracene
604 Dfcenzothiopnene-da
704 Dibenrothiophene
528 Carbazote
621 n-eicoaane-d42
721 noicoaine
268 DMi-butyl phthalate-d4
368 DUvbutyl phthalate
239 Fluoranthene-dIO
339 Ruoranlhane
284 Pyrene-d10
364 Pyrene
205 BenzWine-dB
305 Benzkjine
522 n-docosane
623 n-tetracosane-dSO
723 rvtetracosane
067 Butylbenryl phthalate
276 Chrysene-d12 _ „..
376 Chrysene
272 Bento(a)anlhracene-d12
372 Benzo(a)anthraotne — -..
226 3,3*-4iichlorobenzidil)e-dB
328 Sl'-dchtorobenadlne
266 B!*<2-etnylhexyl) phthalate-d4
366 Bis(2-etnylheityO phthalate
524 n-hexacosane
269 dwvoctyl phthalate^M
369 dMvocV phthalate
525 n-octacosane
274 Benzo(b)fluoranthene-d12
354 Benzo(b)fluoranthene
275 8enzo(k)tluaranthene-d12
375 BenzoOOfluoranthene
273 Benzo(a)pyrene-d12
373 Benzo(a)pyrene
626 N-triacontane-d62
726 N-triaoontane
063 lndeno<1.2.3-cd)pyrene
062 Dibonzo(t, n)AnttVKdn0
279 Benzo(ghiXierylene-d12
379 BenzotghOperylene
1003
1005
1006
1147
1142
1185
1200
1203
1205
1195
1211
1216
1265
1247
1269
1273
1283
1300
1298
1304
1331
1335
1368
1371
1395
1401
1406
1409
1409
1414
1447
1469
1359
1344
1433
1439
1437
1439
1447
1464
1498
1521
1522
1578
1580
1583
1588
1592
1559
1564
1650
1655
1677
1719
1723
1813
1817
1844
1652
1854
1853
1889
1997
2025
2060
2081
2083
2062
2090
2088
2086
2123
2124
2147
2239
2240
2272
2281
2293
2287
2293
2351
2350
2384
2429
2650
2660
2741
2750
164
164
252
164
253
164
220
164
164
612
164
608
164
277
164
271
164
236
164
201
164
605
164
602
164
281
164
240
164
270
164
619
164
235
164
237
164
607
164
262
164
164
209
164
164
281
164
278
164
604
164
164
621
164
268
164
239
164
284
164
205
164
164
612
164
164
276
164
272
164
228
164
266
164
164
269
164
164
274
164
275
164
273
164
626
164
164
164
279
n<
0.856-0.671
0.999-1.002
0.975-0986
0.999-1.001
1.014-1.024
0.997-1.007
ra
1.016-1.027
1.001-1.006
1.036-1.047
0.997-1.009
1.080-1.095
1.000-1.004
1.083-1.102
0.998-1.005
1.090-1.112
1.001-1.005
1.107-1.125
0.999-1.009
1.134-1.155
0.998-1.007
1.163-1.189
0.996-1.007
1.185-1.214
0.999-1.008
1.194-1.223
0.990-1.015
1.197-1.229
0.996-1.006
1.010-1.478
1.013-1.020
1.152-1.181
1.000-1.002
1.216-1.248
0.999-1.009
1.213-1.249
1.000-1.007
1.225-1.252
1.000-1.002
1.271-1.307
1.288-1.327
0.999-1.001
1.334-1.380
ns
1.000-1.005
1.342-1.368
0.998-1.008
1.314-1.361
1.000-1.006
ns
1.184-1.662
1.010-1.021
1.446-1.510
1.000-1.003
1.522-1.596
1.000-1.004
1.523-1.644
1.001-1.003
1.549-1.632
1.000-1.002
ns
1.671-1.764
1.012-1.015
ns
1.743-1.837
1.000-1.004
1.735-1.846
0.999-1.007
1.744-1.848
1.000-1.001
1.771-1.880
1.000-1.002
ns
1.867-1.982
1.000-1.002
ns
1.902-2.025
1.000-1.005
1.906-2.033
1.000-1.005
1.954-2.088
1.000-1.004
1.972-2.127
1.011-1.028
2.187-2.524
1.001-1.006
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
10
10
20
20
20
20
20
20
10
10
10
10
10
10
. 10
10
10
to
20
10
10
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20
20
20
-------
192
Federal Register / Vol. 49. No. 209 / Friday. October 26, 1984 / Rules and Regulations
4
landard m
•Mi 0. 1 or 5 MkMa a pollutant quanatiad by Vw mama) standard mathod: rataranca numbare bagmng vnth 2 or 8 mOicata a I
g with 3 or 7 ntoata • poftMnt quantrfwd by isotopa Ofabon
Th» • • minimum lavai it wnfch Mi arum GC/MS ayatam must gwa racogmzaMa man ipactra (background contend) and acopubto cakbrafton ponta.
«OatacMd n Ofchanylamlna
nt < spacdicatton not avataMa at Km* of ralaaaa ryl«n< eluln.
Gat »«tocity: 30 ±5 cm/«ac.
TABLE 4.—GAS CHROMATOGRAPHV OF ACID EXTRACTABLE COMPOUNDS
EGO
No. '
164
224
324
257
357
231
222
221
530
359
256
356
264
2.?-dMuorotoiptwny1 (in* vtd) . . . .
2-cNoraptMnoMM' '
2l
2 3 6>fricMoraphtnol
2 4-dMtropntnol
jJj^baSiawr/li^
2-mtthyM 6-dinikaphtnol
Ptntichtefophtnol
MMndcc)
1163
701
705
696
900
944
947
1086
1091
1162
1165
1170
1195
1323
1325
1349
1354
1433
1435
1559
1561
RMMionttn
EGO flat
1(4
1«4
224
1(4
257
1(4
231
1(4
222
1(4
221
1(4
1(4
1(4
2M
1(4
25>
1(4
2W
1(4
2(4
•
fldativ*
1 000-1 000
0.5(7-0(1(
OM7-1 010
0.7(1-07(3
09(4-1009
0(02-0122
0 997-1 006
0930-0943
0999-1003
0.994-1 005
0999-1004
1000-1005
1 147-1 175
0 997-1 OM
1.2K-1.249
1 000-1 002
1 320-1 3(3
0998-1 002
Dalaction Urnrt
•fW/L)
10
10
10
20
20
10
10
10
10
10
10
10
10
50
50
50
20
20
50
50
' Oalaranca numbara bagjnring with 0. 1 or 5 Mfcitt « poUuUnt quanWiad by Ida imam*) itandard mathod: rafaranca nun
quantifiad by Via Mamtl standard mathod; ratoranca numbara bagmning with 3 or 7 Mlcata • poKuUnt quanttwd by «otapa dkitton.
'Trm •> a mMmum lava) al wructi Via antira GC/MS ayttam mutt grv» racognaaWa mas* ipactra (background corractad) and |-
) 2 Of 6
(•oowo compound
ra.apacMcallon not avaHaMa «Ikra of rataasa of maViod.
Column: 30±2mxO^S±0.02mm id. 94% malhyl, 4% phanyl, 1% vinyl bondad phaaa tuaad iWea capillaiy.
Tamparatura program; 5 min at 30 T; 30—250 'C or untt pantacntoraphanol akitat.
Gas vakxaty 30±5 cm/sae
TABLE 5.—OfTPP MASS INTENSITY
SPECIFICATIONS
Intanaily raquiad
TABLE 6.—BASE/NEUTRAL EXTRACTABLE COM-
POUND CHARACTERISTIC MASSES—Contin-
ued
TABLE 6.—BASE/NEUTRAL EXTRACTABLE
POUND CHARACTERISTIC MASSES—Contin-^
ued
51
M
70
127
197
199
275
441
442
443
30-10 parcant of maaa 19(.
LaM than 2 parcant of man 9».
Laas V«n 2 parcant of maaa 89.
30-80 paroam of maaa 198.
Laas Vian 1 parcanl of maaa 198.
5-9 parcant of maaa 198.
10-30 parcant of ran 198.
Laas V«n ma** 443.
40-100 pareant of maaa 198.
17-23 parcanl of maaa 442.
TABLE 6.—BASE/NEUTRAL EXTRACTABLE
COMPOUND CHARACTERISTIC MASSES
Compound
AcanapnVwna
§aiiiid»ia
Damolajpyrana
Blphanyl
Bla|2-athylhaxvl) pMhataM
Butyl bantyl phViaMa
(vCIO „_
ivC12
rvC14
ivCK
Labalad
analog
010
08
010
09
012
012
012
012
012
010
08
012
d4
022
028
034
Prinmy
m/z
154/184
152/180
178/188
184/192
228/240
252/2(4
252/284
252/284
278/288
154/184
93/101
93
121/131
149/153
248
149
56/88
56788
55
56788
Compound
rvCK
n-C22
rvC24
rvC28
IVC28
(VC30
Carbarn*
J crtoronanhthalan*
Chryaana
Dibanzofuran .
Dtjanzothiophana
Dhrvbutyl pMhajata
l,2«dtehtorobanMna
i 34cNorobaraana
3 X-tfcNorobanrMkia
DknaVyl pMMM
2.4-oMtcwajana
OMvoctyl pMhaMa
D^rianylanwia
Dlphanyl athar
Rooranthana ..
H«-*«-t«..
tdanofl «2^od)pyiana
Ubalad
analog
d42
dSO
d82
d(
07
d12
dl4
di
d8
d4
d4
d4
d(
d4
d3
d4
d3
83
d4
d10
d10
d10
d10
13C8
13C4
13C
13C4
dB
Pnmary
m/z
55
567(8
55
5(7(8
55
55
55/88
187/175
182/189
228/240
114/130
278
168/176
1(4/192
149/153
146/152
149/152
252/258
149/153
122/125
183/187
184/188
185/187
149/153
189/179
170/180
202/212
1(8/178
284/292
201/204
237/241
278
(2/88
Compound
D-naipnVlytsWIWW..
Nft
M-f
Pyrana .
Styrana
1,2.3-tnehkxobanzana...
1.2,4-lhehlorobanzana...
d10
dS
d7
d10
05
03
(33
03
m/z
128/138
143/150
123/128
74
70
1897175
178/188
94/71
93/100
202/212
104/109
59/82
1(0/1(3
190/183
'Datactad as aujfianiana.
"Datactad a* dfrhanylamlna.
TABLE 7.—Acio EXTRACTABLE COMPOUND
CHARACTERISTIC MASSES
Compound
4-crHoro-3-ma*iylphanol
2-cntarophanol
2.44teNorophanol
2.44nmphanol
2-mathyM.e-drttrophanol..
2-r
ParNacnkxophanol
2.3,6-tlchlorophanol—
Z4*WcNorophanol
2.4.8«oHoraptianol.—
04
03
03
02
04
04
13C8
02
02
02
m/z
107/108
128/132
182/187
1*4/187
198/200
138/143
139/143
288/272
198/200
1987200
198/200
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 193
TABLE 8.—ACCEPTANCE CRITERIA TOR PERFORMANCE TESTS
EGO No. '
301
201
377
277
378
278
305
205
372
272
374
274
375
275
373
273
379
279
712
612
318
218
043
342
242
366
266
041
067
717
617
708
606
518
719
619
520
721
621
522
723
623
524
525
726
626
528
320
220
322
222
324
224
340
240
376
276
713
613
082
705
605
704
604
368
268
325
225
326
226
327
227
328
228
331
231
370
270
334
234
371
271
359
259
335
235
336
236
Compound
Benzidine-d8
Biphenyl-d12
Bis{2-chloroetnyl) ether-d8
rvCl 0-422
n-Cl2-d26
n-014 (Appendix C)*
n-Cl6-d34
R-C18 (Appendix C)*
rvC20-d42
n-C24-d50
n-C28 (Appendix C)"
n-C30 (Appendix C)
n-C30-d62
Carbuole (4c)*
4-cnkxophenyl pnenyl ether-dS
Dibenzothiophene (Syntuel)
Oi-n-6uty1 phthalat0-d4
3 3'-dichkxob«nzidtne-d6
2 4-dichkxoohenoW3
2 4-dimethylpneno*-d3
2 4-dinrtroto4uene-d3 •
2.6-dinrtroto*uene-d3
Acceptance criteria
Initial precision ind accuracy
section 8.2.3 («J/U
>
21
38
38
31
41
49
119
269
20
41
183
168
26
114
26
24
21
45
41
43
34
33
27
17
27
31
29
44
31
51
70
74
S3
109
33
46
39
59
34
31
11
28
35
35
32
41
38
100
41
37
111
13
24
42
52
51
69
18
67
55
20
31
31
31
15
23
17
35
43
48
42
48
26
80
12
28
44
78
13
22
36
108
18
66
18
37
30
59
X
79-134
38-147
69-186
38-146
58-174
31-194
16-518
ns-ns
65-168
25-298
32-545
11-577
59-143
15-514
62-195
35-181
72-160
29-268
75-148
28-165
55-196
29-196
43-153
81-138
35-149
69-220
32-205
44-140
19-233
24-195
ns-298
35-369
ns-331
ns-985
80-162
37-162
42-131
53-263
34-172
45-152
80-139
27-21 1
35-193
35-193
61-200
27-242
36-165
46-357
30-168
76-131
30-174
79-135
36-162
75-166
40-161
59-186
33-219
76-140
ns-359
23-299
85-136
47-136
79-150
48-130
76-165
23-195
73-146
14-212
63-201
13-203
61-194
15-193
68-174
ns-562
85-131
38-164
75-196
TO- 260
62-153
15-228
74-188
ns-640
72-134
22-308
75-158
22-245
80-141
44-184
... tfrmfmi
recovery sec. 6.3
and 14.2 P
(percent)
20-270
23-239
14-419
ns-ns
12-605
ns-ns
ns-ns
21-290
14-529
na-ns
15-372
20-260
18-364
ns-ns
ns-ns
18-308
19-306
15-376
13-479
15-324
ns-613
23-255
19-325
13-512
ns-ns
28-220
29-215
13-346
ns-494
ns-550
ns-474
ns-ns
24-260
ns-ns
ns-449
ns-ns
ns-ns
10-514
17-442
Calibration
verification sac.
12.5 (jig/ml)
80-125
71-141
60-166
66-152
60-168
56-171
34-296
ns-ns
70-142
28-357
61-164
14-ns
13-ns
13-ns
78-129
12-ns
69-145
13-ns
58-171
52-192
61-164
52-194
44-228
67-148
44-229
76-131
43-232
52-193
22-450
42-235
44-227
60-166
41-242
37-268
72-138
54-186
40-249
54-184
62-162
40-249
65-154
50-199
26-392
26-392
66-152
24-423
44-227
58-171
72-139
85-115
68-147
78-129
55-180
71-142
57-175
70-142
24-411
79-127
66-152
13-761
73-136
66-150
72-140
69-145
71-142
52-192
74-135
61-164
65-154
52-192
62-161
65-153
77-130
18-558
67-149
64-157
74-135
47-21 1
67-150
58-172
73-137
50-201
75-133
39-256
79-127
53-187
55-183
36-278
Ovjoing accuracy
sac. 11.6 R Oig/L)
72-144
30-180
61-207
33-168
50-199
23-242
11-672
ns-ns
62-176
22-329
20-ns
ns-ns
53-155
na-«85
59-206
32-194
58-168
25-303
62-176
17-267
50-213
25-222
39-166
77-145
30-169
64-232
28-224
35-172
35-170
19-237
ns-504
29-424
ns-408
ns-ns
71-181
28-202
35-167
46-301
29-198
39-195
78-142
25-229
31-212
31-212
56-215
23-274
31-188
35-442
24-204
62-159
14-314
76-138
33-176
63-194
29-212
48-221
23-290
72-147
ns-468
19-340
79-146
39-160
70-168
40-156
74-169
22-209
70-152
11-247
55-225
ns-260
53-219
11-245
64-185
ns-ns
83-135
34-182
65-222
ns-ns
60-156
14-242
67-207
ns-ns
68-141
17-378
72-164
19-275
70-159
31-250
-------
194
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 8.—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS—Continued
EGO No.1
209
607
706
608
337
339
239
360
290
309
209
352
063
254
3^0
355
255
702
602
356
256
357
257
356
256
061
063
262
364
264
361
365
265
703
603
394
294
710
610
606
529
306
206
531
321
221
Compound
Pi n mjyi ijiifialalt i)4
Otyhtnyt e9ier (Appendn C)
O^ftenyl Mn*r-d10
FhjonMhana-dlO
UMMhlnmhuiMM
I't'if^Hoirtitnitnt-'K*
hM«**Wtad«fMM
ktettf 1 7 3-^kwranf"
Jri|""ioriedO
2-iMtiyM tdl
nTMihtthtoMniM /Auu-noTii Cl
2-nlVopnenot-d4
4-nMophtnoMM
Jj^*""*Jj'*5|*j"^
N-nlbntodpntnylamnt-d*
nI|ifmrlii.rf1uUL|Jt1»l-1 •tf»
*T"*T!!^^?^TWT
Dhanol-dO
a-pmflnt (%nfutl)
OWana^
pyrtnt-dlO
TjjMfcNambamnt (4c)*
1.2.4 ktchtoraaennne
2 4 5-tncNofophtrol (4c)"
2 4 A-McNoropntnoHtt
MMpma
WCHOI
1
16
46
45
42
19
37
73
35
33
35
29
43
16
61
56
63
227
77
15
60
55
25
23
19
•4
20
39
49
33
25
29
15
23
42
186
199
199
45
37
21
49
13
40
36
161
36
136
19
29
42
49
44
4|
69
19
57
30
30
57
47
•on md accuracy
n 9.2.3 04/1)
X
77-191
12-363
56-205
27-206
62-136
36-155
49-309
31-173
71-177
36-161
61-132
51-131
90-124
39-229
51-251
nt-316
21-ra
nt-400
69-144
23-299
76-156
49-133
77-133
36-247
60-139
29-157
10-flt
nt-m
69-161
19-265
76-140
41-145
62-146
14-399
21-472
21-472
65-142
54-126
76-140
37-212
93-119
45-130
77-127
21-210
59-149
11-360
76-152
32-176
53-221
nt-261
42-234
22-292
15-229
92-139
15-212
56-137
56-137
59-205
43-193
Aootptanct cm*
Lafttttd compound
rvcovwy we. 9.3
(pvcara)
11-466
19-261
17-316
20-276
27-236
13-595
33-193
16-527
14-305
27-217
rw-nt
26-256
16-412
24-241
nt-nt
nt-flt
16-303
nt-nt
nt-672
nt-562
21-363
lit
Catbrtkon
125 (M9/n*J
21-467
50-160
63-120
77-129
75-134
67-146
47-215
74-135
61-164
76-128
36-266
74-130
66-146
47-212
47-211
13*761
70-142
52-194
69-145
56-177
73-137
71-141
39-256
44-230
65-115
46-219
77-129
61-163
55-163
35-267
40-249
40-249
59-170
77-130
42-237
75-133
67-149
66*156
46-206
60-166
31-324
76-132
46-210
66-159
44-226
54*166
20-602
60-167
76-126
61-163
56-160
56-180
61-123
69-144
On-going aocmcy
MC. 11.6 A (pg/U
10-433
21-249
77-144
29-186
40-360
64-194
30-167
70-151
36-172
65-132
23-321
43-267
nt-413
44-147
75-149
22-192
65-169
15-314
75-149
37-156
51-175
m-m
12-807
12-607
40-1961
71-fl
zi-m
87-1 2T
34-166
62-154
50-174
n> 6Q6
72*156
29-166
46-244
nt-346
16-339
11-297
77-144
10-282
51-153
51-153
46-244
34-226
0, 1 or 5 Indict* a pdutant quantified by tht mtwnal Mandard method; rttorenca number* beyreng wrtfi 2 or 6 ndttatt a labaltd compound
id imMhod: nriaranoa numbara beginning with 3 or 7 indicate a pollutant quantrted by ieotooe dilution.
quantified by the inttmal «ar
• Maaturad by Mtmai Mandtr* tptcl6ca»on danVtd trom related compound.
na-no apaclfcallon; MM it ouWdt *• range ttiat can be maaturtd reSabry.
90 COM 6690-60-M
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26,1984 / Rules and Regulations
195
10-
> vo
0.1-
I I I I I 1
2 10 20 50 100 200
CONCENTRATION (ug/niL)
(3C>
AREA * 49200
AfltA.43800
= 46300
FIGURE 1 Relative Response Calibration Curve
for Phenol The Dotted Lines Enclose a * 10
Percent Error Window.
FIGURE 3 Extracted Ion Current Profiles (or (3A)
Unlabeled Compound, (36> Labeled Compound,
and (3C) Equal Mixture of Unlabeled and Labeled
Compounds.
FIGURE 2 Extracted Ion Current Profiles foi
Chromatographically Resolved Labeled
-------
196 Federal Register / Vol. 49, No. 209 / Friday, October 26,1984 / Rules and Regulations
I10.1.1J
110.14
HO.UI
110.1.4)
(104
110.3)
(10.4J)
(10.4.10.8)
111.4)
•3TANOAHD
SAMPLE
V
ORG
N
1 L REAGENT
WATER
1
1 L REAGENT
WATER
1L ALIQUOT
1 1
SPIKE 500 JIL
OFaOOjjg/mL
ISOTOPES
i
SPIKE 1.0 ml
OF STANDARDS
1
STIR AND
EQUILIBRATE
SPIKE 500 uL
OF 200 uo/mL
ISOTOPES
^
SPIKE 900 UL
OF2DOuO/mL
ISOTOPES
e *
STIR AND
EQUILIBRATE
rANOAHO Oft BLANK
EXTRACT BASE/
NEUTRAL
WC 1
CONCENTRATE
TO 2-4 ml
L
J,
AQ
IEOUS
X
EXTRACT ACID
i
CONCENTRATE
TO 2-4 mL
1
CONCENTRATE
TO 1.0 ml
1
ADD INTERNAL
STANDARD
X
INJECT
ORG
}
»
STIR AND
EQUILIBRATE
X
EXTRACT BASE/
NEUTRAL
A 4IC AO
f
CONCtNTRATE
TO 1 0 mL
i
^
ECUS
J,
EXTRACT ACID
»
i r
CONCENTRATE
TOLOmL
i
ADD INTERNAL
STANDARD
^
ADD INTERNAL
STANDARD
1
INJtl.f
INJECT
FIGURE 4 Flow Chart tor Extraction/Concentration ol Precision and Recovery Standard, Blank,
and Sample by Method 1029. Numbers In Brackets | ) Refer to Section Numbers In the Method.
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 197
i
K
2 ~> m, .
IIIIIJIII
ANTHRACENE 0-
• ~ --*•
• * " *
1 1 1 1 1 I 1 I 1
1J345t78»10
ANALYSIS NUMBER
o
g no
z
^ I
i;
0.90
ANTHRACENE
6/1 «M W2 « M M 6M 8/S
DATE ANALYZED
FIGURE 5 Quality Control Charts Showing Area (top graph) and
Relative Response of Anthracene to Anthracene-d,0 (lower graph)
Plotted as a Function ol Time or Analysis Number.
(Mini i«Nlbiilb II SC>*6 I
Mill -*
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198 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
Appendix B to Part 136—Definition and
Procedure for the Determination of the
Method Detection Limit—Revision 1.11
Definition
The method detection limit (MDL) is
defined at the minimum concentration of a
substance that can be measured and reported
with 99% confidence that the analyte
concentration is greater than zero and is
determined from analysis of a sample in a
given matrix containing the analyte.
Scope and Application
This procedure is designed for applicability
to a wide variety of sample types ranging
from reagent (blank) water containing
analyte to wastewater containing analyte.
The MDL for an analytical procedure may
vary as a function of sample type. The
procedure requires a complete, specific, and
well defined analytical method. It is essential
that all sample processing steps of the
analytical method be included in the
determination of the method detection limit.
The MDL obtained by this procedure is
used to judge the significance of a single
measurement of a future sample.
The MDL procedure was designed for
applicability to a broad variety of physical
and chemical methods. To accomplish this.
the procedure was made device- or
instrument-independent.
Procedure
1. Make an estimate of the detection limit
using one of the following:
(a) The concentration value that
corresponds to an instrument signal/noise in
the range of 2.5 to 5.
(b) The concentration equivalent of three
times the standard deviation of replicate
instrumental measurements of the analyte in
reagent water.
(c) That region of the standard curve where
there it a significant change in sensitivity,
i.e.. a break in the slope of the standard
curve.
(d) Instrumental limitations.
It is recognized that the experience of the
analyst is important to this process.
However, the analyst must include the above
considerations in the initial estimate of the
detection limit.
2. Prepare reagent (blank) water that is as
free of analyte as possible. Reagent or
interference free water is defined as a water
sample in which analyte and interferent
concentrations are not detected at the
method detection limit of each analyte of
interest. Interferences are defined aa
systematic errors in the measured analytical
signal of an established procedure caused by
the presence of interfering species
(interferent). The interferent concentration is
presupposed to be normally distributed in
representative samples of a given matrix.
3. (a) If the MDL is to be determined in
reagent (blank) water, prepare a laboratory
standard (analyte in reagent water) at a
concentration which is at least equal to or in
the same concentration range as the
estimated method detection Unit.
(Recommend between 1 and S times the
estimated method detection limit.) Proceed to
Step 4.
(b) If the MDL is to be determined in
another sample matrix, analyze the sample. If
the measured level of the analyte is in the
recommended range of one to five times the
estimated detection limit, proceed to Step 4.
If the measured level of analyte is less than
the estimated detection limit, add a known
amount of analyte to bring the level of
analyte between one and five times the
estimated detection limit.
If the measured level of analyte is greater
than five times the estimated detection limit.
there are two options.
(1) Obtain another sample with a lower
level of analyte in the same matrix if
possible.
(2) The sample may be used as is for
determining the method detection limit if the
analyte level does not exceed 10 times the
MDL of the analyte in reagent water. The
variance of the analytical method changes as
the analyte concentration increases from the
MDL, hence the MDL determined under these
circumstances may not truly reflect method
variance at lower analyte concentrations.
4. (a) Take a minimum of seven aliquots of
the sample to be used to calculate the method
detection limit and process each through the
entire analytical method. Make all
computations according to the defined
method with final results in the method
reporting units. If a blank measurement is
required to calculate the measured level of
analyte, obtain a separate blank
measurement for each sample aliquot
analyzed. The average blank measurement is
subtracted from the respective sample
measurements.
(b) It may be economically and technically
desirable to evaluate the estimated method
detection limit before proceeding with 4a.
This will: (1) Prevent repeating this entire
procedure when the costs of analyses are
high and (2) insure that the procedure is being
conducted at the correct concentration. It is
quite possible that an inflated MDL will be
calculated from data obtained at many times
the real MDL even though the level of analyte
is less than five times the calculated method
detection limit. To insure that the estimate of
the method detection limit is a good estimate,
it is necessary to determine that a lower
concentration of analyte will not result in a
significantly lower method detection limit.
Take two aliquots of the sample to be used to
calculate the method detection limit and
process each through the entire method.
including blank measurements as described
above in 4a. Evaluate these data:
(1) If these measurements indicate the
sample Is in desirable range for
determination of the MDL, take five
additional aliquots and proceed. Use all
seven measurements for calculation of the
MDL
(2) If these measurements indicate the
sample is not in correct range, reestimate the
MDL. obtain new sample as in 3 and repeat
either 4a or 4b.
5. Calculate the variance (S») and standard
deviation (S) of the replicate measurements.
as follows:
-a ••)'/•]
(V*>»«
where:
X,; i=l to n. = are the analytical results in
the final method reporting units obtained
from the n sample aliquots and X refers
to the sum of the X values from i = l to n.
6. (a) Compute the MDL as follows:
MDL
t(n 1.1 « - 01*) (S)
where:
MDL = the method detection limit
(<„ i.i « - t»> = the students' t value
appropriate for a 99% confidence level
and a standard deviation estimate with
n-l degrees of freedom. See Table.
S = standard deviation of the replicate
analyses.
(b) The 95% confidence interval estimates
for the MDL derived in 6a are computed
according to the following equations derived
from percentiles of the chi square over
degrees of freedom distribution d'/df).
LCL = 0.64 MDL
UCL = 2.20 MDL
where: LCL and UCL are the lower and
upper 95% confidence limits respectively
based on seven aliquots.
7. Optional iterative procedure to verify the
reasonableness of the estimate of the MDL
and subsequent MDL determinations.
(a) If this is the initial attempt to compute
MDL based on the estimate of MDL
formulated in Step 1. take the MDL as
calculated in Step 0, spike in the matrix at the
calculated MDL and proceed through the
procedure starting with Step 4.
(b) If this is the second or later iteration of
the MDL calculation, use S* from the current
MDL calculation and S' from the previous
MDL calculation to compute the F-ratio. The
F-ratio is calculated by substituting the larger
S* into the numerator S*, and the others into
the denominator S V The computed F-ratio is
then compared with the F-ratio found in the
table which is 3.05 as follows: if S'J
S\<3.05, then compute the pooled standard
deviation by the following equation:
r eA+e ,
I 12 J
if S*A/SS>346. respike at the most recent
calculated MDL and process the samples
through the procedure starting with Step 4. If
the most recent calculated MDL does not
permit qualitative identification when
samples are spiked at that level, report the
MDL as a concentration between the current
and previous MDL which permits qualitative
identification.
(c) Use the Smu •• calculated in 7b to
compute the final MDL according to the
following equation:
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Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Rules and Regulations
199
MDL = 2.681
where 2.681 is equal to tot. i-o =.99).
(d) The 95% confidence limits for MDL
derived in 7c are computed according to the
following equations derived from precentiles
of the chi squared over degrees of freedom
distribution.
LCL=0.72MDL
UCL = 1.65 MDL
where LCL and UCL are the lower and upper
95% confidence limits respectively based on
14 aliquots.
TABLES OF STUDENTS' t VALUES AT THE 99
PERCENT CONFIDENCE LEVEL
Number of replicates
7
Q
g
10
11
18
21
26
61
00
Degrees
0*
freedom
(n-D
6
7
8
9
10
10
20
25
30
60
00
Ui. «>
3.143
2.998
2.896
2.821
2.764
2.602
2.528
2.485
2.457
2.390
2.326
Reporting
The analytical method used must be
specifically identified by number or title and
the MDL for each analyte expressed in the
appropriate method reporting units. If the
analytical method permits options which
affect the method detection limit, these
conditions must be specified with the MDL
value. The sample matrix used to determine
the MDL must also be identified with MDL
value. Report the mean analyte level with the
MDL and indicate if the MDL procedure was
iterated. If a laboratory standard or a sample
that contained a known amount analyte was
used for this determination, also report the
mean recovery.
If the level of analyte in the sample was
below the determined MDL or does not
exceed 10 times the MDL of the analyte in
reagent water, do not report a value for the
MDL
Appendix C to Part 136—Inductively
Coupled Plasma—Atomic Emission
Spectrometric Method for Trace Element
Analysis of Water and Wastes Method
200.7
1. Scope and Application
1.1 This method may be used for the
determination of dissolved, suspended, or
total elements in drinking water, surface
water, and domestic and industrial
wastewaters.
1.2 Dissolved elements are determined in
filtered and acidified samples. Appropriate
steps must be taken in all analyses to ensure
that potential interferences are taken into
account. This is especially true when
dissolved solids exceed 1500 mg/L. (See
section 5.)
1.3 Total elements are determined after
appropriate digestion procedures are
performed. Since digestion techniques
increase the dissolved solids content of the
samples, appropriate steps must be taken to
correct for potential interference effects. (See
section 5.)
1.4 Table 1 lists elements for which this
method applies along with recommended
wavelengths and typical estimated
instrumental detection limits using
conventional pneumatic nebulization. Actual
working detection limits are sample
dependent and as the sample matrix varies,
these concentrations may also vary. In time,
other elements may be added as more
information becomes available and as
required.
1.5 Because of the differences between
various makes and models of satisfactory
instruments, no detailed instrumental
operating instructions can be provided.
Instead, the analyst is referred to the
instruction provided by the manufacturer of
the particular instrument.
2. Summary of Method
2.1 The method describes a technique for
the simultaneous or sequential multielement
determination of trace elements in solution.
The basis of the method is the measurement
of atomic emission by an optical
spectroscopic technique. Samples are
nebulized and the aerosol that is produced is
transported to the plasma torch where
excitation occurs. Characteristic atomic-line
emission spectra are produced by a radio-
frequency inductively coupled plasma (ICP).
The spectra are dispersed by a grating
spectrometer and the intensities of the lines
are monitored by photomultiplier rubes. The
photocurrents from the photomultiplier tubes
are processed and controlled by a computer
system. A background correction technique is
required to compensate for variable
background contribution to the determination
of trace elements. Background must be
measured adjacent to analyte lines on
samples during analysis. The position
selected for the background intensity
measurement, on either or both sides of the
analytical line, will be determined by the
complexity of the spectrum adjacent to the
analyte line. The position used must be free
of spectral interference and reflect the same
change in background intensity as occurs at
the analyte wavelength measured.
Background correction is not required in
cases of line broadening where a background
correction measurement would actually
degrade the analytical result. The possibility
of additional interferences named in 5.1 (and
tests for their presence as described in 5.2)
should also be recognized and appropriate
corrections made.
3. Definitions
3.1 Dissolved—Those elements which
will pass through a 0.45 /im membrane filter.
3.2 Suspended—Those elements which
are retained by a 0.45 jim membrane filter.
3.3 Total—The concentration determined
on an unaltered sample following vigorous
digestion (Section 9.3), or the sum of the
dissolved plus suspended concentrations.
(Section 9.1 plus 9.2).
3.4 TotoJ recoverable—The concentration
determined on an unaltered sample following
treatment with hot, dilute mineral acid
(Section 9.4).
3.5 Instrumental detection limit—The
concentration equivalent to a signal, due to
the analyte. which is equal to three times the
standard deviation of a series of ten replicate
measurements of a reagent blank signal at
the same wavelength.
3.6 Sensitivity—The slope of the
analytical curve, i.e. functional relationship
between emission intensity and
concentration.
3.7 Instrument check standard—A
multielement standard of known
concentrations prepared by the analyst to
monitor and verify instrument performance
on a daily basis. (See 7.6.1)
3.8 interference check sample—A
solution containing both interfering and
analyte elements of known concentration
that can be used to verify background and
interelement correction factors. (See 7.6.2.)
3.9 Quality control sample—A solution
obtained from an outside source having
known, concentration values to be used to
verify the calibration standards. (See 7.6.3)
3.10 Calibration standards—A series of
known standard solutions used by the
analyst for calibration of the instrument (i.e..
preparation of the analytical curve). (See 7.4)
3.11 Linear dynamic range—The
concentration range over which the
analytical curve remains linear.
3.12 Reagent blank—A volume of
deionized, distilled water containing the
same acid matrix as the calibration standards
carried through the entire analytical scheme.
(See 7.5.2)
3.13 Calibration blank—A volume of
deionized, distilled water acidified with
HNQ, and HC1. (See 7.5.1)
3.14 Method of standard addition—The
standard addition technique involves the use
of the unknown and the unknown plus a
known amount of standard. (See 10.6.1.)
4. Safety
4.1 The toxicity of carcinogenicity of each
reagent used in this method has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. From this viewpoint exposure
to these chemicals must be reduced to the
lowest possible level by whatever means
available. The laboratory is repsonsible 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 data
handling sheets should also be made
available to all personnel involved in the
chemical analysis. Additional references to
laboratory safety are available and have
been identified "*'• "••"" "•• for the
information of the analyst.
5. Interferences
5.1 Several types of interference effects
may contribute to inaccuracies in the
determination of trace elements. They can be
summarized as follows:
5.1.1 Spectral interferences can be
categorized as (1) overlap of a spectral line
from another element; (2) unresolved overlap
of molecular band spectra; (3) background
contribution from continuous or
recombination phenomena; and (4)
background contribution from stray light from
-------
200 Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
the line emission of high concentration
elements. The first of these effects can be
compensated by utilizing a computer
correction of the raw data, requiring the
monitoring and measurement of the
interfering element. The second effect may
require selection of an alternate wavelength.
The third and fourth effects can usually be
compensated by a background correction
adjacent to the analyte line. In addition, users
of simultaneous multi-element
instrumentation must assume the
responsibility of verifying the absence of
spectral interferences from an element that
could occur in a sample but for which there is
no channel in the instrument array. Listed in
Table 2 are some interference effects for the
recommended wavelengths given in Table 1.
The data in Table 2 are intended for use only
as • rudimentary guide for the indication of
potential spectral interferences. For this
purpose, linear relations between
concentration and intensity for the analytes
and the interferents can be assumed. The
Interference information, which was
collected at the Ames Laboratory,' is
expressed as analyte concentration
equivalents (i.e. false analyte concentrations)
arising from 100 mg/L of the interferent
element. The suggested use of this
information is as follows: Assume that
arsenic (at 193.606 run) is to be determined in
a sample containing approximately 10 mg/L
of aluminum. According to Table 2.100 mg/L
of aluminum would yield a false signal for
arsenic equivalent to approximately 1.3 mg/L.
Therefore, 10 mg/L of aluminum would result
in a false signal for arsenic equivalent to
approximately 0.13 mg/L The reader is
cautioned that other analytical systems may
exhibit somewhat different levels of
interference than those shown in Table 2, and
that the interference effects must be
evaluated for each individual system.
Only those interferents listed were
investigated and the blank spaces in Table 2
indicate that measurable interferences were
not observed for the interferent
concentrations listed in Table 3. Generally,
interferences were discernible if they
produced peaks or background shifts
corresponding to 2-5% of the peaks generated
by the analyte concentrations also listed in
Table 3.
At present, information on the listed silver
and potassium wavelengths are not available
but it has been reported that second order
energy from the magnesium 383.231 nm
wavelength interferes with the listed
potassium line at 766.491 nm.
5.1.2 Physical interference! are generally
considered to be effects associated with the
sample nebulization and transport processes.
Such properties as change in viscosity and
surface tension can cause significant
inaccuracies especially in samples which
may contain high dissolved solids and/or
add concentrations. The use of a peristaltic
pump may lessen these interferences. If these
types of interferences are operative, they
must be reduced by dilution of the sample
and/or utilization of standard addition
techniques. Another problem which can
1 Amn Ubontoiy. USDOE. Iowa Stilt
University. Ames lows 30011.
occur from high dissolved solids is salt
buildup at the tip of the nebulizer. This
affects aersol flow rate causing instrumental
drift. Wetting the argon prior to nebulization.
the use of a tip washer, or sample dilution
have been used to control this problem. Also.
it has been reported that better control of the
argon flow rate improves instrument
performance. This is accomplished with the
use of mass flow controllers.
5.1.3 Chemical Interferences an
characterized by molecular compound
formation, ionization effects and solute
vaporization effects. Normally these effects
are not pronounced with the ICP technique,
however, if observed they can be minimized
by careful selection of operating conditions
(that is, incident power, observation position,
and so forth), by buffering of the sample, by
matrix matching, and by standard addition
procedures. These types of interferences can
be highly dependent on matrix type and the
specific analyte element.
5.2 It is recommended that whenever a
new or unusual sample matrix is
encountered, a series of tests be performed
prior to reporting concentration data for
analyte elements. These tests, as outlined in
5.2.1 through 5.2.4, will ensure the analyst
that neither positive nor negative interference
effects are operative on any of the analyte
elements thereby distorting the accuracy of
the reported values.
5.2.1 Serial dilution—If the analyte
concentration is sufficiently high (minimally a
factor of 10 above the instrumental detection
limit after dilution), an analysis of a dilution
should agree within 5 percent of the original
determination (or within some acceptable
control limit (14.3) that has been established
for that matrix.). If not. a chemical or physical
interference effect should be suspected.
S.2.2 Spike addition— The recovery of a
spike addition added at a minimum level of
10X the instrumental detection Unit
(maximum 100X) to the original
determination should be recovered to within
90 to 110 percent or within the established
control limit for that matrix. If not, a matrix
effect should be suspected. The use of a
standard addition analysis procedure can
usually compensate for this effect.
Caution: The standard addition technique
does not detect coincident spectral overlap. If
suspected, use of computerized
compensation, an alternate wavelength, or
comparison with an alternate method is
recommended (See 5.2.3).
5.2.3 Companion with alternate method
of analysis—When investigating a new
sample matrix, comparison tests may be
performed with other analytical techniques
such as atomic absorption spectrometry, or
other approved methodology.
5.2.4 Wavelength scanning of analyte line
region—-If the appropriate equipment is
available, wavelength scanning can be
performed to detect potential spectral
interferences.
A Apparatus
6.1 Inductively Coupled Plasma-Atomic
Emission Spectrometer.
6.1.1 Computer controlled atomic
emission spectrometer with background
correction.
6.1.2 Radiofrequency generator.
6.1.3 Argon gas supply, welding grade or
better.
6.2 Operating conditions—Because of the
differences between various makes and
models of satisfactory instruments, no
detailed operating instructions can be
provided. Instead, the analyst should follow
the instructions provided by the manufacturer
of the particular instrument. Sensitivity,
instrumental detection limit, precision, linear
dynamic range, and interference effects must
be investigated and established for each
individual analyte line on that particular
instrument. It is the responsibility of the
analyst to verify that the instrument
configuration and operating conditions used
satisfy the analytical requirements and to
maintain quality control data confirming
instrument performance and analytical
results.
7. Reagents and Standards
7.1 Acids used in the preparation of
standards and for sample processing must be
ultra-high purity grade or equivalent.
Redistilled acids are acceptable.
7.1.1 Acetic acid, cone, (sp gr 1.06).
7.1.2 Hydrochloric acid. cone, (sp gr 1.19).
7.1.3 Hydrochloric acid. (1+1): Add 500
mL cone. HC1 (sp gr 1.19) to 400 mL deionized,
distilled water and dilute to 1 liter.
7.1.4 Nitric acid, cone, (sp gr 1.41).
7.1.5 Nitric acid. (1 +1): Add 500 mL cone.
HNOi (sp gr 1.41) to 400 mL deionized,
distilled water and dilute to 1 liter.
7.2 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. The purity of this water must
be equivalent to ASTM Type n reagent water
of Specification D1193 (14.0).
7.3 Standard stock solutions may be
purchased or prepared from ultra high purity
grade chemicals or metals. All salts must be
dried for 1 h at 105 *C unless otherwise
specified.
(CAUTION: Many metal salts are
extremely toxic and may be fatal if
swallowed. Wash hands thoroughly after
handling.)
Typical stock solution preparation
procedures follow:
7.3.1 Aluminum solution, stock, 1 mL«pg
Al: Dissolve 0.100 g of aluminum metal in an
acid mixture of 4 mL of (1+1) HO and 1 mL
of cone. HNO> in a beaker. Warm gently to
effect solution. When solution is complete,
transfer quantitatively to a liter flask add an
additional 10 mL of (1+1) HC1 and dilute to
1,000 mL with deionized, distilled water.
7.3.2 Antimony solution stock, lmL-100
fig Sb: Dissolve 02869 g K(SbO)CJi.O. in
deionized distilled water, add 10 mL (1+1)
HC1 and dilute to 1,000 mL with deionized.
distilled water.
7.3.3 Arsenic solution, stack, 1 mL»100
jig As: Dissolve 0.1320 g of AstO* in 100 mL of
deionized, distilled water containing 0.4 g
NaOH. Acidify the solution with 2 mL cone.
HNO, and dilute to 1.000 mL with deionized,
distilled water.
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Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 201
7.3.4 Barium solution, stock, 1 mL=100 fig
Ba: Dissolve 0.1516 g Bad, (dried at 250 'C
for 2 hrs) in 10 mL deionized. distilled water
with 1 mL (1 + 1) HCI. Add 10.0 mL (1 + 1) HC1
and dilute to 1.000 with mL deionized,
distilled water.
7.3.5 Beryllium solution, stock, 1 mL = 100
fig Be: Do not dry. Dissolve 1.966 g
BeSCMHtO. in deionized, distilled water.
add 10.0 mL cone. HNOj and dilute to 1.000
mL with deionized. distilled water.
7.3.6 Boron solution, stock. 1 mL=100 fig
B: Do not dry. Dissolve 0.5716 g anhydrous
HiBQj in deionized, distilled water and dilute
to 1,000 mL. Use a reagent meeting ACS
specifications, keep the bottle tightly
stoppered and store in a desiccator to
prevent the entrance of atmospheric
moisture.
7.3.7 Cadmium solution, stock, 1 mL=100
^g Cd: Dissolve 0.1142 g CdO in a minimum
amount of (1 + 1) HNOs. Heat to increase rate
of dissolution. Add 10.0 mL cone. HNOj and
dilute to 1.000 mL with deionized, distilled
water.
7.3.8 Calcium solution, stock, 1 mL=100
fig Ca: Suspend 0.2498 g CaCOj dried at 180
*C for 1 h before weighing in deionized.
distilled water and dissolve cautiously with a
minimum amount of (1+1) HNOj. Add 10.0
mL cone. HNOj and dilute to 1,000 mL with
deionized, distilled water.
7.3.9 Chromium solution, stock, 1 mL=100
fig Cr: Dissolve 0.1923 g of CrOj in deionized.
distilled water. When solution is complete,
acidify with 10 mL cone. HNOj and dilute to
1,000 mL with deionized, distilled water.
7.3.10 Cobalt solution, stock, 1 mL=100
fig Co: Dissolve 0.1000 g of cobalt metal in a
minimum amount of (1 + 1) HNOs. Add 10.0
mL (1 + 1) HCI and dilute to 1,000 mL with
deionized, distilled water.
7.3.11 Copper solution, stock, 1 ml=100
fig Cu: Dissolve 0.1252 g CuO in a minimum
amount of (1+1) HNO,. Add 10.0 mL cone.
HNOj and dilute to 1.000 mL with deionized,
distilled water.
7.3.12 Iron solution, stock, 1 mL=100 fig
Fe: Dissolve 0.1430 g Fe»Oj in a warm mixture
of 20 mL (1+1) HCI and 2 mL of cone. HNOj.
Cool, add an additional 5 mL of cone. HNOj
and dilute to 1,000 mL with deionized,
distilled water.
7.3.13 Lead solution, stock, 1 mL=100 fig
Pb: Dissolve 0.1599 g Pb(NOj)a in a minimum
amount of (1 + 1) HNOs. Add 10.0 mL cone.
HNO> and dilute to 1,000 mL with deionized,
distilled water.
7.3.14 Magnesium solution, stock, 1
mL=100 fig Mg: Dissolve 0.1658 g MgO in a
minimum amount of (1 + 1) HNO>. Add 10.0
mL cone. HNO, and dilute to 1,000 mL with
deionized, distilled water.
7.3.15 Manganese solution, stock, 1
mL=100 fig Mn: Dissolve 0.1000 g of
manganese metal in the acid mixture 10 mL
cone. HCI and 1 mL cone. HNCS, and dilute to
1.000 mL with deionized, distilled water.
7.3.16 Molybdenum solution, stock, 1
mL=100 fig Mo: Dissolve 0.2043 g
(NH4jjMoO4 in deionized. distilled water and
dilute to 1,000 mL.
7.3.17 Nickel solution, stock. lmL=100
fig Ni: Dissolve 0.1000 g of nickel metal in 10
mL hot cone. HNOj, cool and dilute to 1.000
mL with deionized, distilled water.
7.3.18 Potassium solution, stock, 1
mL = 100 fig K: Dissolve 0.1907 g KC1. dried at
110 "C, in deionized, distilled water and
dilute to 1,000 mL.
7.3.19 Selenium solution, stock, 1 mL=100
ug Se: Do not dry. Dissolve 0.1727 g HiSeOj
(actual assay 94.6%) in deionized, distilled
water and dilute to 1.000 mL.
7.3.20 5/7/co solution, stock, 1 mL = 100 fig
SiOi: Do not dry. Dissolve 0.4730 g Na,SiOj
•9HiO in deionized, distilled water. Add 10.0
mL cone. HNOj and dilute to 1,000 mL with
deionized, distilled water.
7.3.21 Silver solution, stock, 1 mL=100 fig
Ag: Dissolve 0.1575 g AgNOj in 100 mL of
deionized, distilled water and 10 mL cone.
HNOj. Dilute to 1,000 mL with deionized,
distilled water.
7.3.22 Sodium solution, stock. 1 mL = 100
fig Na: Dissolve 0.2542 g NaCl in deionized.
distilled water. Add 10.0 mL cone. HNOj and
dilute to 1,000 mL with deionized, distilled
water.
7.3.23 Thallium solution, stock. 1 mL=100
fig Tl: Dissolve 0.1303 g T1NO, in deionized,
distilled water. Add 10.0 mL cone. HNOj and
dilute to 1,000 mL with deionized. distilled
water.
7.3.24 Vanadium solution, stock. 1
mL=100 fig V: Dissolve 0.2297 NH^VOj in a
minimum amount of cone. HNOj. Heat to
increase rate of dissolution. Add 10.0 mL
cone. HNOi and dilute to 1,000 mL with
deionized. distilled water.
7.3.25 Zinc solution, stock, 1 mL=100 fig
Zn: Dissolve 0.1245 g ZnO in a minimum
amount of dilute HNOj. Add 10.0 mL cone.
HNOj and dilute to 1,000 mL deionized,
distilled water.
7.4 Mixed calibration standard
solutions—Prepare mixed calibration
standard solutions by combining appropriate
volumes of the stock solutions in volumetric
flasks. (See 7.4.1 thru 7.4.5) Add 2 mL of •
(1 +1) HNOj and 10 mL of (1+1) HCI and
dilute to 100 mL with deionized, distilled
water. (See Notes 1 and 8.) Prior to preparing
the mixed standards, each stock solution
should be analyzed separately to determine
possible spectral interference or the presence
of impurities. Care should be taken when
preparing the mixed standards that the
elements are compatible and stable. Transfer
the mixed standard solutions to a FEP
fluorocarbon or unused polyethylene bottle
for storage. Fresh mixed standards should be
prepared as needed with the realization that
concentration can change on aging.
Calibration standards must be initially
verified using a quality control sample and
monitored weekly for stability (See 7.6.3).
Although not specifically required, some
typical calibration standard combinations
follow when using those specific wavelengths
listed in Table 1.
7.4.1 Mixed standard solution I—
Manganese, beryllium, cadmium, lead, and
zinc.
7.4.2 Mixed standard solution //—Barium.
copper, iron, vanadium, and cobalt.
7.4.3 Mixed standard solution Ill-
Molybdenum, silica, arsenic, and selenium.
7.4.4 Mixed standard solution IV—
Calcium, sodium, potassium, aluminum,
chromium and nickel.
7.4.5 Mixed standard solution V—
Antimony, boron, magnesium, silver, and
thallium.
Note 1.—If the addition of. silver to the
recommended acid combination results in an
initial precipitation, add 15 mL of deionized
distilled water and warm the flask until the
solution clears. Cool and dilute to 100 mL
with deionized. distilled water. For this acid
combination the silver concentration should
be limited to 2 mg/L. Silver under these
conditions is stable in a tap water matrix for
30 days. Higher concentrations of silver
require additional HCI.
7.5 Two types of blanks are required for
the analysis. The calibration blank (3.13) is
used in establishing the analytical curve
while the reagent blank (3.12) is used to
correct for possible contamination resulting
from varying amounts of the acids used in the
sample processing.
7.5.1 The calibration blank is prepared by
diluting 2 mL of (1+1) HNOj and 10 mL of
(1+1) HCI to 100 mL with deionized. distilled
water. (See Note 6.) Prepare a sufficient
quantity to be used to flush the system
between standards and samples.
7.5.2 The reagent blank must contain all
the reagents and in the same volumes as used
in the processing of the samples. The reagent
blank must be carried through the complete
procedure and contain the same acid
concentration in the final solution as the
sample solution used for analysis.
7.6 In addition to the calibration
standards, an instrument check standard
(3.7), an interference check sample (3.8) and a
quality control sample (3.9) are also required
for the analyses.
7.6.1 The instrument check standard is
prepared by the analyst by combining
compatible elements at a concentration
equivalent to the midpoint of their respective
calibration curves. (See 12.1.1.)
7.6.2 The interference check sample is
prepared by the analyst in the following
manner. Select a representative sample
which contains minimal concentrations of the
analytes of interest but known concentration
of interfering elements that will provide an
adequate test of the correction factors. Spike
the sample with the elements of interest at
the approximate concentration of either 100
fig/L or 5 times the estimated detection limits
given in Table 1. (For effluent samples of
expected high concentrations, spike at an
appropriate level.) If the type of samples
analyzed are varied, a synthetically prepared
sample may be used if the above criteria and
intent are met. A limited supply of a synthetic
interference check sample will be available
from the Quality Assurance Branch of EMSL-
Cincinnati. (See 12.1.2).
7.8.3 The quality control sample should
be prepared in the same acid matrix as the
calibration standards at a concentration near
1 mg/L and in accordance with the
instructions provided by the supplier. The
Quality Assurance Branch of EMSL-
Cincinnati will either supply a quality control
sample or information where one of equal
quality can be procured. (See 12.1.3.)
-------
202 Federal Regirter / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
8. Sample Handling and Preservation
8.1 For the determination of trace
elements, contamination and IOM are of
prime concern. Duct in the laboratory
environment, impurities in reagents and
impurities on laboratory apparatus which the
sample contacts are all sources of potential
contamination. Sample containers can
introduce either positive or negative errors in
the measurement of trace elements 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.
Laboratory glassware including the sample
bottle (whether polyethylene, polyproplyene
or FEP-fluorocarbon) should be thoroughly
washed with detergent and tap water rinsed
with (1+1) nitric acid, tap water, (1+1)
hydrochloric acid, tap and finally deionized.
distilled water in that order (See Notes 2 and
3).
Note 2.—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,
NOCHROMOC. available from Godax
Laboratories, 6 Varick St.. New York, NY
10013, may be used in place of chromic acid.
Chromic acid should not be used with plastic
bottles.
Note 3.—If it can be documented through
an active analytical quality control program
using spiked samples and reagent blanks,
that certain steps in the cleaning procedure
are not required for routine samples, those
steps may be eliminated from the procedure.
&2 Before collection of the sample a
decision must be made as to the type of data
desired, that is dissolved, suspended or total.
so that the appropriate preservation and
pretrtatment steps may be accomplished.
Filtration, add preservation, etc.. are to be
performed at the time the sample is collected
or as soon as possible thereafter.
8.2.1 For the determination of dissolved
elements the sample must be filtered through
a 0.45-fim membrane filter as soon as
practical after collection. (Glass or plastic
filtering apparatus 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) HNO»
to a pH of 2 or less. Normally, 3 mL of (1+1)
acid per liter should be sufficient to preserve
the sample.
8,22 For the determination of suspended
elements a measured volume of unpreserved
sample must be filtered through a 0.45-pm
membrane filter as soon as practical after
collection. The filter plus suspended material
should be transferred to a suitable container
for storage and/or shipment. No preservative
is required.
&Z3 For the determination of total or
total recoverable elements, the sample is
acidified with (1+1) HNO, to pH 2 or less as
soon as possible, preferably at the time of
collection. The sample is not filtered before
processing.
9. Sample Preparation
9.1 For the determinations of dissolved
elements, the filtered, preserved sample may
often be analyzed as received. The acid
matrix and concentration of the samples and
calibration standards must be the same. (See
Note 6.) If a precipitate formed upon
acidification of the sample or during transit
or storage, it must be redissolved before the
analysis by adding additional acid and/or by
heat as described in 9.3.
9.2 For the determination of suspended
elements, transfer the membrane filter
containing the insoluble material to a ISO-mL
Griffin beaker and add 4 mL cone. HNO».
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. HNOi. Cover and continue heating until
the digestion is complete, generally indicated
by a light colored digestate. Evaporate to
near dryness (2 mL). cool, and 10 mL HC1
(1+1) and IS mL deionized. distilled water
per 100 mL dilution and warm the beaker
gently for 15 min. to dissolve any precipitated
or residue material. Allow to cool, wash
down the watch glass and beaker walls with
deionized distilled water and filter the
sample to remove insoluble material that
could clog the nebulizer. (See Note 4.) Adjust
the volume based on the expected
concentrations of elements present. This
volume will vary depending on the elements
to be determined (See Note 6). The sample is
now ready for analysis. Concentrations so
determined shall be reported as "suspended."
Note &—In place of filtring. the sample
after diluting and mixing may be centrifuged
or allowed to settle by gravity overnight to
remove insoluble material.
9.3 For the determination of total
elements, choose a measured volume of the
well mixed add preserved sample
appropriate for the expected level of
elements and transfer to a Griffin beaker.
(See Note 5.) Add 3 mL of cone. HNO,. Place
the beaker on a hot plate and evaporate to
near dryness cautiously, making certain that
the sample does not boil and that no area of
the bottom of the beaker is allowed to go dry.
Cool the beaker and add another 5 mL
portion of cone. HNOj. Cover the beaker with
a watch glass and return to the hot plate.
Increase the temperature of the hot plate so
that a gently reflux action occurs. Continue
heating, adding additional add 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 10 mL
of 1+1 Hd and 15 mL of deionized, distilled
water per 100 mL of final solution and warm
the beaker gently for 15 min. to dissolve any
precipitate or residue resulting from
evaporation. Allow to cool, wash down the
beaker walls and watch glass with deionized
distilled water and filter the sample to
remove insoluble material that could dog the
nebulizer. (See Note 4.) Adjust the sample to
a predetermined volume based on the
expected concentrations of elements present
The sample is now ready for analysis (See
Note a). Concentrations so determined shall
be reported as "total."
Note 5.—If low determinations of boron are
critical, quartz glassware should be used.
Note 8.—If the sample analysis solution
has a different acid concentration from that
given in 9.4. but does not introduce a physical
interference or affect the analytical result, the
same calibration standards may be used.
9.4 For the determination of total
recoverable elements, choose a measured
volume of a well mixed, acid preserved
sample appropriate for the expected level of
elements and transfer to a Griffin beaker.
(See Note 5.) Add 2 mL of (1 +1) HNO> and 10
mL of (1+1) HC1 to the sample and heat on a
steam bath or hot plate until the volume has
been reduced to near 25 mL making certain
the sample does not boil. After this treatment.
cool the sample and filter to remove insoluble
material that could clog the nebulizer. (See
Note 4.) Adjust the volume to 100 mL and
mix. The sample is now ready for analysis.
Concentrations so determined shall be
reported as "total."
10. Procedure
10.1 Set up instrument with proper
operating parameters established in Section
6.2. The instrument must be allowed to
become thermally stable before beginning.
This usually requires at least 30 min. of
operation prior to calibration.
10.2 Initiate appropriate operating
configuration of computer.
10.3 Profile and calibrate instrument
according to instrument manufacturer's
recommended procedures, using the typical
mixed calibration standard solutions
described in Section 7.4. Flush the system
with the calibration blank (7.5.1) between
each standard. (See Note 7.) (The use of the
average intensity of multiple exposures for
both standardization and sample analysis
has been found to reduce random error.)
Note 7.—For boron concentrations greater
than 500 jig/L extended flush times of 1 to 2
minutes may be required.
10.4 Before beginning the sample run,
reanalyze the highest mixed calibration
standard as if it were a sample.
Concentration values obtained should not
deviate from the actual values by more than
±5 percent (or the established control limits
whichever is lower). If they do. follow the
recommendations of the instrument
manufacturer to correct for this condition.
10.5 Begin the sample run flushing the
system with the calibration blank solution
(7.5.1) between each sample. (See Note 7.)
Analyze the instrument check standard (7.8.1)
and the calibration blank (7.5.1) each 10
samples.
10.8 If it has been found that methods of
standard addition an required, the following
procedure is recommended.
10.8.1 The standard addition technique
(M.2) involves preparing new standards in
the sample matrix by adding known amounts
of standard to one or more aliquots of the
processed sample solution. This technique
compensates for a sample constituted that
enhances or depresses the analyte signal thus
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations 203
producing a different slope from that of the
calibration standards. It will not correct for
additive interference which causes a baseline
shift. The simplest version of this technique is
the single-addition method. The procedure is
as follows. Two identical aliquots of the
sample solution, each of volume V,. are
taken. To the first (labeled A) is added a
small volume V, of a standard analyte
solution of concentration c,. To the second
(labeled B) is added the same volume V, of
the solvent. The analytical signals of A and B
are measured and corrected for nonanalyte
signals. The unknown sample concentration
c,is calculated:
c,=
SBV.c.
A-SB) V.
where S* and SB are the analytical signals
(corrected for the blank) of solutions A and B,
respectively. V, and c, should be chosen so
that SA is roughly twice SB on the average. It
is best if V. is made much less than V,, and
thus c. is much greater than c,, to avoid
excess dilution of the sample matrix. If a
separation or concentration step is used, the
additions are best made first and carried
through the entire procedure. For the results
from this technique to be valid, the following
limitations must be taken into consideration:
1. The analytical curve must be linear.
2. The chemical form of the analyte added
must respond the same as the analyte in the
sample.
3. The interference effect must be constant
over the working range of concern.
4. The signal must be corrected for any
additive interference.
11. Calculation
11.1 Reagent blanks (7.5.2) should be
subtracted from all samples. This is
particularly important for digested samples
requiring large quantities of acids to complete
the digestion.
11.2 If dilutions were performed, the
appropriate factor must be applied to sample
values.
11.3 Data should be rounded to the
thousandth place and all results should be
reported in mg/L up to three significant
figures.
12. Quality Control (Instrumental)
12.1 Check the instrument standardization
by analyzing appropriate quality control
check standards as follow:
12.1.1 Analyze and appropriate
instrument check standard (7.6.1) containing
the elements of interest at a frequency of 10%.
This check standard is used to determine
instrument drift. If agreement is not within
±5% of the expected values or within the
established control limits, whichever is
lower, the analysis is out of control. The
analysis should be terminated, the problem
corrected, and the instrument recalibrated.
Analyze the calibration blank (7.5.1) at a
frequency of 10%. The result should be within
the established control limits of 2 standard
deviations of the mean value. If not, repeat
the analysis two more times and average the
three results. If the average is not within the
control limit, terminate the analysis, correct
the problem and recalibrate the instrument.
12.1.2 To verify interelement and
background correction factors analyze the
interference check sample (7.6.2) at the
beginning, end, and at periodic intervals
throughout the sample run. Results should fall
within the established control limits of 1.5
times the standard deviation of the mean
value. If not, terminate the analysis, correct
the problem and recalibrate the instrument.
12.1.3 A quality control sample (7.6.3)
obtained from an outside source must first be
used for the initial verification of the
calibration standards. A fresh dilution of this
sample shall be analyzed every week
thereafter to monitor their stability. If the
results are not within ±5% of the true value
listed for the control sample, prepare a new
calibration standard and recalibrate the
instrument. If this does not correct the
problem, prepare a new stock standard and a
new calibration standard and repeat the
calibration.
13. Precision and Accuracy
13.1 In an EPA round robin phase 1 study,
even laboratories applied the ICP technique
to acid-distilled water matrices that had been
dosed with various metal concentrates. Table
4 lists the true value, the mean reported value
and the mean % relative standard deviation.
14. References
14.1 Winge. R.K., V.J. Peterson, and V.A.
Fassel. "Inductively Coupled Plasma-Atomic
Emission Spectroscopy: Prominent Lines,
EPA-600/4-79-017.
14.2 Winefordner, J.D., "Trace Analysis:
Spectroscopic Methods for Elements,"
Chemical Analysis. Vol, 46, pp. 41-42.
14.3 Handbook for Analytical Quality
Control in Water and Wastewater
Laboratories, EPA-000/4-79-019.
14.4 Carbarino, J.R. and Taylor. H.E.. "An
Inductively-Coupled Plasma Atomic Emission
Spectrometric Method for Routine Water
Quality Testing," Applied Spectroscopy 33.
No. 3 (1979).
14.5 "Methods for Chemical Analysis of
Water and Wastes," EPA-600/4-79-020.
14.6 Annual Book of ASTM Standards.
Part 31.
14.7 "Carcinogens—Working With
Carcinogens," Department of Health.
Education, and Welfare. Public Health
Service. Center for Disease Control. National
Institute for Occupational Safety and Health,
Publication No. 77-206. Aug. 1977.
14.8 "OSHA Safety and Health
Standards, General Industry," (29 CFR 1910).
Occupational Safety and Health
Administration, OSHA 2206, (Revised.
January 1976).
14.9 "Safety in Academic Chemistry
Laboratories. American Chemical Society
Publication, Committee on Chemical Safety.
3rd Edition, 1979.
TABLE 1.—RECOMMENDED WAVELENGTHS '
and Estimated Instrumental Detection Limits
Element
Arsenic
Barium.
Boron... . .
Cadmium
Calcium
Cobalt..
Iron
Load
Magnesium
Molybdenum
Nickel
Selenium .
Silica (SO,)
Silver
Sodkjm
Thallium
Vanadun
Zinc
Wave-
length,
nm
308215
193696
206 833
455403
313 042
249 773
226502
317933
267 716
226616
324 754
259940
220353
279 079
257610
202030
231 604
766491
196026
288 156
328068
$93995
190864
292402
213856
Estimated
detection
limn.
cg'L'
53
32
2
03
5
10
7
7
3
42
30
2
g
15
i
75
58
7
29
40
9
2
'The wavelengths listed are recommended because of
their sensitivity and overall acceptance. Other wavelengths
may be substituted H they can provide the needed sensitivity
and are treated with the same corrective techniques tor
spectral interference. (See 5.1.1|.
'The estimated instrumental detection limits as shown are
taken from "Inductively Coupled Plasma-Atomic Emission
Spectroscopy-Prominent Lines." EPA-600/4-79-017. They
are given as a guide for an instrumental Hmil The actual
method detection limits are sample dependent and may vary
as the sample matrix_ varies.
'Highly doponctont on oporstmg conditions &nd
position.
TABLE 1.—ANALYTE CONCENTRATION EQUIVALENTS (MG/L) ARISING FROM INTERFERENTS AT THE 100 MG/L LEVEL
Analyte
Iron
Wave-
length.
nm
308.214
206.833
193.698
455.403
313.042
249.773
226.502
317.933
267716
226.616
324754
259.940
I nt8nW8«lt™"-
A1
0.47
1.3
0.04
Ca
Cr
29
044
0.06
0.03
Cu
Fa
0.08
0.32
0.03
0.01
0.003
0.005
0.003
Mg
0.01
„ ,^.
Mn
0.21
0.04
0.04
0.12
Ni
0.02
0.03
Ti
0.25
0.04
0.03
0.15
0.05
V
1.4
0.45
1.1
0.05
0.03
0.04
0.02
-------
204
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Rules and Regulations
TABLE 1.—ANALYTE CONCENTRATION EQUIVALENTS (MQ/L) ARISING FROM INTERFERENTS AT THE 100 MQ/L LEVEL—Continued
AnriyM
LMd
!!!• l||MmV .... ii MI
NMol
MvHum
SJScon
Soofcin
TMhjm
VvMdktfn
2)ne
WBM
•ngu.
nm
220353
27*079
257.610
231.604
1*6.026
266.156
SM.N5
1M.M4
2*2402
213.66*
A1
0.17
0.005
0.05
0.23
0.30
Cc
002
Cr
0.11
0.01
0.07
005
Cu
014
Intonfl
Ft
013
0.002
0.03
009
0.005
r«nt—
Mg
0.002
Mn
025
M
02*
Tl
007
0.01
002
V
001
TABLE 3. INTERFERED AND ANALYTE ELEMENTAL CONCENTRATIONS USED FOR INTERFERENCE MEASUREMENTS IN TABLE 2
AnolyM
A|
A§
B
B|
Bt
Cj ..
Gd
Co
Cr
Cu
ft
Mg
Mn ...
Mo
No
Ml
Pb
Sb
S*
Si
Tl
y
In
(mg/L)
10
10
10
1
1
1
10
1
1
1
1
1
1
10
10
10
to
10
10
1
10
1
10
Mtftoranti
A)
C»
Cr
Cu
ft
Mg.
Mn
M
Ti
V
(m»/U
1000
1 000
200
200
1000
1 000
200
200
200
200
TABLE 4.—ICP PRECISION AND ACCURACY DATA
Etenonl
Bo
Mn
y
Ao
Cr
Cu
Fo
Al • •
Co1
Co
N| „ „
PB _ _
2n
So .. _
TIM Mb*
rt/L
760
380
750
200
150
250
600
700
SO
500
250
250
200
40
SmptoNa 1
HMD
(•ported
MkMMA.
7J»
346
74*
206
14*
235
5*4
M6
4*
512
246
23*
201
32
MOT
praMRto
u
17
1.6
7.5
3.6
5.1
3.0
5.6
12
10
5.8
16
5.6
21.*
TlMMtat
pg/L
20
15
70
22
10
11
20
60
2.5
20
30
24
16
6
SvnptoNo.2
M«y
rtportKl
itt^ntn.
20
IS
**
1*
10
11
1*
62
2.1
20
2*
30
1*
6.5
Mean
pwaMRBD
M
6.7
2.1
23
1*
40
16
33
1*
4.1
11
32
45
42
Tiwv*k»
Hfl/L
1*0
100
170
(0
SO
70
1*0
160
14
120
60
(0
60
10
Samp* No. J
M«y
MpOTM
Mtong/L
17*
f)
16*
0
SO
67
176
161
13
106
55
60
62
6.5
MM
PMMMD
jj
3J
1 1
17
33
79
6.0
13
16
21
14
14
(4
63
Not iff •
(Doc. 84-28188 Filed 10-23-84; 8:45 un]
-------
Federal Register / Vol. 49, No. 209 / Friday, October 26, 1984 / Proposed Rules
205
ENVIRONMENTAL PROTECTION
AGENCY
40 CFR Part 136
[FRL-2636-6]
Guidelines Establishing Test
Procedures for the Analysis of
Pollutants
Note: This reprint incorporates a
typographical correction which was
published in the Federal Register of Friday,
January 4.1985 on page 697 (third column).
AGENCY: Environmental Protection
Agency (EPA).
ACTION: Proposed regulation.
SUMMARY: Elsewhere in this issue of the
Federal Register, EPA has promulgated
new test procedures under Clean Water
Act (CWA) Section 304(h) for the
analysis of many priority toxic organic
and other pollutants, which are based
upon gas chromatographic instrumental
systems. This proposal would withdraw
approval for outdated test procedures
which had been approved for sixteen
compounds, including chlorinated
organic compounds, benzidine, and for
fourteen pesticide compounds. EPA is
also proposing to approve two methods
(Methods 1624 and 1625) for new
compounds. These methods were
promulgated for other compounds
elsewhere in today's Federal Register.
DATE: Comments on this proposal must
be submitted on or before January 7,
1985.
ADDRESS: Send comments to Dr. Robert
B. Medz, "Proposed 304(h) Guidelines,"
Water and Waste Management
Monitoring Research Division, Office of
Research and Development (RD-680),
401M Street, SW.. Washington. D.C.
20460.
FOR FURTHER INFORMATION CONTACT:
Dr. Robert B. Medz at the address listed
above, or call (202) 382-5788. The record
for the rulemaking is available for
review at Washington, D.C.
SUPPLEMENTARY INFORMATION:
I. Authority and Background
This regulation is proposed under the
authority of sections 301, 304(h) and
501 (a) of the Clean Water Act of 1977
(CWA).
It would amend 40 CFR Part 136 in
two ways. First, it would withdraw
approval of outmoded methods for the
analysis of 30 chemical compounds.
Second, it would expand the scope of
two recently approved methods to
include thirty-two additional compounds
for which no approved methods now
exist.
Elsewhere in today's Federal Register,
EPA has approved new test procedures
for the analysis of 111 priority, toxic
organic pollutants. The analytical test
procedures are based on 12 gas
chromatograph (GC) methods and 5 gas
chromatograph/mass spectrometer (GC/
MS) methods. These test procedures
differ from those previously approved
by the EPA at 40 CFR Part 136. Their
most important improvement is that they
include detailed quality control
requirements and specify control limits
indicating inadequate performance. If an
analyst's performance falls outside
those control limits, his analytical
system is considered to be out-of-control
and data generated with the system is
not reportable for regulatory purposes.
EPA approved analytical methods for
3 classes of organic pollutants on
December 1,1976 (41 FR 25780). These
classes included 30 compounds which
were later included within the scope of
the priority pollutants. The 1976 test
procedures had neither detailed quality
control requirements nor warning limits
within their provisions. In complex
industrial and municipal wastewater
matricies, application of these 1976 test
procedures results in data of poorly
defined quality if the analyst fails to
perform an adequate level of quality
control. The procedures were updated in
1978 but the underlying problems with
quality control were not addressed.
Therefore, for the 30 organic
compounds, there is an inconsistency
between the test procedures approved in
1976 and the test procedures approved
today. This proposal will withdraw
approval for the test procedures as they
apply to these 30 pollutants. In effect,
the old GC procedures will be
superseded by the 15 test procedures
which have been approved today for the
111 priority toxic organic pollutants.
EPA has approved two GC/MS test
procedures (Methods 1624 and 1625)
today. These methods use stable,
isotopically labeled analogs of the
priority pollutants as internal standards.
They have been extensively tested for
their applicability to the analysis of the
32 pollutants included pursuant to
paragraph 4(c) and Appendix C of the
Consent Decree (NRDC v. Train, 8 ERC
2120 (D.D.C. 1976), as modified 12 ERC
1833 (D.D.C. 1979) and by the Court's
Order of October 26,1982, August 2,
1983, January 6,1984, and July 5.1984).
The Consent Decree settled a suit
between the Natural Resources Defense
Council (NRDC) and EPA regarding
regulation under the CWA. In paragraph
4(c), EPA agreed to study pollutants for
possible regulation, including these
specified in Appendix C. Today's
proposal would extend the approved
scope of the two methods to include the
additional paragraph 4(c) and Appendix
C pollutants.
II. Summary and Rationale for Proposed
Amendments
A. Withdrawal of Former Method
Approvals
Thirty compounds which were
approved as parameters in the 1976
amendments to 40 CFR Part 136 were
also included in the 1976 Consent
Decree as priority pollutants. In the 1976
regulation they were carried under the
parameter designations Benzidine,
Pentachlorophenol, Chlorinated organic
compounds (except pesticides), and
Pesticides. Approved test procedures for
these parameters were available from
EPA's Environmental Monitoring and
Support Laboratory in Cincinnati, Ohio.
The method for Benzidine was covered
under "Method for Benzidine and Its
Salts in Wastewater." The remaining
parameters were covered under
"Procedures for Pentachlorophenol,
Chlorinated Organic Compounds, and
Pesticides."
These analytical methods were
updated in 1978 by the EPA publication,
"Methods for Benzidine, Chlorinated
Organic Compounds,
Pentachlorophenol, and Pesticides in
Water and Wastewater," U.S.
Environmental Protection Agency,
September 1978. However, the earlier
methods and the 1978 update do not
require a mandatory level of quality
control and do not stipulate analytical
control limits, outside of which an
analysis would be considered to be out-
of-control. Therefore, the analyst could
generate data which would be unusable
for regulatory purposes. The new
methods being approved today for
priority pollutant analysis do include
these procedures. This leads to a
significant inconsistency between these
new methods and the updated methods
which had not been anticipated at the
time that the priority pollutant test
procedures were proposed on December
3,1979 (44 FR 69464).
The present proposed action will
eliminate this inconsistency and ensure
that these earlier methods will be
superceded by the test procedures being
approved today for the priority
pollutants for the following specific
parameters in Tables 1C and ID of
S 136.3: Table 1C, Benzidine, Carbon
Tetrachloride, Chlorobenzene,
Chloroform, Methylehe chloride, PCB-
1016, PCB-1221, PCB-1232, PCB-1242,
PCB-1248, PCB-1260.
Pentachlorophenol, 1,1,2,2-
Tetrachloroethane, Tetrachloroethene,
1,2,4-Trichlorobenzene, and 1,1,2-
Trichloroethane; and Table ID, Aldrin,
a-BHC, y-BHC, Chlordane, 4,4'-DDD,
4,4'-DDE, 4.4'-DDT. Dieldrin, Endosulfan
-------
206
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Proposed Rules
I, Endosulfan II. Endrin, Heptachlor,
Heptachlor expoxide, and Toxaphene.
The methods approved in 1976 and
updated in 1978 may continue to be the
basis for EPA enforcement action where
proper quality control and quality
assurance are used. Of course. EPA may
also use the new methods where they
are more appropriate or more
economical.
B. Extension of New Methods to
Appendix C Parameters
EPA is proposing to expand Table 1C
of § 136.3 by 32 paragraph 4(c) and
Appendix C parameters (it will now
include 129 parameters). The purge and
trap test procedure, Method 1624, is
approved for four of these purgeable
compounds: Acetone, Diethyl ether, p-
Dioxane, and Methyl ethyl ketone.
Extraction test procedure. Method 1625,
is approved for 28 compounds: Benzoic
acid, Biphenyl, Carbazole, p-Cymene, n-
Decane, Dibenzofuran,
Dibenzothiophene. Diphenylamine,
Diphenyl ether, 1,2-Diphenylhydrazine.
n-Docosane, n-Dodecane, n-Eicosane, n-
Hexacosane, n-Hexadecane, Hexanoic
add, /3-Naphthylamine, n-Octacosane,
n-Octadecane, a-Picoline, Styrene, a-
Terpineol, n-Tetracosane, n-
Tetradecane, n-Triacontane, 1,2,3-
Trichlorobenzene. 2,3,6-Trichlorophenol,
and 2,4.5-Trichlorophenol. These
methods have been extensively applied
to the analysis of these 32 compounds in
industrial wastewaters and the methods
have been validated for their
applicability to analysis of these
compounds by multi-laboratory testing.
C. Other Part 136 Provisions
EPA wishes to make clear that
methods for the 32 new compounds that
will be covered under Part 136 will be
subject to all the existing definitions and
provisions of Part 136. For example, EPA
will be able to approve equivalent
methods for these new compounds, as it
can for any parameter subject to this
Part.
m Regulatory Analysis
(a) Under Executive Order 12291, EPA
must judge whether a regulation is
"major" and, therefore, subject to the
requirement of a "Regulatory Impact
Analysis." This regulation is not major
for the following reasons:
(1) It proposes analytical methods and
sample handling requirements that
ensure a uniform measure of pollutants
across all wastewater discharges within
minimum acceptance criteria for 32
parameters. The purpose is to ensure
that the quality of environmental
monitoring data meets certain minimum
standards. It would withdraw the use of
outdated methods.
(2) The impact of this regulation will
be far less than $100 million.
(a) The regulation affects unit
monitoring costs for other regulatory
programs, e.g.. effluent guidelines
regulations and the implementation
regulations of the National Pollutant
Discharge Elimination System (NPDES),
and the pretreatment program. However,
it does not impose those costs. In fact,
the monitoring costs for other programs
are considered in those other
rulemakings. This is appropriate
because total (rather than unit)
monitoring costs are determined by the
monitoring provisions of those
regulations.
(b) Equivalency provisions will
encourage the development of
innovative analytical methods by the
private sector and to encourage the
competitive viability of the instrument
manufacturing industry. The
equivalency provision also allows
individual dischargers to gain approval
of analytical systems of their own
design that may further reduce their
total monitoring costs.
(3) The empact of compliance with
these regulations will not be
concentrated on any particular sectors
of American industry. ;
This regulation was submitted to fhe
Office of Management and Budget
(OMB) for review as required by
Executive Order 12291. Any comments
from OMB to EPA and any EPA
response to those comments will be
available for public Inspection at the
Public Information Reference Unit
Room M2904 (EPA Library-Rear). TM-
213, Environmental Protection Agency,
401M Street SW., Washington, D.C.
20460. Phone: (202) 382-6926. Office
Hours 8:00 a.m. to 4:30 p.m.
(b) Under the Regulatory Flexibility
Act 5 U.S.C. 601, et seq., EPA is required
to determine whether a regulation will
significantly affect a substantial number
of small entities so as to require a
regulatory analysis. The regulation
requires no new reports beyond those
already now required. The analytical
techniques approved here either can be
handled by small facilities, or are
widely available by contract at a
reasonable price. Therefore, in
accordance with 5 U.S.C 605(b), I
hereby certify that this rule will not
have a significant adverse economic
impact on a substantial number of small
facilities.
(c) Under the Paperwork Reduction
Act of 1980,44 U.S.C 3501 et seq., the
information provisions in this rule
associated with the analytical test
procedures equivalency program, 40
CFR 136.3 (a), (c) and (d). 136.4 and
136.S. and the sample preservation and
holding times variances, 40 CFR 136.3(e).
have been submitted to the Office of
Management and Budget (OMB) as part
of the final and interim-final rule
published elsewhere in today's Federal
Register. All approvals made on the
Final rule will be applicable to this
proposed rule.
(Sect. 301, 304(h). 307 and 501(a). Pub. L. 95-
217.91 Stat. 1566. et aeq. (33 U.S.C. 1251. et
seq.) (the Federal Water Pollution Control
Act Amendments of 1972 as amended by the
Clean Water Act of 1977)).
Dated: September 26,1964.
William O. RucklMhaus,
Administrator.
List of Subjects in 40 CFR Part 136
Water pollution control.
In consideration of the preceding, EPA
proposes to amend Chapter I,
Subchapter D of Title 40. Code of
Federal Regulations, as follows:
Proposed Rule
For the reasons set out in the
Preamble, it is proposed to amend Part
136, Chapter 1. Subchapter D of Title 40
of the Code of Federal Regulations as
follows:
1. In { 136.3, Table 1C is amended by
renumbering to have 129 parameters, ,
and by alphabetically inserting the
following thirty-two new parameters:
Acetone, Benzoic acid, Biphenyl,
Carbazole, p-Cymene, n-Decane,
Dibenzofuran. Dibenzothiophene,
Diethyl ether, p-Dioxane,
Diphenylamine. Diphenyl ether, 1,2-
Diphenylhydrazine, n-Docosane, n-
Dodecane, n-Eicosane, n-Hexacosane, n-
Hexadecane, Hexanoic acid. Methyl
ethyl ketone, /3-Naphthylamine. n-
Octacosane, n-Octadecane, a-Picoline,
Styrene, a-Terpineol, n-Tetracosane. n-
Tetradecane, n-Triacontane, 1,2,3-
Trichlorobenzene. 2,3,6-Trichlorophenol,
and 2,4,5.-Trichlorophenol; by approving
EPA Method 1624 for the analysis of
Acetone, Diethyl ether, p-Dioxane, and
Methyl ethyl ketone; by approving EPA
Method 1625 for the analysis of Benzoic
acid, Biphenyl, Carbazole, p-Cymene, n-
Decane. Dibenzofuran,
Dibenzothiophene. Diphenylamine,
Diphenyl ether. 1 J-Diphenylhydrazine.
n-Docosane. n-Dodecane. n-Eicosane, n-
Hexacosane. n-Hexadecane, Hexanoic
add. 0-Naphthylamine. n-Octacosane,
n-Octadecane, a-Plcoline, Styrene, a-
TerpineaL n-Tetracosane, n-
Tetradecane, n-Triacontane, 1,2,3-
TricbJorobenzene. 2,34-Trichlorophenol.
and 2,45-TrichlorophenoI: by
withdrawing approval of the following
dted test procedures for the following
-------
Federal Register / .. 49. No. 209 / Friday. October 26V1984 / Proposed Rules
207
parameters: Benzidlne, oxidation-
colorimetric procedure, "Method for
Benzidine and Its Salts in Wastewater".
Environmental Monitoring and Support
Laboratory, U.S. Environmental
Protection Agency. Cincinnati, Ohio
(1976); and Carbon tetrachloride.
Chlorobenzene. Chloroform, Methylene
chloride. PCB-1016, PCB-1221. PCB-1232.
PCB-1242, PCB-1246, PCB-1260.
Pentachlorophenol, 1,1.2,2-
Tetrachloroethane, Tetrachloroethene,
1,2,4-Trichlorobenzene and 1.1.2-
Trichlolorethane, gas chromatography,
"Procedures for Pentachlorphenol.
Chlorinated Organic Compounds, and
Pesticides". Environmental Monitoring
and Support Laboratory, U.S.
Environmental Protection Agency.
Cincinnati. Ohio (1976). As proposed to
be revised. } 136.3. Table 1C would read
as follows:
} 136.3 Identification OH T«t Procedures.
TABLE 1C.—LIST OF APPROVED TEST PROCEDURES FOR ORGANIC COMPOUNDS
3 Actttorw
4 AcraMifi "•
5 Acrvtonitnta
0 Anthracene
7 Bennoe •
9 DeflJLud i l m Oinceoe • • • • "•- < •• •"-- • > •
10 Beoiofc Acid ••
12 8*Mizo(b}fluor>n1twnii • — «• •••
19 Bi6J2-cfitorootfiyQ •ttier
20 BttlZ^tfwttwfYO ofiVViiM
£0 4 ClUmu 3 nnthB^)tM"rt " "
79 CNorotxitune
32. CrUoronMinAn*
37 p-CynwK
*j(j n<0*cw i ,.,— - ,,,,,,,,-,,
TO nt^tfiHrfa M«nttumn*«j
50. i l^icMonMthan* - ..._.....-........ _ _ ..„ . . _
31 UlJil 1,2 CHU^ULKITXIM
54 PHtV Tfl*^ --
W p-0to)«nt •
73. EtMbMmrw. ™ .. .. . — «- • • — —
QC
610
610
i
r" 803
j 603
010
602
610
610
blO
610
610
608
611
611
606
801
J 601
1 301
611
i
1 601
1601
604
601.602
601
601
1601
612
604
611
610
610
601
601 602
612
801 602
612
601 802
612
^ ibi"
601
601
801
604
801
801
801
608
804
608
606
608
604
6M
600
602
E
OC/MS
62S 1625
1624
•824 1624
•824 1624
824. 1624
82S. 162S
1625
625. 1625
625 1625
625 1825
625 1625
625. 1625
624. 1624
624 1824
625. 1625
625 1625
625, 1625
1625
1625
1825
1825
824 825
1625
824 625
1825
624 1624
824.1624
624 1624
824 1624
824 1624
624 1824
1824
625, 1625
825. 1625
625, 1825
625, 1625
625, 1625
1624
1625
1625
1625
1625
1625
1625
824. 1824
PAnwthc
HPIC
610
610
610
"
|
"
"
..
'
610
610
"*
l"
«3numO»«'
Otfnr
Note 3 o 1 30' Not* 6, o Si 02
flOU 3 p 13Q- Not* 6 p Si 02
-------
Federal Register / Vol. 49. No. 209 / Friday. October 26. 1984 / Proposed Rules
74.nkOTMiM
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i fcl of Men of tm* MMnodt, AdMtoraHr. MCA MonMy. en «n onmng MM. muM IP** M m*n* 10% i
•or MM •<««•• IMUHMII dM quMr in «cjeoid«m» «Mh MCMM • 3 »nd M o( MM llanoai, WMA M nceyy el
2. In 1130.3. Table ID it amended by
withdrawing approval of the following
cited test procedure! for the following
parameters: Aldrin. o-BHC y-BHC
(Undane). Chlordane. 4.4'-DDD. 4.4'-
DDE. 4.4'-DDT. Dieldrin. Endotulfan I
Endotulfan U, Endrin, Heptachlor,
Heptachlor epoxide, and Toxaphene by
the Gat Chromatography teat
procedures dted in, "Procedures for
PentachlorophenoL Chlorinated Organic
Compounds, and Pesticides."
Environmental Monitoring and Support
Laboratory, U.S. Environmental
Protection Agency. Cincinnati. Ohio
(1976). As proposed to be revised.
i 1384. Table ID would read as foUows:
MenUflmion of Teat Procedures*
-------
Federal Register / Vol. 49. No. 209 / Friday, October 26. 1984 / Proposed Rules
209
TABLE 10.—UST OF APPROVED TEST PROCEDURES FOR PESTICIDES '
DB*MI«^^W
mnMI
1 AkMn
2. Arn*tjyn • .."•••
4 Atntprfl
5 Atrazin*
7 B*/b*n - • •«•
8 a*BHC
9 0-8HC - -
10 &-8HC - -
17 2.4-O
18 4 4'-000 - •
20 4 4'-OOT .- -
7? 0*ni*rrtrjfvS • "
23 Dtuxion —
24 Oidnio* — —
27 OicotOI -
20 DfeMm
31 OkMon - - -'•
32 Endotutfwi t - -
.
33 Endosi4f*n It
34. EndosUlan surfat*
37 Ethion .. — . . -
39 F*nuron - ..-. ,,.,,,-, ,, ,,
39 r*m*on-TCA
40 1 l*um film
™
41 hMUGNor *paibd* ... ..„„, , ,.., ..!....
r
42 Itodhn . . .,r ., , , ..„
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45 M^ltlOCMD . , ....
47 M*Mjf*JPIT* -
40 Mirav. . ...
50 Monwon-TGA . ... ...... ....
51 I4*tiuron ..... L...
S3 Pmnton tihy* . ..
54 PCNB
55 P*i1h*n* . - . —
57 PraTwrm
56 Prupirnrt
62 SMuron
63 Sfcnnfei*
05. 5w*p
M. 2.4.5.T
07 &4 5*TP (SBvn)
Q0 JCMftofM
70 Trifhrtki
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or
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nr.
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or.
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ac
625
606
62S
606
•625
606
625
606
625
606
606
625
606
625
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606
•625
606
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625
606
625
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625
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62S
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Not* 3. p. 63; Not* 6. p S66
Not* 3, p. 94- Not* 6, p S16
Not* 3 p 63' Not* 6 p S68
Not* 3. p. 63: Not* 8, p S66
Not* 3 p. 25- Not* 6 p S51
Not* 3. p. 104; Not* 6 p S64
Not* 3. p. 7
Not* 3 p d4- Not* 6 p S60
Not* 4 p 30' Not* 6 p S73
Not* 3 p 104' Not* 6 p S64
Not* 3 p 115- Not* 4 p 35
Not* 3 p 25* Not* 6 p S51
Not* 3. p. 25- Not* 6 p SS1
Not* 3. p. 25* Not* 4 p 30* Not* 6, P. S51
Not* 3 p. 115
Not* 4. p. 30- Not* 6 p S73
Not* 3 p. 7
Not* 4 p 30' Not* 6 p S73
Not* 3 p. 25' Not* 6 p S51
Not* 3. p. 104.
Not* 4. p. 30.
Not* 3. p. 104; Not* 6. p. S64.
Not* 3. P. 104* Not* 6, p. S64
Not* 4 p. 30- Not* 8. p S73
Not* 3. p. 25; Not* 6. p. S64
Not* 3, p. 25; Not* 4 p 30* Not* 6. p SS1
Not* 3. p. 94; Not* 6. p. S60
Not* 3 p 7* Not* 4 p 30
Not* 3. p. 94' Not* 6. p SAO
Not* 3. p. 7
Not* 3 p 104' Not* 6. p S64
Not* 3 p. 104* Not* 6. p S64
Not* 3 p 104* Not* 6 p S64
Not* 3 p 25* Not* 4 p. 30
Not* 3 p. 25.
Not* 3. P. 7
Not* 3. p. 83; Not* 6. p. S66
Not* 3, p. 83; Not* 6. p. See.
Not* 3 p. 83: Not* 6. p. S66.
Not* 3 p. 63: Nol* 6. p. S68
Nol* 3. p. 104* Not* 6, p. S64
Not* 3 p. 94' Nol* 6. p. S60
NOH3.P. 63- Not* 6. p. 568.
Not* 3, p. 104: Not* 6. p. S64.
NOI* 3. p. 63: Not* 6, p. see.
Nol* 3. p. 7
Not* 3. p. 104; Not* 6, p. S64
Not* 3. p. 115; Noll 4 P- 36.
Not* 3. p. 116.
Nol* 3, p. 83; Nol* 6. p. S68 69
Noto 3. p. 7
•» MX) in IN* OO» ty common nun* tor IK* eamnfnet of •» rMd*. MOtant pmciilii m*y b* kuid mtv T*u* C *t*n *nM** tra MM) by a*mc* ram*.
"Th* tut ton o< MMhodi 608 *nd 625. *ra grv«i *t Appwdh A. -T*M Preo«kn* lor An*M* of Crawfc PoMM*,* ol Mi Put 136. Th* U»na«nto«l Ml prooMf* to b* uMd to
m'm in* mnnod d*t*c«on Km* (MOD lor OM** MM proeMim • gNxn O Appmdbi B. "IMMIon mdPreoidLn lor ttm OcMrfrmllon at th* M*trod OtKiuun Urn*", ol IN* Pvt 136.
• -M*ewo* lor Bwmolr* CNorinMd Orgnic Compound*. P*nbKntaropn*noT *nd PiMiMn in W«Br end Wamnin.- U.S. EiiyimrimOl ProMcDon Agtrcy, S«pttmb«r. 1976. Thk
EPA &****&*> ndudM tfwv(rr*r chronMognpny (TLC) iraVno*.
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210Fed«MJ Rtlttor / Vol. 49. No. 209 / Friday. October 28. 1964 / RU!M and Reg
MWWt. _-«.-,m.m
•Htm W MM. «»*M, 4OTOMMM it Ml iMr •
[FR Doc. M-2AMQ FU«d 10-25-S4; 8:45 am]
O.S. UOV1IUBBM FmRSM UVF1UI t IMS 0 - 465-028
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