METHOD 3600C
CLEANUP
SW846PU33
1.0 SCOPE AND APPLICATION
1.1 Method 3600 provides general guidance on selection of cleanup methods
that are appropriate for the target analytes of interest. Cleanup methods are
applied to the extracts prepared by one of the extraction methods, to eliminate
sample interferences.
1.2 The following table lists the cleanup methods and provides a brief
description of the type of cleanup.
SW-846 CLEANUP METHODS
Method #
3610
3611
3620
3630
3640
3650
3660
3665
Method Name
Alumina Cleanup
Alumina Cleanup & Separation
for Petroleum Waste
Florisil Cleanup
Silica Gel Cleanup
Gel -Permeation Cleanup (GPC)
Acid-Base Partition Cleanup
Sulfur Cleanup
Sulfuric Acid/Permanganate
Cleanup
Cleanup Type
Adsorption
Adsorption
Adsorption
Adsorption
Size-Separation
Acid-Base Partitioning
Oxidation/Reduction
Oxi dat i on/Reducti on
1.3 The purpose of applying a cleanup method to an extract is to remove
interferences and high boiling material that may result in:
• errors in quantitation (data may be biased low because of analyte
adsorption in the injection port or front of the GC column or biased high
because of overlap with an interference peak);
• false positives because of interference peaks falling within the analyte
retention time window;
• false negatives caused by shifting the analyte outside the retention time
window;
• rapid deterioration of expensive capillary columns; and,
• instrument downtime caused by cleaning and rebuilding of detectors and ion
sources.
1.4 The following techniques have been applied to extract purification:
adsorption chromatography; partitioning between immiscible solvents; gel
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permeation chromatography; oxidation of interfering substances with acid, alkali,
or oxidizing agents. These techniques may be used individually or in various
combinations, depending on the extent and nature of the co-extractives.
1.5 Most extracts of soil and waste require some degree of cleanup,
whereas, cleanup for water extracts may be unnecessary. Highly contaminated
extracts (e.g. sample extracts of oily waste or soil containing oily residue)
often require a combination of cleanup methods. For example, when analyzing for
organochlorine pesticides and PCBs, it may be necessary to use gel permeation
chromatography (GPC), to eliminate the high boiling material and a micro alumina
or Florisil column to eliminate interferences with the analyte peaks on the
GC/ECD.
2.0 SUMMARY OF METHOD
Refer to the specific cleanup method for a summary of the procedure.
3.0 INTERFERENCES
3.1 Analytical interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware. All of these
materials must be routinely demonstrated to be free of interferences, under the
conditions of the analysis, by running laboratory reagent blanks.
3.2 More extensive procedures than those outlined in the methods may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
Refer to the specific cleanup method for apparatus and materials needed.
5.0 REAGENTS
Refer to the specific cleanup method for the reagents needed.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Prior to using the cleanup procedures, samples normally undergo
solvent extraction. Chapter Two, Section 2.0, may be used as a guide for
choosing the appropriate extraction procedure based on the physical composition
of the waste and on the analytes of interest in the matrix (see also Method 3500
for a general description of the extraction technique). For some organic
liquids, extraction prior to cleanup may not be necessary.
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7.2 Most soil/sediment and waste sample extracts will require some degree
of cleanup. The extract is then analyzed by one of the determinative methods.
If interferences still preclude analysis for the analytes of interest, additional
cleanup may be required.
7.3 Many of the determinative methods identify cleanup methods that should
be used when determining particular analytes (e.g. Method 8061, gas
chromatography of phthalate esters, recommends using either Method 3610 (Alumina
column cleanup) or Method 3620 (Florisil column cleanup) if interferences prevent
analysis. However, the experience of the analyst may prove invaluable in
determining which cleanup methods are needed. Many matrices may require a
combination of cleanup procedures in order to ensure proper analytical
determinations.
7.4 Specific guidance on each cleanup technique is listed in the
individual cleanup methods that follow. The amount of extract cleanup required
prior to the final determination depends on the concentration of interferences
in the sample, the selectivity of both the extraction procedure and the
determinative method and the required detection limit. The following Sections
give a description of the different cleanup approaches:
7.4.1 Adsorption column chromatography - Alumina (Methods 3610 and
3611), Florisil (Method 3620), and silica gel (Method 3630) are useful for
separating analytes of a relatively narrow polarity range away from
extraneous, interfering peaks of a different polarity. These are
primarily used for cleanup of a specific chemical group of relatively
non-polar analytes, i.e., organochlorine pesticides, polynuclear aromatic
hydrocarbons (PAHs), nitrosamines, etc.. Solid phase extraction
cartridges have been added as an option.
7.4.2 Acid-base partitioning (Method 3650) - Useful for separating
acidic or basic organics from neutral organics. It has been applied to
analytes such as the chlorophenoxy herbicides and phenols. It is very
useful for separating the neutral PAHs from the acidic phenols when
analyzing a site contaminated with creosote and pentachlorophenol.
7.4.3 Gel permeation chromatography (GPC) (Method 3640) - The most
universal cleanup technique for a broad range of semivolatile organics and
pesticides. It is capable of separating high molecular-weight, high
boiling material from the sample analytes. It has been used successfully
for all the semivolatile base, neutral, and acid compounds associated with
the EPA Priority Pollutant and the Superfund Target Compound list prior to
GC/MS analysis for semivolatiles and pesticides. GPC may not be
applicable to elimination of extraneous peaks on a chromatogram which
interfere with the analytes of interest. It is, however, useful for the
removal of high boiling materials which would contaminate injection ports
and column heads, prolonging column life, stabilizing the instrument, and
reducing column reactivity.
7.4.4 Sulfur cleanup (Method 3660) - Useful in eliminating sulfur
from sample extracts, which may cause chromatographic interference with
analytes of interest.
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7.4.5 Sulfuric acid/permanganate cleanup (Method 3665) - Useful for
the rigorous cleanup of sample extracts prior to analysis for
polychlorinated biphenyls (PCBs). This method should be used whenever
elevated baselines or overly complex chromatograms prevent accurate
quantitation of PCBs. This method cannot be used to cleanup extracts for
other target analytes, as it will destroy most organic chemicals including
the pesticides Aldrin, Dieldrin, Endrin, Endosulfan (I and II), and
Endosulfan sulfate.
7.5 Fractionation is a useful technique that can aid in the separation of
complex mixtures of analytes. For instance, an analyst may use Method 3630
(Silica Gel) for separating the PCBs away from most organochlorine pesticides.
Method 3611 (Alumina) may be used for fractionation of aliphatic, aromatic, and
polar analytes. Method 3620 (Florisil) provides for fractionation of the
organochlorine pesticides.
7.6 Cleanup capacity is another factor that must be considered in choosing
a cleanup technique. The adsorption methods (3610, 3620, and 3630) provide the
option of using standard column chromatography techniques or solid phase
extraction cartridges. The decision process in selecting between the different
options available generally depends on the amount of interferences/high boiling
material in the sample extract and the degree of cleanup required by the
determinative method. The solid phase extraction cartridges require less elution
solvent and less time, however, their cleanup capacity is drastically reduced
when comparing a 0.5 g or 1.0 g Florisil cartridge to a 20 g standard Florisil
column. The same factor enters into the choice of the 70 g gel permeation column
specified in Method 3640 versus a high efficiency column.
7.7 Table 1 indicates the recommended cleanup techniques for the indicated
groups of compounds. This information can also be used as guidance for compounds
that are not listed. Compounds that are chemically similar to these groups of
compounds should behave similarly when taken through the cleanup procedure.
However, this must be demonstrated by determining recovery of standards taken
through the method.
7.8 Following cleanup, the sample is concentrated to whatever volume is
listed in the determinative method using the procedures described in the
appropriate 3500 series method. Analysis follows as per the appropriate
determinative procedure.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered by the cleanup technique before the cleanup is applied
to actual samples. For sample extracts that are cleaned up, the associated
quality control samples (e.g. spikes, blanks, replicates, and duplicates) must
also be processed through the same cleanup procedure.
8.3 The analysis using each determinative method (GC, GC/MS, HPLC) lists
instrument calibration procedures using stock standards. It is recommended that
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cleanup also be performed on a series of the same type of standards to validate
chromatographic elution patterns for the compounds of interest and to verify the
absence of interferences from reagents.
9.0 METHOD PERFORMANCE
Refer to the specific cleanup method for performance data.
10.0 REFERENCES
Refer to the specific cleanup method.
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TABLE 1.
RECOMMENDED CLEANUP TECHNIQUES FOR INDICATED GROUPS OF COMPOUNDS
Determinative8 Cleanup
Analyte Group Method Method Options
Phenols 8041 3630b, 3640, 3650, 8040°
Phthalate esters 8061 3610, 3620, 3640
Nitrosamines 8070 3610, 3620, 3640
Organochlorine pesticides 8081 3620, 3640, 3660
PCBs 8082 3620, 3630, 3665
Nitroaromatics and cyclic ketones 8091 3620, 3640
Polynuclear aromatic hydrocarbons 8100/8310 3611, 3630, 3640
Haloethers 8111 3620, 3640
Chlorinated hydrocarbons 8121 3620, 3640
Aniline and derivatives 8131 3620, 3640
Organophosphorus pesticides 8141 3620
Chlorinated herbicides 8151 8151d, 3620
Semivolatile organics 8270 3640, 3650, 3660
Petroleum waste 8270 3611, 3650
PCDDs and PCDFs by LR/MS 8280 8280
PCDDs and PCDFs by HR/MS 8290 8290
N-methyl carbamate pesticides 8318 8318
a The GC/MS Method 8270 is also an appropriate determinative method for all
analyte groups, unless lower detection limits are required.
b Cleanup applicable to derivatized phenols.
c Method 8041 includes a derivatization technique followed by GC/ECD analysis,
if interferences are encountered using GC/FID.
d Method 8151 incorporates an acid-base cleanup step as an integral part of the
method.
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METHOD 3610B
ALUMINA CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Alumina is a highly porous and granular form of aluminum oxide. It
is available in three pH ranges (basic, neutral, and acidic) for use in
chromatographic cleanup procedures. It is used to separate analytes from
interfering compounds of a different chemical polarity.
1.2 Each of the three pH ranges of alumina has different uses and
disadvantages as a cleanup procedure.
1.2.1 Basic alumina has a pH of 9-10. It is used to separate basic
and neutral compounds that are stable to alkali, alcohols, hydrocarbons,
steroids, alkaloids, natural pigments. Its disadvantages are that it can
cause polymerization, condensation, and dehydration reactions, and one
cannot use acetone or ethyl acetate as eluants.
1.2.2 Neutral alumina has a pH of 6-8. It is used to separate
aldehydes, ketones, quinones, esters, lactones, glycoside. Its
disadvantage is that is it considerably less active than the basic form.
1.2.3 Acidic alumina has a pH of 4-5. It is used to separate
acidic pigments (natural and synthetic), and strong acids (that otherwise
chemisorb to neutral and basic alumina). This method does not address the
use of acid alumina.
1.3 Basic, neutral, and acidic alumina can be prepared in various activity
grades (I to V), according to the Brockmann scale, by the addition of water to
Grade 1 (prepared by heating at 400-450°C until no more water is lost). The
Brockmann scale is reproduced below:
Activity grade I II III IV V
Water added (wt. %) 0 36 10 15
RF (p-aminoazobenzene) 0.0 0.13 0.25 0.45 0.55
where RF is the retention factor for p-aminoazobenzene.
1.4 Alumina cleanup may be accomplished using a glass chromatographic
column packed with alumina or using solid-phase extraction cartridges containing
alumina.
1.5 This method includes procedures for cleanup of sample extracts
containing phthalate esters and nitrosamines. See Method 3611, Alumina Column
Cleanup of Petroleum Wastes, for alumina cleanup of petroleum wastes.
1.6 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
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2.0 SUMMARY OF METHOD
2.1 This method describes procedures for alumina cleanup of solvent
extracts of environmental samples. It provides the option of using either
traditional column chromatography techniques to solid-phase extraction
cartridges. Generally, the traditional column chromatography technique uses
larger amounts of adsorbent and, therefore, has a greater cleanup capacity.
2.2 In the column cleanup protocol, the column is packed with the
appropriate amount of adsorbent, topped with a water adsorbent, and then loaded
with the sample extract. Elution of the analytes is effected with a suitable
solvent(s), leaving the interfering compounds on the column. The eluate may be
further concentrated prior to gas chromatographic analysis.
2.3 The cartridge cleanup procedure uses solid-phase extraction
cartridges containing 40 urn particles of alumina (60 A pores). Each cartridge
is washed with solvent immediately prior to use. The sample extract is loaded
onto the cartridge which is then eluted with suitable solvent(s). A vacuum
manifold is needed to obtain reproducible results. The eluate may be further
concentrated prior to gas chromatographic analysis.
2.4 The phthalate esters may be considered either the analytes of
interest or the interferants, depending on which eluant fraction is analyzed.
3.0 INTERFERENCES
3.1 A reagent blank should be prepared and analyzed for the compounds of
interest prior to the use of this method. The level of interferences must be
below the method detection limit before this method is performed on actual
samples.
3.2 The procedures for reagent purification outlined here should be
considered to be the minimum requirements for use of this method. More extensive
procedures may be necessary to achieve acceptable levels of interferences for
some analytes.
4.0 APPARATUS AND MATERIALS
4.1 Chromatography column - 300 mm x 10 mm ID, with a Teflon® stopcock.
NOTE: Columns with fritted glass discs are difficult to clean once the column
has been used to process highly contaminated extracts. Columns without
frits may be purchased, and a small pad of Pyrex® glass wool may be used
to retain the adsorbent. Prewash the glass wool pad with 50 ml of acetone
followed by 50 ml of elution solvent prior to packing the column with
adsorbent.
4.2 Beakers - Appropriates sizes
4.3 Reagent bottle - Appropriate sizes
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tops.
4.4 Muffle furnace - capable of maintaining 400°C.
4.5 Vials - Glass, 2-mL capacity, with Teflon®-lined screw caps or crimp
4.6 Vacuum manifold - VacElute Manifold SPS-24 (Analytichem
International), Visipr'ep (Supelco, Inc.) or equivalent, consisting of glass
vacuum basin, collection rack and funnel, collection vials, replaceable stainless
steel delivery tips, built-in vacuum bleed valve and gauge. The system is
connected to a vacuum pump or water aspirator through a vacuum trap made from a
500-mL sidearm flask fitted with a one-hole stopper and glass tubing. The
manifold is needed for use of the cartridge cleanup protocol.
4.7 Top-loading balance - capable of weighing 0.01 g.
5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Sodium sulfate - Sodium sulfate (granular, anhydrous), Na2S04.
Purify by heating at 400°C for 4 hours in a shallow tray, or by precleaning the
sodium sulfate with methylene chloride. A method blank must be analyzed in order
to demonstrate that there is no interference from the sodium sulfate.
5.3 Eluting solvents - all solvents must be pesticide quality or
equivalent.
5.3.1 Diethyl Ether, C2H5OC2H5. Must be free of peroxides as
indicated by test strips (EM Quant, or equivalent). Procedures for
removal of peroxides are provided with the test strips. After cleanup,
20 ml of ethyl alcohol preservative must be added to each liter of ether.
5.3.2 Methanol, CH3OH
5.3.3 Pentane, CH3(CH2)3CH3
5.3.4 Hexane, C6H14
5.3.5 Methylene chloride, CH2C12
5.3.6 Acetone, CH3COCH3
5.4 Granular alumina, for column cleanup procedure
5.4.1 Neutral alumina, for cleanup of phthalates, activity Super
I, W200 series (ICN Life Sciences Group, No. 404583 or equivalent). To
activate, place 100 g of alumina into a 500-mL beaker and heat for
approximately 16 hr at 400°C. After heating, transfer to a 500-mL reagent
bottle. Tightly seal the bottle and cool to room temperature. When cool,
add 3 mL of organic-free reagent water. Mix thoroughly by shaking or
rolling for 10 min and let it stand for at least 2 hr. The preparation
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should be homogeneous before use. Keep the bottle sealed tightly to
ensure proper activity.
5.4.2 Basic alumina, for cleanup of nitrosamines, activity Super
I, W200 series (ICN Life Sciences Group, No. 404571, or equivalent). To
activate, place 100 g of alumina into a 500-mL reagent bottle and add 2
ml of organic-free reagent water. Mix thoroughly by shaking or rolling
for 10 min and let it stand for at least 2 hr. The preparation should be
homogeneous before use. Keep the bottle sealed tightly to ensure proper
activity.
5.5 Alumina cartridges - 40 pm particles, 60 A pores, for cleanup of
phthalates. The cartridges from which this method were developed consist of 6-mL
serological-grade polypropylene tubes, with the 1 g of alumina held between two
polyethylene or stainless steel frits with 20 ^m pores. Cartridges containing
0.5 g and 2.0 g of alumina are available, however, the compound elution patterns
need to be verified when cartridges containing other than 1 g of alumina are
used.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
The chromatographic separation procedures for the phthalate esters may be
accomplished by either the column cleanup approach (Sec. 7.3) or the cartridge
cleanup approach (Sec. 7.4). The procedure for the nitrosamines includes only
the column cleanup approach (Sec. 7.5). Sec. 7.1 describes the procedures for
assembling and conditioning the alumina cartridges. Sec. 7.2 describes general
procedures for handling sample extracts prior to cleanup.
The column chromatography procedures employ a larger amount of alumina
than the cartridge procedures and, therefore, have a greater cleanup capacity.
Samples that exhibit greater degrees of interferences should be cleaned up using
the column procedures. However, both techniques have limitations on the amount
of interferences that they can remove.
7.1 Cartridge set-up and conditioning
7.1.1 Arrange the cartridges on the manifold in the closed-valve
position.
7.1.2 Turn on the vacuum pump and set the vacuum to 10 in (254 mm)
of Hg. Do not exceed the manufacturer's recommendation for manifold
vacuum. Flow rates may be controlled by opening and closing cartridge
valves.
7.1.3 Condition the cartridges by adding 4 mL of hexane to each
cartridge. Slowly open the cartridge valves to allow hexane to pass
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through the sorbent beds to the lower frits. Allow a few drops per
cartridge to pass through the manifold to remove all air bubbles. Close
the valves and allow the solvent to soak the entire sorbent bed for 5
minutes. Do not turn off the vacuum.
7.1.4 Slowly open cartridge valves to allow the hexane to pass
through the cartridges. Close the cartridge valves when there is still
at least 1 mm of solvent above the sorbent bed. Do not allow cartridges
to become dry. If cartridges go dry, repeat the conditioning step.
7.2 Handling sample extracts
7.2.1 Reduce the sample extract volume to 2 ml (per 3500 series
methods) prior to cleanup. The extract solvent should be hexane for the
phthalate esters and methylene chloride for the nitrosamines.
7.2.2 Allow extract to reach room temperature if it was in cold
storage. Inspect the extract visually to ensure that there are no
particulates or phase separations and that no evaporative loss has taken
place. If crystals of sulfur are visible or if the presence of sulfur is
suspected, proceed with Method 3660.
7.3 Column procedure for phthalate esters
7.3.1 Place approximately 10 g of neutral alumina (Sec. 5.4.1) into
a 10-mm ID chromatographic column. Tap the column to settle the alumina,
and add 1-2 cm of anhydrous sodium sulfate to the top.
7.3.2 Pre-elute 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 (Sec. 7.2) onto the column using an additional 2
ml of hexane to complete the transfer.
7.3.3 Just prior to exposure of the sodium sulfate layer to the
air, add 35 ml of hexane to the column and continue the elution of the
column. Discard this hexane eluate.
7.3.4 Elute the column with 140 ml of ethyl ether/hexane (20/80,
v/v) and collect this fraction in a flask for concentration.
7.3.5 Concentrate the collected fraction to the volume required by
the determinative method (e.g., 2 mL for Method 8061), using the
techniques described in the appropriate 3500 series method. No solvent
exchange is necessary. Compounds that elute in this fraction are:
Bis(2-ethylhexyl) phthalate Diethyl phthalate
Butyl benzyl phthalate Dimethyl phthalate
Di-n-butyl phthalate Di-n-octyl phthalate
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7.4 Cartridge procedure for phthalate esters
NOTE: If organochlorine pesticides are known to be present in the extract,
Florisil cartridges (Method 3620) are recommended instead of Alumina
cartridges.
7.4.1 Using 1-g alumina cartridges, condition the cartridges with
hexane as described in Sec. 7.1.
7.4.2 Transfer the extract (Sec. 7.2) to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.4.3 When the entire extract has passed through the cartridge, but
before the cartridge becomes dry, rinse the sample vial with an additional
0.5 ml of solvent, and add the rinse to the cartridge to complete the
quantitative transfer.
7.4.4 Close the cartridge valve and turn off the vacuum after the
solvent has passed through, ensuring that the cartridge never gets dry.
7.4.5 Place a 5-mL vial or volumetric flask into the sample rack
corresponding to the cartridge position. Attach a solvent-rinsed
stainless steel solvent guide to the manifold cover and align it with the
collection vial.
7.4.6 Add 10 ml of acetone/hexane (20/80, v/v) to the cartridge.
Turn on the vacuum pump and adjust the pump pressure to 10 in (254 mm) of
Hg. Allow the solvent to soak the sorbent bed for 1 minute or less.
Slowly open the cartridge valve and collect the eluate into the collection
vial.
7.4.7 Adjust the final volume of the eluant to the volume listed
in the determinative method, using the techniques described in the
appropriate 3500 series method.
7.5 Column procedure for nitrosamines
7.5.1 Diphenylamine, if present in the original sample extract,
must be separated from the nitrosamines if N-nitrosodiphenylamine is to
be determined by this method.
7.5.2 Place approximately 12 g of basic alumina (Sec. 5.4.2) into
a 10-mm ID chromatographic column. Tap the column to settle the alumina
and add 1-2 cm of anhydrous sodium sulfate to the top.
7.5.3 Pre-elute the column with 10 ml of ethyl ether/pentane
(30/70, 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 (Sec. 7.2, in methylene chloride) onto the column using an
additional 2 ml of pentane to complete the transfer.
7.5.4 Just prior to exposure of the sodium sulfate layer to the
air, add 70 ml of ethyl ether/pentane (30/70, v/v). Discard the first
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10 mL of eluate. Collect the remainder of the eluate in a flask for
concentration.
This fraction contains some N-nitroso-di-n-propylamine, if any is
present in the sample extract.
7.5.5 Elute the column with 60 ml of ethyl ether/pentane (50/50,
v/v), collecting the eluate in a second flask for concentration. Add 15
mL of methanol to the flask.
This fraction will contain N-nitrosodimethylamine, most of the
N-nitroso-di-n-propylamine, and any diphenylamine that is present.
7.5.6 Concentrate both fractions to the final volumes listed in the
appropriate determinative method, using the techniques described in the
appropriate 3500 series method.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered before applying this method to actual samples. This
test applies to both the column cleanup and cartridge cleanup procedures. A
recovery check needs to be performed using standards of the target analytes at
known concentration.
8.2.1 This test should be conducted on each batch of alumina
following its activation (Sec. 5.4).
8.2.2 The efficiency of each lot of the solid-phase extraction
cartridges needs to be verified. Only lots of cartridges from which the
spiked analytes are quantitatively recovered may be used to process the
samples. A check should also be performed at least once on each
individual lot of cartridges and at least once for every 300 cartridges
of a particular lot, whichever frequency is greater.
8.3 The quality control samples associated with sample extracts that are
cleaned up using this method, should also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
Table 1 provides data for the recoveries of phthalate esters obtained from
1-g alumina cartridges.
10.0 REFERENCES
1. U.S. EPA, "Evaluation of Sample Extract Cleanup Using Solid-Phase
Extraction Cartridges", Project Report, December 1989.
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TABLE 1
PERCENT RECOVERIES AND ELUTION PATTERNS FOR
16 PHTHALATE ESTERS FROM ALUMINA CARTRIDGES8
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(Z-methoxyethyl) phthalate
Diamyl phthalate
Bis(Z-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(Z-n-butoxyethyl) phthalate
Bis(Z-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Average %
Recovery
108
129
92.6
107
88.3
92.2
100
101
93.2
113
104
99.5
101
97.2
103
110
Average
RSD
4.6
6.6
7.3
5.6
9.8
5.0
6.4
6.3
13
5.4
3.9
4.7
6.1
6.2
7.5
5.2
aAlumina cartridges (J.T. Baker) were conditioned with 4 mL of hexane. Each
experiment was performed in duplicate at three spiking concentrations (40 /ng,
80 ^g, and 120 p.g per compound, per cartridge). The cartridges were eluted with
5 mL of acetone/hexane (20/80, v/v).
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SPE Cartridge
Cleanup j
7.4.1 Condition
cartridges with
hexane.
7.4.2 - 7.4.3
Open valve,
transfer extract
to cartridge,
rinse sample
vial with CH2CI2.
7.4.4 - 7.4.5
Close valve/turn
off vacuum attach
solvent guide and
collection vial.
7.4.6 Add acetone/
hexane, apply
vacuum, open cart-
ridge valve, collect
eluate in collection
vial.
7.4.7 Concentrate
collected fraction,
adjust volume.
METHOD 3610B
ALUMINA COLUMN CLEANUP
Phthalate
Esters.
Nitrosammes
Column Cleanup I
Column Cleanup
7.2.1 Reduce
volume of
sample extract.
7.3.1 Put
alumina in column,
add anhydrous
sodium sulfate.
7.3.2
Preelute
column with
hexane.
7.3.2 Transfer
sample extract
to column, elute
column with
hexane.
7.3.4 Elute
column with
ethyl ether/
hexane. Collect
eluate in flask.
7.3.5 Concentrate
collected fraction
adjust volume.
Analyze by
appropriate
determinative
method.
7.2.1 Reduce
volume of
sample extract.
7.5.2 Put
alumina in column,
add anhydrous
sodium sulfate
7.5.3 Preelute
column with ethyl
ether/pentane.
Transfer sample
extract to column,
add pentane.
7.5.4 Elute column
with ethyl
ether/pentane.
Collect eluate in
second flask, add
methanol.
7.5.5 Elute
column with
ethyl ether/
pentane. Collect
eluate in second
flask, add
methanol.
7.5.6
Concentrate
both fractions;
adjust volume.
3610B - 9
Revision 2
January 1995
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METHOD 3611B
ALUMINA COLUMN CLEANUP AND SEPARATION OF PETROLEUM WASTES
1.0 SCOPE AND APPLICATION
1.1 Alumina is a highly porous and granular form of aluminum oxide. It
is available in three pH ranges (basic, neutral, and acidic) for use in
chromatographic cleanup procedures. Method 3611 utilizes neutral pH alumina to
separate petroleum wastes into aliphatic, aromatic, and polar fractions.
1.2 Method 3611 was formerly Method 3570 in the Second Edition of this
manual.
1.3 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of adsorbent, topped
with a water adsorbent, and then loaded with the sample to be analyzed. Elution
of the analytes is effected with a suitable solvent(s), leaving the interfering
compounds on the column. The eluate is then concentrated (if necessary).
3.0 INTERFERENCES
3.1 A reagent blank should be performed for the compounds of interest
prior to the use of this method. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
3.3 Caution must be taken to prevent overloading of the chromatographic
column. As the column loading for any of these types of wastes approaches
0.300 g of extractable organics, separation recoveries will suffer. If
overloading is suspected, an aliquot of the base-neutral extract prior to cleanup
may be weighed and then evaporated to dryness. A gravimetric determination on
the aliquot will indicate the weight of extractable organics in the sample.
3.4 Mixtures of petroleum wastes containing predominantly polar solvents,
i.e., chlorinated solvents or oxygenated solvents, are not appropriate for this
method.
4.0 APPARATUS AND MATERIALS
4.1 Chromatography column: 300 mm x 10 mm ID, with Pyrex® glass wool at
bottom and a Teflon® stopcock.
3611B - 1 Revision 2
January 1995
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NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits may
be purchased. Use a small pad of Pyrex® glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed by
50 ml of elution solvent prior to packing the column with adsorbent.
4.2 Beakers: Appropriate sizes.
4.3 Reagent bottle: Appropriate sizes.
4.4 Muffle furnace.
4.5 Water bath: Heated with concentric ring cover, capable of temperature
control (±5°C). The bath should be used in a hood.
4.6 Erlenmeyer flasks: 50 and 250 ml.
5.0 REAGENTS
5.1 Sodium sulfate: (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.2 Eluting solvents:
5.2.1 Methanol, CH3OH - Pesticide quality or equivalent.
5.2.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.2.3 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.3 Alumina: Neutral 80-325 MCB chromatographic grade or equivalent. Dry
alumina overnight at 130°C prior to use.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 It is suggested that Method 3650, Acid-Base Partition Cleanup, be
performed on the sample extract prior to alumina cleanup.
7.2 Place approximately 10 g of alumina into a chromatographic column, tap
to settle the alumina, and add 1 cm of anhydrous sodium sulfate to the top.
7.3 Pre-elute the column with 50 mL of hexane. Discard the eluate and,
just prior to exposure of the sodium sulfate layer to the air, quantitatively
3611B - 2 Revision 2
January 1995
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transfer the 1 ml sample extract onto the column using an additional 1 ml of
hexane to complete the transfer. To avoid overloading the column, it is
suggested that no more than 0.300 g of extractable organics be placed on the
column (see Sec. 3.3).
7.4 Just prior to exposure of the sodium sulfate to the air, elute the
column with a total of 15 ml of hexane. If the extract is in 1 ml of hexane, and
if 1 ml of hexane was used as a rinse, then 13 ml of additional hexane should be
used. Collect the effluent in a 50 ml flask and label this fraction
"base/neutral aliphatics." Adjust the flow rate to 2 mL/min.
7.5 Elute the column with 100 ml of methylene chloride and collect the
effluent in a 250 ml flask. Label this fraction "base/neutral aromatics."
7.6 Elute the column with 100 ml of methanol and collect the effluent in
a 250 ml flask. Label this fraction "base/neutral polars."
7.7 Following cleanup, concentrate the fractions to the final volumes
listed in the appropriate determinative method, using the techniques described
in an appropriate 3500 series method. Analysis follows as specified in the
determinative procedure.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst should demonstrate that the compounds of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
9.1 The precision and accuracy of the method will depend upon the overall
performance of the sample preparation and analysis.
9.2 Rag oil is an emulsion consisting of crude oil, water, and soil
particles. It has a density greater than crude oil and less than water. This
material forms a layer between the crude oil and water when the crude oil is
allowed to gravity separate at the refinery. A rag oil sample was analyzed by
a number of laboratories according to the procedure outlined in this method. The
results of these analyses by GC/MS for selected components in the rag oil are
presented in Table 1. Reconstructed ion chromatograms from the GC/MS analyses
are included as Figures 1 and 2.
3611B - 3 Revision 2
January 1995
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10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3611B - 4 Revision 2
January 1995
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TABLE 1
RESULTS OF ANALYSIS FOR SELECTED COMPONENTS IN RAG OIL
Analyte
Naphthalene
Fluorene
Phenanthrene
2-Methyl naphtha! ene
Dibenzothiophene
Methyl phenanthrene
Methyl dibenzothiophene
Mean
Cone. (mg/kg)a
216
140
614
673
1084
2908
2200
Standard
Deviation
42
66
296
120
286
2014
1017
%RSDb
19
47
18
18
26
69
46
Average Surrogate Recovery
Nitrobenzene-d5 58.6 11
Terphenyl-d14 83.0 2.6
Phenol-d, 80.5 27.6
Naphthalene-d8 64.5 5.0
a Based on five determinations from three laboratories.
b Percent Relative Standard Deviation.
3611B - 5 Revision 2
January 1995
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FIGURE 1
Reconstructed Ion Chromatogram from GC/MS Analysis of the Aromatic Fraction
from Rag Oil
•1C MT* CUM* II . SOWS
lllfftM CM.li 27CM.I II OUT OF
IMC Oil HM.9M. Ill* OIL O.ICMN SMVIC EUCO MO. FMC IOUC 55
RMRZl C 1.3790 LMCLi N •. 4.« OUMh A •, l.t MSEi U M. 3
mraznt
TM 10
-IM.I
3611B - 6
-------
FIGURE 2
Reconstructed Ion Chromatogram from GC/MS Analysis of the Aliphatic Fraction
from Rag Oil
MC Mffci CMML tl
M/l?*l Hlttl« CM.fl 2ICM.I II
SMfflCi MC Cli FVH.M. » If t.lCMM SMftC CM* AMU. FMC I«C «
MHOEi C I.17N UKLi N t. 4.« •MNi A t. !.• MSCt U N. 3
•ff OF
Ml M
Ml ff
IK
3611B - 7
Revision 2
January 1995
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METHOD 3611B
ALUMINA COLUMN CLEANUP AND SEPARATION OF PETROLEUM WASTES
7.1 Cleanup
using Method
3650.
7.2 Add alumina
to chromatographic
column.
7.2 Add anhydrous
sodium sulfate
to top of column.
7.3 Preelute
column with
hexane.
7.3
Quantitatively add
extract to column.
7.4 Elute "base-neutral
aliphatics" fraction with
hexane.
7.5 Elute
"base-neutral
aromatics"
fraction with CH2CI2 •
7.6 Elute
"base-neutral
polars"
fraction with methanol.
7.7
Concentrate
extracts.
Analyze using
appropriate
determinative
Method.
3611B - 8
Revision 2
January 1995
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METHOD 3620B
FLORISIL CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Florisil, a registered trade name of the Floridin Co., is a magnesium
silicate with acidic properties. It is used to separate analytes from
interfering compounds prior to sample analysis by gas chromatography.
1.2 Florisil has been used for the cleanup of pesticide residues and other
chlorinated hydrocarbons; the separation of nitrogen compounds from hydrocarbons;
the separation of aromatic compounds from aliphatic-aromatic mixtures; and
similar applications for use with fats, oils, and waxes (Floridin).
Additionally, Florisil is considered good for separations with steroids, esters,
ketones, glycerides, alkaloids, and some carbohydrates.
1.3 Florisil cleanup may be accomplished using a glass chromatographic
column packed with Florisil or using solid-phase extraction cartridges containing
Florisil.
1.4 This method includes procedures for cleanup of sample extracts
containing the following analyte groups:
Phthalate esters Chlorinated hydrocarbons
Nitrosamines Organochlorine pesticides
Nitroaromatics Organophosphates
Haloethers Organophosphorus pesticides
Aniline and aniline derivatives
1.5 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 This method describes procedures for Florisil cleanup of solvent
extracts of environmental samples. It provides the option of using either
traditional column chromatography techniques to solid-phase extraction
cartridges. Generally, the traditional column chromatography technique uses
larger amounts of adsorbent and, therefore, has a greater cleanup capacity.
2.2 In the column cleanup protocol, the column is packed with the required
amount of adsorbent, topped with a water adsorbent, and then loaded with the
sample extract. Elution of the analytes is effected with a suitable solvent(s),
leaving the interfering compounds on the column. The eluate may be further
concentrated prior to gas chromatographic analysis.
2.3 The cartridge cleanup protocol uses solid-phase extraction cartridges
containing 40 urn particles of Florisil (60 A pores). Each cartridge is washed
with solvent immediately prior to use. The sample extract is loaded onto the
3620B - 1 Revision 2
January 1995
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cartridge which is then eluted with suitable solvent(s). A vacuum manifold is
required to obtain reproducible results. The eluate may be further concentrated
prior to gas chromatographic analysis.
3.0 INTERFERENCES
3.1 A reagent blank should be prepared and analyzed for the compounds of
interest prior to the use of this method. The level of interferences must be
below the method detection limit before this method is performed on actual
samples.
3.2 The procedures for reagent purification outlined here should be
considered to be the minimum requirements for use of this method. More extensive
procedures may be necessary to achieve acceptable levels of interferences for
some analytes. However, during the evaluation of the cartridge cleanup
procedure, phthalate esters were detected in the Florisil cartridge method blanks
at concentrations up to 400 ng per cartridge. Therefore, complete removal of the
phthalate esters from Florisil cartridges may not be possible.
4.0 APPARATUS AND MATERIALS
4.1 Chromatography column - 300 mm x 10 mm ID, with a Teflon® stopcock.
NOTE: Columns with fritted glass discs are difficult to clean once the column
has been used to process highly contaminated extracts. Columns without
frits may be purchased, and a small pad of Pyrex® glass wool may be used
to retain the adsorbent. Prewash the glass wool pad with 50 ml of acetone
followed by 50 ml of elution solvent prior to packing the column with
adsorbent.
4.2 Beakers - Appropriate sizes.
4.3 Reagent bottle - Appropriate sizes.
4.4 Muffle furnace - capable of maintaining 400°C.
4.5 Vials - Glass, 10-mL and 25-mL capacity, with Teflon®-!ined screw caps
or crimp tops.
4.6 Vacuum manifold - VacElute Manifold SPS-24 (Analytichem
International), Visiprep (Supelco, Inc.) or equivalent, consisting of glass
vacuum basin, collection rack and funnel, collection vials, replaceable stainless
steel delivery tips, built-in vacuum bleed valve and gauge. The system is
connected to a vacuum pump or water aspirator through a vacuum trap made from a
500-mL sidearm flask fitted with a one-hole stopper and glass tubing. The
manifold is required for use of the cartridge cleanup protocol.
4.7 Top-loading balance - capable of weighing to 0.01 g.
3620B - 2 Revision 2
January 1995
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5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Granular Florisil - for column cleanup procedure. Pesticide residue
(PR) grade (60/100 mesh). Purchase Florisil activated at 675eC, store in glass
containers with ground-glass stoppers or foil-lined screw caps.
5.3 Laurie acid - reagent grade. Used for the standardization of the
Florisil activity. Weigh 10.00 g of lauric acid in a 500-mL volumetric flask.
Add 50 mL of hexane to the flask to dissolve the lauric acid. Swirl the flask
gently until the lauric acid is dissolved, then dilute the solution in the flask
to 500 mL with additional hexane.
5.4 Phenolphthalein Indicator - Dissolve 1 g of phenolphthalein in ethanol
and dilute to 100 mL in a 100-mL volumetric flask.
5.5 Sodium hydroxide - Weigh out 20 g of NaOH (pellets, reagent grade) in
a 500-mL volumetric flask. Dissolve in organic-free reagent water and dilute to
500 mL to make a IN solution. Dilute 25 mL of the IN NaOH to 500 mL with water
in a second 500-mL volumetric flask, yielding a 0.05N solution. The NaOH
solution must be standardized against lauric acid, as follows.
5.5.1 Weigh 100 - 200 mg of lauric acid to the nearest 1 mg in a
125-mL Erlenmeyer flask. Add 50 mL of ethanol to the flask and swirl to
dissolve the lauric acid.
5.5.2 Add 3 drops of phenolphthalein indicator to the flask, and
titrate with the 0.05 N NaOH solution to a permanent endpoint (i.e., the
indicator color does not disappear when the solution is allowed to stand
for 1 min).
5.5.3 Calculate the "strength" of the NaOH solution as the mg of
lauric acid neutralized per mL of NaOH solution.
5.6 Deactivation/Activation of Florisil
5.6.1 Deactivation of Florisil - for cleanup of phthalate esters.
To prepare for use, place 100 ± 10 g of Florisil into a 500-mL beaker and
heat to 40°C for approximately 16 h. After heating, transfer to a 500-mL
reagent bottle. Tightly seal and cool to room temperature. When cool,
add 3 ± 0.1 mL of organic-free reagent water. Mix thoroughly by shaking
or rolling for 10 min and let stand for at least 2 h. Keep the bottle
sealed tightly.
5.6.2 Activation of Florisil - for cleanup of nitrosamines,
organochlorine pesticides and PCBs, nitroaromatics, haloethers,
chlorinated hydrocarbons, organophosphorus pesticides, and chlorophenoxy
acid herbicides. Just before use, activate each batch at least 16 h at
130°C in a glass container loosely covered with aluminum foil.
Alternatively, store the Florisil in an oven at 130°C. Cool the Florisil
in a desiccator before use.
3620B - 3 Revision 2
January 1995
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5.6.3 Florisil from different batches or sources may vary in
adsorptive capacity. To standardize the amount of Florisil which is used,
use the 1 auric acid value, described below. The procedure determines the
adsorption from a hexane solution of lauric acid (mg) per g of Florisil.
5.6.3.1 Weigh 2.000 g of Florisil in a 25-mL glass-stoppered
Erlenmeyer flask. Cover loosely with aluminum foil and heat
overnight at 130°C. Stopper the flask and cool to room
temperature.
5.6.3.2 Add 20.0 ml of the lauric acid solution to the flask,
stopper, and shake occasionally for 15 min.
5.6.3.3 Let the Florisil settle and using a volumetric pipet,
transfer 10.0 ml of supernatant liquid into a 125-mL Erlenmeyer
flask. Avoid inclusion of any Florisil.
5.6.3.4 Add 60 ml of ethanol and 3 drops of the
phenolphthalein indicator solution to the flask.
5.6.3.5 Titrate the solution in the flask with the 0.05N NaOH
solution until a permanent end point is reached (i.e., the indicator
color does not disappear when the solution is allowed to stand for
1 min).
5.6.3.6 The lauric acid value is calculated as follows:
lauric acid value = 200 - (Titration volume in ml of NaOH x Strength of NaOH)
where the strength of the NaOH is measured in Sec. 7.5.3 as the mg
of lauric acid neutralized per ml of NaOH solution.
5.6.3.7 Use the following equation to obtain an equivalent
quantity of any batch of Florisil.
lauric acid value on n . . . , . - r, . .,
x 20 g = Required weight of Florisil
110
5.7 Sodium sulfate (granular, anhydrous), Na2S04 - Purify by heating at
400"C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. A method blank must be analyzed in order to demonstrate that
there is no interference from the sodium sulfate.
5.8 Florisil cartridges - 40 jum particles, 60 A pores. The cartridges
from which this method were developed consist of 6-mL serological- grade
polypropylene tubes, with the 1 g of Florisil held between two polyethylene or
stainless steel frits with 20 /xm pores. Cartridges containing 0.5 g and 2.0 g
of Florisil are available, however, the compound elution patterns must be
verified when cartridges containing other than 1 g of Florisil are used.
3620B - 4 Revision 2
January 1995
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5.9 Eluting solvents - all solvents must be pesticide quality or
equivalent.
5.9.1 Diethyl Ether, C2H5OC2H5. Must be free of peroxides as
indicated by test strips (EM Quani, or equivalent). Procedures for
removal of peroxides are provided with the test strips. After cleanup, 20
ml of ethyl alcohol preservative must be added to each liter of ether.
5.9.2 Pentane, CH3(CH2)3CH3
5.9.3 Hexane, C6HU
5.9.4 Methylene chloride, CH2C12
5.9.5 Acetone, CH3COCH3
5.9.6 Petroleum ether (boiling range 30-60°C)
5.9.7 Toluene, C6H5CH3
5.9.8 Isopropanol, (CH3)2CHOH
5.10 Florisil cartridge phenol check solution (for the organochlorine
pesticide technique) - Prepare a solution of 2,4,5-Trichlorophenol in acetone at
a concentration of 100 mg/L.
5.11 Florisil cartridge pesticide check solution - Prepare a solution
containing the following analytes in hexane:
a-BHC 5 mg/L
Heptachlor 5 mg/L
Y-BHC 5 mg/L
Endosulfan I 5 mg/L
Dieldrin 10 mg/L
Endrin 10 mg/L
4,4'-ODD 10 mg/L
4,4'-DDT 10 mg/L
Methoxychlor 50 mg/L
Tetrachloro-m-xylene 20 mg/L
Decachlorobiphenyl 20 mg/L
5.12 Chlorophenoxy acid herbicide check solution - Prepare a solution
containing 2,4,5-T methyl ester at 100 mg/L, Pentachlorophenyl methyl ester at
50 mg/L, and Picloram methyl ester at 200 mg/L.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this chapter, Organic Analytes,
Section 4.1.
3620B - 5 Revision 2
January 1995
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7.0 PROCEDURE
Sec. 7.1 describes the procedures for assembling and conditioning the
alumina cartridges. Sec. 7.2 describes general procedures for handling sample
extracts prior to cleanup. Sees. 7.3 - 7.13 describe the column and cartridge
procedures for phthalate esters; nitrosamines; organochlorine pesticides,
haloethers, and organophosphorus pesticides; nitroaromatics and isophorone;
chlorinated hydrocarbons; aniline and aniline derivatives; organophosphates;
and derivatized chlorophenoxy acid herbicides.
The column chromatography procedures employ a larger amount of Florisil
than the cartridge procedures and, therefore, have a greater cleanup capacity.
Samples that exhibit greater degrees of interferences should be cleaned up using
the column procedures. However, both techniques have limitations on the amount
of interferences that they can remove.
If the interference is caused by high boiling materials, then Method 3640
should be employed prior to Florisil cleanup. If the interference is caused by
relatively polar compounds in the same boiling range as the analytes of interest,
then multiple column or cartridge cleanups may be required. For additional
cleanup of organochlorine pesticides and Aroclors, see Method 3665. If crystals
of sulfur are present in the extract, then Method 3660 should be employed prior
to Florisil cleanup.
7.1 Cartridge set-up and conditioning
7.1.1 Arrange the cartridges on the manifold in the closed-valve
position.
7.1.2 Turn on the vacuum pump and set the vacuum to 10 in (254 mm)
of Hg. Do not exceed the manufacturer's recommendation for manifold
vacuum. Flow rates may be controlled by opening and closing cartridge
valves.
7.1.3 Condition the cartridges by adding 4 ml of hexane to each
cartridge. Slowly open the cartridge valves to allow hexane to pass
through the sorbent beds to the lower frits. Allow a few drops per
cartridge to pass through the manifold to remove all air bubbles. Close
the valves and allow the solvent to soak the entire sorbent bed for 5
minutes. Do not turn off the vacuum.
7.1.4 Slowly open cartridge valves to allow the hexane to pass
through the cartridges. Close the cartridge valves when there is still at
least 1 mm of solvent above the sorbent bed. Do not allow cartridges to
become dry. If cartridges go dry, repeat the conditioning step.
7.2 Handling sample extracts
7.2.1 Reduce the sample extract volume to 2 ml prior to cleanup
for:
Phthalate esters Chlorinated hydrocarbons
Nitrosamines Chlorophenoxy acid herbicides
Nitroaromatics and Isophorone Aniline and aniline derivatives
3620B - 6 Revision 2
January 1995
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The extract solvent should be hexane for the phthalate esters,
nitroaromatics, chlorinated hydrocarbons, and chlorophenoxy acid
herbicides, and methylene chloride for the nitrosamines and aniline and
aniline derivatives.
7.2.2 Reduce the sample extract volume to 10 ml prior to cleanup
for:
Organochlorine pesticides
Haloethers
Organophosphorus pesticides
Organophosphates
The extract solvent should be hexane for these analytes.
7.2.3 Allow the extract to reach room temperature if it was in cold
storage. Inspect the extract visually to ensure that there are no
particulates or phase separations and that no evaporative loss has taken
place. If crystals of sulfur are visible or if the presence of sulfur is
suspected, proceed with Method 3660.
7.3 Column procedure for phthalate esters
7.3.1 Place approximately 10 g of deactivated Florisil (Sec. 5.2.1)
into a 10 mm ID chromatographic column. Tap the column to settle the
Florisil and add approximately 1 cm of anhydrous sodium sulfate to the
top.
7.3.2 Pre-elute 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.
7.3.3 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.
7.3.4 Elute the column with 100 mL of ethyl ether/hexane (20/80,
v/v) and collect this fraction in a flask (e.g. a 500 mL K-D flask
equipped with a clean 10 mL concentrator tube). Concentrate the collected
fraction to the volume required by the determinative method (e.g., 2 mL
for Method 8061). No solvent exchange is necessary. Compounds that elute
in this fraction are:
Bis(2-ethylhexyl) phthalate Diethyl phthalate
Butyl benzyl phthalate Dimethyl phthalate
Di-n-butyl phthalate Di-n-octyl phthalate
7.4 Cartridge procedure for phthalate esters
7.4.1 Using 1-g Florisil cartridges, condition the cartridges with
hexane as described in Sec. 7.1.
3620B - 7 Revision 2
January 1995
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7.4.2 Transfer the extract (Sec. 7.2) to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.4.3 When the entire extract has passed through the cartridges,
but before the cartridge becomes dry, rinse the sample vials with an
additional 0.5 mL of solvent, and add the rinse to the cartridges to
complete the quantitative transfer.
7.4.4 Close the cartridge valve and turn off the vacuum after the
solvent has passed through, ensuring that the cartridge never gets dry.
7.4.5 Place a 5-mL vial or volumetric flask into the sample rack
corresponding to the cartridge position. Attach a solvent-rinsed
stainless steel solvent guide to the manifold cover and align it with the
collection vial.
7.4.6 If the sample is suspected to contain organochlorine
pesticides, elute the cartridge with methylene chloride/hexane (20/80,
v/v). Turn on the vacuum pump and adjust the pump pressure to 10 inches
(254 mm) of Hg. Allow the solvent to soak the sorbent bed for 1 minute or
less. Slowly open the cartridge valve, and collect the eluate (this
fraction contains the organochlorine pesticides, and should be discarded).
7.4.7 Close the cartridge valve, replace collection vials, and add
10 ml of acetone/hexane (10/90, v/v) to the cartridge. Slowly open the
cartridge valve and collect the eluate into the collection vial. This
fraction contains the phthalate esters, and should be retained for
analysis.
7.4.8 Adjust the final volume of the eluant to the volume specified
in the determinative method.
7.5 Column procedure for nitrosamines
7.5.1 Add a weight of activated Florisil (nominally 22 g)
predetermined by calibration (Sec. 5.6.3.7) into a 20 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 5 mm of anhydrous sodium sulfate to the top.
7.5.2 Pre-elute 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 (Sec. 7.2) onto the column using an additional 2 ml of pentane to
complete the transfer.
7.5.3 Just prior to the exposure of the sodium sulfate layer to the
air, elute the column with 90 ml of ethyl ether/pentane (15/85, v/v).
Discard the eluate. This fraction will contain and diphenylamine present
in the extract.
7.5.4 Elute the column with 100 mL of acetone/ethyl ether (5/95,
v/v), collecting the eluate in a flask (e.g. a 500 mL K-D flask equipped
3620B - 8 Revision 2
January 1995
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with a clean 10 ml concentrator tube). This fraction will contain all of
the nitrosamines listed in the scope of the method.
7.5.5 Add 15 ml of methanol to the collected fraction, and
concentrate this fraction to the volume listed in the determinative
method, using the techniques described in the appropriate 3500 series
method.
7.6 Column procedure for organochlorine pesticides, haloethers, and
organophosphorus pesticides (see Table 2 for fractionation patterns of
organophosphorus pesticides)
7.6.1 Add a weight of activated Florisil (nominally 20 g),
predetermined by calibration (Sec. 5.6.3.7), to a 20 mm ID chromatographic
column. Settle the Florisil by tapping the column. Add anhydrous sodium
sulfate to the top of the Florisil to form a layer 1 to 2 cm deep.
7.6.2 Pre-elute the column with 60 ml of hexane and discard the
eluate. Just prior to exposure of the sodium sulfate to air,
quantitatively transfer the 10-mL sample extract (Sec. 7.2) onto the
column, completing the transfer with two 1-2 mL rinses with hexane.
7.6.3 Place a flask (e.g. a 500 ml K-D flask equipped with a 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 ethyl ether/hexane (6/94, v/v) using a drip rate
of about 5 mL/min. This is Fraction 1, and all of the haloethers are in
this fraction. Remove the flask and set aside for later concentration.
7.6.4 Elute the column again, using 200 ml of ethyl ether/hexane
(15/85, v/v), into a second flask. This is Fraction 2.
7.6.5 Perform a third elution using 200 ml of diethyl ether/hexane
(50/50, v/v), collecting the eluate in a third flask. This is Fraction 3.
7.6.6 Perform a final elution with 200 ml of 100% ethyl ether,
collecting the eluate in a fourth flask. This is Fraction 4.
7.6.7 Concentrate the four eluates to the volume listed in the
determinative method, using the techniques described in the appropriate
3500 series method.
7.7 Cartridge procedure for organochlorine pesticides and Aroclors
7.7.1 Using 1-g Florisil cartridges, condition the cartridges with
hexane, as described in Sec. 7.1.
7.7.2 Transfer the extract (Sec. 7.2) to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.7.3 When the entire extract has passed through the cartridge, but
before the cartridge becomes dry, rinse the sample vial with an additional
3620B - 9 Revision 2
January 1995
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0.5 mL of hexane, and add the rinse to the cartridge to complete the
quantitative transfer.
7.7.4 Close the cartridge valve and turn off the vacuum after the
solvent has passed through, ensuring that the cartridge never goes dry.
7.7.5 Place a 5-mL vial or volumetric flask into the sample rack
corresponding to the cartridge position. Attach a solvent-rinsed
stainless steel solvent guide to the manifold cover and align with the
collection vial.
7.7.6 If there is no need to separate the organochlorine pesticides
from the Aroclors, then add 9 mL of acetone/hexane (10/90, v/v) to the
cartridge. Turn on the vacuum pump and adjust the pump pressure to 10
inches (254 mm) of Hg. Allow the solvent to soak the sorbent bed for 1
minute or less. Slowly open the cartridge valve and collect, the eluate
into the collection vial.
7.7.7 In order to separate the organochlorine pesticides from the
Aroclors, add 3 ml of hexane to the cartridge. Turn on the vacuum pump
and adjust the pump pressure to 10 inches (254 mm) of Hg. Allow the
solvent to soak the sorbent bed for 1 minute or less. Slowly open the
cartridge valve and collect the eluate into the collection vial. This is
Fraction 1 and it will contain most of the organochlorine pesticides.
Proceed with'Sec. 7.7.8.
7.7.8 Close the cartridge valve, replace the collection vial, and
add 5 ml of methylene chloride/hexane (74/26, v/v) to the cartridge.
Slowly open the cartridge valve and collect the eluate into the collection
vial. This is Fraction 2 and it will contain additional pesticides. The
eluants from Fraction 1 and Fraction 2 may be combined for analysis. If
the Aroclors are analytes of interest, proceed with Sec. 7.7.9.
7.7.9 Close the cartridge valve, replace collection vials, and add
5 ml of acetone/hexane (10/90, v/v) to the cartridge. Slowly open the
cartridge valve and collect the eluate into the collection vial. This is
Fraction 3 and it will contain the Aroclors.
7.7.10 Adjust the final volume of the eluant to the volume listed
in the determinative method, using the techniques described in the
appropriate 3500 series method.
7.8 Column procedure for nitroaromatics and isophorone
7.8.1 Add a weight of activated Florisil (nominally 10 g)
predetermined by calibration (Sec. 5.6.3.7) into a 10 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 1 cm of anhydrous sodium sulfate to the top.
7.8.2 Pre-elute the column with methylene chloride/hexane (10/90,
v/v) at 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 (Sec. 7.2) onto the column using an additional 2 ml of
hexane to complete the transfer.
3620B - 10 Revision 2
January 1995
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7.8.3 Just prior to exposure of the sodium sulfate layer to the
air, add 30 ml of methylene chloride/hexane (10/90, v/v) and continue the
elution of the column. Discard the eluate.
7.8.4 Elute the column with 90 ml of ethyl ether/pentane (15/85,
v/v) and discard the eluate. This fraction will contain any diphenylamine
present in the extract.
7.8.5 Elute the column with 100 ml of acetone/ethyl ether (5/95,
v/v), and collect the eluate in a flask (e.g. 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.
7.8.6 Add 15 ml of methanol to the collected fraction, and
concentrate to the volume listed in the determinative method, using the
techniques described in the appropriate 3500 series method.
7.8.7 Elute the column with 30 ml of acetone/methylene chloride
(10/90, v/v), and collect the eluate in a flask (e.g. a 500-mL K-D flask
equipped with a 10-mL concentrator tube). Concentrate the collected
fraction to the volume listed in the determinative method, using the
techniques described in the appropriate 3500 series method, and exchanging
the solvent to hexane. Compounds that elute in this fraction are:
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Isophorone
Nitrobenzene
7.9 Column procedure for chlorinated hydrocarbons
7.9.1 Add a weight of activated Florisil (nominally 12 g)
predetermined by calibration (Sec. 5.6.3.7) into a 10 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 1 to 2 cm of anhydrous sodium sulfate to the top.
7.9.2 Pre-elute 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 (Sec. 7.2) to the column
by decantation and subsequent petroleum ether washings. Discard the
eluate.
7.9.3 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 flask (e.g. a 500-mL K-D flask equipped with a 10-mL
concentrator tube).
This fraction should contain the following chlorinated hydrocarbons:
2-Chloronaphthalene Hexachlorobenzene
1,2-Di chlorobenzene Hexachlorobutadi ene
1,3-Dichlorobenzene Hexachlorocyclopentadiene
1,4-Di chlorobenzene Hexachloroethane
1,2,4-Trichlorobenzene
3620B - 11 Revision 2
January 1995
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7.9.4 Concentrate the collected fraction to the volume listed in
the determinative method, using the techniques described in the
appropriate 3500 series method.
7.10 Cartridge procedure for chlorinated hydrocarbons
7.10.1 Using 1-g Florisil cartridges, condition the cartridges with
5 mL of acetone/hexane (10/90, v/v) as described in Sec. 7.1.
7.10.2 Transfer the extract (Sec. 7.2) to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.10.3 When the entire extract has passed through the cartridges,
but before the cartridge becomes dry, rinse the sample vial with an
additional 0.5 ml of acetone/hexane (10/90), and add the rinse to the
cartridges to complete the quantitative transfer.
7.10.4 Close the cartridge valve and turn off the vacuum after the
solvent has passed through, ensuring that the cartridge never gets dry.
7.10.5 Place a 5-mL vial or volumetric flask into the sample rack
corresponding to the cartridge position. Attach a solvent-rinsed
stainless steel solvent guide to the manifold cover and align it with the
collection vial.
7.10.6 Add 10 ml of acetone/hexane (10/90, v/v) to the cartridge.
Turn on the vacuum pump and adjust the pump pressure to 10 inches (254
mm) of Hg. Allow the solvent to soak the sorbent bed for 1 minute or
less. Slowly open the cartridge valve and collect the eluate into the
collection vial.
7.10.7 Adjust the final volume of the eluant to the volume listed
in the determinative method, using the techniques described in the
appropriate 3500 series method.
7.11 Column procedure for Aniline and Aniline derivatives (see Table 4 for
elution patterns)
7.11.1 Add a weight of activated Florisil predetermined by
calibration (Sec. 5.6.3.7) into a 20 mm ID chromatographic column. Tap
the column to settle the Florisil.
7.11.2 Pre-elute the column with 100 ml of isopropanol/methylene
chloride (5/95, v/v), followed by 100 ml of hexane/methylene chloride
(50/50, v/v), followed by 100 ml of hexane. Discard the eluate and leave
a column of about 5 cm of hexane above the Florisil.
7.11.3 Quantitatively transfer the 2-mL sample extract (Sec. 7.2)
onto 2.0 g of activated Florisil in a 50-mL beaker, using a small volume
of methylene chloride, and dry under a gentle stream of nitrogen.
3620B - 12 Revision 2
January 1995
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7.11.4 Place the dried Florisil containing the sample extract onto
the chromatographic column, and wash the beaker which contained the
Florisil with 75 ml of hexane, adding this wash to the reservoir.
7.11.5 Elute the hexane from the column and discard. Stop the
column flow just prior to the exposure of the Florisil to air.
7.11.6 Elute the column with 50 ml of methylene chloride/hexane
(50/50, v/v), using a drip rate of about 5 mL/minute, and collect the
eluate in a flask (e.g. a 500-mL K-D flask equipped with a 10-mL
concentrator tube). This is Fraction 1.
7.11.7 Elute the column with 50 ml of isopropanol/hexane (5/95,
v/v), and collect the eluate in a second flask. This is Fraction 2.
7.11.8 Elute the column a third time using 50 ml of methanol/hexane
(5/95, v/v). Collect the eluate in a third flask. This is Fraction 3.
Frequently, it will prove useful to combine the three fractions prior to
analysis. However, in some situations, analysis of each separate fraction
may be required. Refer to Method 8131.
7.11.9 Concentrate the collected fractions to the volume listed in
the determinative method, using the techniques described in the
appropriate 3500 series method.
7.12 Column procedure for organophosphates
7.12.1 Add a weight of activated Florisil, predetermined by
calibration (Sec. 5.6.3.7), to a 20 mm ID chromatographic column. Settle
the Florisil by tapping the column. Add anhydrous sodium sulfate to the
top of the Florisil to form a layer 1 to 2 cm deep.
7.12.2 Pre-elute the column with 50-60 ml of hexane. Discard the
eluate and just prior to exposure of the sulfate layer to air,
quantitatively transfer the 10-mL sample extract (Sec. 7.2) onto the
column using a hexane wash to complete the transfer.
7.12.3 Just as the sample reaches the sodium sulfate, elute the
column with 100 mL of diethyl ether/hexane (10/90, v/v). Discard the
eluate.
7.12.4 Just prior to exposure of the sodium sulfate to air, elute
the column with 200 ml of diethyl ether/hexane (30/70, v/v). This
fraction contains all of the target analytes except for
tris(2,3-dibromopropyl) phosphate.
7.12.5 Elute the column with 200 mL of diethyl ether/hexane (40/60,
v/v). This fraction contains tris(2,3-dibromopropyl) phosphate.
7.12.6 Concentrate the collected fraction to the volume required by
the determinative method, using the techniques described in the
appropriate 3500 series method.
3620B - 13 Revision 2
January 1995
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7.13 Column procedure for derivatized chlorophenoxy acid herbicides
7.13.1 Add a weight of activated Florisil (nominally 4 g)
predetermined by calibration (Sec. 5.6.3.7) into a 20 mm ID
chromatographic column. Tap the column to settle the Florisil and add
approximately 5 mm of anhydrous sodium sulfate to the top.
7.13.2 Pre-elute the column with 15 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 to air, quantitatively transfer the 2-mL
sample extract (Sec. 7.2) onto the column, using an additional 2 ml of
hexane to complete the transfer.
7.13.3 Just prior to the exposure of the sodium sulfate layer to
the air, elute the column with 35 ml of methylene chloride/hexane (20/80,
v/v), collecting the eluate in a clean flask (e.g. a 500 ml K-D flask
equipped with a concentrator tube). This is Fraction 1, and will contain
any pentachlorophenyl methyl ester that is present.
7.13.4 Elute the column with 60 ml of methylene
chloride/acetonitrile/hexane (50/0.035/49.65, v/v/v), collecting the
eluate in a second flask. This is Fraction 2.
7.13.5 If Picloram is to be determined, perform a third elution
with the volume of diethyl ether determined from the Florisil check in
Sec. 8.2.4, collecting this eluate in a third flask. This is Fraction 3,
and will contain the Picloram.
7.13.6 The three fractions may be combined for analysis.
Concentrate the combined fractions to the volume listed in the
determinative method, using the techniques described in the appropriate
3500 series method.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered before applying this method to actual samples. This
test applies to both the column cleanup and cartridge cleanup procedures. A
recovery check must be performed using standards of the target analytes at known
concentration.
8.2.1 This test must be conducted on each batch of Florisil
following its activation (Sec. 5.4).
8.2.2 The efficiency of each lot of the solid-phase extraction
cartridges must be verified. Only lots of cartridges from which the
spiked analytes are quantitatively recovered may be used to process the
samples. A check should also be performed at least once on each
individual lot of cartridges and at least once for every 300 cartridges of
a particular lot, whichever frequency is greater.
3620B - 14 Revision 2
January 1995
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8.2.3 Organochlorine pesticides - To check each new lot of Florisil
cartridges before use, perform the following in duplicate:
8.2.3.1 Combine 0.5 ml of the 2,4,5-trichlorophenol solution
in Sec. 5.10, 1.0 ml of the pesticide solution in Sec. 5.11, and 0.5
ml of hexane in a vial.
8.2.3.2 Condition the cartridge as described in Sec. 7.2 and
then perform the cartridge cleanup starting with Sec. 7.11.
8.2.3.3 Elute the cartridge with 9 ml of acetone/hexane
(10/90, v/v) only. Reduce the volume to 1.0 ml and analyze by
Method 8081.
8.2.3.4 The lot of Florisil cartridges is acceptable if all
pesticides are recovered at 80 to 110 %, if the recovery of
trichlorophenol is less than 5 %, and if no peaks interfering with
the target analytes are detected.
8.2.4 Chlorophenoxy acid herbicides - To check each new lot of
granular Florisil perform the following:
8.2.4.1 Add 5 ml of the chlorophenoxy acid herbicide check
solution (Sec. 5.12) to a Florisil column packed and washed as in
Sec. 7.13.2.
8.2.4.2 Elute Fractions 1 and 2 as described in Sees. 7.13.3
and 7.13.4, collecting each in a separate flask.
8.2.4.3 Elute the column with approximately 100 ml diethyl
ether and collect ten separate 10-mL fractions.
8.2.4.4 Concentrate Fraction 1 and Fraction separately and
concentration each of the ten 10-mL diethyl ether fractions to 5 ml.
8.2.4.5 Analyze each of the 12 eluates by GC/ECD and
calculate the recovery of each analyte. Pentachlorophenyl methyl
ether should be found in Fraction 1. 2,4,5-T methyl ester (and the
methyl esters of the other chlorophenoxy acids) should be found in
Fraction 2. Determine the volume of diethyl ether that is required
to elute picloram methyl ester.
8.2.4.6 The lot of Florisil is acceptable is the target
analytes are quantitatively recovered and if the recovery of
trichlorophenol is less than 5%. No interferences should be
detected in any of these eluates.
8.3 The quality control samples associated with sample extracts that are
cleaned up using this method must also be processed through this cleanup method.
3620B - 15 Revision 2
January 1995
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9.0 METHOD PERFORMANCE
9.1 Table 1 provides recoveries of phthalate esters obtained from the
Florisil column procedure.
9.2 Table 2 provides recoveries of phthalate esters obtained from the
Florisil cartridge procedure.
9.3 Table 3 provides the distribution of organochlorine pesticides and
Aroclors from the Florisil column procedure.
9.4 Table 4 provides recoveries of Aroclors from the Florisil cartridge
procedure.
9.5 Table 5 provides the distribution of organochlorine pesticides from
the Florisil column procedure.
9.6 Table 6 provides the distribution of organophosphorus pesticides from
the Florisil column procedure.
9.7 Table 7 provides recoveries of chlorinated hydrocarbons obtained from
the Florisil cartridge procedure.
9.8 Table 8 provides the elution patterns for aniline compounds from the
Florisil column procedure.
10.0 REFERENCES
1. Gordon, A.J. and R.A. Ford, The Chemist's Companion: A Handbook of
Practical Data, Techniques, and References (New York: John Wiley & Sons,
Inc.), pp. 372, 374, and 375, 1972.
2. Floridin of ITT System, Florisil: Properties, Application, Bibliography,
Pittsburgh, Pennsylvania, 5M381DW.
3. 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.
4. U.S. Food and Drug Association, Pesticides Analytical Manual (Volume 1),
July 1985.
5. Lopez-Avila, V., Milanes, J., Dodhiwala, N.S., and Beckert, W.F., "Cleanup
of Environmental Sample Extracts Using Florisil Solid-Phase Extraction
Cartridges," J. Chrom. Sci. 27, 209-215, 1989.
6. US EPA "Evaluation of Sample Extract Cleanup Using Solid-Phase Extraction
Cartridges," Project Report, December 1989.
7. US EPA Method 650, Aniline and Selected Substituted Derivatives.
3620B - 16 Revision 2
January 1995
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8. Beckert, W.F., and Lopez-Avila, V., "Evaluation of SW-46 Method 8060 for
Phthalate Esters," Proceedings of the Fifth Annual Waste Testing and
Quality Assurance Symposium, 1989, pp. 144-156.
9. US EPA Method 608, Organochlorine Pesticides and PCBs.
3620B - 17 Revision 2
January 1995
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TABLE 1
AVERAGE RECOVERIES OF 16 PHTHALATE ESTERS FROM THE FLORISIL COLUMN PROCEDURE8
Average %
Compound Recovery
Dimethyl phthalate 40
Diethyl phthalate 57
Diisobutyl phthalate 80
Di-n-butyl phthalate 85
Bis(4-methyl-2-pentyl) phthalate 84
Bis(2-methoxyethyl) phthalate 0
Diamyl phthalate 82
Bis(2-ethoxyethyl) phthalate 0
Hexyl 2-ethylhexyl phthalate 105
Dihexyl phthalate 74
Benzyl butyl phthalate 90
Bis(2-n-butoxyethyl) phthalate 0
Bis(2-ethylhexyl) phthalate 82
Dicyclohexyl phthalate 84
Di-n-octyl phthalate 115
Dinonyl phthalate 72
aAverage recovery from two determinations, data from Reference 8.
3620B - 18 Revision 2
January 1995
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TABLE 2
AVERAGE RECOVERIES OF 16 PHTHALATE ESTERS FROM FLORISIL CARTRIDGES8
Average %
Compound Recovery
Dimethyl phthalate 89
Diethyl phthalate 97
Diisobutyl phthalate 92
Di-n-butyl phthalate 102
Bis(4-methyl-2-pentyl) phthalate 105
Bis(2-methoxyethyl) phthalate 78
Diamyl phthalate 94
Bis(2-ethoxyethyl) phthalate 94
Hexyl 2-ethylhexyl phthalate 96
Dihexyl phthalate 97
Benzyl butyl phthalate 99
Bis(2-n-butoxyethyl) phthalate 92
Bis(2-ethylhexyl) phthalate 98
Dicyclohexyl phthalate 90
Di-n-octyl phthalate 97
Dinonyl phthalate 105
aAverage recovery from two determinations, data from Reference 6.
3620B - 19 Revision 2
January 1995
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TABLE 3
DISTRIBUTION OF ORGANOCHLORINE PESTICIDES AND PCBs
IN FLORISIL COLUMN FRACTIONS
Compound
Percent Recovery by Fraction3
1 2 3
Aldrin
a-BHC
/3-BHC
6-BHC
Y-BHC
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Toxaphene
Aroclor 1016
Aroclor 1221
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1260
100
100
97
98
100
100
99
98
100
0 100
37 64
0 7
0 0
4 96
0 68
100
100
96
97
97
95 4
97
103
90
95
91
106
26
aEluant composition:
Data from Reference 9.
Fraction 1 - 200 mL of 6% ethyl ether in hexane
Fraction 2 - 200 mL of 15% ethyl ether in hexane
Fraction 3 - 200 mL of 50% ethyl ether in hexane
3620B - 20
Revision 2
January 1995
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TABLE 4
AVERAGE RECOVERIES OF AROCLORS FROM FLORISIL CARTRIDGES3
Average %
Compound Recovery
Aroclor 1016 105
Aroclor 1221 76
Aroclor 1232 90
Aroclor 1242 94
Aroclor 1248 97
Aroclor 1254 95
Aroclor 1260 90
al-g Florisil cartridges were loaded with 10 jug of each Aroclor and eluted with
3 ml of hexane. Data from Reference 6.
3620B - 21 Revision 2
January 1995
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TABLE 5
ELUTION PATTERNS AND RECOVERIES OF ORGANOCHLORINE PESTICIDES
FROM FLORISIL CARTRIDGES3
Compound
Fraction 1
Rec. RSD
Fraction 2
Rec. RSD
Fraction 3
Rec. RSD
a-BHC
0-BHC
S-BHC
Heptachlor
Y-BHC
Aldrin
Heptachlor epoxide
Chlordane
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
4,4'-DDD
Endosulfan II
Endrin aldehyde
4,4'-DDTb
' i
Endosulfan sulfate
Methoxychlor
83
36
0
94
0
93
0
48
0
94
0
0
38
0
0
50
0
3.4 26
13 78
102
2.3 0
100
2.0 0
102
9.8 66
101
1.7 0
101
57
12 69
58
36
1.9 11
96
11
3.1
2.3
1.4
2.4
3.5
2.7
2.9
7.5
2.9
13
7.2
16
3.4
0
0
0
0
0
0
0
0
0
0
0
0
0
61
79
59
12
9.9
2.6
3.1
3.0
al-g Florisil cartridges spiked with 0.5 M9 of each compound.
Eluant composition:
Fraction 1 - 3 mL of hexane
Fraction 2 - 5 ml of methylene chloride/hexane
(26/74, v/v)
Fraction 3 -5 ml of acetone/hexane (10/90, v/v)
bThese two compounds coelute on the DB-5 capillary column.
3620B - 22
Revision 2
January 1995
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TABLE 6
DISTRIBUTION OF ORGANOPHOSPHORUS PESTICIDES
IN FLORISIL CLEANUP FRACTIONS
Compound
Percent Recovery by Fraction3
1234
Azinphos methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Monochrotophos
Naled
Parathion
Parathion methyl
Phorate
Ronnel
Stirophos (Tetrachlorvinphos)
Sulfotepp
TEPP
Tokuthion (Prothiofos)
Trichloronate
aEluant composition: Fraction 1
Fraction 2
Fraction 3
Fraction 4
ND
>80
NR
100
NR
ND
25-40
V
ND
R
V
ND
ND
NR
0-62
>80
ND
V
ND
>80
>80
ND
NR
100
NR
ND
>80
V
ND
R
5
V
ND
ND
NR
100
100
ND
V
ND
20
ND
NR
NR
ND
V
ND
95
V
ND
ND
NR
ND
ND
- 200 mL of 6% ethyl ether in
- 200 mL of 15%
- 200 mL of 50%
- 200 mL of 100%
ethyl
ethyl
ethyl
ether in
ether in
ether
80
ND
ND
ND
ND
ND
ND
ND
hexane
hexane
hexane
R = Recovered (no percent recovery data provided by U.S. FDA)
NR = Not recovered (U. S. FDA)
V = Variable recovery (U. S. FDA)
ND = Not determined
3620B - 23
Revision 2
January 1995
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TABLE 7
PERCENT RECOVERIES AND ELUTION PATTERNS FOR 22
CHLORINATED HYDROCARBONS FROM 1-g FLORISIL CARTRIDGES3
Fraction 2
Average % Average
Compound Recovery RSD
Hexachloroethane
1,3-Dichlorobenzene
1, 4 -Di chl orobenzene
1,2-Dichlorobenzene
Benzyl chloride
1, 3, 5-Tri chl orobenzene
Hexachlorobutadiene
Benzal chloride
1, 2, 4-Tri chl orobenzene
Benzotrichloride
1,2, 3 -Tri chl orobenzene
Hexachlorocyclopentadiene
1,2,4 , 5-Tetrachl orobenzene
1,2,3 , 5-Tetrachl orobenzene
1,2, 3, 4 -Tetrachl orobenzene
2 -Chl oronaphthal ene
Pentachl orobenzene
Hexachl orobenzene
alpha-BHC
gamma -BHC
beta-BHC
delta-BHC
95
101
100
102
101
98
95
99
99
90
97
103
98
98
99
95
104
78
100
99
95
97
2.0
2.3
2.3
1.6
1.5
2.2
2.0
0.8
0.8
6.5
2.0
3.3
2.3
2.3
1.3
1.4
1.5
1.1
0.4
0.7
1.8
2.7
aFlorisil cartridges (Supelco, Inc.) were conditioned with 4 mL of hexane. Five
replicate experiments were performed. The cartridges were spiked with 1.0 /jg
per cartridge for hexachloroethane, hexachlorobutadiene, hexachloropentadiene,
pentachlorobenzne, and hexachlorobenzene. The trichlorobenzenes,
tetrachlorobenzenes, benzal chloride, benzotrichloride, and the BHCs were spiked
at 10 jug per cartridge. The dichlorobenzenes and benzyl chloride were spiked
at 100 jug per cartridge, and 2-chloronaphthalene was spiked at 200 /j,g per
cartridge. The cartridges were eluted with 5 mL of acetone/hexane (10/90, v/v).
3620B - 24 Revision 2
January 1995
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TABLE 8
DISTRIBUTION OF ANILINES IN FLORISIL CLEANUP FRACTIONS
Compound
Percent Recovery by Fraction
1 2 3
Aniline
2-Chloroaniline
3-Chloroanil ine
4-Chloroanil ine
4-Bromoanil ine
3,4-Dichloroaniline
2,4,6-Trichloroanil ine
2,4,5-Trichloroanil ine
2-Nitroanil ine
3-Nitroanil ine
4-Nitroanil ine
2,4-Dinitroanil ine
4-Chloro-2-nitroanil ine
2-Chl.oro-4-nitroanil ine
2,6-Dichloro-4-nitroanil ine
2,6-Dibromo-4-nitroanil ine
2-Bromo-6-chloro-4-nitroanil ine
2-Chloro-4,6-dinitroanil ine
2-Bromo-4,6-dinitroaniline
Eluant composition: Fraction 1
Fraction 2
Fraction 3
41 52
71 10
78 4
7 56 13
71 10
83 1
70 14
35 53
91 9
89 11
67 30
75
84
71 10
89 9
89 9
88 16
76
100
- 50% methyl ene chloride in hexane
- 5% isopropanol in hexane
- 5% methanol in hexane
3620B - 25
Revision 2
January 1995
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METHOD 3620B
FLORISIL CLEANUP
Column Cleanup
I
Reduce volume of
sample extract.
Place Florisil into
chromatographic column;
add anydrous sodium
sulfate to top of column.
Preelute column with
appropriate solvent.
Transfer sample
extract to column.
Elute column with
appropriate solvent
mixture(s).
Collect and concentrate
fractions to specified
final volumes.
Analyze by appropriate
determinative Method.
SPE Cartridge Cleanup
I
Condition cartridges.
Quantitatively transfer
sample extract to
cartridge.
Close valve/turn off
vacuum to cartridge.
Attach solvent guide
and collection vial.
Add appropriate solvent,
apply vacuum, open
cartridge valve, collect
eluate in collection vial.
Concentrate fractions to
specified final volumes.
Analyze by appropriata
determinative method.
Note: Select specific procedures provided in the method depending
on the type(s) of analytes of interest. See the method for details
regarding the appropriate elution and collection procedures.
3620B - 26
Revision 2
January 1995
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METHOD 3630C
SILICA GEL CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Silica gel is a regenerative adsorbent of amorphous silica with
weakly acidic properties. It is produced from sodium silicate and sulfuric acid.
Silica gel can be used in column chromatography for the separation of analytes
from interfering compounds of a different chemical polarity. It may be used
activated, after heating to 150 - 160°C, or deactivated with up to 10% water.
1.2 This method includes guidance for standard column cleanup of sample
extracts containing polynuclear aromatic hydrocarbons, derivatized phenolic
compounds, organochlorine pesticides, and PCBs as Aroclors.
1.3 This method also provides cleanup procedures using solid-phase
extraction cartridges for pentafluorobenzyl bromide-derivatized phenols,
organochlorine pesticides, and PCBs as Aroclors. This technique also provides
the best separation of PCBs from most single component organochlorine pesticides.
When only PCBs are to be measured, this method can be used in conjunction with
sulfuric acid/permanganate cleanup (Method 3665).
1.4 Other analytes may be cleaned up using this method if the analyte
recovery meets the criteria specified in Sec. 8.0.
1.5 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 This method provides the option of using either standard column
chromatography techniques or solid-phase extraction cartridges. Generally, the
standard column chromatography techniques use larger amounts of adsorbent and,
therefore, have a greater cleanup capacity.
2.2 In the standard column cleanup protocol, the column is packed with
the required amount of adsorbent, topped with a water adsorbent, and then loaded
with the sample to be analyzed. Elution of the analytes is accomplished with a
suitable solvent(s) that leaves the interfering compounds on the column. The
eluate is then concentrated (if necessary).
2.3 The cartridge cleanup protocol uses silica solid-phase extraction
cartridges packed with 1 g or 2 g of adsorbent. Each cartridge is solvent washed
immediately prior to use. Aliquots of sample extracts are loaded onto the
cartridges, which are then eluted with suitable solvent(s). A vacuum manifold
is required to obtain reproducible results. The collected fractions may be
further concentrated prior to gas chromatographic analysis.
3630C - 1 Revision 3
January 1995
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2.4 The appropriate gas chromatographic method is listed at the end of
each technique. Analysis may also be performed by gas chromatography/mass
spectrometry (Method 8270).
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
See Sec. 8 for guidance on a reagent blank check.
3.2 Phthalate ester contamination may be a problem with certain
cartridges The more inert the column and/or cartridge material (i.e., glass or
Teflon®), the less problem with phthalates. Phthalates create interference
problems for all method analytes, not just the phthalate esters themselves.
3.3 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Chromatographic column - 250 mm long x 10 mm ID; with Pyrex® glass
wool at bottom and a Teflon® stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits may
be purchased. Use a small pad of Pyrex® glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed by
50 ml of elution solvent prior to packing the column with adsorbent.
4.2 Beakers - appropriate sizes.
4.3 Vials - 2, 10, 25 ml, glass with Teflon® lined screw-caps or crimp
tops.
4.4 Muffle furnace.
4.5 Reagent bottle - appropriate sizes.
4.6 Erlenmeyer flasks - 50 and 250 ml.
4.7 Vacuum manifold: VacElute Manifold SPS-24 (Analytichem
International), Visiprep (Supelco, Inc.) or equivalent, consisting of glass
vacuum basin, collection rack and funnel, collection vials, replaceable stainless
steel delivery tips, built-in vacuum bleed valve and gauge. The system is
connected to a vacuum pump or water aspirator through a vacuum trap made from a
500 ml sidearm flask fitted with a one-hole stopper and glass tubing.
3630C - 2 Revision 3
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5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Silica gel for chromatography columns.
5.3.1 Silica Gel for Phenols and Polynuclear Aromatic Hydrocarbons:
100/200 mesh desiccant (Davison Chemical grade 923 or equivalent). Before
use, activate for at least 16 hr. at 130°C in a shallow glass tray,
loosely covered with foil.
5.3.2 Silica Gel for Organochlorine pesticides/PCBs: 100/200 mesh
desiccant (Davison Chemical grade 923 or equivalent). Before use,
activate for at least 16 hr. at 130°C in a shallow glass tray, loosely
covered with foil. Deactivate it to 3.3% with reagent water in a 500 ml
glass jar. Mix the contents thoroughly and allow to equilibrate for 6
hours. Store the deactivated silica gel in a sealed glass jar inside a
desiccator.
5.4 Silica cartridges: 40 jum particles, 60 A pores. The cartridges with
which this method was developed consist of 6 ml serological-grade polypropylene
tubes, with the 1 g of silica held between two polyethylene or stainless steel
frits with 20 jum pores. 2 g silica cartridges are also used in this method, and
0.5 g cartridges are available. The compound elution patterns must be verified
when cartridges other than the specified size are used.
5.5 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. A method blank must be analyzed in order to demonstrate that
there is no interference from the sodium sulfate.
5.6 Eluting solvents
5.6.1 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.6.4 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.6.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.6 Pentane, C5H12 - Pesticide quality or equivalent.
3630C - 3 Revision 3
January 1995
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5.6.7 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.6.8 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 mL of ethanol preservative must be added
to each liter of ether.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 General Guidance
7.1.1 The procedure contains two cleanup options for the derivatized
phenols and organochlorine pesticides/Aroclors, but only one technique for
the polynuclear aromatic hydrocarbons (PAHs) (standard column
chromatography). Cleanup techniques by standard column chromatography for
all analytes are found in Sec. 7.2. Cleanup techniques by solid-phase
cartridges for derivatized phenols and PAHs are found in Sec. 7.3. The
standard column chromatography techniques are packed with a greater amount
of silica gel adsorbent and, therefore, have a greater cleanup capacity.
A rule of thumb relating to cleanup capacity is that 1 g of sorbent
material will remove 10 to 30 mg of total interferences. (However,
capacity is also dependent on the sorbent retentiveness of the
interferences.) Therefore, samples that exhibit a greater degree of
sample interference should be cleaned up by the standard column technique.
However, both techniques have limits on the amount of interference that
can be removed. If the interference is caused by high boiling material,
then Method 3640 should be used prior to this method. If the interference
is caused by relatively polar compounds of the same boiling range as the
analytes, then multiple column or cartridge cleanups may be required. If
crystals of sulfur are noted in the extract, then Method 3660 should be
utilized prior to this method. The cartridge cleanup techniques are often
faster and use less solvent, however they have less cleanup capacity.
7.1.2 Allow the extract to reach room temperature if it was in cold
storage. Inspect the extracts visually to ensure that there are no
particulates or phase separations and that the volume is as stated in the
accompanying documents. Verify that the solvent is compatible with the
cleanup procedures. If crystals of sulfur are visible or if the presence
of sulfur is suspected, proceed with Method 3660.
7.1.3 If the extract solvent is methylene chloride, for most cleanup
techniques, it must be exchanged to hexane. (For the PAHs, exchange to
cyclohexane as per Sec. 7.2.1). Follow one of the standard concentration
techniques provided in each extraction method. The volume of methylene
chloride should have been reduced to 1-2 mL. Add 40 mL of hexane, a fresh
3630C - 4 Revision 3
January 1995
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boiling chip and repeat the concentration as written. The final volume
required for the cleanup techniques is normally 2 ml.
7.2 Standard Column Cleanup Techniques
7.2.1 Polynuclear aromatic hydrocarbons
7.2.1.1 Before the silica gel cleanup technique can be
utilized, the extract solvent must be exchanged to cyclohexane. The
exchange is performed by adding 4 ml of cyclohexane following
reduction of the sample extract to 1-2 ml using an appropriate
concentration technique (e.g., K-D using two-ball micro-snyder
column) found in the 3500 series methods. The final extract volume
is 2.0 ml.
CAUTION: When the volume of solvent is reduced below 1 ml, semivolatile
analytes may be lost. If the extract goes to dryness, the
extraction must be repeated.
7.2.1.2 Prepare a slurry of 10 g of activated silica gel
(Sec. 5.3.1) in methylene chloride and place this into a 10 mm ID
chromatographic column. Tap the column to settle the silica gel and
elute the methylene chloride. Add 1 to 2 cm of anhydrous sodium
sulfate to the top of the silica gel.
7.2.1.3 Pre-elute the column with 40 ml of pentane. The
rate for all elutions should be about 2 mL/min. Discard the eluate
and, just prior to exposure of the sodium sulfate layer to the air,
transfer the 2 ml cyclohexane sample extract onto the column using
an additional 2 mL cyclohexane to complete the transfer. Just prior
to exposure of the sodium sulfate layer to the air, add 25 ml of
pentane and continue the elution of the column. Discard this
pentane eluate.
7.2.1.4 Next, elute the column with 25 ml of methylene
chloride/pentane (2:3)(v/v) into a flask for concentration.
Concentrate the collected fraction to whatever volume is required
(1-10 mL). Proceed with HPLC (Method 8310) or GC analysis (Method
8100). Validated components that elute in this fraction are:
Acenaphthene Chrysene
Acenaphthylene Dibenzo(a,h)anthracene
Anthracene Fluoranthene
Benzo(a)anthracene Fluorene
Benzo(a)pyrene Indeno(l,2,3-cd)pyrene
Benzo(b)fluoranthene Naphthalene
Benzo(g,h,i)perylene Phenanthrene
Benzo(k)fluoranthene Pyrene
7.2.2 Derivatized Phenols
7.2.2.1 This silica gel cleanup procedure is performed on
sample extracts that have undergone pentafluorobenzyl bromide
3630C - 5 Revision 3
January 1995
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derivatization, as described in Method 8041. The sample extract
must be in 2 ml of hexane at this point.
7.2.2.2 Place 4.0 g of activated silica gel (Sec. 5.3.1)
into a 10 mm ID chromatographic column. Tap the column to settle
the silica gel and add about 2 g of anhydrous sodium sulfate to the
top of the silica gel.
7.2.2.3 Pre-elute the column with 6 ml of hexane. The
rate for all elutions should be about 2 mL/min. Discard the eluate
and, just prior to exposure of the sodium sulfate layer to the air,
pipet onto the column 2 ml of the hexane solution that contains the
derivatized sample or standard. Elute the column with 10.0 ml of
hexane and discard the eluate.
7.2.2.4 Elute the column, in order, with 10.0 ml of 15%
toluene in hexane (Fraction 1); 10.0 ml of 40% toluene in hexane
(Fraction 2); 10.0 ml of 75% toluene in hexane (Fraction 3); and
10.0 ml of 15% 2-propanol in toluene (Fraction 4). All elution
mixtures are prepared on a volume:volume basis. Elution patterns
for the phenolic derivatives are shown in Table 1. Fractions may be
combined, as desired, depending upon the specific phenols of
interest or level of interferences. Proceed with GC analysis.
7.2.3 Organochlorine Pesticides and Aroclors
7.2.3.1 Transfer a 3 g portion of deactivated silica gel
(Sec. 5.3.2) into a 10 mm ID glass chromatographic column and top it
with 2 to 3 cm of anhydrous sodium sulfate.
7.2.3.2 Add 10 mL of hexane to the top of the column to
wet and rinse the sodium sulfate and silica gel. Just prior to
exposure of the sodium sulfate layer to air, stop the hexane eluate
flow by closing the stopcock on the chromatographic column. Discard
the eluate.
7.2.3.3 Transfer the sample extract (2 mL in hexane) onto
the column. Rinse the extract vial twice with 1 to 2 ml of hexane
and add each rinse to the column. Elute the column with 80 mL of
hexane (Fraction I) at a rate of about 5 mL/min. Remove the
collection flask and set it aside for later concentration. Elute
the column with 50 mL of hexane (Fraction II) and collect the
eluate. Perform a third elution with 15 mL of methylene chloride
(Fraction III). The elution patterns for the organochlorine
pesticides, Aroclor-1016, and Aroclor-1260 are shown in Table 2.
7.2.3.4 Prior to gas chromatographic analysis, the
extraction solvent must be exchanged to hexane. Fractions may be
combined, as desired, depending upon the specific
pesticides/Aroclors of interest or level of interferences. Analyze
Fraction I containing Aroclors separated from most pesticides by
Method 8082. Use Method 8081 to analyze for organochlorine
pesticides.
3630C - 6 Revision 3
January 1995
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7.3 Cartridge Cleanup Techniques
7.3.1 Cartridge Set-up and Conditioning
7.3.1.1 Arrange the 1 g silica cartridges (2 g for phenol
cleanup) on the manifold in the closed-valve position. Other size
cartridges may be used, however the data presented in the Tables are
all based on 1 g cartridges for pesticides/Aroclors and 2 g
cartridges for phenols. Therefore, supporting recovery data must be
developed for other sizes. Larger cartridges will probably require
larger volumes of elution solvents.
7.3.1.2 Turn on the vacuum pump and set pump vacuum to 10
inches (254 mm) of Hg. Do not exceed the manufacturer's
recommendation for manifold vacuum. Flow rates can be controlled by
opening and closing cartridge valves.
7.3.1.3 Condition the cartridges by adding 4 ml of hexane
to each cartridge. Slowly open the cartridge valves to allow hexane
to pass through the sorbent beds to the lower frits. Allow a few
drops per cartridge to pass through the manifold to remove all air
bubbles. Close the valves and allow the solvent to soak the entire
sorbent bed for 5 minutes. Do not turn off the vacuum.
7.3.1.4 Slowly open cartridge valves to allow the hexane
to pass through the cartridges. Close the cartridge valves when
there is still at least 1 mm of solvent above the sorbent bed. Do
not allow cartridges to become dry. If cartridges go dry, repeat
the conditioning step.
7.3.2 Derivatized Phenols
7.3.2.1 Reduce the sample extract volume to 2 ml prior to
cleanup. The extract solvent must be hexane and the phenols must
have undergone derivatization by pentafluorobenzyl bromide, as per
the appropriate method.
7.3.2.2 Transfer the extract to the 2 g cartridge that has
been conditioned as described in Sec. 7.3.1. Open the cartridge
valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.3.2.3 When the entire extract has passed through the
cartridges, but before the cartridge becomes dry, rinse the sample
vials with an additional 0.5 ml of hexane, and add the rinse to the
cartridges to complete the quantitative transfer.
7.3.2.4 Close the cartridge valve and turn off the vacuum
after the solvent has passed through, ensuring that the cartridge
never gets dry.
7.3.2.5 Place a 5 ml vial or volumetric flask into the
sample rack corresponding to the cartridge position. Attach a
3630C - 7 Revision 3
January 1995
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solvent-rinsed
and align with
stainless steel solvent guide to the manifold cover
the collection vial.
7.3.2.6 Add 5 ml of hexane to the cartridge. Turn on the
vacuum pump and adjust the pump pressure to 10 inches (254 mm) of
Hg. Allow the solvent to soak the sorbent bed for 1 minute or less.
Slowly open the cartridge valve, and collect the eluate (this is
Fraction 1, and should be discarded).
NOTE: If cartridges smaller than 2 g are used, then Fraction 1 cannot be
discarded, since it contains some of the phenols.
7.3.2.7 Close the cartridge valve, replace the collection
vial, and add 5 ml of toluene/hexane (25/75, v/v) to the cartridge.
Slowly open the cartridge valve and collect the eluate into the
collection vial. This is Fraction 2, and should be retained for
analysis.
7.3.2.8 Adjust the final volume of the eluant to a known
volume which will result in analyte concentrations appropriate for
the project requirements (normally 1 - 10 ml) using techniques
described in an appropriate 3500 series method. Table 3 shows
compound recoveries for 2 g silica cartridges. The cleaned up
extracts are ready for analysis by Method 8041.
7.3.3 Organochlorine Pesticides/Aroclors
NOTE: The silica cartridge procedure is appropriate when polychlorinated
biphenyls are known to be present.
7.3.3.1 Reduce the sample extract volume to 2 ml prior to
cleanup. The extract solvent must be hexane.
7.3.3.2
Sec. 7.3.1.
Use the 1 g cartridges conditioned as described in
7.3.3.3 Transfer the extract to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge
bed at approximately 2 mL/minute.
7.3.3.4 When the entire extract has passed through the
cartridges, but before the cartridge becomes dry, rinse the sample
vials with an additional 0.5 ml of solvent, and add the rinse to the
cartridges to complete the quantitative transfer.
7.3.3.5 Close the cartridge valve and turn off the vacuum
after the solvent has passed through, ensuring that the cartridge
never goes dry.
7.3.3.6 Place a 5 ml vial or volumetric flask into the
sample rack corresponding to the cartridge position. Attach a
solvent-rinsed stainless steel solvent guide to the manifold cover
and align with the collection vial.
3630C - 8
Revision 3
January 1995
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7.3.3.7 Add 5 ml of hexane to the cartridge. Turn on the
vacuum pump and adjust the pump pressure to 10 inches (254 mm) of
Hg. Allow the solvent to soak the sorbent bed for 1 minute or less.
Slowly open the cartridge valve and collect the eluate into the
collection vial (Fraction 1).
7.3.3.8 Close the cartridge valve, replace the collection
vial, and add 5 ml of diethyl ether/hexane (50/50, v/v) to the
cartridge. Slowly open the cartridge valve and collect the eluate
into the collection vial (Fraction 2).
7.3.3.9 Adjust the final volume of each of the two
fractions to a known volume which will result in analyte
concentrations appropriate for the project requirements (normally 1
- 10 mL) using techniques described in an appropriate 3500 series
method. The fractions may be combined prior to final adjustment of
volume, if analyte fractionation is not required. Table 4 shows
compound recoveries for 1 g silica cartridges. The cleaned up
extracts are ready for analysis by Methods 8081 for OC pesticides or
8082 for Aroclors.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 A reagent blank (consisting of the elution solvents) must be passed
through the column or cartridge and checked for the compounds of interest, prior
to the use of this method. This same performance check is required with each new
lot of adsorbent or cartridges. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
8.3 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered before applying this method to actual samples. See the
attached Tables for acceptable recovery data. For compounds that have not been
tested, recovery must be > 85%.
8.3.1 Before any samples are processed using the solid-phase
extraction cartridges, the efficiency of the cartridge must be verified.
A recovery check must be performed using standards of the target analytes
at known concentration. Only lots of cartridges that meet the recovery
criteria for the spiked compounds can be used to process the samples.
8.3.2 A check should also be performed on each individual lot of
cartridges and for every 300 cartridges of a particular lot.
8.4 For sample extracts that are cleaned up using this method, the
associated quality control samples should also be processed through this cleanup
method.
3630C - 9 Revision 3
January 1995
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9.0 METHOD PERFORMANCE
9.1 Table 1 provides performance information on the fractionation of
phenolic derivatives using standard column chromatography.
9.2 Table 2 provides performance information on the fractionation of
organochlorine pesticides/Aroclors using standard column chromatography.
9.3 Table 3 shows recoveries of derivatized phenols obtained using 2 g
silica cartridges.
9.4 Table 4 shows recoveries and fractionation of organochlorine
pesticides obtained using 1 g silica cartridges.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
2. U.S EPA "Evaluation of Sample Extract Cleanup Using Solid-Phase Extraction
Cartridges," Project Report, December 1989.
3630C - 10 Revision 3
January 1995
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TABLE 1
SILICA GEL FRACTIONATION OF PFBB DERIVATIVES
Percent Recovery by Fraction8
Parameter "123
2-Chlorophenol
2-Nitrophenol
Phenol
2,4-Dimethylphenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
4-Chloro-3-methyl phenol
Pentachlorophenol
4-Nitrophenol
90
90
95
95
50 50
84
75 20
1
9
10
7
1
14
1
90
90
" Eluant composition:
Fraction 1 - 15% toluene in hexane.
Fraction 2 - 40% toluene in hexane.
Fraction 3 - 75% toluene in hexane.
Fraction 4 - 15% 2-propanol in toluene.
Data from Reference 1 (Method 604)
3630C - 11 Revision 3
January 1995
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TABLE 3
PERCENT RECOVERIES AND ELUTION PATTERNS FOR 18
PHENOLS FROM 2 g SILICA CARTRIDGES8
Compound
Fraction 2
Average Percent
Recovery RSD
Phenol
2-Methyl phenol
3-Methylphenol
4-Methyl phenol
2,4-Dimethylphenol
2-Chlorophenol
2,6-Dichlorophenol
4-Chloro-3-methyl phenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
2,3,6-Trichlorophenol
2,4,5-Trichlorophenol
2,3,5-Trichlorophenol
2,3,5,6-Tetrachlorophenol
2,3,4 , 6-Tetrachl orophenol
2,3,4-Trichlorophenol
2,3,4 , 5-Tetrachl orophenol
Pentachl orophenol
74.1
84.8
86.4
82.7
91.8
88.5
90.4
94.4
94.5
97.8
95.6
92.3
92.3
97.5
97.0
72.3
95.1
96.2
5.2
5.2
4.4
5.0
5.6
5.0
4.4
7.1
7.0
6.6
7.1
8.2
8.2
5.3
6.1
8.7
6.8
8.8
a Silica cartridges (Supelco, Inc.) were used; each cartridge was
conditioned with 4 mL of hexane prior to use. Each experiment was
performed in duplicate at three spiking concentrations (0.05 /zg, 0.2 /^g,
and 0.4 p,g per compound per cartridge). Fraction 1 was eluted with 5 mL
hexane and was discarded. Fraction 2 was eluted with 5 mL toluene/hexane
(25/75, v/v).
Data from Reference 2
3630C - 14
Revision 3
January 1995
-------
TABLE 4
PERCENT RECOVERIES AND ELUTION PATTERNS FOR 17 ORGANOCHLORINE
PESTICIDES AND AROCLORS FROM 1 g SILICA CARTRIDGES8
Compound
Fraction 1
Average Percent
Recovery RSD
Fraction 2
Average Percent
Recovery RSD
alpha-BHC
gamma-BHC
beta-BHC
Heptachlor
delta-BHC
Aldrin
Heptachlor epoxide
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
4,4'-DDD
Endosulfan II
4,4'-DDT
Endrin aldehyde
Endosulfan sulfate
4,4'-Methoxychlor
Aroclor 1016
Aroclor 1221
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1264
0
0
0
97.3 1.3
0
95.9 1.0
0
0
99.9 1.7
0
0
10.7 41
0
94.1 2.0
0
0
0
124
93.5
118
116
114
108
112
98.7
94.8
94.3
0
90.8
0
97.9
102
0
92.3
117
92.4
96.0
0
59.7
97.8
98.0
2.3
1.9
3.0
2.5
2.1
2.3
2.0
2.6
3.3
2.2
2.6
2.1
2.4
a Silica cartridges (Supelco, Inc. lot SP0161) were used; each cartridge was
conditioned with 4 mL hexane prior to use. The organochlorine pesticides were
tested separately from PCBs. Each organochlorine pesticides experiment was
performed in duplicate, at three spiking concentrations (0.2 ng, 1.0 jug, and
2.0 jug per compound per cartridge). Fraction 1 was eluted with 5 mL of
hexane, Fraction 2 with 5 mL of diethyl ether/hexane (50/50, v/v). PCBs were
spiked at 10 /ug per cartridge and were eluted with 3 mL of hexane. The values
given for PCBs are the percent recoveries for a single determination.
Data from Reference 2
3630C - 15
Revision 3
January 1995
-------
METHOD 3630C
SILICA GEL CLEANUP
Column Cleanup
Reduce volume of
sample extract;
exchange solvent as
appropriate.
Place silica gel into
chromatographic column;
add anydrous sodium
sulfate to top of column.
Preelute column with
appropriate solvent.
Transfer sample
extract to column.
Elute column with
appropriate solvent
mixture(s).
Collect and concentrate
fractions to specified
final volumes; exchange
solvent, if necessary, for
determinative analysis.
Analyze by appropriate
determinative Method.
SPE Cartridge Cleanup
I
Condition cartridges.
Quantitatively transfer
sample extract to
cartridge.
Close valve/turn off
vacuum to cartridge.
Attach solvent guide
and collection vial.
Add appropriate solvent,
apply vacuum, open
cartridge valve, collect
eluate in collection vial.
Concentrate fractions to
specified final volumes.
Analyze by appropriate
determinative Method.
Note: Select specific procedures provided in the method depending
on the type(s) of analytes of interest. See the method for details
regarding the appropriate elution and collection procedures.
3630C - 16
Revision 3
January 1995
-------
METHOD 3650B
ACfD-BASE PARTITION CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Method 3650 is a liquid-liquid partitioning cleanup method to
separate acid analytes, e.g. organic acids and phenols, from base/neutral
analytes, e.g. amines, aromatic hydrocarbons, and halogenated organic compounds,
using pH adjustment. It may be used for cleanup of petroleum waste prior to
analysis or further cleanup (e.g., alumina cleanup). The following compounds can
be separated by this method:
Compound Name
Benz(a)anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Chlordane
Chlorinated dibenzodioxins
2-Chlorophenol
Chrysene
Creosote
Cresol (s)
Dichlorobenzene(s)
Dichlorophenoxyacetic acid
2,4-Dimethylphenol
Dinitrobenzene
4,6-Dinitro-o-cresol
2,4-Dinitrotoluene
Heptachlor
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Hexachlorocyclopentadiene
Naphthalene
Nitrobenzene
4-Nitrophenol
Pentachlorophenol
Phenol
Phorate
2-Picol ine
Pyridine
Tetrachlorobenzene(s)
Tetrachlorophenol (s)
Toxaphene
Trichlorophenol (s)
2,4,5-TP (Silvex)
CAS No.a
56-55-3
50-32-8
205-99-2
57-74-9
95-57-8
218-01-9
8001-58-9
94-75-7
105-67-9
25154-54-5
534-52-1
121-14-2
76-44-8
118-74-1
87-68-3
67-72-1
77-47-4
91-20-3
98-95-3
100-02-7
87-86-5
108-95-2
298-02-2
109-06-8
110-86-1
8001-35-2
93-72-1
Fraction
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Base-neutral
Base-neutral and Acid
Acid
Base-neutral
Acid
Acid
Base-neutral
Acid
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Acid
Acid
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Base-neutral
Acid
Acid
Chemical Abstract Services Registry Number.
3650B - 1
Revision 2
January 1995
-------
1.2 Method 3650 was formerly Method 3530 in the second edition of this
manual.
•
1.3 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 The solvent extract from a prior solvent extraction method is shaken
with water that is strongly basic. The acid analytes partition into the aqueous
layer, whereas, the basic and neutral compounds stay in the organic solvent. The
base/neutral fraction is concentrated and is then ready for further cleanup, if
necessary, or analysis. The aqueous layer is acidified and extracted with an
organic solvent. This extract is concentrated (if necessary) and is then ready
for analysis of the acid analytes.
3.0 INTERFERENCES
3.1 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
3.2 A method blank must be run for the compounds of interest prior to
use of the method. The interferences must be below the method detection limit
before this method is applied to actual samples.
4.0 APPARATUS AND MATERIALS
4.1 Drying column - 20 mm ID Pyrex® chromatographic column with Pyrex®
glass wool at bottom, or equivalent.
NOTE: Fritted glass discs are difficult to clean after highly contaminated
extracts have been passed through them. Columns without frits are
recommended. Use a small pad of Pyrex® glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed by
50 ml of elution solvent prior to packing the column with adsorbent.
4.2 Vials - Glass, 2 mL capacity with Teflon® lined screw-caps or crimp
tops.
4.3 Water bath - Heated, concentric ring cover, temperature control of
± 2°C. Use this bath in a hood.
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 pH indicator paper - pH range including the desired extraction pH.
4.6 Separatory funnel - 125 mL.
4.7 Erlenmeyer flask - 125 mL.
3650B - 2 Revision 2
January 1995
-------
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium hydroxide, NaOH, (ION) - Dissolve 40 g of sodium hydroxide
in 100 ml of organic-free reagent water.
5.4 Sulfuric acid, H2S04, (1:1 v/v in water) - Slowly add 50 mL H2S04 to
50 ml of organic-free reagent water.
5.5 Sodium sulfate (granular, anhydrous), Na2S04 - Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.6 Solvents:
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.6.3 Methanol, CH3OH - Pesticide quality or equivalent.
5.6.4 Diethyl Ether, C2H5OC2H? - Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 mL of ethyl alcohol preservative must be
added to each liter of ether.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Place 10 mL of the solvent extract from a prior extraction procedure
into a 125 mL separatory funnel.
7.2 Add 20 mL of methylene chloride to the separatory funnel.
7.3 Slowly add 20 mL of prechilled organic-free reagent water which has
been previously adjusted to a pH of 12-13 with ION sodium hydroxide.
3650B - 3 Revision 2
January 1995
-------
7.4 Seal and shake the separatory funnel for at least 2 minutes with
periodic venting to release excess pressure.
NOTE: Methylene chloride creates excessive pressure very rapidly; therefore,
initial venting should be done immediately after the separatory funnel has
been sealed and shaken once. The separatory funnel should be vented into
a hood to prevent unnecessary exposure of the analyst to the organic
vapor.
7.5 Allow the organic layer to separate from the aqueous phase for a
minimum of 10 minutes. If the emulsion interface between layers is more than
one-third the size of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. The optimum technique depends upon
the sample, and may include stirring, filtration of the emulsion through glass
wool, centrifugation, or other physical methods.
7.6 Separate the aqueous phase and transfer it to a 125 ml Erlenmeyer
flask. Repeat the extraction two more times using 20 ml aliquots of dilute
sodium hydroxide (pH 12-13). Combine the aqueous extracts.
7.7 Water soluble organic acids and phenols will be primarily in the
aqueous phase. Base/neutral analytes will be in the methylene chloride. If the
analytes of interest are only in the aqueous phase, discard the methylene
chloride and proceed to Sec. 7.8. If the analytes of interest are only in the
methylene chloride, discard the aqueous phase and proceed to Sec. 7.10.
7.8 Externally cool the 125 ml Erlenmeyer flask with ice while adjusting
the aqueous phase to a pH of 1-2 with sulfuric acid (1:1). Quantitatively
transfer the cool aqueous phase to a clean 125 ml separatory funnel. Add 20 ml
of methylene chloride to the separatory funnel and shake for at least 2 minutes.
Allow the methylene chloride to separate from the aqueous phase and collect the
methylene chloride in an Erlenmeyer flask.
7.9 Add 20 mL of methylene chloride to the separatory funnel and extract
at pH 1-2 a second time. Perform a third extraction in the same manner combining
the extracts in the Erlenmeyer flask.
7.10 Prepare a concentration apparatus (if necessary). Refer to the 3500
series methods for guidance regarding concentration of samples.
7.11 Dry both acid and base/neutral fractions by passing them through a
drying column containing about 10 cm of anhydrous sodium sulfate. Collect the
dried fractions in concentrator container. Rinse the Erlenmeyer flasks which
contained the solvents and the columns with 20 ml of methylene chloride to
complete the quantitative transfer.
7.12 The acid fraction is now ready for analysis. If the base/neutral
fraction requires further cleanup by the alumina column cleanup for petroleum
waste (Method 3611), the solvent may have to be changed to hexane. If a solvent
exchange is required, add approximately 5 mL of the exchange solvent to the
fraction before concentration. Concentrate the fractions to the final volume
(usually 1 mL) listed in the appropriate determinative method using the
techniques described in an appropriate 3500 series method. If no further cleanup
of the base/neutral extract is required, the fraction is ready for analysis.
3650B - 4 Revision 2
January 1995
-------
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for general quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered before applying this method to actual samples.
8.3 For samples that are cleaned using this method, the associated
quality control samples must be processed through this cleanup method.
9.0 METHOD PERFORMANCE
Refer to the determinative methods for performance data.
10.0 REFERENCES
1. Test Methods: Methods for Organic Chemical Analysis of Municipal and
Industrial Wastewater; U.S. Environmental Protection Agency. Office of
Research and Development. Environmental Monitoring and Support Laboratory.
ORD Publication Offices of Center for Environmental Research Information:
Cincinnati, OH, 1982; EPA-600/4-82-057.
3650B - 5 Revision 2
January 1995
-------
METHOD 36508
ACID-BASE PARTITION CLEANUP
>
r
7.1 Place extract
or organic liquid
waste into
separatory funnel.
P 1
r
7.2 Add methylene
chloride.
•^
r
7.3 Add prechilled
• dilute sodium
hydroxide.
>
r
7.4 Seal and shake
separatory funnel.
• i
r
7.5 Allow
separation of
organic layer from
aqueous phase.
i>
7.5 Complete phase
separation with
mechanical
techniques.
7.6 Transfer aqueous
phase to flask; repeat
extraction twice;
combine aqueous
extracts.
7.7 Discard aqueous
phase.
7.7 Discard organic
phase.
7.8 Adjust pH with
sulfuric acid; trans-
fer aqueous phase to
clean separatory funnel,
add methylene chloride;
shake; allow phase
separation; collect sol-
vent phase in flask.
7.10 Assemble
concentration
apparatus.
7.9 Perform 2 more
extractions; combine
all extracts.
3650B - 6
Revision 2
January 1995
-------
METHOD 36508
ACID-BASE PARTITION CLEANUP (Continued)
7.11 Dry extracts;
collect extracts in
concentrator; rinse
flask with methylene
chloride.
i-
.'»
*
7.12 Concentrate
both fractions.
/ 7.12 Is \
further cleanup\ Yes
needed for ) — >
base/neutral/
\ extract?/
7.1 2 Exchange
solvent,
perform additional
cleanup.
No
Analyze
fractions by
appropriate
determinative
method.
3650B - 7
Revision 2
January 1995"
-------
METHOD 3660B
SULFUR CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Elemental sulfur is encountered in many sediment samples (generally
specific to different areas in the country), marine algae, and some industrial
wastes. The solubility of sulfur in various solvents is very similar to the
organochlorine and organophosphorus pesticides. Therefore, the sulfur
interference follows along with the pesticides through the normal extraction and
cleanup techniques. In general, sulfur will usually elute entirely in Fraction
1 of the Florisil cleanup (Method 3620).
1.2 Sulfur will be quite evident in gas chromatograms obtained from
electron capture detectors, flame photometric detectors operated in the sulfur
or phosphorous mode, and Coulson electrolytic conductivity detectors in the
sulfur mode. If the gas chromatograph is operated at the normal conditions for
pesticide analysis, the sulfur interference can completely mask the region from
the solvent peak through Aldrin.
1.3 Three techniques for the elimination of sulfur are detailed within
this method: (1) the use of copper powder; and (2) the use of tetrabutylammonium
sulfite. Tetrabutylammonium sulfite causes the least amount of degradation of
a broad range of pesticides and organic compounds, while copper may degrade
organophosphorus and some organochlorine pesticides.
1.4 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 The sample to undergo cleanup is mixed with either copper or
tetrabutylammonium (TBA) sulfite. The mixture is shaken and the extract is
removed from the sulfur cleanup reagent.
3.0 INTERFERENCES
3.1 Removal of sulfur using copper:
3.1.1 The copper must be very reactive. Therefore, all oxides of
copper must be removed so that the copper has a shiny, bright appearance.
3.1.2 The sample extract must be vigorously agitated with the
reactive copper for at least one minute.
3660B - 1 Revision 2
January 1995
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4.0 APPARATUS AND MATERIALS
4.1 Mechanical shaker or mixer - Vortex Genie or equivalent.
4.2 Pipets, disposable - Pasteur type.
4.3 Centrifuge tubes, calibrated - 12 ml.
4.4 Glass bottles or vials - 10 ml and 50 ml, with Teflon®-lined screw
caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Nitric acid, HN03, dilute.
5.4 Solvents
5.4.1 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.4.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.4.3 2-Propanol, CH3CH(OH)CH3 - Pesticide quality or equivalent.
5.5 Copper powder - Remove oxides by treating with dilute nitric acid,
rinse with organic-free reagent water to remove all traces of acid, rinse with
acetone and dry under a stream of nitrogen. (Copper, fine granular Mallinckrodt
4649 or equivalent).
5.6 Tetrabutylammonium (TBA) sulfite reagent
5.6.1 Tetrabutylammonium hydrogen sulfate, [CH3(CH2)3]4NHS04.
5.6.2 Sodium sulfite, Na2S03.
5.6.3 Prepare reagent by dissolving 3.39 g tetrabutylammonium
hydrogen sulfate in 100 ml organic-free reagent water. To remove
impurities, extract this solution three times with 20 ml portions of
hexane. Discard the hexane extracts, and add 25 g sodium sulfite to the
water solution. Store the resulting solution, which is saturated with
sodium sulfite, in an amber bottle with a Teflon®-!ined screw cap. This
solution can be stored at room temperature for at least one month.
3660B - 2 Revision 2
January 1995
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7.2.4 Add 5 ml organic free reag...
min. Allow the sample to stand for 5-10
(top) to a concentrator tube and conceptr
1-° "L man tun tnibi.1 '"
at
. vi Ob I
Transfer the hexane
I •••!• - '
-------
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Removal of sulfur using copper
7.1.1 Concentrate the sample to exactly 1.0 mL or other known
volume. Perform concentration using the techniques described in the
appropriate 3500 series method.
CAUTION: When the volume of solvent is reduced below 1 mL, semi volatile
analytes may be lost.
7.1.2 If the sulfur concentration is such that crystallization
occurs, centrifuge to settle the crystals, and carefully draw off the
sample extract with a disposable pipet leaving the excess sulfur in the
concentration vessel. Transfer 1.0 mL of the extract to a calibrated
centrifuge tube.
7.1.3 Add approximately 2 g of cleaned copper powder (to the 0.5 mL
mark) to the centrifuge tube. Mix for at least 1 min on the mechanical
shaker.
7.1.4 Separate the extract from the copper by drawing off the
extract with a disposable pipet and transfer to a clean vial. The volume
remaining still represents 1.0 mL of extract.
NOTE: This separation is necessary to prevent further degradation of the
pesticides.
7.2 Removal of sulfur using TBA sulfite
7.2.1 Concentrate the sample extract to exactly 1.0 mL or other
known volume. Perform concentration using the techniques described in the
appropriate 3500 series method.
CAUTION: When the volume of solvent is reduced below 1 mL, semi volatile
analytes may be lost.
7.2.2 Transfer 1.0 mL of the extract to a 50 mL clear glass bottle
or vial with a Teflon®-!ined screw-cap. Rinse the concentrator tube with
1 mL of hexane, adding the rinsings to the 50 mL bottle.
7.2.3 Add 1.0 mL TBA sulfite reagent and 2 mL 2-propanol, cap the
bottle, and shake for at least 1 min. If the sample is colorless or if
the initial color is unchanged, and if clear crystals (precipitated sodium
sulfite) are observed, sufficient sodium sulfite is present. If the
precipitated sodium sulfite disappears, add more crystalline sodium
sulfite in approximately 0.100 g portions until a solid residue remains
after repeated shaking.
3660B - 3 Revision 2
January 1995
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7.2.4 Add 5 ml organic free reagent water and shake for at least 1
min. Allow the sample to stand for 5-10 min. Transfer the hexane layer
(top) to a concentrator tube and concentrate the extract to approximately
1.0 ml using the techniques described in the appropriate 3500 series
method. Record the actual volume of the final extract.
7.3 Analyze the cleaned up extracts by gas chromatography (see the
determinative methods, Section 4.3 of this chapter).
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 All reagents should be checked prior to use to verify that
interferences do not exist.
9.0 METHOD PERFORMANCE
9.1 Table 1 indicates the effect of using copper to remove sulfur on the
recovery of certain pesticides.
10.0 REFERENCES
1. Loy, E.W., private communication.
2. Goerlitz, D.F. and L.M. Law, Bulletin for Environmental Contamination and
Toxicology, 6, 9 (1971).
3. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, Revision, July 1985.
3660B - 4 Revision 2
January 1995
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TABLE 1
EFFECT OF COPPER ON PESTICIDES
Percent Recovery"
Pesticide Using Copper
Aroclor 1254 104.26
Lindane 94.83
Heptachlor 5.39
Aldrin 93.29
Heptachlor epoxide 96.55
DDE 102.91
DDT 85.10
BHC 98.08
Dieldrin 94.90
Endrin 89.26
Chiorobenzilate 0.00
Malathion 0.00
Diazinon 0.00
Parathion 0.00
Ethion 0.00
Trithion 0.00
Percent recoveries cited are averages based on duplicate analyses for all
compounds other than for Aldrin and BHC. For Aldrin, four and three
determinations were averaged to obtain the result for copper. Recovery of BHC
using copper is based on one analysis.
3660B - 5 Revision 2
January 1995
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METHOD 3660B
SULFUR CLEANUP
TBA-sulfite
7.1.1
Concentrate
sample extract.
7.1.2 Di
cyrstallization
occur'
7.1.2
Centrifuge
and draw off
sample extract.
7.1.2
Transfer
extract to
centrifuge tube.
7.1 .3 Add
copper powder
mix.
7.1 .4 Separate
extract from
copper.
7.2.1
Concentrate
sample extract.
7.2.2
Transfer
extract to
bottle or tube.
7.2.3 Add
TBA-sulfite
and 2-propanol,
agitate.
7.2.3
Sample
colorless or
color unchanged
and crystals of
sodium sulfite
present?
7.2.4 Add organic
free water, shake,
allow to settle.
Transfer hexane
layer to concentrator.
Add additional
crystalline
sodium sulfite
until residue
remains after
shaking.
3660B - 6
Revision 2
January 1995
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METHOD 3665A
SULFURIC ACID/PERMANGANATE CLEANUP
1.0 SCOPE AND APPLICATION
1.1 This method is suitable for the rigorous cleanup of sample extracts
prior to analysis for polychlorinated biphenyls. This method should be used
whenever elevated baselines or overly complex chromatograms prevent accurate
quantitation of PCBs. This method cannot be used to cleanup extracts for other
target analytes, as it will destroy most organic chemicals including the
pesticides Aldrin, Dieldrin, Endrin, Endosulfan (I and II), and Endosulfan
sulfate.
1.2 This method is restricted to use by or under the supervision of
trained analysts. Each analyst must demonstrate the ability to generate
acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 An extract is solvent exchanged to hexane, then the hexane is
sequentially treated with (1) concentrated sulfuric acid and, if necessary, (2)
5% aqueous potassium permanganate. Appropriate caution must be taken with these
corrosive reagents.
2.2 Blanks and replicate analysis samples must be subjected to the same
cleanup as the samples associated with them.
2.3 It is important that all the extracts be exchanged to hexane before
initiating the following treatments.
3.0 INTERFERENCES
3.1 This technique will not destroy chlorinated benzenes, chlorinated
naphthalenes (Halowaxes), and a number of chlorinated pesticides.
4.0 APPARATUS
4.1 Syringe or Class A volumetric pipet, glass; 1.0, 2.0 and 5.0 mL.
4.2 Vials -1,2 and 10 mL, glass with Teflon® lined screw caps or crimp
tops.
4.3 Vortex mixer.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
3665A - 1 Revision 1
January 1995
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specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sulfuric acid/Water, H2S04/H20, (1:1, v/v).
5.4 Hexane, C6HU - Pesticide grade or equivalent.
5.5 Potassium permanganate, KMn04, 5 percent aqueous solution (w/v).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sulfuric acid cleanup
7.1.1 Using a syringe or a volumetric pipet, transfer 1.0 or 2.0 mL
of the hexane extract to a 10 mL vial and, in a fume hood, carefully add
5 mL of the 1:1 sulfuric acid/water solution.
7.1.2 The volume of hexane extract used depends on the requirements
of the GC autosampler used by the laboratory. If the autosampler
functions reliably with 1 mL of sample volume, 1.0 mL of extract should be
used. If the autosampler requires more than 1 mL of sample volume, 2.0 mL
of extract should be used.
CAUTION: Make sure that there is no exothermic reaction nor
evolution of gas prior to proceeding.
7.1.3 Cap the vial tightly and vortex for one minute. A vortex must
be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks, AVOID
SKIN CONTACT, SULFURIC ACID BURNS.
7.1.4 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer; it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.1.5 If a clean phase separation is achieved, proceed to
Sec. 7.1.8.
7.1.6 If the hexane layer is colored or the emulsion persists for
several minutes, remove the sulfuric acid layer from the vial and dispose
of it properly. Add another 5 mL of the clean 1:1 sulfuric acid/water.
3665A - 2 Revision 1
January 1995
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NOTE: Do not remove any hexane at this stage of the procedure.
7.1.7 Vortex the sample for one minute and allow the phases to
separate.
7.1.8 Transfer the hexane layer to a clean 10 ml vial.
7.1.9 Add an additional 1 ml of hexane to the sulfuric acid layer,
cap and shake. This second extraction is done to ensure quantitative
transfer of the PCBs and Toxaphene.
7.1.10 Remove the second hexane layer and combine with the
hexane from Sec. 7.1.8.
7.2 Permanganate cleanup
7.2.1 Add 5 ml of the 5 percent aqueous potassium permanganate
solution to the combined hexane fractions from 7.1.10.
CAUTION: Make sure that there is no exothermic reaction nor
evolution of gas prior to proceeding.
7.2.2 Cap the vial tightly and vortex for 1 minute. A vortex must
be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks. AVOID
SKIN CONTACT, POTASSIUM PERMANGANATE BURNS.
7.2.3 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer, it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.2.4 If a clean phase separation is achieved, proceed to
Sec. 7.2.7.
7.2.5 If the hexane layer is colored or the emulsion persists for
several minutes, remove the permanganate solution from the vial via a
glass pipette and dispose of it properly. Add another 5 ml of the clean
aqueous permanganate solution.
NOTE: Do not remove any hexane at this stage of the procedure.
7.2.6 Vortex the sample and allow the phases to separate.
7.2.7 Transfer the hexane layer to a clean 10 ml vial.
7.2.8 Add an additional 1 ml of hexane to the permanganate layer,
cap the vial securely and shake. This second extraction is done to ensure
quantitative transfer of the PCBs and Toxaphene.
7.2.9 Remove the second hexane layer and combine with the hexane
from Sec. 7.2.7.
3665A - 3 Revision 1
January 1995
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7.3 Final preparation
7.3.1 Reduce the volume of the combined hexane layers to the
original volume (1 or 2 ml) using an appropriate concentration technique,
refer to the 3500 series methods.
7.3.2 Remove any remaining organochlorine pesticides from the
extracts using Florisil Column Cleanup (Method 3620) or Silica Gel Cleanup
(Method 3630).
7.3.3 The extracts obtained may now be analyzed for the target
analytes using the appropriate organic technique(s) (see Sec. 4.3 of this
Chapter). If analysis of the extract will not be performed immediately,
stopper the concentrator tube and store in a refrigerator. If the extract
will be stored longer than 2 days, it should be transferred to a vial with
a Teflon® lined screw cap or crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 No performance data are currently available.
10.0 REFERENCES
None required.
3665A - 4 Revision 1
January 1995
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METHOD 3665
SULFURIC ACID/PERMANGANATE CLEANUP
7.1.1 Combine
hexane
with 1:1
H2S04/H20
solution.
7.1.2 Transfer
the appropriate
volume to
vial.
7.1.3 - 7.1.4
Cap, vortex,
and allow
phase
separation.
7.1.5 Is
phase
separation
clean?
7.1.8
Transfer
hexane layer
to clean vial.
7.1.6 Remove
and dispose
HaS04 solution,
add clean h^SO
HjO solution.
7.1.7 Cap,
vertax, and
allow phase
separation.
7.1.9 Add
hexane to
H2S04 layer,
cap and shake.
7.1.10 Combine
hexane layers.
7.2.1 Add
KMn04
solution.
7.2.2 - 7.2.3
Cap, vortex,
and allow phase
separation.
7.2.4 Is
phase
separation
clean?
7.2.7
Transfer
hexane layer
to clean vial.
7.2.8 Add
hexane to
KMn04 layer,
cap and shake.
7.2.9 Combine
two hexane
layers.
3665A - 5
7.2.5 Remove
and dispose
KMnO4 solution,
add clean KMnCU
solution.
7.2.6 Cap,
vortex, and
allow phase
separation.
7.3.1 - 7.3.3
Reduce volume
using concentra-
tion technique.
7.3.4 Use
Method 3620 or
3630 to further
remove
contaminants.
7.3.5 Stopper
and refrigerate
for further
analysis.
Stop
Revision 1
January 1995
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.1 GAS CHROMATOGRAPHIC METHODS
The following methods are included in this section:
Method 8000B:
Method 8011:
Method 8015B:
Method 8021B:
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
8031:
8032A:
8033:
8041:
8061A:
8070A:
8081A:
8082:
8091:
8100:
8111:
8121:
8131:
8141A:
8151A:
Determinative Chromatographic Separations
1,2-Dibromoethane and l,2-Dibromo-3-chloropropane
by Microextraction and Gas Chromatography
Nonhalogenated Organics Using GC/FID
Halogenated Volatiles by Gas Chromatography Using
Photoionization and Electrolytic Conductivity
Detectors in Series: Capillary Column Technique
Acrylonitrile by Gas Chromatography
Acrylamide by Gas Chromatography
Acetonitrile by Gas Chromatography with Nitrogen-
Phosphorus Detection
Phenols by Gas Chromatography: Capillary Column
Technique
Phthalate Esters by Capillary Gas Chromatography
with Electron Capture Detection (GC/ECD)
Nitrosamines by Gas Chromatography
Organochlorine Pesticides by Capillary Column Gas
Chromatography
Polychlorinated Biphenyls (PCBs) by Capillary
Column Gas Chromatography
Nitroaromatics and Cyclic Ketones: Capillary
Column Technique
Polynuclear Aromatic Hydrocarbons
Haloethers: Capillary Column Technique
Chlorinated Hydrocarbons by Gas Chromatography:
Capillary Column Technique
Aniline and Selected Derivatives by GC:
Capillary Column Technique
Organophosphorus Compounds by Gas Chromatography:
Capillary Column Technique
Chlorinated Herbicides by GC Using Methylation or
Pentafluorobenzylation Derivatization: Capillary
Column Technique
FOUR - 10
Revision 3
January 1995
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METHOD 8000B
DETERMINATIVE CHROMATOGRAPHIC SEPARATIONS
1.0 SCOPE AND APPLICATION
1.1 Method 8000 is not a determinative method but instead provides
guidance on analytical chromatography and describes calibration and quality
control requirements that are common to all SW-846 chromatographic methods.
Method 8000 is to be applied in conjunction with the determinative
chromatographic methods listed below.
SW-846 CHROMATOGRAPHIC DETERMINATIVE METHODS
Method
Number Analytes
Chromatographic
Technique
Detector
8011 EDB, DBCP
8015 Non-halogenated volatiles
8021 Volatiles
8031 Acrylonitrile
8032 Acrylamide
8033 Acetonitrile
8041 Phenols
8061 Phthalates
8070 Nitrosamines
8081 Organochlorine pesticides
8082 Polychlorinated biphenyls
8091 Nitroaromatics and cyclic
ketones
GC, capillary
column
GC, packed &
capillary column
GC, capillary
column
GC, packed column
GC, packed column
GC, capillary
column
Underivatized or
Derivatized, GC,
capillary column
GC, capillary
column
GC, packed column
GC, capillary
column
GC, capillary
column
GC, capillary
column
ECD
FID
PID, ELCD
NPD
ECD
NPD
FID, ECD
ECD
NPD, ELCD, TED
ECD, ELCD
ECD, ELCD
ECD
8000B - 1
Revision 2
January 1995
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SW-846 CHROMATOGRAPHIC DETERMINATIVE METHODS (continued)
Method
Number Analytes
Chromatographic
Technique
Detector
8100 PAHs
8111 Haloethers
8121 Chlorinated hydrocarbons
8131 Aniline and selected
derivatives
8141 Organophosphorus
pesticides
8151 Acid herbicides
8260 Volatiles
8270 Semivolatiles
8275 Semivolatiles
8280 Dioxins/Dibenzofurans
8290 Dioxins/Dibenzofurans
8310 PAHs
8315 Carbonyl compounds
8316 Acrylamide,
acrylonitrile, acrolein
8318 N-Methyl Carbamates
8321 Extractable non-volatiles
8325 Extractable non-volatiles
8330 Nitroaromatics and
nitramines
GC, packed column
GC, capillary
column
GC, capillary
column
GC, capillary
column
GC, capillary
column
Derivatize; GC,
capillary column
GC, capillary
column
GC, capillary
column
Thermal
extraction/GC
GC, capillary
column
GC, capillary
column
HPLC, reverse phase
Derivatize; HPLC
HPLC, reverse phase
FID
ECD
ECD
NPD
FPD, NPD, ELCD
ECD
MS
MS
MS
Low resolution MS
High resolution MS
UV, Fluorescence
Fluorescence
UV
Derivatize; HPLC Fluorescence
HPLC, reverse phase TS/MS, UV
HPLC, reverse phase PB/MS, UV
HPLC, reverse phase UV
8000B - 2
Revision 2
January 1995
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SW-846 CHROMATOGRAPHIC DETERMINATIVE METHODS (continued)
Method
Number Analytes
Chromatographic
Technique
Detector
8331 Tetrazene
8332 Nitroglycerine
8410 Semivolatiles
8430 bis(Z-Chloroethyl) ether
hydrolysis products
HPLC, ion pair,
reverse phase
UV
HPLC, reverse phase UV
FT-IR
GC, capillary
column
GC, capillary
column
FT-IR
where:
DBCP
ECD
EDB
ELCD
no
FPD
FT-IR =
GC
HPLC
Dibromochloropropane MS
Electron capture detector NPO
Ethylene dibromide PAHs
Electrolytic conductivity detector PB/MS
Flame ionization detector PIO
Flame photometric detector TED
Fourier transform-infrared TS/MS
Gas chromatography UV
High performance liquid chromatography
Mass spectrometry
Nitrogen/phosphorous detector
Polynuclear aromatic hydrocarbons
Particle beam mass spectrometry
Photoionization detector
Thermionic emission detector
Thermospray mass spectrometry
Ultraviolet
1.2 Analytical chromatography is used to separate target analytes from
co-extracted interferences in samples. Chromatographic methods can be divided
into two major categories: gas chromatography (GC) and high performance liquid
chromatography (HPLC).
1.2.1 Gas chromatography (more properly called gas-liquid
chromatography) is the separation technique of choice for organic compounds
which can be volatilized without being decomposed or chemically rearranged.
1.2.2 High performance liquid chromatography (HPLC) is a separation
technique useful for semivolatile and nonvolatile chemicals or for analytes
that decompose upon heating. Successful liquid Chromatographic separation
requires that the analyte(s) of interest be soluble in the solvent(s)
selected for use as the mobile phase. Because the solvents are delivered
under pressure, the technique was originally designated as high pressure
liquid chromatography, but now is commonly referred to as high performance
liquid chromatography (HPLC).
1.3 All Chromatographic processes achieve separation by passing a mobile
phase over a stationary phase. Constituents in a mixture are separated because
they partition differently between the mobile and stationary phases and thus have
different retention times. Compounds that interact strongly with the stationary
phase elute slowly (i.e., long retention time), while compounds that remain in
the mobile phase elute quickly (i.e., short retention time).
8000B - 3
Revision 2
January 1995
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1.3.1 The mobile phase for GC is an inert gas, usually helium, and
the stationary phase is generally a silicone oil or similar material.
1.3.2 In "normal phase" HPLC, the mobile phase is less polar than
the stationary phase. In "reverse phase" HPLC, the converse is true.
Reverse phase HPLC is the technique of choice for environmental and waste
analyses of non-volatile organic targets.
1.4 A number of specific GC and LC techniques are used for environmental
and waste analyses. The specific techniques are distinguished by the
chromatographic hardware or by the chemical mechanisms used to achieve
separations.
1.4.1 SW-846 GC methods are categorized on the basis of hardware.
1.4.1.1 Packed columns are typically made from glass columns
that are 1.5 -3m long with a 2 - 4 mm ID, and filled with small
particles (60-100 mesh diatomaceous earth or carbon) coated with a
liquid phase.
1.4.1.2 Capillary columns are typically made from open
tubular glass capillary columns that are 15 - 100 m long with a
0.2 - 0.75 mm ID, and coated with a liquid phase. Most capillary
columns are now made of fused silica, although glass columns are still
sold for the analysis of volatiles. Capillary columns are inherently
more efficient than packed columns and have replaced packed columns
for most SW-846 applications.
1.4.2 SW-846 HPLC methods are categorized on the basis of the
mechanism of separation.
1.4.2.1 Partition chromatography is the basis of reverse
phase HPLC separations. Analytes are separated on a hydrophobic
column using a polar mobile phase pumped at high pressure (800 - 4000
psi) through a stainless steel column 10 - 25 cm long with a 2 - 4 mm
ID and packed with 3 - 10 /zm silica or divinyl benzene-styrene
particles.
1.4.2.2 Ion exchange chromatography is used to separate
ionic species.
1.5 SW-846 methods describe columns and conditions that have been
demonstrated to provide optimum separation of all or most target analytes listed
in that specific procedure. Most often, those columns were the ones used by EPA
during method development and testing. Analysts may change those columns and
conditions (with certain exceptions) provided that they demonstrate adequate
performance. This is especially true when limited groups of analytes are to be
monitored (i.e., if only a subset of the list of target analytes in a method are
required, then the chromatographic conditions and columns can be optimized for
those analytes).
1.5.1 Chromatographic performance is demonstrated by the resolution
of standards and the linearity of the detector during calibration, and by
the accuracy, precision, frequency of false positives, and frequency of
8000B - 4 Revision 2
January 1995
-------
false negatives during analysis. A laboratory must demonstrate that an
alternate chromatographic procedure provides performance at least as good
as those conditions presented in a method, or that satisfy the analytical
requirements of the specific application for which it is being used.
1.5.2 In addition, laboratories must be cautious whenever the use
of two dissimilar columns is included in a method for confirmation of
identification. For instance, a DB-5 column cannot be used for
confirmation of results obtained using an SPB-5 column because the
stationary phases are not sufficiently dissimilar and the changes in
elution order (if any) will not provide adequate confirmation.
1.6 When gas chromatographic conditions are changed, retention times and
analytical separations are often affected. For example, increasing the GC oven
temperature changes the partitioning between the mobile and stationary phases,
leading to shorter retention times. GC retention times can also be changed by
selecting a column with a different length, stationary-phase loading (i.e.,
capillary film thickness or percent loading for packed columns), or alternate
liquid phase. As a result, two critical aspects of any SW-846 chromatographic
method are the determination and/or verification of retention times and analyte
separation.
1.7 HPLC retention times and analytical separations are also affected by
changes in the mobile and stationary phases. The HPLC mobile phase is easily
changed by adjusting the composition of the solvent mixture being pumped through
the column. In reverse phase HPLC, increasing the ratio of methanol (or
acetonitrile) to water shortens retention times. HPLC retention times can also
be changed by selecting a column with (1) a different length, (2) an alternate
bonded phase, or (3) a different particle size (e.g., smaller particles generally
increase column resolution). SW-846 methods provide conditions that have been
demonstrated to provide good HPLC separations using specific instruments to
analyze a limited number of samples. Analysts (particularly those using HPLC/MS)
may need to tailor the chromatographic conditions listed in the method for their
specific application and/or instrument. HPLC methods are particularly sensitive
to small changes in chromatographic conditions, including temperature. HPLC
column temperature control ovens should be used to maintain constant retention
times since ambient laboratory temperatures often fluctuate throughout the course
of a day.
1.8 Chromatographic methods can be used to produce quality data for the
analysis of environmental and waste samples. These methods are recommended for
use only by, or under the close supervision of, experienced analysts. Many
difficulties observed in the performance of SW-846 methods for the analysis of
RCRA wastes can be attributed to the lack of skill and training of the analyst.
2.0 SUMMARY OF METHOD
Method 8000 describes general considerations in achieving chromatographic
separations and performing calibrations. Method 8000 is to be used in
conjunction with each of the methods listed in Sec. 1.1. Each of these
chromatographic methods recommends appropriate procedures for sample preparation,
extraction, cleanup, and/or derivatization. Consult the specific procedures for
additional information on these crucial steps in the analytical process.
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2.1 Sec. 3.1 of this method provides general guidance on minimizing
contamination, including cross-contamination between samples. Sample screening
procedures are strongly recommended, and discussed in Sec. 3.2.
2.2 Before any sample or blank is introduced into a chromatographic
system, the appropriate resolution criteria and calibration procedure(s)
described in Method 8000 must be satisfied (see Sees. 3.3 and 8.3).
2.3 Sees. 3.4 and 3.5 provide information on the effects of
chromatographic interferences.
2.4 Sec 4.0 of this method contains generalized specifications for the
components of both GC and HPLC systems used in SW-846 analyses.
2.5 Calibration of the analytical system is another critical step in the
generation of quality data. Sec. 7.5 discusses specific procedures and
calculations for both linear and non-linear calibration relationships. The
continued use of any chromatographic procedure requires a verification of the
calibration relationship, and procedures for such verifications are described in
this method as well (see Sec. 7.7).
2.6 The identification of target compounds by any chromatographic
procedure is based, at least in part, on retention times. Sec. 7.6 provides
procedures for the determination of retention times and retention time windows
to be used with the specific methods listed in Sec. 1.1.
2.7 The calculations necessary to derive sample-specific concentration
results from the instrument responses are common to most of the analytical
methods listed in Sec. 1.1. Therefore, Sec. 7.10 of Method 8000 contains a
summary of the commonly used calculations.
2.8 Preventive maintenance and corrective actions are essential to the
generation of quality data in a routine laboratory setting. Suggestions for such
procedures are found in Sec. 7.11.
2.9 Most of the methods listed in Sec. 1.1 employ a common approach to
quality control (QC). While some of the overall procedures are described in
Chapter One, Sec. 8.0 describes routinely used procedures for calibration
verification, instrument performance checks, demonstrating acceptable
performance, etc.
2.10 Before performing specific analyses, analysts should determine
acceptable recovery ranges and limits of detection for all target analytes of
interest in the matrices to be tested. These procedures are described in Sec.
8.5.
3.0 INTERFERENCES/CHROMATOGRAPHIC PERFORMANCE
3.1 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are analyzed in sequence. To reduce the potential for
carryover, the sample syringe or purging device must be rinsed out between
samples with an appropriate solvent. Whenever an unusually concentrated sample
is encountered, it should be followed by injection of a solvent blank or purging
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of organic-free reagent water to check for cross-contamination. Whenever samples
contain large amounts of water-soluble materials, oils or resins, suspended
solids, high boiling compounds, or organohalide compounds, it may be necessary
to wash out the sample introduction device.
Clean purging vessels by washing them with a detergent solution, rinse with
distilled water, and then dry in a 105°C oven between analyses. Clean syringes
and autosamplers by flushing all surfaces that contact samples using appropriate
solvents.
3.2 In addition to carryover of compounds from one sample to the next, the
analysis of high-concentration samples can lead to contamination of the
analytical instrument itself. This is particularly true for GC/MS. Eliminating
this contamination can require significant time and effort in cleaning the
instruments, time that cannot be spent analyzing samples. The most reliable
procedure for ensuring minimum down time during the GC/MS analysis of samples is
to screen samples by some other technique. Samples to be analyzed for volatiles
can be screened using an automated headspace sampler (Method 5021) connected to
a GC/PID/ELCD detector (Method 8021). Samples to be analyzed for semivolatiles
can be screened using GC/FID. Other screening methods are also acceptable. The
analyst should use the screening results to choose an appropriate dilution factor
for the GC/MS analysis that will prevent system contamination yet still provide
adequate sensitivity for the major constituents of the sample.
3.3 One of the most important measures of chromatographic performance is
resolution, the separation of chromatographic peaks (peak separation/average peak
width). Peak separations are facilitated by good column efficiency (i.e., narrow
peak widths) and good column selectivity (i.e., analytes partition differently
between the mobile and stationary phases).
3.3.1 The goal of analytical chromatography is to separate sample
constituents within a reasonable time. Baseline resolution of each target
analyte from co-extracted materials provides the best quantitative results,
but is not always possible to achieve.
3.3.2 The ability to resolve individual compounds is generally the
limiting factor for the number of analytes that can be measured using a
single procedure. Some procedures, particularly Method 8081
(Organochlorine Pesticides), Method 8082 (PCBs as Aroclors), and Method
8141 (Organophosphorus Pesticides), list analytes that may not all be
resolved from one another. Therefore, while each of these methods is
suitable for the listed compounds, they may not be suitable to measure the
entire list in a single analysis. Laboratories should demonstrate that all
reported target analytes are resolved during calibration and satisfy the
requirements in Sec. 8.3. Methods that utilize mass spectrometry for
detection are affected less by resolution problems because overlapping
peaks may still be mass-resolved.
NOTE: It is highly recommended that HPLC software packages be utilized to
optimize separations and minimize solvent usage. Each laboratory
is also encouraged to recycle solvents by purification of the
solvents.
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3.4 Elevated chromatographic baselines shorld be minimized or eliminated
during these analyses. Baseline humps can usually be reduced or eliminated by
the application of appropriate sample clean-up (see Method 3600), extract
dilution, the use of pre-columns and/or inserts, or use of a selective detector.
Integration of "hump-o-grams" can result in significant quantitative errors.
When elevated baselines are observed during the analysis of blanks and standards,
the chromatographic system should be considered contaminated. This contamination
may be the result of impure carrier gas, inadequate gas conditioning, septum
bleed, column oxidation, and/or pyrolysis products in the injector or column.
Such contamination is unacceptable and should be addressed through a program of
preventive maintenance and correction actions.
3.5 GC preventive maintenance and corrective action
Poor GC performance may be expected whenever a chromatographic system is
contaminated with high-boiling materials, particularly in the injector. Analysts
should perform routine maintenance including replacement of septa, cleaning and
deactivating injector liners, and removing as much as 0.5 - 1 m from the injector
side of a capillary column.
If chromatographic performance or ghost peaks are still a problem, cleaning
of the metallic surfaces of the injection port itself may be necessary.
Capillary columns are reliable and easy to use but several specific actions are
necessary to ensure good performance.
3.5.1 Instruments should be checked daily to ensure that capillary
columns do not touch the oven walls.
3.5.2 Care should be taken to keep oxygen out of capillary columns.
3.5.3 Septa should only be changed after the oven has cooled.
3.5.4 Columns should be flushed with carrier gas for 10 minutes
before reheating the oven.
3.5.5 Carrier gas should be scrubbed to remove traces of oxygen and
scrubbers should be changed regularly.
3.5.6 Carrier gas should always be passed through the column
whenever the oven is heated.
3.6 HPLC preventive maintenance and corrective action
HPLC band broadening results from improper instrument setup or maintenance.
Band broadening results whenever there is dead volume between the injector and
the detector. Therefore, plumbing connections should be of minimum length and
diameter, and ferrules should be properly positioned on the tubing to minimize
dead volume.
3.6.1 Columns should not be subjected to sudden physical stress
(e.g., dropping) or solvent shocks (e.g., changing solvents without a
gradient).
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3.6.2 Columns can become contaminated with participates or insoluble
materials. Guard columns should be used when dirty samples are analyzed.
3.6.3 High quality columns are packed uniformly with small uniform
diameter particles with a minimum number of free silol groups. Use of such
columns will result in optimum chromatographic performance.
3.6.4 Columns should be replaced when performance degrades (e.g.,
significant band broadening, peak splitting, loss of chromatographic
resolution).
3.6.5 Pumping systems should deliver reproducible gradients at a
uniform flow rate. Rates can be checked by collecting solvent into a
graduated cylinder.
3.6.6 Column temperatures should be regulated by the use of column
temperature control ovens to ensure reproducibility of retention times.
3.6.7 Small changes in the composition or pH of the mobile phase can
have a significant effect on retention times.
4.0 APPARATUS AND MATERIALS
4.1 GC inlet systems
4.1.1 Volatile organics
Volatile organic analytes are introduced into a GC through a
purge-and-trap system, by direct injection, or by other devices. The
purge-and-trap apparatus is described in Method 5030 for water samples and
in Method 5035 for soil and other solid samples. See Method 5000 for
guidance on all forms of sample introduction of volatiles into the GC and
GC/MS system.
4.1.2 Semivolatile organics
Sample extracts containing semivolatile organic compounds are
introduced into a GC with a syringe that passes through a septum into an
injection port. The injection port allows the sample extract to be
vaporized prior to being flushed onto the GC column, hence the term "gas"
chromatography. Correct set up and maintenance of the injector port is
necessary to achieve acceptable performance with GC methods. Septa should
be changed frequently enough to prevent retention time shifts of target
analytes and peak tailing. The schedule for such septa changes is
dependent on the quality of the septa, the sharpness of the needle, and the
operation of the injection system. Appropriate injector liners should be
installed, and liners should be cleaned and deactivated (with
dichlorodimethylsilane) regularly.
4.1.3 Injector difficulties include the destruction of labile
analytes and discrimination against high boiling compounds in capillary
injectors.
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4.1.3.1 Packed columns and wide-bore capillary columns
(> 0.50 mm ID) should be mounted in 1/4-inch injectors. An injector
liner is needed for capillary columns.
4.1.3.2 Narrow-bore capillary columns (< 0.32 mm ID) should
be mounted in split/splitless (Grob-type) injectors. Split/splitless
injectors require automated valve closures that direct most of the
flow (and sample) onto the head of the analytical column. After 30
- 45 seconds, the split valve is opened, so that most of the flow is
vented during analysis, thus eliminating the solvent tail, and
maintaining proper flow through the column. The initial oven
temperature should be below the boiling point of the injection solvent
if the solvent front interferes with early eluting analytes or if the
solvent effect is needed to resolve difficult-to-separate analytes.
4.1.3.3 Cool on-column injection allows the analysis of
labile compounds that degrade on packed columns and in spl it/splitless
injectors.
4.2 GC flow control
Precise control of the gas mobile phase is necessary to achieve
reproducible GC retention times. Flow controllers within any GC used for SW-846
analyses must deliver a precisely metered gas flow at a rate appropriate for the
GC column mounted in the instrument.
4.2.1 Most GCs have restrictors built into flow controllers. These
restrictors are used to provide precise flow at the carrier gas flow rate
specified in the method (e.g., use <20 mL/min restrictors for wide-bore
capillary methods). Carrier gas flow rates should be checked regularly
(with both the injector and the oven heated) using a bubble meter.
4.2.2 Cylinder pressures should also be regulated properly.
Manifold pressures must be sufficiently large that a change in the head
pressure of an individual instrument does not affect the flow through all
instruments. Toggle valves that allow instruments to be isolated are
recommended for all multi-instrument gas delivery systems. Analysts should
spend time each week conducting preventative maintenance in order to ensure
that proper flow control is maintained. One needs to search for leaks
using a helium tester or soap solution at each connector in the gas
delivery system.
4.2.3 Carrier gas should be of high purity and should be conditioned
between the cylinder and the GC to remove traces of water and oxygen.
Scrubbers should be changed according to manufacturers recommendations.
Gas regulators should contain stainless steel diaphragms. Neoprene
diaphragms are a potential source of gas contamination, and should not be
used.
4.3 Gas chromatographic columns
Each determinative method in SW-846 provides a description of a
chromatographic column or columns with associated performance data. Other packed
or capillary (open-tubular) columns may be substituted in SW-846 methods to
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improve performance if (1) the requirements of Sees. 8.3 and 8.4 are satisfied,
and (2) target analytes are sufficiently resolved from one another and from
co-extracted interferences.
4.3.1 Narrower columns are more efficient (i.e., can resolve more
analytes) but have a lower capacity (i.e., can accept less sample without
peak distortion).
4.3.2 Longer columns can resolve more analytes, as resolution
increases as a function of the square root of column length.
4.3.3 Increasing column film thickness or column loading increases
column capacity and retention times.
4.3.4 Use of capillary columns has become standard practice in
environmental and waste analysis. Capillary columns have an inherently
greater ability to separate analytes than packed columns.
4.3.5 All columns used for SW-846 analyses should be installed
properly. Column ends should be cut square. Contaminated ends should be
trimmed off, and columns should be placed through ferrules before they are
trimmed. Columns should never touch the walls of the GC oven during
analysis, and the manufacturer's column temperature limits should not be
exceeded.
4.3.6 Septa should be changed regularly and septum nuts should not
be overtightened. Oxygen should not be introduced into a hot column and
carrier gas should be passed through a column whenever it is heated. New
columns, particularly packed columns, should be conditioned prior to
analyzing samples.
4.4 GC detectors
Detectors are the transducers that respond to components that elute from
a GC column and produce the electrical signal that is used for quantitative
determinations. SW-846 analyses are conducted using selective detectors or mass
spectrometers listed in Sec. 1.1. Detectors should be maintained at least 20°C
above the highest oven temperature employed to prevent condensation and detector
contamination.
4.5 HPLC injectors
Liquids are essentially non-compressible, so a mechanical device is
necessary that allows introduction of the sample into a high pressure flow
without significant disruption in the flow rate and hydraulic pressure.
Normally, a 6-port valve is used for this purpose. A sample loop (generally
10-100 juL) is isolated from the flow of the mobile phase and filled with a
sample extract. (Larger sample loops may be used to increase sensitivity,
however, they may degrade chromatographic performance). The extract is then
injected by turning the valve so that the mobile phase flows through the loop.
This procedure virtually eliminates dead volume in the injector and is fully
compatible with automated operation.
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4.5.1 When the extract is highly viscous, a pressure spike results
which can automatically shut off the HPLC pump.
4.5.2 Contamination of subsequent injections may occur when the
extract contains material that is not soluble in the mobile phase.
4.5.3 Injection loops are easily changed but analysts must ensure
that the compression fittings are properly installed to prevent leaks.
Injectors require maintenance, as the surfaces that turn past each other
do wear down.
4.6 HPLC pumps
The mobile phase used for HPLC must be accurately pressurized before it
enters the injector. HPLC pumps are generally capable of delivering solvent at
5000 psi with excellent precision. The rate of delivery depends on the column
that is used for the separation. Most environmental methods recommend flow rates
of 0.25-1.0 mL/min. Flow rates should be checked by collecting column effluent
in a graduated cylinder.
Most pumping systems are capable of changing solvent concentration during
an analysis (i.e., gradient elution). Gradients are generated by either high
pressure mixing of two streams between the pump and the injector or by
proportional mixing of the solvents before they are pumped. In either case,
solvent mixing can cause changes in the solubility of dissolved gases, the
formation of bubbles in the mobile phase, or non-reproducible gradients.
4.6.1 Air bubbles result in erratic baseline and, in the case of low
pressure mixing, bubbles can cause the pump to cavitate. Therefore, HPLC
solvents should be degassed prior to use.
4.6.2 Non-reproducible gradients can result in significant changes
in retention times from run to run.
4.6.3 HPLC solvents should be filtered to remove particles that
cause pump piston wear. HPLC pump maintenance includes replacing seals
regularly. (Use of strong buffers or solvents like tetrahydrofuran can
significantly shorten the lifetime of pump seals.) Pumps should deliver
solvent with minimal pulsation.
4.7 HPLC Columns
These columns must be constructed with minimum dead volume and a narrow
particle size distribution. HPLC columns are generally constructed of stainless
steel tubing and are sealed with compression fittings. Manufacturers provide
columns that are bonded with different alkyl groups (e.g., C18, cyano, TMS), have
different percent carbon loading, are packed with different particle sizes (3-10
jum), and are packed with particles of different pore size (smaller pores mean
greater surface area), or are of different dimensions.
4.7.1 Columns with higher percent loading have the capacity to
analyze somewhat larger samples, but extremely high loadings may contribute
to problems with the particle beam MS interface.
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4.7.2 Columns with free silol groups show less tailing of polar
materials (e.g., amines).
4.7.3 A smaller particle (and pore) size generally gives better
resolution, higher back pressure, and smaller sample capacity. Columns
with 3 jum particle size may have short lifetimes when they are used for the
analysis of complex waste extracts.
4.7.4 Improvements in column packing have resulted in 10 and 15 cm
columns that provide the separating power necessary for most environmental
and waste analyses.
4.7.5 Internal diameters of columns used for environmental and waste
analysis are generally 2-5 mm. Narrower columns are called microbore
columns. While they provide better separations, they become fouled more
easily.
4,7.6 The lifetime and performance of HPLC columns can be improved
through proper maintenance. Analysts should filter sample extracts, use
compatible guard columns, check for clogged frits and for column voids.
Columns should not be stored dry or containing strong buffers.
4.8 HPLC column temperature control ovens
HPLC retention times are much more reproducible if the column is held at
a constant temperature. Temperature control ovens capable of maintaining the
HPLC column at ± 0.1°C should be utilized to maintain consistent retention times
throughout the course of an HPLC analysis. Normal oven operating temperature
would be 3-5°C above ambient laboratory temperature.
4.9 HPLC detectors
Detectors are the transducers that respond to components that elute from
a HPLC column and produce the electrical signal that is used for quantitative
determinations. SW-846 analyses are conducted using selective detectors or mass
spectrometers listed in Sec. 1.1. HPLC/MS requires the use of a sophisticated
interface that separates target analytes from the aqueous mobile phase. Example
interfaces include the thermospray (TSP), electrospray (ESP), and the particle
beam (PB) interfaces.
4.10 Data systems
Raw chromatographic data have to be reduced in order to provide the
quantitative information required by analysts. The use of sophisticated data
systems are strongly recommended for SW-846 chromatographic methods. The ability
to store and replot chromatographic data is invaluable during data reduction and
review. Organizations should establish their priorities and select the system
that is most suitable for their applications.
4.11 Supplies
Chromatographers require a variety of supplies. The specific items that
should be stocked depend on laboratory instrumentation and the analyses
performed. At a minimum, laboratories need PTFE tape, stainless steel
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regulators, acid-washed copper tubing, and syringes, and replacement parts for
instruments.
4.11.1 Laboratories performing GC analyses also require high purity
gases, scrubbers for gas conditioning, gas-tight fittings, capillary
cutters, magnifying glasses, septa with proper temperature limits,
appropriate ferrules, dichlorodimethylsilane (for deactivating surfaces),
pyrex wool, spare columns, and injection port liners.
4.11.2 Laboratories performing HPLC analyses require high purity
solvents, column packing material, frits, 1/16-inch tubing, appropriate
ferrules, solvent filtration apparatus, and solvent degassing apparatus.
5.0 REAGENTS
See the specific extraction and determinative methods for the reagents
needed.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
Refer to Chapter 4, Organic Analytes, Sec. 4.1, for information on sample
collection, preservation and handling procedures.
7.0 PROCEDURE
Extraction and cleanup are critical for the successful analyses of
environmental samples and wastes. Analysts should pay particular attention to
selection of sample preparation procedures to obtain reliable measurements.
7.1 Extraction
The methods for organic analytes in SW-846 usually recommend appropriate
sample extraction procedures. General guidance on semivolatile extraction
procedures can be found in Method 3500. Guidance on volatile procedures can be
found in Method 5000.
7.2 Cleanup and separation
The methods for organic analytes in SW-846 usually recommend appropriate
cleanup procedures. General guidance on cleanup procedures can be found in
Method 3600. While some relatively clean matrices (such as ground water samples)
may not require extensive cleanups, the analyst should carefully balance the time
savings gained by skipping cleanups against the potential increases in down time
and loss of data quality that can occur as a result.
7.3 Recommended chromatographic columns and instrument conditions are
described in each determinative method. As noted earlier, these columns and
conditions are typically those used during the development and testing of the
method. However, other chromatographic systems may have somewhat different
characteristics. In addition, analytical instrumentation continues to evolve.
Therefore, SW-846 methods allow analysts some flexibility to change these
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conditions (with certain exceptions), as long as they demonstrate adequate
performance.
Chromatographic performance is demonstrated by: resolution of standards
and detector linearity during calibration; accuracy; precision; and avoidance of
false positives/false negatives during analysis. If a laboratory employs an
alternative Chromatographic procedure or conditions, then the laboratory must
demonstrate that the performance is at least as good as the performance which can
be achieved using the conditions presented in the SW-846 method, or that the
performance satisfies the analytical requirements of the specific application for
which the alternative procedure is being used.
7.4 Initial Calibration
Calibration of an analytical instrument involves the delineation of the
relationship between the response of the instrument to the amount or
concentration of an analyte introduced into the instrument. The graphical
depiction of this relationship is often referred to as the calibration curve.
In order to perform quantitative measurements, this relationship must be
established prior to the analysis of any samples, and thus, is termed initial
cal ibration.
The initial calibration for SW-846 Chromatographic methods involves the
analysis of standards containing the target compounds at a minimum of five
concentrations covering the working range of the instrument. In order to produce
acceptable sample results, the response of the instrument must be within the
working range established by the initial calibration.
7.4.1 Calibration standards are prepared using the procedures
indicated in Sec. 5.0 of the determinative method of interest. However,
the general procedure is described here.
7.4.1.1 For each analyte and surrogate of interest, prepare
calibration standards at a minimum of five concentrations by adding
volumes of one or more stock standards to a volumetric flask and
diluting to volume with an appropriate solvent.
7.4.1.2 The lowest concentration calibration standard should
be at a concentration equal to the method quantitation limit (based
on the concentration in the final volume listed in the preparation
method, with no dilutions).
7.4.1.3 The other concentrations should define the working
range of the detector or correspond to the expected range of
concentrations found in actual samples that are also within the
working range of the detector.
7.4.1.4 At least one of the calibration solutions should
correspond to a sample concentration at or below any regulatory or
action limit associated with a target compound.
7.4.1.5 Given the number of target compounds addressed by
some of the methods listed in Sec. 1.1, it may be necessary to prepare
several sets of calibration standards, each set consisting of five
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solutions. The initial calibration will then involve the analysis of
each of these sets of five standards.
7.4.1.6 Once the standards have been prepared, the initial
calibration begins by establishing chromatographic operating
parameters that provide instrument performance equivalent to that
documented in Sec. 7.0 of the determinative method of interest.
7.4.2 External standard and internal standard calibration techniques
The chromatographic system may be calibrated using either the external
standard or the internal standard techniques described below. General
calibration criteria are provided in this section for GC and HPLC
procedures using non-MS detection. The applicable calibration procedures
for GC/MS (e.g., Methods 8260, 8270, 8280, and 8290), HPLC/MS (e.g.,
Methods 8321 and 8325), and GC/FT-IR (e.g., Method 8410) are described in
those methods. Some determinative methods may provide special guidance on
calibration that is specific to that method.
Regardless of whether external standard or internal standard
calibration will be used, introduce each calibration standard into the
instrument using the same technique that will be used to introduce the
actual samples into the gas chromatograph (e.g., 1-3 p,l injections for GC
methods, 10-100 ^L injections for HPLC methods, purge-and-trap techniques
for volatiles, etc.). Tabulate peak area or height responses against the
Tiass or concentration injected, as described below.
7.4.2.1 External standard calibration procedure
External standard calibration involves comparison of
instrument responses from the sample to the responses from the target
compounds in the calibration standards. Sample peak areas (or peak
heights) are compared to peak areas (or heights) of the standards.
The ratio of the detector response to the amount (mass) of analyte in
the calibration standard is defined as the calibration factor (CF).
A CF is calculated for each analyte and surrogate at each
initial calibration standard concentration, according to the equation
below.
r[: _ Peak Area (or Height) of the Compound in the Standard
Mass of the Compound Injected (in nanograms)
For multi-component analytes, the total area (or heights) of several
peaks is used for quantitation.
The CF can also be calculated using the concentration of the
standard rather than the mass in the denominator of the equation
above. However, the use of concentrations in CFs will require some
changes in the way that sample concentrations are calculated (see Sec.
7.10).
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7.4.2.2 Internal standard calibration procedure
Internal standard calibration involves the comparison of
instrument responses from the target compounds in the sample to the
responses of specific standards added to the sample or sample extract
prior to injection. The ratio of the peak area (or height) of the
target compound in the sample or sample extract to the peak area (or
height) of the internal standard in the sample or sample extract is
compared to a similar ratio derived for each calibration standard.
The ratio is termed the response factor (RF), and may also be known
as a relative response factor in other methods.
In many cases, internal standards are recommended in SW-846
methods. These recommended internal standards are often brominated,
fluorinated, or stable isotopically labeled analogs of specific target
compounds, or are closely related compounds whose presence in
environmental samples is highly unlikely. If internal standards are
not recommended in the method, then the analyst needs to select one
or more internal standards that are similar in analytical behavior to
the compounds of interest, and not expected to be found in the samples
otherwise.
Whichever internal standards are employed, the analyst needs
to demonstrate that the measurement of the internal standard is not
affected by method analytes and surrogates or by matrix interferences.
In general, internal standard calibration is not as useful for GC and
HPLC methods with non-MS detectors because of the inability to
chromatographically resolve many internal standards from the target
compounds. The use of MS detectors makes internal standard
calibration practical because the masses of the internal standards can
be resolved from those of the target compounds even when
chromatographic resolution cannot be achieved.
When preparing calibration standards for use with internal
standard calibration, add a constant amount of one or more internal
standards to each calibration standard and dilute to volume with an
appropriate solvent. This same amount of the internal standard is
added to each sample extract immediately prior to injection into the
instrument.
For each of the initial calibration standards, calculate the
RF values for each target compound relative to one of the internal
standards as follows:
A x C
RF = - "
A x C
where: IS s
As = Peak area (or height) of the analyte or surrogate.
A,s = Peak area (or height) of the internal standard.
Cs = Concentration of the analyte or surrogate, in M9/L.
Cis = Concentration of the internal standard, in M9/L-
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Note that in the equation above, RF is unitless, i.e., the
units from the two area terms and the two concentration terms cancel
out. Therefore, units other than p.g/1 may be used for the
concentrations of the analyte, surrogate, and internal standard,
provided that both Cs and Cis are expressed in the same units. The
mass of the analyte and internal standard may also be used in
calculating the RF value.
7.5 Calibration linearity
Historically, many analytical methods have relied on linear models of the
calibration relationship, where the instrument response is directly proportional
to the amount of a target compound. The linear model has many advantages, among
them, simplicity and ease of use. Unfortunately, given the advent of new
detection techniques and the fact that many techniques cannot be optimized for
all of the analytes to which they may be applied, the analyst is increasingly
likely to encounter situations where the linear model neither applies nor is
appropriate.
Therefore, SW-846 chromatographic methods allow the use of both linear and
non-linear models for the calibration data, as described below. Given the
limitations in instrument data systems, it is likely that the analyst will have
to choose one model for all analytes in a particular method. Both models can be
applied to either external or internal standard calibration data.
Note: The option for non-linear calibration may be necessary to achieve low
detection limits or to address specific instrumental techniques.
However, it is not EPA's intent to allow non-linear calibration to be
used to compensate for detector saturation at higher concentrations or to
avoid proper instrument maintenance.
Whichever calibration model is employed, a unique analyte or surrogate
concentration must fall within the calibration range. Sample with concentrations
that exceed the calibration range must be diluted to fall within the range.
7.5.1 Linear calibration through the origin
When calculated as described above, both calibration factors and
response factors are a measure of the slope of the calibration relationship
and assume that the curve passes through the origin. Under ideal
conditions, the factors will not vary with the concentration of the
standard that is injected into the instrument. In practice, some variation
is to be expected. However, when the variation, measured as the relative
standard deviation (RSD), is less than or equal to 20%, the use of the
linear model is appropriate, and the calibration curve can be assumed to
be linear and to pass through the origin.
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Therefore, to evaluate the linearity of the initial calibration,
calculate the mean CF (external standard calibration) or RF (internal
standard calibration), the standard deviation (SD), and the RSD as follows:
E CF. £ RF.
mean CF = CF = -^ _ '- mean RF = RF" = ^ _ '
SD =
E(CR-CF):
n-1
SD =
1=1
n-1
RSD = .55 x 100 RSD = I? x 100
CF RF
where n is the number of calibration standards and RSD is expressed as a
percentage (%).
If the RSD of the calibration or response factors is less than or
equal to 20% over the calibration range, then linearity through the origin
may be assumed, and the average calibration or response factor may be used
to determine sample concentrations.
7.5.2 Other calibration techniques
If the RSD of the calibration or response factors is greater than 20%
over the calibration range, then linearity through the origin cannot be
assumed. If this is the case, then the analyst has four options of how to
proceed. They are to:
• Adjust the instrument and/or perform instrument maintenance until
the RSD of the calibration or response factors meets the 20% QC
limit.
• Narrow the calibration range until the response is linear.
• Use a linear calibration that does not pass through the origin,
and adjust the quantitation limits accordingly.
• Use a calibration curve or non-linear calibration model.
The options above are listed in order of increasing difficulty, and should
be attempted in that order, that is, trying the simplest solutions first
and evaluating the results before proceeding with the more difficult
options. These options are discussed in greater detail in the following
sections.
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7.5.2,1 The first option for addressing the calibration
1inearity difficulties is to check the instrument operating conditions
and make adjustments to achieve a linear calibration. The suggested
maintenance procedures in Sec. 7.11 may be useful in guiding such
adjustments. This option will apply in those instances where a linear
instrument response is expected. It may involve some trade-offs to
optimize performance across all target analytes. For instance,
changes to the operating conditions necessary to achieve linearity for
problem compounds may cause the RSD for other compounds to increase,
but as long as all analytes meet the RSD limits for linearity, the
calibration is acceptable.
7.5.2.2 The second option is to narrow the calibration
range. This process usually involves an examination of the
calibration curve itself, i.e., a graph of the instrument response
versus concentration. If the data indicate that the response is
leveling off at the upper end of the calibration, then recalculate the
RSD without the data for the highest calibration standard, and see if
the RSD meets the QC limit. If so, prepare a new calibration standard
at a concentration between the existing fourth and fifth calibration
standards, analyze it, and calculate the RSD with all five points.
Similarly, if the non-linearity appears to be at the low end
of the calibration range, eliminate the lowest standard, recalculate
the RSD, and prepare a new lowest standard for testing.
If linearity can be achieved using a narrower calibration
range, document the calibration linearity, and proceed with analyses.
The changes to the upper end of the calibration range will affect need
to dilute samples above the range, while changes to the lower end will
affect the overall sensitivity of the method. Consider the regulatory
limits associated with the target analytes when adjusting the lower
end of the range.
7.5.2.3 The third option is to use a linear calibration line
that does not pass through the origin. This is most easily achieved
by performing a linear regression of the instrument response versus
the concentration of the standards. Make certain that the instrument
response is treated as the dependent variable (y) and the
concentration as the independent variable (x).
The regression will produce the slope and intercept terms for
a linear equation in the form:
y = ax + b
where:
y = Instrument response
a = Slope of the line (also called the coefficient of x)
x = Concentration of the calibration standard
b = The intercept
The analyst should not force the line through the origin, but
have the intercept calculated from the five data points. Otherwise,
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the problems noted with the RSD value will occur, i.e., a line through
the origin will not meet the QC specifications. In addition, do not
include the origin (0,0) as a sixth calibration point. The regression
calculation will generate a correlation coefficient (R2) that is a
measure of the "goodness of fit" of the regression line to the data.
A value of 1.00 indicates a perfect fit. In order to be used for
quantitative purposes, R2 must be greater than or equal to 0.99.
The calculated intercept value needs to be evaluated before
reporting sample results. A positive value for the intercept
indicates that there is some threshold instrument response which is
the limiting factor in establishing linearity. A negative intercept
value can be transformed into an x-intercept value that represents a
threshold concentration which is the limitation. If the intercept is
positive, then, as a general rule, results where the instrument
response is less than three times (3x) the intercept value may be
unreliable. This will afford some protection against false positive
results. If the intercept is negative, results below the
concentration of the lowest concentration calibration standard may be
unreliable. These adjustments to the quantisation limits will apply
to all samples analyzed using the regression line.
In calculating sample concentrations, the regression equation
is rearranged to solve for the concentration (x), as shown below.
7.5.2.4 The fourth option is to use a non-linear calibration
model. This option should only be used after exhausting the other
three options, or in situations where the analyst knows that the
instrument response does not follow a linear model over a sufficiently
wide working range.
When using a calibration curve or a non-linear calibration
model (e.g., a polynomial fit) for quantitation, the curve must
produce a unique value over the calibration range, and the polynomial
equation may be no more than third order, i.e.,
y = ax3 + bx2 + ex + d
In developing a polynomial fit for the calibration data, the
instrumental response (y) must be treated as the dependent variable,
and the concentration of the calibration standard (x) must be the
independent variable. Do not force the line through the origin, i.e.,
do not set the intercept as 0, and do not include (0,0) as a
calibration point.
The statistical considerations in developing a non-linear
calibration model require more data than the more traditional linear
approaches described above. As a result, the analyst has two options
for how to perform the calibration. The first option is to employ at
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least three replicate 5-point calibrations. The instrument operating
conditions must remain the same during all three calibration runs.
The second option is to perform at least a single 10-point
calibration. The purpose of the additional five calibration points
is to better define the shape for the calibration curve. The
concentrations of most of the additional five standards should fall
between the concentrations of the original 5-point calibration in Sec.
7.4.1. As necessary to describe the curve, some standards may be
added above or below the concentration range of the original 5-point
curve, depending on the working range of interest.
Most curve fitting programs will use some form of least
squares minimization to adjust the coefficients of the polynomial
(a,b,c, and d, above) to obtain the polynomial that best fits the
data. The "goodness of fit" of the polynomial equation is evaluated
by calculating the weighted coefficient of the determination (COD).
n ( i
i —\2 n - i
(yobs -y) - -
COD =
where:
yobs = Observed response (area) for each concentration from each
initial calibration point (i.e., 10 observed responses for
the 10-point curve, and 15 observed responses for the three
replicate 5-point curves)
y = Mean observed response from the 10-point calibration or from
all three 5-point calibrations
Y, = Calculated (or predicted) response at each concentration from
the initial calibration(s)
n = Total number of calibration points (i.e., 10, for a single
10-point calibration, and 15, for three 5-point calibrations)
p = Number of adjustable parameters in the polynomial equation
(i.e., 3 for a third order; 2 for a second order polynomial)
Under ideal conditions, with a "perfect" fit of the model to
the data, the coefficient of the determination will equal 1.0. In
order to be an acceptable non-linear calibration, the COD must be
greater than or equal to 0.99.
As noted in Sec. 7.5, whichever of these options is employed,
a unique analyte or surrogate concentration must fall within the
calibration range. Samples with concentrations that exceed the
calibration range must be diluted to fall within the range.
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7.6 Retention time windows
Retention time windows are crucial to the identification of target
compounds. Absolute retention times are used for compound identification in all
GC and HPLC methods that do not employ internal standard calibration. Retention
time windows are established to compensate for minor shifts in absolute retention
times as a result of sample loadings and normal chromatographic variability. The
width of the retention time window should be carefully established to minimize
the occurrence of both false positive and false negative results. Tight
retention time windows may result in false negatives and/or may cause unnecessary
reanalysis of samples when surrogates or spiked compounds are erroneously not
identified. Overly wide retention time windows may result in false positive
results that cannot be confirmed upon further analysis.
7.6.1 Before establishing retention time windows, make sure that the
chromatographic system is operating reliably and that the system conditions
have been optimized for the target analytes and surrogates in the sample
matrix to be analyzed. Make three injections of all single component
standard mixtures and multi-component analytes (such as PCBs) over the
course of a 72-hour period. Serial injections or injections over a period
of less than 72 hours may result in retention time windows that are too
tight.
NOTE: The criteria listed in Sec. 7.6 are provided for GC and HPLC
procedures using non-MS or FTIR detection. Identification procedures
are different for GC/MS (e.g., Methods 8260 and 8270), HPLC/MS (e.g.,
Methods 8321 and 8325), and GC/FT-IR (e.g., Method 8410).
7.6.2 Calculate the mean and standard deviation of the three
absolute retention times for each single component analyte and surrogate.
For multi-component analytes, choose three to five major peaks (generally
listed in the determinative method) and calculate the mean and standard
deviation of those peaks.
7.6.3 If the standard deviation of the retention times for a target
compound is 0.00 (i.e., no difference between the absolute retention
times), then the laboratory must inject 10 sample extracts during the
72-hour period to demonstrate that absolute retention times are not
affected by co-extracted interferences or instrument instability.
7.6.4 The width of the retention time window for each analyte,
surrogate, and major constituent in multi-component analytes is defined as
plus and minus three times the standard deviation of the mean absolute
retention time established during the 72-hour period.
7.6.5 Establish the midpoint of the retention time window for each
analyte and surrogate by using the absolute retention time for each analyte
and surrogate from the mid-concentration standard of the initial
calibration. The absolute retention time window equals the midpoint ±3 SD,
where the standard deviation is determined as described in Sec. 7.6.4.
7.6.6 The retention times of all target analytes and surrogates in
the calibration verification standard analyzed at the beginning of the
analytical shift must fall within the absolute retention time windows
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calculated in Sec. 7.6.4. The purpose of this check is to ensure that
retention times do not continually drift further and further from those
used to establish the widths of the retention time windows.
If the retention time of any analyte in the standard at the beginning
of the analytical shift does not fall within the ±3 SD window, then a new
initial calibration is necessary unless system maintenance (Sec. 7.11)
corrects the problem.
In addition, the retention times of all analytes in the subsequent
calibration verification standards analyzed during the analytical shift
(see Sec. 7.7) must fall within the absolute retention time windows
established in Sec. 7.6.5.
7.6.7 The laboratory must calculate absolute retention time windows
for each analyte and surrogate on each chromatographic column. The
retention time windows must be recalculated (beginning with Sec. 7.6.1)
whenever significant column maintenance is performed. New retention time
windows must be established when a new GC column is installed. The
retention time windows should be reported with the analysis results in
support of the identifications made.
7.7 Calibration verification
The calibration relationship established during the initial calibration
(Sec. 7.5) must be verified at periodic intervals. The process of calibration
verification applies to both external standard and internal standard calibration
techniques, as well as to linear and non-linear calibration models.
As a general rule, the initial calibration in an SW-846 method must be
verified at the beginning of each 12-hour analytical shift during which samples
are analyzed. (Some methods may specify more frequent verifications). The
verification process involves analysis of a single calibration standard, usually
the mid-point standard from the initial calibration. The response of each
analyte in this standard is evaluated in one of two ways. The first approach
compares the calibration or response factor calculated from the single standard
to the average factor calculated for the initial calibration. The second
approach treats the standard as an "unknown" and calculates a concentration for
each analyte, using the initial calibration. This calculated concentration is
then compared to the nominal or theoretical concentration of the standard.
If the response (or calculated concentration) for an analyte is within
±15% of the response obtained during the initial calibration, then the initial
calibration is considered still valid, and the analyst may continue to use the
CF or RF values from the initial calibration to quantitate sample results. The
±15% criterion may be superseded in certain determinative methods.
Except where the determinative method contains alternative calibration
verification criteria, if the response (or calculated concentration) for any
analyte varies from the response obtained during the initial calibration by more
than ±15%, then the initial calibration relationship may no longer be valid.
When this occurs, check the instrument operating conditions and/or perform
instrument maintenance (see Sec. 7.11), and inject another aliquot of the
calibration verification standard. If the response for the analyte is still not
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within ±15%, then a new initial calibration must be prepared for that compound.
Note: The process of calibration verification is fundamentally different from
the approach called "continuing calibration" in some methods from other
sources. As described in those methods, the calibration factors or
response factors calculated during continuing calibration are used to
update the calibration factors or response factors used for sample
quantitation. This approach, while employed in other EPA programs,
amounts to a daily single-point calibration, and is not appropriate nor
permitted in SW-846 chromatographic procedures.
7.7.1 Verification of linear calibrations
Calibration verification for linear calibrations involves the
calculation of the percent drift or the percent difference of the
instrument response between the initial calibration and each subsequent
analysis of the verification standard. Use the equations below to
calculate % Drift or % Difference, depending on the procedure specified in
the determinative method.
„. _. . _. Calculated concentration - Nominal concentration 1rtrt
% Drift = x 100
Theoretical concentration
where the calculated concentration is determined using the calibration
factors or response factors from the initial calibration and the
theoretical concentration is the concentration at which the standard was
prepared.
CF - CF RF - RF
% Difference - v x 100 or = _ v x 100
CF RF
where CFV and RFV are the calibration factor and the response factor
(whichever applies) from the analysis of the verification standard, and CF
and RF are the mean calibration factor and mean response factor from the
initial calibration. Except where superseded in certain determinative
methods, the % difference or % drift calculated for the calibration
verification standard must be less than or equal to 15% before any sample
analyses may take place.
7.7.2 Verification of a non-linear calibration
Calibration verification of a non-linear calibration is performed
using the percent drift calculation described in Sec. 7.7.1, above. Except
where superseded in certain determinative methods, the % drift calculated
for the calibration verification standard must be less than or equal to 15%
before any sample analyses may take place.
7.7.3 Regardless of whether a linear or non-linear calibration model
is used, if either the percent drift or percent difference criterion is not
met, then no sample analyses may take place until the calibration has been
verified or a new initial calibration is performed that meets the
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specifications in Sec. 7.5 and those in the determinative method. If the
calibration cannot be verified after the analysis of a single verification
standard, then adjust the instrument operating conditions and/or perform
instrument maintenance (see Sec. 7.11), and analyze another aliquot of the
verification standard. If the calibration cannot be verified with the
second standard, then a new initial calibration must be performed.
7.7.4 All target analytes and surrogates, including those reported
as non-detects, must be included in a periodic calibration for purposes of
retention time confirmation and to demonstrate that calibration
verification criteria are being met. The frequency of this periodic
calibration is project-, method-, and analyte-specific.
7.7.5 Calibration verification should be performed using both high
and low concentration standards from time to time. This is particularly
true when the ECD or ELCD is used. These detectors drift and are not as
stable as FID or FPD, and periodic use of the high and low concentration
standards serves as a further check on the initial calibration.
7.7.6 Additional analyses of the midpoint calibration standard
during a 12-hour analytical shift are strongly recommended for methods
involving external standard calibration. If the response for any analyte
varies from the average initial calibration response by more than ±15% in
these additional determinations, corrective action should be taken (see
Sec. 7.11) to restore the system or a new calibration curve should be
prepared for that compound.
The frequency of verification necessary to ensure accurate measurement
is dependent on the detector and the sample matrix. Very sensitive
detectors that operate in the sub-nanogram range are generally more
susceptible to changes in response caused by column contamination and
sample carryover. Therefore, more frequent verification of calibration
(i.e., after every 10 samples) may be necessary for the electron capture,
electrochemical conductivity, photoionization, and fluorescence detectors.
Sec. 8.2.2 specifies that samples analyzed using external standards
must be bracketed by periodic analyses of standards that meet the QC
acceptance criteria (e.g., calibration and retention time). Therefore,
more frequent analyses of standards will minimize the number of sample
extracts that would have to be reinjected if the QC limits are violated for
the standard analysis.
7.7.7 Solvent blanks and any method blanks specified in the
preparative methods (Methods 3500 and 3600) should be run immediately after
the calibration verification analyses to confirm that laboratory
contamination does not cause false positive results (method blank) and that
there is no carryover from standards to samples (solvent blank).
7.8 Chromatographic analysis of samples
7.8.1 Introduction of sample extracts into the chromatograph varies,
depending on the volatility of the compound. Volatile organics are
primarily introduced by purge-and-trap techniques (Method 5030, water and
Method 5035, soils). However, use of Methods 3810, 5021, or an automated
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headspace technique may be advisable for screening volatiles in some sample
matrices to prevent overloading and contamination of the purge-and-trap
system. Semivolatile and non-volatile analytes are introduced by direct
or split/splitless injection.
7.8.1.1 Manual injection (GC)
Inject 1-5 /il_ of the sample extract. The use of the solvent
flush technique is necessary for packed columns. Use 1-2 juL of
sample extract for capillary columns.
7.8.1.2 Automated injection (GC)
Using automated injection, smaller volumes (i.e., 1 nl) may
be injected, and the solvent flush technique is not necessary.
Laboratories should demonstrate that the injection volume is
reproducible.
7.8.1.3 Purge-and-trap
Refer to Methods 5000, 5030, or 5035 for details.
7.8.1.4 Manual injection (HPLC)
Inject 10-100 juL. This is generally accomplished by over-
filling the injection loop of a zero-dead-volume injector. Larger
volumes may be injected if better sensitivity is required, however,
chromatographic performance may be affected.
7.8.1.5 Automated injection (HPLC)
Inject 10-100 /iL. Laboratories should demonstrate that the
injection volume is reproducible. Larger volumes may be injected if
better sensitivity is required, however, chromatographic performance
may be affected.
7.8.2 Samples are analyzed during an analysis sequence. The
sequence begins with instrument calibration, which is followed by the
analysis of sample extracts. Verification of calibration and retention
times is necessary no less than once every 12-hour analytical shift. The
sequence ends when the set of samples has been injected or when qualitative
and/or quantitative QC criteria are exceeded. As noted in Sees. 7.7.6 and
8.2.2, when employing external standard calibration, it is necessary that
a calibration verification standard be run at the end of the sequence to
bracket the sample analyses. Acceptance criteria for the initial
calibration and calibration verification are described in Sees. 7.5 - 7.7.
Analysis of mid-level standards every 10 samples is strongly
recommended, especially for the highly sensitive GC and HPLC detectors that
detect sub-nanogram concentrations. Frequent analysis of calibration check
solutions helps ensure that chromatographic systems are performing
acceptably and that false positives, false negatives and poor quantitation
are minimized. Samples analyzed using external standard calibration must
be bracketed by the analyses standards that meet the QC limits for
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verification of calibration and retention times. If criteria are exceeded,
corrective action must be taken (see Sec. 7.11) to restore the system
and/or a new calibration curve must be prepared for that compound and the
samples must be reanalyzed.
Certain methods may also include QC checks on column resolution,
analyte degradation, mass calibration, etc., at the beginning of a 12-hour
analytical shift.
7.8.3 Sample concentrations are calculated by comparing sample
responses with the initial calibration of the system (Sec. 7.5). If sample
response exceeds the limits of the initial calibration range, dilute the
extract (or sample) and reanalyze. Extracts should be diluted so that all
peaks are on scale, as overlapping peaks are not always evident when peaks
are off scale. Computer reproduction of chromatograms, manipulated to
ensure all peaks are on scale over a 100-fold range, is acceptable, as long
as calibration limits are not exceeded. When overlapping peaks cause
errors in peak area integration, the use of peak height measurements is
recommended.
7.8.4 If chromatographic peaks are masked by the presence of
interferences, further sample cleanup is necessary. See Method 3600 for
guidance.
7.8.5 When there are a large number of target analytes, it may be
difficult to fully resolve these compounds. Examples of chromatograms for
the compounds of interest are provided in many determinative methods.
7.9 Compound Identification
Tentative identification of an analyte occurs when a peak from a sample
extract falls within the daily retention time window. Normally, confirmation is
necessary. Confirmation techniques include analysis on a second column with
dissimilar stationary phase, by GC/MS (full scan or SIM) or HPLC/MS (if
concentration permits), HPLC/UV data at two different wavelengths, GC or HPLC
data from two different detectors, or by other recognized confirmation
techniques. For HPLC/UV methods, the ability to generate UV spectra with a diode
array detector may provide confirmation data from a single analysis, provided
that the laboratory can demonstrate this ability for typical sample extracts (not
standards) by comparison to another recognized confirmation technique.
When confirmation is made on a second column, that analysis should meet all
of the QC criteria described above for calibration, retention times, etc.
Confirmation is not required for GC/MS and HPLC/MS methods.
Confirmation may not be necessary if the composition of the sample matrix
is well established by prior analyses, for instance, when a pesticide known to
be produced or used in a facility is found in a sample from that facility.
When using GC/MS for confirmation, ensure that GC/MS analysis is performed
on an extract at the appropriate pH for the analyte(s) being confirmed, i.e., do
not look for base/neutral analytes in an acid extract. Certain analytes,
especially pesticides, may be degraded if extraction was either strongly acid
and/or strongly basic.
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Many chromatographic interferences result from co-elution of one or more
compounds with the analyte of interest, or may be the result of the presence of
a non-analyte peak in the retention time window of an analyte. Such co-elution
problems affect quantitation as well as identification, and may result in poor
agreement between the quantitative results from two dissimilar columns.
Therefore, even when the compound identification has been confirmed on a
dissimilar column, the analyst should evaluate the agreement of the quantitative
results on both columns, as described in Sec. 7.10.4.
7.10 Calculations
The calculation of sample results depends on the type of calibration
(external or internal standard) and the calibration model employed (linear or
non-linear). The following sections describe the calculations used in each
instance. Specific determinative methods may contain additional information.
7.10.1 External standard calibration - linear calibration
The concentration of each analyte in the sample is determined by
comparing the detector response (peak area or height) to the response for
that analyte in the initial calibration. The concentration of a specific
analyte may be calculated as follows, depending on the sample matrix:
7.10.1.1 Aqueous samples
(A.)(Vt)(D)
Concentration (ug/L) =
where:
As = Area (or height) of the peak for the analyte in the sample.
Vt = Total volume of the concentrated extract (/jL). For
purge-and-trap analysis, Vt is not applicable and therefore is
set at 1.
D = Dilution factor, if the sample or extract was diluted prior
to analysis. If no dilution was made, D = 1. The dilution
factor is always dimensionless.
CF = Mean calibration factor from the initial calibration (area
per ng).
V, = Volume of the extract injected (/^L). The nominal injection
volume for samples and calibration standards must be the
same. For purge-and-trap analysis, V, is not applicable and
therefore is set at 1. If concentration units are used in
calculating the calibration factor (see Sec. 7.4.2.1), then
Vj is not used in this equation.
Vs = Volume of the aqueous sample extracted or purged in ml. If
units of liters are used for this term, multiply the results
by 1000.
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Using the units specified here for these terms will result in a
concentration in units of ng/mL, which is equivalent to
7.10.1.2 Nonaqueous samples
Concentration (ug/kg) = ——i-
(CF)(V.)(Ws)
where As, Vt, D, CF, and Vj are as described in 7.10.1.1, and
Ws = Weight of sample extracted or purged (g). Either the wet
weight or dry weight may be used, depending upon the specific
application of the data. If units of kilograms are used for
this term, multiply the results by 1000.
Using the units specified here for these terms will result in a
concentration in units of ng/g, which is equivalent to ^g/kg.
For purge-and-trap analyses where a volume of methanol extract
is added to reagent water and purged, Vt is the total volume of the
methanol extract and V, is the volume of methanol extract that is
added to the 5 ml of reagent water.
7.10.1.3 If a linear calibration that does not pass through
the origin has been employed, then the regression equation is
rearranged as shown in Sec. 7.5.2.3, and the concentration of the
analyte is calculated from the area response (y), the slope (a), and
the intercept (b). When using this form of linear calibration, it is
the laboratory's responsibility to ensure that the calculations take
into account the volume or weight of the original sample, the dilution
factor (if any), and dry weight (as applicable). One approach to this
calculation is to perform the original linear regression using the
concentration of the analyte in the final extract volume or the volume
purged. The concentration of the analyte in the sample may then be
calculated as follows:
(V )
where: s
Cs = Concentration in the sample
Cex = Concentration in the final extract
Vt = Total volume of the concentrated extract
Vs = Volume of the sample extracted or purged
For solid samples, substitute the weight of the sample, Ws, for V
For purge-and-trap analyses, the concentration of the analyte
in the volume of the sample that is purged will be the same as in the
original sample, except when dilutions are performed.
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7.10.2 Internal standard calibration - linear calibration
The concentration of each analyte in the sample is calculated using
the results of the initial calibration, according to one of the following
sections, depending on the sample matrix:
7.10.2.1 Aqueous samples
(As)(C.s)(D)(V)
Concentration (ug/L) =
(AJ(RF)(VB)(1000)
where:
As = Area (or height) of the peak for the analyte in the sample.
A,s = Area (or height) of the peak for the internal standard.
C = Concentration of the internal standard in the concentrated
sample extract or volume purged in
is
Dilution factor, if the sample or extract was diluted prior
to analysis. If no dilution was made, D = 1. The dilution
factor is always dimensionless.
Vj = Volume of the extract injected (juL)- The nominal injection
volume for samples and calibration standards must be the
same. For purge-and-trap analysis, V; is not applicable and
is set at 1.
RF = Mean response factor from the initial calibration. Unlike
calibration factors for external standard calibration, the
response factor is dimensionless (see Sec. 7.5).
Vs = Volume of the aqueous sample extracted or purged (ml). If
units of liters are used for this term, multiply the results
by 1000.
The 1000 in the denominator represents the number of /jL in 1 ml. If
the injection (V,) is expressed in ml, then the 1000 may be omitted.
Using the units specified here for these terms will result in a
concentration in units of ng/mL, which is equivalent to M9/L-
7.10.2.2 Nonaqueous samples
... , /, x (A.i
Concentration (ug/kg) = - L— '
(Ais)(RF)(WJ(1000)
where: As, Ais, CIS, D, and RF are the same as for aqueous samples, and
Ws = Weight of sample extracted (g). Either a dry weight or wet
weight may be used, depending upon the specific application
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of the data. If units of kilograms are used for this term,
multiply the results by 1000.
The 1000 in the denominator represents the number of /zL in 1 ml. If
the injection (V|) is expressed in ml, then the 1000 may be omitted.
Using the units specified here for these terms will result in
a concentration in units of ng/g, which is equivalent to /xg/kg.
7.10.2.3 If a linear calibration that does not pass through
the origin has been employed, then the regression equation is
rearranged in a fashion similar to that described in Sec. 7.10.1.3.
7.10.3 Calculations for a non-linear calibration curve
When a non-linear curve has been employed, the non-linear model is
rearranged to solve for the concentration of the analyte in the extract or
purge volume, and the extract concentration is converted to a sample
concentration in a fashion similar to that described in Sec. 7.10.1.3.
When non-linear calibrations are employed, it is essential that the
laboratory clearly document the calculation of analyte concentrations.
Example calculations should be reported that clearly indicate how the
instrument response (area) was converted into a sample result.
7.10.4 Comparison between results from different columns or detectors
When sample results are confirmed using two dissimilar columns or with
two dissimilar detectors, the agreement between the quantitative results
should be evaluated after the identification has been confirmed. Calculate
the relative percent difference (RPD) between the two results using the
formula below.
I R, - R, I
RPD = _!—1 2— x 100
-, + R2 1
where R, and R2 are the results on the two columns and the vertical bars in
the equation above indicate the absolute value of the difference.
Therefore, the RPD is always a positive value.
7.10.4.1 If one result is significantly higher (e.g., >40%),
check the chromatograms to see if an obviously overlapping peak is
causing an erroneously high result. If no overlapping peaks are
noted, examine the baseline parameters established by the instrument
data system (or operator) during peak integration.
7.10.4.2 If no anomalies are noted, review the
chromatographic conditions. It may be necessary to adjust the
chromatographic conditions to better resolve the compound of interest
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from potential interferents. It may even be necessary to employ a
different chromatographic column to overcome the problem.
7.11 Suggested chromatography system maintenance
Corrective measures may involve any one or more of the following remedial
actions. This list is by no means comprehensive and analysts should develop
expertise in troubleshooting their specific instruments and analytical
procedures. The manufacturers of chromatographic instruments, detectors,
columns, and accessories generally provide detailed information regarding the
proper operation and limiting factors associated with their products. The
importance of reading and reviewing this information cannot be over-emphasized.
7.11.1 Capillary GC columns
Routine maintenance may compel the analyst to clean and deactivate the
glass injection port insert or replace it with a fresh insert that has been
cleaned and deactivated with dichlorodimethylsilane. Cut off 0.5 - 1.0 m
of the injector end of the column using a 90° cut). Place ferrule onto the
column before cutting.
Exceptional maintenance may compel the analyst to replace gas traps
and backflush the column with solvent according to the manufacturer's
instructions. If these procedures fail to eliminate the degradation
problem, it may be necessary to deactivate the metal injector body and/or
replace the column.
7.11.2 Packed GC columns
For instruments with injection port traps, replace the demister trap,
clean, and deactivate the glass injection port insert or replace with a
cleaned and deactivated insert. Inspect the injection end of the column
and remove any foreign material (broken glass from the rim of the column
or pieces of septa). Replace the glass wool with fresh deactivated glass
wool. It may also be necessary to remove the first few millimeters of the
packing material if any discoloration is noted. Whenever packing material
is removed, swab out the inside walls of the column if any residue remains.
If these procedures fail to eliminate the degradation problem, it may be
necessary to deactivate the metal injector body (described in Sec. 7.11.3)
and/or repack/replace the column.
7.11.3 Metal (GC) injector body
Turn off the oven and remove the analytical column when the oven has
cooled. Remove the glass injection port insert. Lower the injection port
temperature to room temperature. Inspect the injection port and remove any
noticeable foreign material.
Place a beaker beneath the injector port inside the GC oven. Using
a wash bottle, serially rinse the entire inside of the injector port with
acetone and then toluene, catching the rinsate in the beaker.
Prepare a solution of deactivating agent (dichlorodimethylsilane)
following manufacturer's directions. After all metal surfaces inside the
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injector body have been thoroughly coated with the deactivation solution,
serially rinse the injector body with toluene, methanol, acetone, and
hexane. Reassemble the injector and replace the GC column.
7.11.4 HPLC columns
Examine the system and check for drips that are indicative of plumbing
leaks. Check that tubing connectors are of the shortest possible length
to minimize dead volumes and reduce band broadening. Compatible guard
columns should be installed to protect analytical columns.
If degradation of resolution or changes in back pressure are observed,
first action should be to replace the guard column if one is installed.
Secondly, temporarily reverse the flow through the column to dislodge
contamination in the frit with the column disconnected from the detector.
If this does not correct the problem, place the analytical column in a
vise, remove the inlet compression fitting and examine the column.
Analysts should establish that no void volume has developed, that the
column packing has not become contaminated, and that the frit is not
clogged. Void volumes can be filled with compatible packing and frits
replaced.
Columns must eventually be replaced as the bonding and end-capping
groups used to modify the silica are lost with time. Loss of these groups
will result in chromatographic tailing and changes in analyte retention
times. Retention times may also change because of differences in column
temperature or because the composition of the solvent gradient is not
completely reproducible.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. Each
laboratory using SW-846 methods should maintain a formal quality assurance
program. The use of instrument-specific QC limits is encouraged, provided such
limits are at least as stringent as those described in the specific determinative
method being used. In general, the following QC requirements pertain to all the
determinative methods listed in Sec. 1.1 unless superseded by specific
requirements provided in the determinative method.
8.2 Evaluating chromatographic performance
The analyst's expertise in performing chromatography is a critical element
in the successful performance of chromatographic methods. Successful generation
of data requires selection of suitable preparation and analysis methods and an
experienced staff to use these methods.
8.2.1 For each 12-hour period during which analysis is performed,
the performance of the entire analytical system should be checked. These
checks should be part of a formal quality control program that includes the
analysis of blanks, calibration standards, matrix spikes, laboratory
control samples and replicate samples.
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8.2.2 Ongoing data quality checks are compared with established
performance criteria to determine if the results of analyses meet the
performance characteristics of the method. Therefore, all sample analyses
performed using external standard calibration must be bracketed with
acceptable data quality analyses (e.g., calibration standards and blanks).
8.2.3 In addition to the quantitative measures of comparison
described below and in the individual methods, analysts should evaluate
chromatograms and instrument operation. Questions that should be asked
include:
Do the peaks look normal (Gaussian)?
Is the response obtained comparable to the response from previous
calibrations?
Do the column fittings need tightening?
Are non-target peaks present in calibration analyses?
Are contaminants present in the blanks?
Is the injector leaking (e.g., does the GC injector septum need
replacing)?
Does the HPLC guard column need replacement?
8.2.4 Significant peak tailing, leaks, changes in detector response
and laboratory contamination should be corrected. Tailing problems are
generally traceable to active sites on the column, cold spots in a GC,
improper choice of HPLC mobile phase, the detector inlet, or leaks in the
system.
8.2.5 Whenever significant differences in instrument performance are
observed or hardware changes are made (e.g., column changed), recalibration
of the system must take place.
8.2.6 The analysis of method blanks is critical to the provision of
meaningful sample results. Consult the appropriate 3500 or 5000 series
method for the specifics of the preparation of method blanks. The
following general guidelines apply to the interpretation of method blank
results.
8.2.6.1 Method blanks should be prepared at a frequency of
at least 5%, that is, one method blank for each group of up to 20
samples prepared at the same time, by the same procedures. For
samples analyzed for volatiles by the purge-and-trap technique, the
preparation is equivalent to the analysis. Therefore, one method
blank must be analyzed with each group of up to 20 samples analyzed
during the same analytical shift.
8.6.2.2 When samples that are extracted together are
analyzed on separate instruments or on separate analytical shifts, the
method blank associated with those samples (e.g., extracted with the
samples) should be analyzed on all the instruments used for sample
analyses, and on all the analytical shifts.
8.6.2.3 Unless otherwise described in a determinative
method, the method blank should be analyzed immediately after the
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calibration verification standard, to ensure that there is no
carryover from the standard.
8.2.6.4 When sample extracts are subjected to cleanup
procedures, the associated method blank must also be subjected to the
same cleanup procedures.
8.2.6.5 As described in Chapter One, the results of the
method blank should be:
8.2.6.5.1 Less than the MDL for the analyte.
8.2.6.5.2 Less than 5% of the regulatory limit
associated with an analyte.
8.2.6.5.3 Or less than 5% of the sample result for
the same analyte, whichever is greater.
8.2.6.5.4 If the method blank results do not meet
the acceptance criteria above, then the laboratory should
take corrective action to locate and reduce the source of the
contamination and to re-extract and reanalyze any samples
associated with the contaminated method blank.
8.2.6.6 The laboratory should not subtract the results of
the method blank from those of any associated samples. Such "blank
subtraction" is inappropriate for the GC and HPLC methods addressed
here, and often leads to negative sample results. If the method blank
results do not meet the acceptance criteria in 8.2.6.5 and reanalysis
is not practical, then the data user should be provided with the
sample results, the method blank results, and a discussion of the
corrective actions undertaken by the laboratory.
8.3 Summary of required instrument QC
The following criteria primarily pertain to GC and HPLC methods with non-MS
or FTIR detectors, and may be superseded by criteria specified in individual
determinative methods (e.g., Methods 8410, 8260, and 8270).
8.3.1 The criteria for linearity of the initial calibration curve
is an RSD of < 20%.
8.3.2 For non-linear calibration curves, the coefficient of the
determination (COD) must be greater than or equal to 0.99 (see Sec. 7.5.2).
8.3.3 Retention time (RT) windows (±3 SD around the mean retention
time) must be established for the identification of target analytes. See
Sec. 7.6 for guidance on establishing the absolute RT window.
8.3.4 The retention times of all analytes in all verification
standards must fall within the absolute RT windows. If an analyte falls
outside the RT window in a calibration verification standard, new absolute
RT windows must be calculated, unless instrument maintenance corrects the
problem.
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8.3.5 The calibration verification results must be within ± 15% of
the response calculated using the initial calibration. If the limit is
exceeded, a new standard curve must be prepared unless instrument
maintenance corrects the problem.
8.4 Initial demonstration of proficiency
Each laboratory must demonstrate initial proficiency with each combination
of sample preparation and determinative methods that it utilizes, by generating
data of acceptable accuracy and precision for a reference sample containing the
target analytes in a clean matrix. The laboratory must also repeat this
demonstration whenever new staff are trained or significant changes in
instrumentation are made.
8.4.1 The reference samples are prepared from a spiking solution
containing each analyte of interest. The reference sample concentrate
(spiking solution) may be prepared from pure standard materials, or
purchased as certified solutions. If prepared by the laboratory, the
reference sample concentrate must be made using stock standards prepared
independently from those used for calibration.
Preparation of the reference sample concentrate is dependent upon the
method being evaluated. Guidance for reference sample concentrations for
certain methods are listed in Sec. 8.0 of Methods 3500 and 5000. In other
cases, the determinative methods contain guidance on preparing the
reference sample concentrate and the reference sample. If no guidance is
provided, prepare a reference sample concentrate in methanol (or any water
miscible solvent) at a concentration such that the spike will provide a
concentration in the clean matrix that is 10 - 50 times the MDL for each
analyte in that matrix.
The concentration of target analytes in the reference sample may be
adjusted to more accurately reflect the concentrations that will be
analyzed by the laboratory. If the concentration of an analyte is being
evaluated relative to a regulatory limit, see Sec. 8.5.1 for information
on selecting an appropriate spiking level.
8.4.2 To evaluate the performance of the total analytical process,
the reference samples must be handled in exactly the same manner as actual
samples. Use a clean matrix for spiking purposes (one that does not have
any target or interference compounds), e.g., organic-free reagent water for
the aqueous matrix and organic-free sand or soil for the solid matrix.
8.4.3 Preparation of reference samples
8.4.3.1 Volatile organic analytes
Prepare the reference sample by adding 200 juL of the
reference sample concentrate (Sec. 8.4.1) to 100 ml of organic-free
reagent water. Transfer this solution immediately to a 20- or 25-mL
(or four 5-mL) gas-tight syringe(s) when validating water analysis
performance by Method 5030. See Method 5000 (Sec. 8.0) for guidance
on other preparative methods and matrices.
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8.4.3.2 Semivolatile and nonvolatile organic analytes
Prepare the reference sample by adding 1.0 ml of the reference
sample concentrate (Sec. 8.4.1) to each of four 1-L aliquots of
organic-free reagent water. See Method 3500 (Sec. 8.0) for other
matrices.
8.4.4 Analyze at least four replicate aliquots of the well-mixed
reference samples by the same procedures used to analyze actual samples
(Sec. 7.0 of each of the methods). This will include a combination of the
sample preparation method (usually a 3500 series method for extractable
organics or a 5000 series method for volatile organics) and the
determinative method (an 8000 series method).
8.4.5 Calculate the average recovery (x) in ng/l, and the standard
deviation of the recovery (s) in M9/U for each analyte of interest using
the four results.
8.4.6 When acceptance criteria are presented in the determinative
method, compare s and x for each analyte with the corresponding acceptance
criteria for precision and accuracy given in the QC acceptance criteria
table at the end of the determinative method. If s and x for all analytes
of interest meet the acceptance criteria, then the system performance is
acceptable and analysis of actual samples can begin. _ If any individual s
value exceeds the precision limit or any individual x value falls outside
the range for accuracy, then the system performance is unacceptable for
that analyte.
NOTE: The large number of analytes in each of the methods presents a
substantial probability that one or more analyte will fail at least
one of the acceptance criteria when all analytes of a given method
are determined.
When one or more of the analytes fail at least one of the acceptance
criteria, the analyst should proceed according to Sec. 8.4.6.1 or 8.4.6.2.
8.4.6.1 Locate and correct the source of the problem and
repeat the test for all analytes of interest, beginning at Sec. 8.4.2.
8.4.6.2 Beginning at Sec. 8.4.2, repeat the test only for
those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning at Sec. 8.4.2.
8.4.7 The acceptance criteria in many of the methods are based on
single laboratory performance data. As a result, the criteria in those
methods should be used as guidance when evaluating laboratory performance.
When comparing your laboratory data to acceptance criteria developed from
single laboratory data, certain analytes may be outside the limits,
however, the majority should be within the acceptance limits.
8.4.8 Where the results of the initial demonstration of proficiency
frequently fall outside of single laboratory acceptance criteria included
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in a particular method, the laboratory should consider developing in-house
acceptance limits, using the general considerations described in Sec. 8.7.
8.4.9 In the absence of method-specific acceptance criteria for the
initial demonstration of proficiency, the laboratory should use recoveries
of 70 - 130% as guidance in evaluating the results. Given that the initial
demonstration is performed in a clean matrix, the average recoveries of
analyte from the four replicates should generally fall within this range.
In addition, since the laboratory will repeat the initial demonstration of
proficiency whenever new staff are trained or significant changes in
instrumentation are made, the resulting data should be used to develop in-
house acceptance criteria, as described in Sec. 8.7.
8.5 Matrix spike and laboratory control samples
The laboratory must also have procedures for documenting the effect of the
matrix on method performance (precision, accuracy, and detection limit). At a
minimum, this will include the analysis of at least one matrix spike and one
duplicate unspiked sample or one matrix spike/matrix spike duplicate (MS/MSD)
pair per analytical batch. If samples are expected to contain target analytes,
then laboratories may use one matrix spike and a duplicate analysis of an
unspiked field sample (see Sec. 8.5.3).
In addition, a Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control) matrix
similar to the sample matrix and of the same weight or volume. The LCS is spiked
with the same analytes at the same concentrations as the matrix spike. When the
results of the matrix spike analysis indicates a potential problem due to the
sample matrix itself, the LCS results are used to verify that the laboratory can
perform the analysis in a clean matrix.
The concentration of the matrix spike sample and/or the LCS should be
determined as described in Sees. 8.5.1 and 8.5.2.
8.5.1 If, as in compliance monitoring, the concentration of a
specific analyte in the sample is being checked against a regulatory
concentration limit, the spike should be at or below the regulatory limit,
or 1 - 5 times the background concentration (if historical data are
available), whichever concentration is higher.
If historical data are not available, it is suggested that a
background sample of the same matrix from the site be submitted for matrix
spiking purposes to ensure that high concentrations of target analytes
and/or interferences will not prevent calculation of recoveries.
8.5.2 If the concentration of a specific analyte in a sample is not
being checked against a limit specific to that analyte, then the spike
should be at the same concentration as the reference sample (Sec. 8.4.1)
or 20 times the estimated quantitation limit (EQL) in the matrix of
interest. It is again suggested that a background sample of the same
matrix from the site be submitted as a sample for matrix spiking purposes.
8.5.3 To develop precision and accuracy data for each of the spiked
compounds, the analyst has two choices: analyze the original sample, and
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an MS/MSD pair; or analyze the original saipple, a duplicate sample, and
one spiked sample. If samples are expected to contain target analytes,
then laboratories may use one matrix spike and a duplicate analysis of an
unspiked field sample. If samples are not expected to contain target
analytes, the laboratories should use a matrix spike and matrix spike
duplicate pair.
Begin by analyzing one sample aliquot to determine the background
concentration of each analyte. Prepare a matrix spike concentrate
according to one of the options specified in Sec. 8.5.1 or 8.5.2.
Prepare a matrix spike sample by adding the appropriate volume of the
matrix spike concentrate to another aliquot of the sample to yield the
desired concentration (see Sees. 8.5.1 and 8.5.2). Prepare a matrix spike
duplicate sample from a third aliquot of the sample.
Analyze the MS/MSD samples using the same procedures employed for the
original sample, and calculate the concentration of each analyte in the
matrix spike and matrix spike duplicate. Likewise, analyze the LCS samples
using the same procedures employed for the original sample, and calculate
the concentration of each analyte in the LCS.
8.5.3.1 Calculation of recovery
Accuracy is estimated from the recovery of spiked analytes
from the matrix of interest. Laboratory performance in a clean matrix
is estimated from the recovery of analytes in the LCS. Calculate the
recovery of each spiked analyte in the matrix spike, matrix spike
duplicate (if performed) and LCS according to the following formula.
C - C
Recovery = %R = -? u- x 100
C
where: n
Cs = Measured concentration of the spiked sample aliquot
Cu = Measured concentration of the unspiked sample aliquot (use 0 for
the LCS)
Cn = Nominal (theoretical) concentration of the spiked sample aliquot
8.5.3.2 Calculation of precision
Precision is estimated from the relative percent difference
(RPD) of the concentrations (not the recoveries) measured for matrix
spike/matrix spike duplicate pairs, or for duplicate analyses of
unspiked samples. Calculate the RPD according to the formula below.
I C - C I
RPD = J ] 2— x 100
K +C2 1
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where:
C, = Measured concentration of the first sample aliquot
C2 = Measured concentration of the second sample aliquot
8.5.4 QC acceptance criteria for matrix spike samples and LCS
It is necessary for the laboratory to develop single-laboratory
performance data for accuracy and precision in the matrices of interest
(see Sec. 8.7). In addition, laboratories should monitor method
performance in each matrix, through the use of control charts and other
techniques.
Many methods may not contain acceptance criteria for LCS results. The
laboratory should use 70 - 130% as interim acceptance criteria for
recoveries of spiked analytes, until in-house LCS limits are developed (see
Sec. 8.7). Where in-house limits have been developed for matrix spike
recoveries, the LCS results should fall within those limits, as the LCS is
prepared in a clean matrix.
Even where the determinative methods provide QC acceptance criteria
for matrix spikes and LCS, it is necessary for laboratories to develop in-
house performance criteria and compare them to those in the methods. The
development of in-house performance criteria is discussed in Sec. 8.7.
As a general rule, the recoveries of most compounds spiked into
samples should fall within the range of 70 - 130%, and this range should
be used as a guide in evaluating in-house performance. However, as
described in Sec. 8.5.4.1, matrix spike recoveries and LCS recoveries may
be affected by the spike-to-background ratio.
Where methods do contain acceptance criteria for the matrix of
interest, use Sees. 8.5.4.1 and 8.5.4.2 as guidance in evaluating data
generated by the laboratory.
8.5.4.1 When acceptance criteria for the matrix of interest
are provided in the determinative method, compare the percent recovery
(%R) for each analyte in a water sample with the QC acceptance
criteria. These acceptance criteria are generally based on multi-
laboratory studies. Therefore, they should be met in almost all
laboratories. The acceptance criteria include an allowance for error
in measurement of both the background and spike concentrations, and
assume a spike-to-background ratio of 5:1. If spiking was performed
at a concentration lower than that used for the reference sample (Sec.
8.4), the analyst may use either the QC acceptance criteria presented
in the tables, or laboratory-generated QC acceptance criteria
calculated for the specific spike concentration.
8.5.4.2 When the sample was spiked at a spike-to-background
ratio other than 5:1, the laboratory should calculate acceptance
criteria for the recovery of an analyte. Some determinative methods
contain a table entitled "Method Accuracy and Precision as a Function
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of Concentration." This table gives equations for calculating
accuracy and precision as a function of the spiking concentration.
The equations are the result of linear regression analyses of
the performance data from a multiple laboratory study. The equations
are of the form:
Accuracy = x' = (a)C + b
where a is a number less than 1.0, b is a value greater than 0.0, and
C is the test concentration (or true value).
Acceptance criteria for accuracy may be calculated from these
equations by substituting the spiking concentration used by the
laboratory in place of "C," and using the values of a and b given in
the table for each analyte.
Acceptance criteria for precision are calculated in a similar
fashion, using the a and b values for precision given in the table for
each analyte. Precision may be calculated as single analyst
precision, or overall precision, using the appropriate equations from
the table. An acceptance range may be calculated for each analyte as:
Acceptance range (ug/L) = Accuracy ± (2.44)Precision
8.5.5 Also compare the recovery data from the matrix spike with the
LCS data (use the average recovery if a matrix spike and matrix spike
duplicate were analyzed). If any individual percent recovery in the matrix
spike (or matrix spike duplicate) falls outside the designated range for
recovery, the laboratory should determine if there is a matrix effect or
a laboratory performance problem. A matrix effect is indicated if the LCS
data are within limits but the matrix spike data exceed the limits. The
surrogate recovery data (Sec. 8.6) should also be used to evaluate the
data. Recoveries of both matrix spike compounds and surrogates that are
outside of the acceptance limits suggest more pervasive analytical problems
then problems with the recoveries of either matrix spikes or surrogates
alone.
8.6 Surrogate recoveries
8.6.1 It is necessary that the laboratory evaluate surrogate
recovery data from individual samples versus surrogates recovery limits
developed in the laboratory. The general considerations for developing in-
house acceptance criteria for surrogate recoveries are described in Sec.
8.7.
8.6.2 Surrogate recovery is calculated as:
n ,,,> Concentration (or amount) found ...
Recovery (%) = - x 100
Concentration (or amount) added
If recovery is not within in-house surrogate recovery limits, the
following procedures are necessary.
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8.6.2.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly. Examine chromatograms
for interfering peaks and integrated peak areas.
8.6.2.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract (or re-analyze the sample for volatiles).
8.6.2.3 If no problem is found, re-extract and re-analyze
the sample (or re-analyze the sample for volatiles).
8.6.2.4 If, upon re-analysis (in either 8.6.2.2 or 8.6.2.3),
the recovery is again not within limits, flag the data as "estimated
concentration". If the recovery is within the limits in the re-
analysis, provide the re-analysis data to the data user. If the
holding time for the method has expired prior to the re-analysis,
provide both the original and re-analysis results to the data user,
and note the holding time problem.
8.7 Generating performance criteria for matrix spike recoveries, surrogate
recoveries, initial demonstration of proficiency, and laboratory
control sample recoveries
It is essential that laboratories calculate in-house performance
criteria for matrix spike recoveries and surrogate recoveries. It may also
be useful to calculate such in-house criteria for laboratory control sample
(LCS) recoveries and for the initial demonstration of proficiency when
experience indicates that method-specific criteria are frequently missed
for some analytes or matrices. The development of in-house performance
criteria and the use of control charts or similar procedures to track
laboratory performance cannot be over-emphasized. Many data systems and
commercially-available software packages support the use of control charts.
The procedures for the calculation of in-house performance criteria
for matrix spike recovery and surrogate recovery are provided below. These
procedures may also be applied to the development of in-house criteria for
the initial demonstration of proficiency and for LCS recoveries.
8.7.1 For each matrix spike sample analyzed, calculate the percent
recovery of each matrix spike compound added to the sample, in a fashion
similar to that described in Sec. 8.5.3.3. For each field sample,
calculate the percent recovery of each surrogate as described in Sec. 8.6.
8.7.2 Calculate the average percent recovery (p) and the standard
deviation (s) for each of the matrix spike compounds after analysis of 15-
20 matrix spike samples of the same matrix, using the equations in Sec.
7.5.1, as guidance. Calculate the average percent recovery (p) and the
standard deviation (s) for each of the surrogates after analysis of 15-20
field samples of the same matrix, in a similar fashion.
8.7.3 After the analysis of 15-20 matrix spike samples of a
particular matrix (or matrix spike limits) or 15-20 field samples (for
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surrogate limits), calculate upper and lower control limit for each matrix
spike or surrogate compound:
Upper control limit = p + 3s
Lower control limit = p - 3s
Calculate warning limits as:
Upper warning limit = p + 2s
Lower warning limit = p - 2s
For laboratories employing statistical software to determine these
limits, the control limits represent a 99% confidence interval around the
mean recovery, while the warning limits represent a 95% confidence
interval.
8.7.4 Any matrix spike or surrogate results outside of the control
limits require corrective action by the laboratory, including, but not
limited to the review of the sample results, inspection of the
chromatographic system, and re-analysis of the sample.
The laboratory should use the warning limits to guide internal
evaluations of method performance, track the performance of individual
analysts, and monitor the effects of changes to the analytical procedures.
Repeated results outside of the warning limits should result in corrective
actions.
8.7.5 Once established, control 1 imits and warning 1 imits for matrix
spike compounds should be updated after every 10-20 matrix spike samples
of the same matrix, or at least quarterly. Control limits and warning
limits for surrogates should be updated after every 20-30 field samples of
the same matrix, or at least quarterly. The laboratory should track trends
in both performance and in the control limits themselves.
8.7.6 For methods and matrices with very 1 imi ted data (e.g., unusual
matrices not analyzed often), interim limits should be established using
available data or by analogy to similar methods or matrices.
8.7.7 Results used to develop acceptance criteria should meet all
other QC criteria associated with the determinative method. For instance,
matrix spike recoveries from a GC/MS procedure should be generated from
samples analyzed after a valid GC/MS tune and a valid initial calibration
that includes the matrix spike compounds. Another example is that analytes
in GC or HPLC methods must fall within the established retention time
windows in order to be used to develop acceptance criteria.
8.7.8 Laboratories are advised to consider the effects of the
spiking concentration on matrix spike performance criteria, and to avoid
censoring of data. As noted in Sec. 8.5.4, the acceptance criteria for
matrix spike recovery and precision are often a function of the spike
concentration used. Therefore, use caution when pooling matrix
spike/matrix spike duplicate data for use in establishing acceptance
criteria. Not only should the results all be from the same (or very
8000B - 44 Revision 2
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similar) matrix, but the spiking levels should also be approximately the
same (within a factor of 2).
Similarly, the matrix spike and surrogate results should all be
generated using the same set of extraction, cleanup, and analysis
techniques. For example, do not mix results from solid samples extracted
by ultrasonic extraction with those extracted by Soxhlet.
8.7.9 Another common error in developing acceptance criteria is to
discard data that do not meet a preconceived notion of acceptable
performance. This results in a censored data set, which, when used to
develop acceptance criteria, will lead to unrealistically narrow criteria.
Remember that for a 95% confidence interval, 1 out of every 20 observations
likely will still fall outside the limits.
While professional judgement is important in evaluating data to be
used to develop acceptance criteria, do not discard specific results simply
because they do not meet one's expectations. Rather, employ a statistical
test for outlier values, or at least calculate the acceptance limits both
with and without the results that are considered suspect and observe the
effect of deleting suspect data.
8.8 It is recommended that the laboratory adopt additional quality
assurance practices for use with these methods. The specific practices that are
most productive depend upon the needs of the laboratory, the nature of the
samples, and project-specific requirements. 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 (selected ion monitoring or full scan) must be used. Whenever
possible, the laboratory should analyze standard reference materials and
participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined 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 the
SW-846 analytical methods generally were obtained using organic-free reagent
water. Similar results were achieved using representative wastewaters. The MDL
actually achieved in a given analysis will vary depending on instrument
sensitivity and matrix effects. See Chapter One for more guidance on
determination of laboratory-specific MDLs.
9.2 Refer to the determinative methods for method performance information.
10.0 REFERENCES
For further information regarding these methods, review Methods 3500, 3600,
5000, and Chapter One.
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METHOD 8015B
NONHALOGENATED ORGANICS USING GC/FID
1.0 SCOPE AND APPLICATION
1.1 Method 8015 is used to determine the concentration of various
nonhalogenated volatile organic compounds, semivolatile organic compounds, and
petroleum hydrocarbons including gasoline range organics (GROs), diesel range
organics (DROs), and jet fuel by gas chromatography. The following compounds and
mixtures can be determined quantitatively by this method:
Compound Name
CAS No.'
Appropriate Technique
Purge-and-
Trap
Direct Solvent
Injection Extraction
Acetone 67-64-1
Acetonitrile 75-05-8
Acrolein 107-02-8
Acrylonitrile 107-13-1
Ally! alcohol 107-18-6
1-Butanol (n-Butyl alcohol) 71-36-3
t-Butyl alcohol 75-65-0
2-Chloroacrylonitrile (I.S.) 920-37-6
Crotonaldehyde 123-73-9
Diethyl ether 60-29-7
1,4-Dioxane 123-91-1
Ethanol 64-17-5
Ethyl acetate 141-78-6
Ethylene glycol 107-21-1
Ethylene oxide 75-21-8
Hexafluoro-2-propanol (I.S.) 920-66-1
Hexaf1uoro-2-methyl-
2-propanol (I.S.) 515-14-6
Isobutyl alcohol 78-83-1
Isopropyl alcohol 67-63-0
Methanol 67-56-1
Methyl ethyl ketone (MEK) 78-93-3
Methyl isobutyl ketone (MIBK) 108-10-1
N-Nitroso-di-n-butyl amine 924-16-3
Paraldehyde 123-63-7
2-Pentanone 107-87-9
2-Picoline 109-06-8
1-Propanol 71-23-8
Propionitrile 107-12-0
Pyridine 110-86-1
o-Toluidine 95-53-4
Gasoline range organics (GROs) NA
Diesel range organics (DROs) NA
Jet fuel NA
PP
PP
PP
PP
ht
ht
PP
NA
PP
b
PP
i
i
i
i
NA
NA
PP
PP
i
PP
PP
PP
PP
PP
PP
PP
ht
i
i
b
i
i
b,d
b,d
b,d
b,d
b,d
b,d
b,d
d
b,d
b
b,d
b,d
b,d
b
b,d
d
d
b,d
b,d
b,d
b,d
b,d
b,d
b,d
b,d
b,d
b,d
d
b,d
b,d
i
b
b
i
i
i
i
i
i
i
NA
i
i
i
i
i
i
i
NA
NA
i
i
i
i
i
b
i
i
i
i
i
b
b
i
b
b
8015B - 1
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a Chemical Abstract Services Registry Number.
b Adequate response using this technique
d Amenable to concentration by azeotropic distillation (Method 5031)
ht Method analyte only when purged at 80°C
i Inappropriate technique for this analyte
pp Poor purging efficiency, resulting in high EQLs
NA Not available
I.S. Internal standard appropriate for Method 5031
1.2 When the method is used for the determination of petroleum
hydrocarbons, analysts should use the fuel contaminating the site for
quantitation. Retention times of n-alkane mixtures are also to be measured to
facilitate identification of the fuel type and the degree of environmental
degradation of the fuel. Capillary columns are needed for petroleum hydrocarbon
analyses. Only capillary columns provide the resolving power necessary to
separate the complex mixtures of hydrocarbons found in fuels.
1.3 This method is restricted for use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. In addition, if this method is used for the
analysis of petroleum hydrocarbons, it is limited to analysts experienced in the
interpretation of hydrocarbon data. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.4 The method can also be used as a screening tool (for both volatile and
semivolatile organics) to obtain semiquantitative data for the prevention of
sample overload during quantitative analysis on a GC/MS system. This may be
accomplished using either a manual (Method 3810) or an automated (Method 5021)
headspace method or by direct injection if a solvent extraction method has been
utilized for sample preparation. Single point calibration would be acceptable
in this situation. Performance data are not provided for screening.
2.0 SUMMARY OF METHOD
2.1 Method 8015 provides gas chromatographic conditions for the detection
of certain nonhalogenated volatile and semivolatile organic compounds.
2.1.1 Samples may be introduced into the GC:
• following solvent extraction (Methods 3510, 3520, 3540, 3541, 3545,
or 3560);
• by direct injection (aqueous samples) including analyte
concentration by azeotropic distillation (Method 5031);
• by purge-and-trap (Methods 5030 or 5035); or,
• by vacuum distillation (Method 5032).
2.1.2 Ground or surface water samples must generally be analyzed in
conjunction with Methods 5030, 5031, 5032, 3510, 3520, or other
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appropriate preparatory methods to obtain the necessary quantitation
limits.
2.1.3 Diesel range organics (DROs) and jet fuel may be prepared by
one of the solvent extraction methods.
2.1.4 Gasoline range organics (GROs) may be introduced into the
GC/FID by purge-and-trap, automated headspace, vacuum distillation, or
other appropriate technique.
2.2 An appropriate column and temperature program is used in the gas
chromatograph to separate the organic compounds. Detection is achieved by a
flame ionization detector (FID).
2.3 The method allows the use of packed or capillary columns for the
analysis and confirmation of the non-halogenated individual analytes. Columns
and conditions listed have been demonstrated to provide separation of those
target analytes. Analysts may change these conditions as long as they
demonstrate adequate performance.
2.4 Fused silica capillary columns are needed for the analysis of
petroleum hydrocarbons.
3.0 INTERFERENCES
3.1 When analyzing for volatile organics, samples can be contaminated by
diffusion of volatile organics (particularly chlorofluorocarbons and methylene
chloride) through the sample container septum during shipment and storage. A
trip blank prepared from organic-free reagent water and carried through sampling
and subsequent storage and handling must serve as a check on such contamination.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are analyzed in sequence. To reduce the potential for
carryover, the sample syringe or purging device must be rinsed out between
samples with an appropriate solvent. Whenever an unusually concentrated sample
is encountered, it should be followed by injection of a solvent blank to check
for cross contamination.
3.2.1 Clean purging vessels with a detergent solution, rinse with
distilled water, and then dry in a 105°C oven between analyses. Clean
syringes or autosamplers by flushing all surfaces that contact samples
using appropriate solvents.
3.2.2 All glassware must be scrupulously cleaned. Clean all
glassware as soon as possible after use by rinsing with the last solvent
used. This should be followed by detergent washing with hot water, and
rinses with tap water and organic-free reagent water. Drain the glassware
and dry in an oven at 130°C for several hours or rinse with methanol and
drain. Store dry glassware in a clean environment.
3.3 The flame ionization
There is a potential for many
interfere with this analysis.
detector (FID) is a non-selective detector.
non-target compounds present in samples to
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for solvent injections or purge-and-trap sample
introduction and all required accessories, including detectors, column
supplies, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Recommended GC Columns:
4.1.2.1 Column 1 -
column packed with 1%
equivalent.
8 ft x 0.1 in. id stainless steel
SP-1000 on Carbopack-B 60/80
or glass
mesh or
4.1.2.2 Column 2 - 6 ft x 0.1 in. id stainless steel or glass
column packed with n-octane on Porasil-C 100/120 mesh (Durapak) or
equivalent.
4.1.2.3 Column 3 - 30 m x 0.53 mm id fused silica capillary
column bonded with DB-Wax (or equivalent), 1 /xm film thickness.
4.1.2.4 Column 4 - 30 m x 0.53 mm id fused silica capillary
column chemically bonded with 5% methyl silicone (DB-5, SPB-5, RTx,
or equivalent), 1.5 /xm film thickness.
4.1.2.4.1 Capillary columns are needed for petroleum
hydrocarbon analyses. Laboratories may use other capillary
columns (e.g. 0.25-0.32 mm id capillary columns) if they
document method performance data (e.g. chromatographic
resolution, and MDLs) equal to or better than that provided
with the method.
4.1.2.4.2 Wide-bore columns should be installed in 1/4
inch injectors, with deactivated liners designed specifically
for use with these columns.
4.1.3 Detector - Flame ionization (FID).
4.2 Sample introduction and preparation apparatus
4.2.1 Refer to Method 5021 for the appropriate equipment for
automated headspace analysis of soils and other solid matrices.
4.2.2 Refer to Method 5030 for the appropriate equipment for
purge-and-trap analysis of aqueous samples.
4.2.3 Refer to Method 5031 for the appropriate equipment for
azeotropic distillation analysis of aqueous and solid matrices.
4.2.4 Refer to Method 5032 for the appropriate equipment for vacuum
distillation analysis of aqueous, solid matrices and tissue.
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4.2.5 Refer to Method 5035 for the appropriate equipment for
purge-and-trap analysis of solid samples.
4.2.6 GC injection port for the analysis of solvent extracts or
aqueous samples by direct injection.
4.3 Syringes:
4.3.1 A 5-mL Luer-Lok glass hypodermic and a 5 ml gas-tight syringe
with shutoff valve for volatile analytes.
4.3.2 Microsyringes - 10 and 25 p.L with a 0.006 in. ID needle
(Hamilton 702N or equivalent) and 100 p,l.
4.4 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
4.5 Analytical balance - 0 - 160 g capacity, capable of measuring
differences of 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used whenever possible. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. Store away from
other solvents.
5.4 Fuels, e.g. gasoline, diesel or jet fuel. Purchase from a commercial
source. Low boiling components in fuel evaporate quickly. If available, obtain
fuel from the leaking tank on site.
5.5 Alkane standard. A standard containing a homologous series of
n-alkanes for establishing retention times (e.g. C10-C32 for diesel and jet fuel).
5.6 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
methanol using assayed liquids:
5.6.1 Place about 9.8 ml of methanol in a 10 ml tared, ground-glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all solvent wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
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5.6.2 Using a 100 /iL syringe, immediately add two or more drops of
assayed reference material to the flask; then reweigh. The liquid must
fall directly into the alcohol without contacting the neck of the flask.
5.6.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.6.4 Transfer the stock standard solution into a bottle with a
Teflon®-!ined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.6.5 Standards must be replaced after 6 months, or sooner if
comparison with check standards indicates a problem.
5.7 Secondary dilution standards - Using stock standard solutions, prepare
secondary dilution standards, as needed, that contain the compounds of interest,
either singly or mixed together. The secondary dilution standards should be
prepared at concentrations such that the aqueous calibration standards prepared
in Sec. 5.6 will bracket the working range of the analytical system. Secondary
dilution standards should be stored with minimal headspace for volatiles and
should be checked frequently for signs of degradation or evaporation, especially
just prior to preparing calibration standards from them.
5.8 Calibration standards - Calibration standards at a minimum of five
concentrations are prepared in water (purge-and-trap or direct injection) or in
methylene chloride (solvent injection) from the secondary dilution of the stock
standards. One of the concentrations should be at a concentration near, but
above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples or
should define the working range of the GC. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Sec. 1.1 may be included). Volatile organic standards are prepared in
organic-free reagent water. In order to prepare accurate aqueous standard
solutions, the following precautions must be observed:
5.8.1 Do not inject more than 20 /iL of methanolic standards into
100 ml of water.
5.8.2 Use a 25 juL Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.8.3 Rapidly inject the primary standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.8.4 Mix diluted standards by inverting the flask three times
only.
8015B - 6 Revision 2
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5.8.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.8.6 Never use pipets to dilute or transfer samples or aqueous
standards when diluting volatile organic standards.
5.8.7 Aqueous standards are not stable and should be discarded
after 1 hour, unless properly sealed and stored. The aqueous standards
can be stored up to 24 hours, if held in sealed vials with zero headspace.
5.9 Internal standards (if internal standard calibration is used) - To use
this approach, the analyst must select one or more internal standards that are
similar in analytical behavior to the compounds of interest. The analyst must
further demonstrate that the measurement of the internal standard is not affected
by method or matrix interferences. Because of these limitations, no internal
standard can be suggested that is applicable to all samples. The following
internal standards are recommended when preparing samples by azeotropic
distillation: 2-chloroacrylonitrile, hexafluoro-2-propanol and
hexafluoro-2-methyl-2-propanol.
5.10 Surrogate standards - Whenever possible, 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 blank
with one or two surrogate compounds which are not affected by method
interferences.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this Chapter, Organic Analytes, Sec. 4.1.
7.0 PROCEDURE
7.1 Introduction/preparation methods: Various alternate methods are
provided for sample introduction. All internal standards, surrogates, and matrix
spikes (when applicable) must be added to samples before introduction into the
GC/FID system. Follow the introduction method on when to add standards.
7.1.1 Direct injection - This involves direct syringe injection
into the GC injection port.
7.1.1.1 Volatile organics (includes gasoline range organics
[GROs]): This may involve injection of an aqueous sample containing
a very high concentration of analytes; injection of aqueous
concentrates from Method 5031 (azeotropic distillation for
nonpurgeable volatile organics); and injection of an organic solvent
waste. Direct injection of aqueous samples (non-concentrated) has
very limited applications. It is only permitted for the
determination of volatiles at the toxicity characteristic (TC)
regulatory limits or at concentrations in excess of 10,000 /xg/L.
It may also be used in conjunction with the test for ignitability in
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aqueous samples (along with Methods 1010 and 1020) to determine if
alcohol is present at > 24%.
7.1.1.2 Semivolatile organics (includes diesel range organics
[DROs] and jet fuel): This may involve syringe injection of
extracts of aqueous samples prepared by Methods 3510 or 3520 or
extracts of soil/solids prepared by Methods 3540, 3541, 3545, 3550
or 3560.
WARNING: Ultrasonic extraction (Method 3550) is not as rugged a method as the
other extraction methods for soil/solids. This means it is very
critical that the method be followed explicitly to achieve
extraction efficiency which approaches that of Soxhlet extraction.
This requires that:
• the necessary equipment must be used (a 3/4" horn and a
minimum of 300 watts of power);
• the horn is properly maintained (tuned prior to use according
to manufacturers instructions and that the tip of the horn is
not worn);
• the samples are properly prepared (the sample is thoroughly
mixed with anhydrous sodium sulfate so that it exists as a
free flowing powder prior to the addition of solvent);
• the correct extraction procedure is followed (three
extractions are performed with the proper solvent, the
sonication is performed in the specified pulse mode and the
tip is positioned just below the solvent surface but above the
sample); and,
• there is visible observation of a very active mixing of the
sample throughout the solvent when the energy pulse is on.
7.1.2 Purge and trap - this includes purge and trap for aqueous
samples (Method 5030) and purge and trap for solid samples (Method 5035).
Method 5035 also provides techniques for extraction of solid and oily
waste samples by methanol (and other water miscible solvents) with
subsequent purge and trap from an aqueous matrix using Method 5030.
Normally purge and trap for aqueous samples is performed at ambient
temperatures while soil/solid samples utilize a 40°C purge to improve
extraction efficiency. Occasionally, there may be a need to perform a
heated purge for aqueous samples to push detection limits lower, however,
a 25 ml sample should provide the sensitivity needed in most situations.
7.1.3 Vacuum distillation - this is a device for the introduction
of volatile organics from aqueous, solid or tissue samples (Method 5032)
into the GC/FID system.
7.1.4 Automated static headspace - this is a device for the
introduction of volatile organics from solid samples (Method 5021) into
the GC/FID system.
8015B - 8 Revision 2
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7.2 Chromatographic conditions (recommended)
7.2.1 Column 1
Carrier gas (Helium) flow rate: 40 mL/min
Temperature program:
Initial temperature: 45"C, hold for 3 minutes
Program: 45°C to 220°C at 8°C/min
Final temperature: 220°C, hold for 15 minutes.
7.2.2 Column 2
Carrier gas (Helium) flow rate: 40 mL/min
Temperature program:
Initial temperature: 50°C, hold for 3 minutes
Program: 50°C to 170°C at 6'C/min
Final temperature: 170°C, hold for 4 minutes.
7.2.3 Column 3
Carrier gas (Helium) flow rate: 15 mL/min
Temperature program:
Initial temperature: 45'C, hold for 4 minutes
Program: 45eC to 220°C at 12°C/min
Final temperature: 220°C, hold for 3 minutes.
7.2.4 Column 4 (DROs and jet fuel)
Carrier gas (Helium) flow rate: 5-7 mL/minute
Makeup gas (Helium) flow rate: 30 mL/min
Injector temperature: 200"C
Detector temperature: 340°C
Temperature program:
Initial temperature: 45°C, hold 3 minute
Program: 45°C to 275°C at 12°C/min
Final temperature: 275°C, hold 12 min
7.2.5 Column 4 (GROs)
Carrier gas (Helium) flow rate: 5-7 mL/minute
Makeup gas (Helium) flow rate: 30 mL/min
Injector temperature: 200°C
Detector temperature: 340°C
Temperature program:
Initial temperature: 45'C, hold 1 minute
Program: 45°C to 100°C at 5°C/min
Final temperature: 100°C to 275°C, at 8°C/min
Final hold: 5 min
7.3 Initial calibration
7.3.1 Set up the sample introduction system as outlined in the
method of choice (see Sec. 7.1). A different calibration curve is
necessary for each sample introduction mode because of the differences in
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conditions and equipment. Establish chromatographic operating parameters
that provide instrument performance equivalent to that documented in this
method. Prepare calibration standards using the procedures described
above (Sec. 5.8). The external standard technique is described below.
Analysts wishing to use the internal standard technique are referred to
Method 8000. Recommended internal standards for the non-purgeable
volatiles include hexafluoro-2-propanol, hexafluoro-2-methyl-2-propanol,
and 2-chloroacrylonitrile.
7.3.2 External standard calibration procedure
7.3.2.1 For each analyte and surrogate of interest or for
each type of fuel, prepare calibration standards at a minimum of
five concentrations by adding volumes of one or more stock standards
to a volumetric flask and diluting to volume with an appropriate
solvent. One of the external standards should be at a concentration
equal to the method quantitation limit (based on the concentration
in the final volume specified in the preparation method, with no
dilutions). The other concentrations should correspond to the
expected range of concentrations found in real samples or should
define the working range of the detector.
NOTE: When the method is used for the determination of petroleum hydrocarbons,
analysts should use the fuel contaminating the site for quantitation.
7.3.2.2 Introduce each calibration standard for individual
analytes using the technique that will be used to introduce the
actual samples into the gas chromatograph. Tabulate peak height or
area responses against the mass injected.
7.3.2.2.1 The ratio of the response to the amount of
analyte introduced to the detector response is defined as the
calibration factor (CF).
7.3.2.2.2 A CF is calculated for each single component
analyte and surrogate at each standard concentration.
Area or Height of Peak
Calibration Factor =
Mass injected (in nanograms)
7.3.2.3 Introduce each calibration standard for fuels
(e.g., JP-4, diesel, gasoline) using the technique that will
be used to introduce the actual samples into the gas
chromatograph. Tabulate the sum of the peak heights or total
area responses under the resolved peaks against the mass
injected.
7.3.2.3.1 A CF can be calculated for each fuel or
hydrocarbon mixture using the total area or the sum of the
peak heights within the retention time range of the specific
fuel.
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Sum (Area or Height of Peaks)
Calibration Factor =
Mass injected (in nanograms)
7.3.2.4 When fuels are analyzed, also analyze a hydrocarbon
retention time standard containing a homologous series of n-alkanes.
The peak heights of these alkanes should be greater than the height
of the mid-point calibration curve. This standard facilitates
identification of the fuel type and the degree of environmental
degradation of the fuel.
7.3.2.5 Linear Calibration: If the percent relative standard
deviation (%RSD) of the calibration factor is less than 20% over the
working range, linearity through the origin can be assumed, and the
average calibration factor can be used in place of a calibration
curve.
7.3.2.6 Non-Linear Calibration: If the % RSD is more than
20% over the working range, linearity through the origin cannot be
assumed. See the discussion on "calibration linearity" in Sec. 7.0
of Method 8000 for options that may be selected.
7.4 Retention time windows
7.4.1 Before establishing retention time windows, make sure that
the chromatographic system is functioning reliably and that the operating
parameters have been optimized for the target analytes and surrogates in
the sample matrix to be analyzed. Establish the retention time windows
for single component target analytes using the procedure described in Sec.
7.0 of Method 8000.
7.4.2 A retention time range for a fuel is defined during initial
calibration. The retention time range is the period between the mean
retention time of the initial rise of the first major eluting peak and the
mean of the final descent of last major eluting peak in the fuel pattern.
Major peaks are at least 10% of the height of the largest peak in the fuel
pattern.
7.4.3 Retention time windows are established for the individual
n-alkane standards using the procedure described in Method 8000.
7.5 Calibration verification
7.5.1 The working calibration curve, and retention times must be
verified at the beginning of each 12-hour work shift as a minimum
requirement. Verification is accomplished by the measurement of one or
more calibration standards (normally mid-concentration) that contain all
of the target analytes and surrogates when individual target analytes are
being analyzed. Verification is accomplished by the measurement of the
fuel standard and the hydrocarbon retention time standard when petroleum
hydrocarbons are being analyzed. Additional analyses of the verification
standard(s) throughout a 12-hour shift are strongly recommended,
especially for samples that contain visible concentrations of oily
8015B - 11 Revision 2
January 1995
-------
material. See Sec. 7.0 "calibration verification" of Method 8000 for more
detailed information.
7.5.2 Calculate the % difference as detailed in Sec. 7.0
"calibration verification" of Method 8000. If the response (or calculated
concentration) for any analyte is within ±15% of the response obtained
during the initial calibration, then the initial calibration is considered
still valid, and analyst may continue to use the mean CF or RF values from
the initial calibration to quantitate sample results. If the response for
any analyte varies from the predicted response by more than ±15% in these
additional determinations, corrective action must be taken to restore the
system or a new calibration curve must be prepared for that compound.
7.5.3 All target analytes and surrogates or n-alkanes in the
calibration verification analyses must fall within previously established
retention time windows. If the retention time of any analyte does not
fall within the ± 3a window, corrective action must be taken to restore
the system or a new calibration curve must be prepared for that compound.
7.5.4 Solvent blanks and any method blanks should be run with
calibration verification analyses to confirm that laboratory contamination
does not cause false positives.
7.6 Gas chromatographic analysis
7.6.1 Samples are analyzed in a set referred to as an analysis
sequence. The sequence begins with calibration verification followed by
sample extract analyses. Additional analyses of the verification
standard(s) throughout a 12-hour shift are strongly recommended,
especially for samples that contain visible concentrations of oily
material. A verification standard is also necessary at the end of a set.
The sequence ends when the set of samples has been injected or when
retention time and/or % difference QC criteria are exceeded.
If the criteria are exceeded, inspect the gas chromatographic system
to determine the cause and perform whatever maintenance is necessary
before recalibrating and proceeding with sample analysis. All sample
analyses performed using external standard calibration must be bracketed
with acceptable data quality analyses (e.g., calibration and retention
time criteria). Therefore, all samples must be reanalyzed that fall
within the standard that exceeded criteria and the last standard that was
acceptable.
7.6.2 Samples are analyzed with the same instrument configuration
as is used during calibration. Analysts are cautioned that opening a
sample vial or drawing an aliquot from a sealed vial (thus creating
headspace) will compromise samples analyzed for volatiles. It is
recommended that analysts prepare two samples for analysis. The second
sample can be stored for 24 hours to ensure that an uncompromised sample
is available for analysis or dilution, if the analysis of the first sample
is unsuccessful or if results exceed the calibration range of the
instrument.
8015B - 12 Revision 2
January 1995
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7.6.3 Sample concentrations are calculated by comparing sample
response data with the initial calibration of the system (Sec. 7.3).
Therefore, if sample response exceeds the limits of the initial
calibration range, a dilution of the sample must be analyzed. For
volatile organic aqueous samples, the dilution must be performed on a
second aliquot of the sample which has been properly sealed and stored
prior to use and reanalysis. Extracts should be diluted so that all peaks
are on scale, as overlapping peaks are not always evident when peaks are
off scale. Computer reproduction of chromatograms, manipulated to ensure
all peaks are on scale over a 100-fold range, are acceptable as long as
calibration limits are not exceeded. Peak height measurements are
recommended over peak area integration when overlapping peaks cause errors
in area integration.
7.6.4 Tentative identification of a single component analyte occurs
when a peak from a sample extract falls within the daily retention time
window. Confirmation is required on a second column or by GC/MS. Since
the flame ionization detector is non-specific, it is highly recommended
that GC/MS confirmation be performed on single component analytes unless
historical data is available to support the identification(s).
7.6.5 Second column confirmation is generally not necessary for
petroleum hydrocarbon analysis. However, if analytical interferences are
indicated, analysis using the second GC column is required. Also, the
analyst must ensure that the sample hydrocarbons fall within the retention
time range established during the initial calibration.
NOTE: Identification of fuels, especially gasoline, is complicated by their
inherent volatility. The early eluting compounds in fuels are obviously
the most volatile and the most likely to have weathered unless sampled
immediately following a spill. The most highly volatile fraction of
gasoline constitutes 50% of the total peak area of a gasoline
chromatogram. This fraction is least likely to be present in an
environmental sample or present in only very low concentration in relation
to the remainder of a gasoline chromatogram.
7.6.6 The performance of the entire analytical system should be
checked every 12 hours, using data gathered from analyses of blanks,
standards, and replicate samples. Significant peak tailing must be
corrected. Tailing problems are generally traceable to active sites on
the column, cold spots in a GC, the detector operation, or leaks in the
system. See Sec. 7.9 for GC/FID system maintenance. Follow
manufacturer's instructions for maintenance of the introduction device.
7.7 Calculations
7.7.1 The concentration of each analyte in the sample may be
determined by calculating the amount of standard purged or injected, from
the peak response, using the calibration curve or the mean CF or RF from
the initial curve.
7.7.2 Analysts should recognize that co-extracted materials may
produce elevated baselines or humps in chromatograms. This is a
particular problem in petroleum hydrocarbon analysis. Therefore,
8015B - 13 Revision 2
January 1995
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integration of the total area under the peaks from the beginning of the
range to the end of the range is required.
7.7.3 Refer to Method 8000, Sec. 7.0 for calculation formulae. The
formulae cover external and internal standard calibration, aqueous and
non-aqueous samples and linear and non-linear calibration curves.
7.8 Screening
7.8.1 Method 8015 with single-point calibration can also be used
for GC/FID screening in order to reduce instrument down-time when highly
contaminated samples are analyzed using GC/MS (e.g., Methods 8260 and
8270).
7.8.2 The same configuration of introduction device interfaced to
the GC/MS may be utilized for the GC/FID or alternative configurations are
acceptable.
7.8.3 Establish that the system response and chromatographic
retention times are stable. Analyze the high-point GC/MS calibration
standard.
7.8.4 Analyze samples or sample extracts. Compare peak heights in
the sample chromatograms with the high-point standard to establish that no
compound with the same retention time as a target analyte exceeds the
calibration range. However, the FID is much less sensitive to halogenated
compounds than the GC/MS system, therefore, the above comparison is not an
absolute certainty.
7.8.5 It is recommended that the high-point standard should be run
at least every 12 hours to confirm the stability of instrument response
and chromatographic retention times. However, there is no QC requirement
for screening.
7.9 Instrument Maintenance:
7.9.1 Injection of sample extracts from waste sites often leaves a
high boiling residue in: the injection port area, splitters when used, and
the injection port end of the chromatographic column. This residue
effects chromatography in many ways (i.e., peak tailing, retention time
shifts, analyte degradation, etc.) and, therefore, instrument maintenance
is very important. Residue buildup in a splitter may limit flow through
one leg and therefore change the split ratios. If this occurs during an
analytical run, the quantitative data may be incorrect. Proper cleanup
techniques will minimize the problem and instrument QC will indicate when
instrument maintenance is required.
7.9.2 Suggested chromatograph maintenance: Corrective measures may
require any one or more of the following remedial actions. Also see Sec.
7.0 in Method 8000 for additional guidance on corrective action for
capillary columns and the injection port.
7.9.2.1 Splitter connections: For dual columns which are
connected using a press-fit Y-shaped glass splitter or a Y-shaped
8015B - 14 Revision 2
January 1995
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fused-silica connector, clean and deactivate the splitter or replace
with a cleaned and deactivated splitter. Break off the first few
inches (up to one foot) of the injection port side of the column.
Remove the columns and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate
the degradation problem, it may be necessary to deactivate the metal
injector body and/or replace the columns.
7.9.2.2 Column rinsing: The column should be rinsed with
several column volumes of an appropriate solvent. Both polar and
nonpolar solvents are recommended. Depending on the nature of the
sample residues expected, the first rinse might be water, followed
by methanol and acetone; methylene chloride is a satisfactory final
rinse and in some cases may be the only solvent required. The
column should then be filled with methylene chloride and allowed to
remain flooded overnight to allow materials within the stationary
phase to migrate into the solvent. The column is then flushed with
fresh methylene chloride, drained, and dried at room temperature
with a stream of ultrapure nitrogen passing through the column.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
8015B - 15 Revision 2
January 1995
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If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicate a potential problem due to the sample matrix itself, the
LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL actually achieved in a given
analysis, accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and calibration procedures used.
9.2 Specific method performance information for non-purgeable volatiles
prepared using the azeotropic microdistillation technique from Method 5031 is
included in Tables 1, 3 and 4 for aqueous matrices and in Tables 2 and 5 for
solid matrices.
9.3 Specific method performance information is provided for diesel fuel
spiked into soil in Tables 6, 7, 8, 9, and 10.
8015B - 16 Revision 2
January 1995
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10.0 REFERENCES
1. Bellar, T.A., and J.J. Lichtenberg. "Determining Volatile Organics at
Microgram-per-Liter Levels by Gas Chromatography", J. Amer. Water Works
Assoc., 66(12), pp. 739-744 (1974).
2. Bellar, T.A., and J.J. Lichtenberg. "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds", in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
3. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters: Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane, Report for EPA
Contract 68-03-2635.
4. Bruce, M.L., R.P. Lee, and M.W. Stevens. "Concentration of Water Soluble
Volatile Organic Compounds from Aqueous Samples by Azeotropic
Microdistillation", Environ. Sci. Technol. 1992, 26, 160-163.
5. Tsang, S.F., N. Chau, P.J. Marsden, and K.R. Carter. "Evaluation of the
EnSys PETRO RISc kit for TPH", Report for Ensys, Inc., Research Triangle
Park, NC, 27709, 1992.
8015B - 17 Revision 2
January 1995
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TABLE 1
METHOD DETECTION LIMITS FOR NON-PURGEABLE VOLATILE COMPOUNDS
IN AQUEOUS MATRICES BY AZEOTROPIC MICRODISTILLATION (METHOD 5031)
MDL
Analyte Reagent Water
Acetoneb
Acetonitrile
Acrolein
Acrylonitrile
1-Butanol
t-Butyl alcohol
1,4-Dioxane
Ethanol
Ethyl acetate
Ethyl ene oxide
Isobutyl alcohol
Methanol
Methyl ethyl ketone
Methyl isobutyl ketone
2-Pentanone
1-Propanol
2-Propanol (Isopropyl ale.)
Propionitrile
Pyridine
48
15
13
8
14
8
12
18
9
8
11
21
4
4
2
--
18
10
11
Ground Water
16
6
15
9
8
7
15
12
8
9
8
21
5
2
2
7
17
6
9
TCLP Leachate
63
14
7
14
7
17
16
13
16
10
4
22
9
8
7
--
7
13
21
Produced by analysis of 7 aliquots of water spiked at 25 M9/U using
internal standard calibration.
Problematic due to transient laboratory contamination.
8015B - 18 Revision 2
January 1995
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TABLE 2
METHOD DETECTION LIMITS FOR NON-PURGEABLE VOLATILE COMPOUNDS
IN SOLID MATRICES BY AZEOTROPIC MICRODISTILLATION (METHOD 5031)
MDL (mq/kq)
Analyte Incinerator Ash Kaolin
Acrylonitrile
1-Butanol
t-Butyl alcohol
1,4-Dioxane
Ethanol
Ethyl acetate
Isopropyl alcohol
Methanol
Methyl ethyl ketone
Methyl isobutyl ketone
2-Pentanone
Pyridine
0.42
0.23
0.34
0.31
0.47
0.18
0.40
0.46
0.27
0.12
0.16
0.20
0.09
0.09
0.13
0.16
0.19
0.07
0.19
0.31
0.12
0.05
0.07
0.08
NOTE: The MDLs calculated for this table were produced by the analysis of 7
replicates spiked at 0.50 mg/kg, using internal standard calibration.
8015B - 19 Revision 2
January 1995
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TABLE 3
METHOD PERFORMANCE DATA FOR NON-PURGEABLE VOLATILES IN GROUND
WATER BY AZEOTROPIC MICRODISTILLATION (METHOD 5031)
Compound
Acetone"
Acetonitrile
Acrolein
Acrylonitrile
1-Butanol
t-Butyl alcohol
1,4-Dioxane
Ethanol
Ethyl Acetate
Ethyl ene oxide
Isobutyl alcohol
Isopropyl alcohol
Methanol
Methyl ethyl ketone
Methyl isobutyl ketone
2-Pentanone
1-Propanol
Propionitrile
Pyridine
8 25 jug/L spikes,
b 100 p.g/1 spikes,
0 750 jug/L spikes,
Low Cone."
Averaged
%Rec %RSD
126 17
147 5
146 13
179 7
127 8
122 7
124 16
152 10
142 7
114 10
122 8
167 13
166 14
105 6
66 4
94 3
N/A
135 5
92 12
using internal
using internal
using internal
Medium Conc.b High Cone.0
Average51
Averaged
%Rec %RSD %Rec %RSD
N/A
105 8
120 27
143 28
86 8
N/A
96 10
N/A
135 33
N/A
87 13
N/A
94 9
N/A
N/A
N/A
91 7
102 14
N/A
calibration.
calibration.
calibration.
N/A
92
80
94
90
N/A
99
N/A
92
N/A
89
N/A
95
N/A
N/A
N/A
91
90
N/A
— -
9
20
21
9
--
8
--
25
--
13
--
7
--
--
--
7
14
- -
d Average of 7 replicates
" Problematic due
to transient laboratory contamination.
N/A Data not available
8015B - 20
Revision 2
January 1995
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TABLE 4
METHOD PERFORMANCE DATA FOR NON-PURGEABLE VOLATILES IN TCLP
LEACHATE BY AZEOTROPIC MICRODISTILLATION (METHOD 5031)
Compound
Acetone"
Acetonitrile
Acrolein
Acrylonitrile
1-Butanol
t-Butyl alcohol
1,4-Dioxane
Ethanol
Ethyl Acetate
Ethyl ene oxide
Isobutyl alcohol
Isopropyl alcohol
Methanol
Methyl ethyl ketone
Methyl isobutyl ketone
2-Pentanone
1-Propanol
Propionitrile
Pyridine
8 25 M9/L spikes,
b 100 jug/L spikes,
c 750 /ng/L spikes,
Low Cone."
Averaged
%Rec %RSD
99 91
107 17
88 10
133 13
119 7
70 31
103 20
122 13
164 12
111 12
115 4
114 8
107 10
87 13
78 13
101 8
N/A
100 16
46 59
using internal
using internal
using internal
Medium Conc.b
Averaged
%Rec %RSD
N/A
111
109
123
89
N/A
103
N/A
119
N/A
86
N/A
102
N/A
N/A
N/A
98
100
N/A
calibration.
calibration
calibration
_ _
10
29
29
12
--
16
--
29
--
13
--
6
--
--
--
10
11
— —
High
Cone.0
Averaged
%Rec
N/A
95
87
103
86
N/A
102
N/A
107
N/A
82
N/A
N/A
N/A
N/A
N/A
89
90
N/A
%RSD
_ _
11
41
38
8
--
7
--
41
--
13
--
--
--
--
--
7
17
d Average of 7 replicates
" Problematic due
to transient laboratory contamination.
N/A Data not available
8015B - 21
Revision 2
January 1995
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TABLE 5
METHOD PERFORMANCE DATA FOR NON-PURGEABLE VOLATILE COMPOUNDS
IN SOLID MATRICES BY AZEOTROPIC MICRODISTILLATION (METHOD 5031)
Incinerator Ash
Low Cone.8
Average0
Acrylonitrile
1-Butanol
t-Butyl alcohol
1,4-Dioxane
Ethanol
Ethyl acetate
Isopropyl alcohol
Methanol
Methyl ethyl ketone
Methyl isobutyl ketone
2-Pentanone
Pyridine
%Rec
50
105
101
106
117
62
119
55
81
68
79
52
%RSD
53
14
21
19
25
19
21
53
21
11
13
24
High
Conc.D
Average0
%Rec
10
61
60
48
52
39
61
33
40
57
54
44
%RSD
31
12
13
18
20
12
15
28
12
14
10
20
Kaolin
Low Cone.3
Average0
%Rec
102
108
97
105
108
90
108
117
91
71
91
50
%RSD
6
5
9
10
11
5
11
17
8
5
5
10
High Cone."
Average0
%Rec
12
58
59
48
48
41
58
37
42
55
54
49
%RSD
52
25
23
25
24
25
24
22
20
23
19
31
a 0.5 mg/kg spikes, using internal calibration.
b 25 mg/kg spikes, using internal calibration.
Average of 7 replicates
8015B - 22
Revision 2
January 1995
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TABLE 6
RESULTS FROM ANALYSIS8 OF LOW
AROMATIC DIESEL6 BY GC/FID
(5 replicates/test)
Spike
Concentration
12.5 ppm
75 ppm
105 ppm
150 ppm
1000 ppm
Analysis Results
ND
54 ± 7 ppm
90 ± 15 ppm
125 ± 12 ppm
960 ± 105 ppm
8 Samples were prepared using 2 g aliquots of sandy loam soil spiked
with known amounts of low aromatic diesel. Extractions were
accomplished using methylene chloride as a solvent (Method 3550,
high concentration option).
b Low aromatic diesel is sold in California (Section 2256, CCR). For
this study it was purchased at a gas station in San Diego,
California.
TABLE 7
INITIAL 5-POINT CALIBRATION8 OF LOW
AROMATIC DIESEL-2 (5/21/92)
Concentration (mg/L)
10
50
100
500
1000
Mean =
Std. Dev. =
%RSD =
Response Factor
0.002287
0.002414
0.002080
0.001991
0.001980
0.002150
0.000192
8.9
10 peaks were used for quantitation.
8015B - 23
Revision 2
January 1995
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TABLE 8
SECOND 5-POINT CALIBRATION" OF LOW
AROMATIC DIESEL-2 (5/26/92)
Concentration (mg/L)
10
50
100
500
1000
Mean =
Std. Dev. =
%RSD =
Response Factor
0.002356
0.002226
0.001929
0.001988
0.001961
0.002092
0.000188
9.0
10 peaks were used for quantitation.
8015B - 24
Revision 2
January 1995
-------
TABLE 9
RESULTS FROM ANALYSIS8 OF
REGULAR #2 DIESEL BY GC/FID
(4 replicates/test)
Spike
Concentration
25 ppm
75 ppm
125 ppm
150 ppm
Analysis Results
51.2 ± 6.4 ppm
75.9 ± 7.8 ppm
98.9 ± 5.2 ppm
162 ± 10.4 ppm
Samples were prepared using 10 g aliquots of sandy loam soil spiked
with known amounts of regular #2 diesel purchased at a gas station
in Northern Virginia. Extractions were accomplished using methylene
chloride as a solvent (Method 3550).
TABLE 10
6-POINT CALIBRATION" OF
NORTHERN VIRGINIA REGULAR #2 DIESEL
Concentration (mg/L)
5
10
50
100
500
1000
Mean =
Std. Dev. =
%RSD =
Response Factor
0.00143
0.00158
0.00179
0.00173
0.00175
0.00167
0.00166
0.00013
8.1
10 peaks were used for quantitation.
8015B - 25
Revision 2
January 1995
-------
FIGURE 1
CHROMATOGRAM OF C10 - C32 HYDROCARBON STANDARD USING COLUMN 4
ia
0
n
r\ w
'O *"
r> _
^
,
1
w
n
Q
n
n
n
Q
Q
n
n
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A
0
n
0
C
i '
01
n
Q
0
n
V*
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000
000
000
000
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\
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n
n
n
n
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n
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r\
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IT
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-5.874
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"
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,08
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36.218
40.222
524
11.351
Data File Name : C:\HPCHEM\1\DATA\0615\O02P0101.D
Operator : fai
Instrument : ANALYZERi
Sample Name : 25ppm n-alkane
8015B - 26
Revision 2
January 1995
-------
FIGURE 2
CHROMATOGRAM OF JP-4 USING COLUMN 4
(500 ppm standard)
Data File
Operator
Instrument
Sample Name
Name
.819
261
.956
-5.514
7.510
C:\HPCHKM\1\DAIA\0519\008P0101.D
PAI
ANALYZKR1
jp-4 SOOppm
8015B - 27
Revision 2
January 1995
-------
FIGURE 3
CHROMATOGRAM OF LOW AROMATIC DIESEL USING COLUMN 4
(500 ppm standard)
0
0
0
1
OOO9
03
0
0
0
i i
M M-
C M
(D 'D
, t , , , f .
M
. , f
M I— ^
/^ ^n
iJJ gu
fH (D-
W
0
fl
, , f
— — ^_
w
0
—15.210
16.376
-17.479
Data File
Operator
Instrument
Sample Name
C:\HPCHBM\1\DATA\0615\032F0101.D
fai
AKALYZKR1
CA diesel SOOppn
8015B - 28
Revision 2
January 1995
-------
FIGURE 4
CHROMATOGRAM OF NORMAL DIESEL USING COLUMN 4
(500 ppm standard)
^
0
0
0
1
^-^= 1-BW
0)
0
0
0
, 1 ,
CD
0
0
0
M l-k
0 M
(D (I
\ V
.A L^
P
(D
f
1 ' L,,.- i_
H
OJ
(H
f
M
CD
(H
f
L 1 1 I 1
W
0
(5
, f
r-oo*
(0
0
-12.639
13.969
15.212
-16.379
-17.482
18.524
-19.514
20.457
-21.351
.
Data File Nan*
Operator
Instrument
Sample Name
C:\HPCHBM\1\DATA\0615\033P0101.D
fai
ANALYZBR1
VA diesel SOOppm
8015B - 29
Revision 2
January 1995
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FIGURE 5
CHROMATOGRAM OF SEVERAL NON-PURGEABLE VOLATILE COMPOUNDS IN
SPIKED REAGENT WATER USING AZEOTROPIC MICRODISTILLATION (METHOD 5031)
J
ouedojdosjOJonijBxeq 'S'l
|oue»nq-1
9UEXO|p-t'
|OUBL|ja
euojaoe
Mix 1:
Conditions:
Analytes distilled at 0.25mg/L, Internal Stds. at 2.5 mg/L
J&W DB-Wax column with 0.53 ID
Temperature program: 30°C for 2 min.
3°C/min. to 100°C and held for 0 min.
25°C/min. to 200°C and held for 4 min
8015B - 30
Revision 2
January 1995
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FIGURE 6
CHROMATOGRAM OF SEVERAL NON-PURGEABLE VOLATILE COMPOUNDS IN
SPIKED REAGENT WATER USING AZEOTROPIC MICRODISTILLATION (METHOD 5031)
loiredcudosiojonijexeii '
aufpuAd.
|ouedojdos;|Ameuiojon|jexai| 'S'l
9|U}!UO|AjOEOJO|l|0-2 'S'l
_o
Mix 2: Analytes distilled at 0.25mg/L, Internal Stds. at 2.5 mg/L
Conditions: J&W DB-Wax column with 0.53 ID
Temperature program: 30"C for 2 min.
3°C/min. to 100°C and held for 0 min.
25°C/min. to 200°C and held for 4 min
8015B - 31
Revision 2
January 1995
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METHOD 8015B
NONHALOGENATED ORGANICS USING GC/FID
7 2 Set chromatographic conditions
I
7 4 Initial calibration
Internal
7 4 1 See Method 8000 for
internal std technique
External
7 4 2.1 Prepare at least 5 concentrations
of calibration stds using stock stds.
7 4 2.3 Introduce
each std using
technique to be used
for actual samples
Calculate CF
I
Individual
Analysis for \ analytes
individual analytes
or fuels?
7 4 2.2 Introduce
each std into GC
using technique to be
used for actual
samples Calculate
CF
7424 Analyze
hydrocarbon retention
time std
< 20% over
What is ^working range
percent relative
std deviation
ofCF->
>20% over
working range
7 4.2.5 Use linear
calibration.
7 4 2.6 Use non-linear
calibration
8015B - 32
Revision 2
January 1995
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METHOD 3015B
(continued)
7 5 Establish retention time windows
7.7 Perform chromatographic analysis
772
Does sample
response exceed
limits of initial
calibration''
772 Analyze dilution
of sample
7.72
Are all peaks
7,7.2 Analyze dilution
of sample.
7.7 3 Confirm id of analyte on 2nd column
7 7.4 Are analytical
interferences suspected'7
774 Analyze sample
with 2nd column
8015B - 33
Revision 2
January 1995
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METHOD 802IB
HALOGENATED VOLATILES BY GAS CHROMATOGRAPHY USING
PHOTOIONIZATION AND ELECTROLYTIC CONDUCTIVITY DETECTORS
IN SERIES: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8021 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Analyte
CAS No."
Appropriate Technique
Purge-and Direct Vac Head
-Trap Injection Distln Space
Ally! chloride 107-05-1
Benzene 71-43-2
Benzyl chloride 100-44-7
Bis(2-chloroisopropyl) ether 39638-32-9
Bromoacetone 598-31-2
Bromobenzene 108-86-1
Bromochloromethane 74-97-5
Bromodichloromethane 75-27-4
Bromoform 75-25-2
Bromomethane 74-83-9
Carbon tetrachloride 56-23-5
Chlorobenzene 108-90-7
Chlorodibromomethane 124-48-1
Chloroethane 75-00-3
2-Chloroethanol 107-07-03
2-Chloroethyl vinyl ether 110-75-8
Chloroform 67-66-3
Chloromethyl methyl ether 107-30-2
Chloroprene 126-99-8
Chloromethane 74-87-3
4-Chlorotoluene 106-43-4
l,2-Dibromo-3-chloropropane 96-12-8
1,2-Dibromoethane 106-93-4
Dibromomethane 74-95-3
1,2-Dichlorobenzene 95-50-1
1,3-Dichlorobenzene 541-73-1
1,4-Dichlorobenzene 106-46-7
Dichlorodifluoromethane 75-71-8
b
b
PP
b
PP
b
b
b
b
b
b
b
b
b
PP
b
b
PP
b
b
b
PP
b
b
b
b
b
b
b
b
b
b
b
nd
b
b
b
b
b
b
b
b
b
b
b
pc
nd
b
b
b
nd
b
nd
nd
nd
b
nd
b
nd
nd
nd
nd
nd
b
b
b
b
b
b
b
nd
b
b
nd
nd
b
nd
nd
nd
b
nd
nd
nd
b
nd
b
nd
nd
nd
nd
b
b
b
b
b
b
b
b
nd
nd
b
nd
nd
b
nd
b
b
b
b
b
b
b
8021B - 1
Revision 2
January 1995
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Appropriate
Purge- and Direct
Analyte
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans -1,2-Dichloroethene
1,2-Dichloropropane
l,3-Dichloro-2-propanol
cis-l,3-dichloropropene
trans-l,3-dichloropropene
Epichlorhydrin
Ethyl benzene
Hexachlorobutadiene
Methylene chloride
Naphthalene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,4-Trichlorobenzene
1,1,1 -Tri chl oroethane
1,1, 2 -Trichl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
CAS No." -Trap Injection
75-34-3
107-06-2
75-35-4
156-59-4
156-60-5
78-87-5
96-23-1
10061-01-5
10061-02-6
106-89-8
100-41-4
87-68-3
75-09-2
91-20-3
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
120-82-1
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
75-01-4
95-47-6
108-38-3
106-42-3
b
b
b
b
b
b
PP
b
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
nd
b
nd
b
b
b
b
b
nd
b
nd
b
nd
b
b
b
nd
b
b
b
b
b
b
b
b
b
Technique
Vac
Distln
b
b
b
nd
b
b
nd
b
b
nd
b
nd
b
nd
b
nd
b
b
b
nd
b
b
b
b
b
b
b
b
b
Head
Space
b
b
b
nd
b
b
nd
nd
nd
nd
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
a Chemical Abstract Services Registry Number.
b Adequate response by
i Inappropriate techni
nd Not Determined
pc Poor chromatographic
this technique.
que for this analyte.
behavior.
pp Poor purging efficiency resulting in high
EQLs.
is
1.2 Method detection limits (MDLs) are compound dependent and vary with
purging efficiency and concentration. The MDLs for selected analytes are
presented in Table 1. The applicable concentration range of this method
compound and instrument dependent but is approximately 0.1 to 200
Analytes that are inefficiently purged from water will not be detected when
present at low concentrations, but they can be measured with acceptable accuracy
and precision when present in sufficient amounts. Determination of some
structural isomers (i.e. xylenes) may be hampered by coelution.
8021B - 2
Revision 2
January 1995
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1.3 The estimated quantitation limit (EQL) of Method 8021A for an
individual compound is approximately 1 M9/kg (wet weight) for soil/sediment
samples, 0.1 mg/kg (wet weight) for wastes, and 1 p.g/1 for ground water (see
Table 3). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 This method is restricted for use by, or under the supervision of,
analysts experienced in the use of gas chromatographs for measurement of
purgeable organics at low ^g/L concentrations and skilled in the interpretation
of gas chromatograms. Each analyst must demonstrate the ability to generate
acceptable results with this method.
1.5 The toxicity or carcinogenicity of chemicals used in this method has
not been precisely defined. Each chemical should be treated as a potential
health hazard, and exposure to these chemicals should be minimized. Each
laboratory is responsible for maintaining awareness of OSHA regulations regarding
safe handling of chemicals used in this method. Additional references to
laboratory safety are available for the information of the analyst (References
4 and 6).
1.6 The following method analytes have been tentatively classified as
known or suspected human or mammalian carcinogens: benzene, carbon tetrachloride,
1,4-dichlorobenzene, 1,2-dichloroethane, hexachlorobutadiene, 1,1,2,2-
tetrachloroethane, 1,1,2-trichloroethane, chloroform, 1,2-dibromoethane,
tetrachloroethene, trichloroethene, and vinyl chloride. Pure standard materials
and stock standard solutions of these compounds should be handled in a hood. A
NIOSH/MESA approved toxic gas respirator should be worn when the analyst handles
high concentrations of these toxic compounds.
1.7 Other non-RCRA compounds which are amenable to analysis by Method
8021 are:
Analyte CAS No.a
n-Butylbenzene104-51-8
sec-Butyl benzene 135-98-8
tert-Butylbenzene 98-06-6
2-Chlorotoluene 95-49-8
1,3-Dichloropropane 142-28-9
2,2-Dichloropropane 590-20-7
1,1-Dichloropropene 563-58-6
Isopropylbenzene 98-82-8
p-Isopropyltoluene 99-87-6
n-Propylbenzene 103-65-1
1,2,3-Trichlorobenzene 87-61-6
1,2,4-Trimethylbenzene 95-63-6
1,3,5-Trimethylbenzene 108-67-8
8021B - 3 Revision 2
January 1995
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2.0 SUMMARY OF METHOD
2.1 Method 8021 provides gas chromatographic conditions for the
detection of halogenated and aromatic volatile organic compounds. Samples can
be analyzed using direct injection (Method 3585 for oily matrices) or purge-and-
trap (Method 5030/5035), headspace (Method 5021), or vacuum distillation (Method
5032). Groundwater samples may be analyzed using Method 5030, Method 5021, or
Method 5032. A temperature program is used in the gas chromatograph to separate
the organic compounds. Detection is achieved by a photoionization detector (PID)
and an electrolytic conductivity detector (HECD) in series. The GC system may
also be set up to use a single detector when an analyst is looking for only
halogenated compounds (HECD) or aromatic compounds (PID).
2.2 Tentative identifications are obtained by analyzing standards under
the same conditions used for samples and comparing resultant GC retention times.
Confirmatory information can be gained by comparing the relative response from
the two detectors. Concentrations of the identified components are measured by
relating the response produced for that compound to the response produced by a
compound that is used as an internal standard.
3.0 INTERFERENCES
3.1 Refer to the appropriate 5000 series method and Method 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
3.3 Sulfur dioxide is a potential interferant in the analysis for vinyl
chloride.
4.0 APPARATUS AND MATERIALS
4.1 Sample introduction apparatus - Refer to Sec. 4.0 of the appropriate
5000 series method for a listing of the equipment for each sample introduction
technique.
4.2 Gas Chromatograph - capable of temperature programming; equipped
with variable-constant differential flow controllers, subambient oven controller,
photoionization and electrolytic conductivity detectors connected with a short
piece of uncoated capillary tubing, 0.32-0.5 mm ID, and data system.
4.2.1 Column - 60 m x 0.75 mm ID VOCOL wide-bore capillary column
with 1.5 )Ltm film thickness (Supelco Inc., or equivalent).
4.2.2 Photoionization detector (PID) (Tracor Model 703, or
equivalent).
4.2.3 Electrolytic conductivity detector (HECD) (Tracor Hall Model
700-A, or equivalent).
8021B - 4 Revision 2
January 1995
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4.3 Syringes - 5 ml glass hypodermic with Luer-Lok tips.
4.4 Syringe valves - 2-way with Luer ends (Teflon® or Kel-F).
4.5 Microsyringe - 25 p.1 with a 2 in. x 0.006 in. ID, 22" bevel needle
(Hamilton #702N or equivalent).
4.6 Microsyringes - 10, 100 /iL.
4.7 Syringes - 0.5, 1.0, and 5 ml, gas tight with shut-off valve.
4.8 Bottles - 15 ml, Teflon®-!ined with screw-cap or crimp top.
4.9 Analytical balance - 0.0001 g.
4.10 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all inorganic reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to
be free of analytes. Store away from other solvents.
5.4 Vinyl chloride, (99.9% pure), CH2=CHC1. Vinyl chloride is available
from Ideal Gas Products, Inc., Edison, New Jersey and from Matheson, East
Rutherford, New Jersey, as well as from other sources. Certified mixtures of
vinyl chloride in nitrogen at 1.0 and 10.0 ppm (v/v) are available from several
sources.
5.5 Stock standards - Stock solutions may either be prepared from pure
standard materials or purchased as certified solutions. Prepare stock standards
in methanol using assayed liquids or gases, as appropriate. Because of the
toxicity of some of the organohalides, primary dilutions of these materials of
the toxicity should be prepared in a hood.
NOTE: If direct injection is used, the solvent system of standards must match
that of the sample. It is not necessary to prepare high concentration
aqueous mixed standards when using direct injection.
5.5.1 Place about 9.8 ml of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
8021B - 5 Revision 2
January 1995
-------
about 10 minutes until all alcohol-wetted surfaces have dried. Weigh the
flask to the nearest 0.1 mg.
5.5.2 Add the assayed reference material, as described below.
5.5.2.1 Liquids: Using a 100 /ul_ syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.5.2.2 Gases: To prepare standards for any compounds that
boil below 30'C (e.g. bromomethane, chloroethane, chloromethane,
dichlorodifluoromethane, trichlorofluoromethane, vinyl chloride),
fill a 5 ml valved gas-tight syringe with the reference standard to
the 5.0 ml mark. Lower the needle to 5 mm above the methanol
meniscus. Slowly introduce the reference standard above the surface
of the liquid. The heavy gas rapidly dissolves in the methanol.
This may also be accomplished by using a lecture bottle equipped
with a septum. Attach Teflon® tubing to the side-arm relief valve
and direct a gentle stream of gas into the methanol meniscus.
5.5.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.5.4 Transfer the stock standard solution into a bottle with a
Teflon®-!ined screw-cap or crimp top. Store, with minimal headspace, at
-10°C to -20°C and protect from light.
5.5.5 Prepare fresh stock standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months. Both
gas and liquid standards must be monitored closely by comparison to the
initial calibration curve and by comparison to QC check standards. It may
be necessary to replace the standards more frequently if either check
exceeds a 20% drift.
5.6 Prepare secondary dilution standards, using stock standard
solutions, in methanol, as needed, that contain the compounds of interest, either
singly or mixed together. The secondary dilution standards should be prepared
at concentrations such that the aqueous calibration standards prepared in Sec.
5.7 will bracket the working range of the analytical system. Secondary dilution
standards should be stored with minimal headspace for volatiles and should be
checked frequently for signs of degradation or evaporation, especially just prior
to preparing calibration standards from them.
5.7 Calibration standards, at a minimum of five concentrations are
prepared in organic-free reagent water from the secondary dilution of the stock
standards. One of the concentrations should be at a concentration near, but
8021B - 6 Revision 2
January 1995
-------
above, the method detection limit. The remaining concentrations should
correspond to the expected range of the concentrations found in real samples or
should define the working range of the GC. Standards (one or more) should
contain each analyte for detection by this method. In order to prepare accurate
aqueous standard solutions, the following precautions must be observed.
NOTE: Prepare calibration solutions for use with direct injection analyses in
water at the concentrations required.
5.7.1 Do not inject more than 20 pi of alcoholic standards into
100 mL of water.
5.7.2 Use a 25 /il_ Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.7.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.7.4 Mix aqueous standards by inverting the flask three times.
5.7.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.7.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.7.7 Aqueous standards are not stable and should be discarded after
one hour, unless properly sealed and stored. The aqueous standards can
be stored up to 12 hours, if held in sealed vials with zero headspace.
5.7.8 Optionally calibration using a certified gaseous mixture can
be accomplished daily utilizing commercially available gaseous analyte
mixture of bromomethane, chloromethane, chloroethane, vinyl chloride,
dichlorodifluoromethane and trichlorofluoromethane in nitrogen. These
mixtures of documented quality are stable for as long as six months
without refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.8 Internal standards - Prepare a spiking solution containing
fluorobenzene and 2-bromo-l-chloropropane in methanol, using the procedures
described in Sees. 5.5 and 5.6. It is recommended that the secondary dilution
standard be prepared at a concentration of 5 mg/L of each internal standard
compound. The addition of 10 fj.1 of such a standard to 5.0 mL of sample or
calibration standard would be equivalent to 10 M9/L-
5.9 Surrogate standards - The analyst should monitor both the
performance of the analytical system and the effectiveness of the method in
dealing with each sample matrix by spiking each sample, standard, and reagent
blank with two or more surrogate compounds. A combination of 1,4-dichlorobutane
and bromochlorobenzene is recommended to encompass the range of the temperature
program used in this method. From stock standard solutions prepared as in Sec.
5.5, add a volume to give 750 jug of each surrogate to 45 ml of organic-free
8021B - 7 Revision 2
January 1995
-------
reagent water contained in a 50 ml volumetric flask, mix, and dilute to volume
for a concentration of 15 ng//iL. Add 10 /xL of this surrogate spiking solution
directly into the 5 ml syringe with every sample and reference standard analyzed.
If the internal standard calibration procedure is used, the surrogate compounds
may be added directly to the internal standard spiking solution (Sec. 5.8).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection (Method 3585 for oily matrices) or purge-and-trap (Methods
5030/5035), headspace (Method 5021), or by vacuum distillation (Method 5032).
Methods 5030, 5021, or 5032 may be used directly on groundwater samples. Methods
5035, 5021, or 5032 may be used for low-concentration contaminated soils and
sediments. For high-concentration soils or sediments (>200 M9Ag), methanolic
extraction, as described in Method 5035, may be necessary prior to purge-and-trap
analysis. For guidance on the dilution of oily waste samples for direct
injection refer to Method 3585.
7.2 Gas chromatography conditions (Recommended)
7.2.1 Set up the gas chromatograph system so that the
photoionization detector (PID) is in series with the electrolytic
conductivity detector (HECD). It may be helpful to contact the
manufacturer of the GC for guidance on the proper installation of dual
detector systems.
NOTE: Use of the dual detector system is not a requirement of the method. The
GC system may also be set up to use a single detector when the analyst is
looking for just halogenated compounds (using the HECD) or for just
aromatic compounds (using the PID).
7.2.2 Oven settings:
Carrier gas (Helium) Flow rate: 6 mL/min.
Temperature program
Initial temperature: 10°C, hold for 8 minutes at
Program: 10°C to 180°C at 4°C/min
Final temperature: 180"C, hold until all expected
compounds have eluted.
7.2.3 The carrier gas flow is augmented with an additional 24 mL of
helium flow before entering the photoionization detector. This make-up
gas is necessary to ensure optimal response from both detectors.
7.2.4 These halogen-specific systems eliminate misidentifications
due to non-organohalides which are coextracted during the purge step. A
Tracer Hall Model 700-A detector was used to gather the single laboratory
8021B - 8 Revision 2
January 1995
-------
accuracy and precision data presented in Table 2. The operating
conditions used to collect these data are:
Reactor tube: Nickel, 1/16 in OD
Reactor temperature: 810°C
Reactor base temperature: 250°C
Electrolyte: 100% n-Propyl alcohol
Electrolyte flow rate: 0.8 mL/min
Reaction gas: Hydrogen at 40 mL/min
Carrier gas plus make-up gas: Helium at 30 mL/min
7.2.5 A sample chromatogram obtained with this column is presented
in Figure 1. This column was used to develop the method performance
statements in Sec. 9.0. Estimated retention times and MDLs that can be
achieved under these conditions are given in Table 1. Other columns or
element specific detectors may be used if the requirements of Sec. 8.0 are
met.
7.3 Calibration - Refer to Method 8000 for proper calibration
techniques. Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Sec. 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Methods 5030/5035 (purge-and-trap method) or the direct injection
method (see Sec. 7.4.1.1), by Method 5021 (headspace) or by Method 5032
(vacuum distillation). If the internal standard calibration technique is
used, add 10 jiiL of internal standard to the sample prior to purging.
7.4.1.1 Direct injection - In very limited applications (e.g.
aqueous process wastes) direct injection of the sample into the GC
system with a 10 /nL syringe may be appropriate. The detection limit
is very high (approximately 10,000 M9/L), therefore, it is only
permitted where concentrations in excess of 10,000 jug/L are expected
or for water-soluble compounds that do not purge. The system must
be calibrated by direct injection (bypassing the purge-and-trap
device).
7.4.1.2 Refer to Method 3585 for guidance on the dilution and
direct injection of oily waste samples.
7.4.2 Follow Sec. 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria. Include a mid-concentration
standard after each group of 10 samples in the analysis sequence.
8021B - 9 Revision 2
January 1995
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7.4.3 Table 1 summarizes the estimated retention times on the two
detectors for a number of organic compounds analyzable using this method.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using a second GC column is recommended.
7.4.7 If the response for a peak is off-scale, i.e., beyond the
calibration range of the standards, prepare a dilution of the sample with
organic-free reagent water. The dilution must be performed on a second
aliquot of the sample which has been properly sealed and stored prior to
use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
(QC) procedures. Quality control procedures to ensure the proper operation of
the various sample preparation and/or sample introduction techniques can be found
in Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample
or one matrix spike/matrix spike duplicate pair. The decision on whether
to prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
8021B - 10 Revision 2
January 1995
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If samples are not expected to contain target analytes, laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicate a potential problem due to the sample matrix itself, the
LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate
recovery data from individual samples versus the surrogate control limits
developed by the laboratory. See Method 8000, Sec. 8.0 for information on
evaluating surrogate data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Method detection limits for these analytes have been calculated from
data collected by spiking organic-free reagent water at 0.1 M9/L- These data
are presented in Table 1.
9.2 This method was tested in a single laboratory using organic-free
reagent water spiked at 10 M9/L. Single laboratory precision and accuracy data
for each detector are presented for the method analytes in Table 2.
10.0 REFERENCES
1. "Volatile Organic Compounds in Water by Purge-and-Trap Capillary Column
Gas Chromatography with Photoionization and Electrolytic Conductivity
Detectors in Series", Method 502.2, Rev. 2.0 (1989); Methods for the
Determination of Organic Compounds in Drinking Water", U.S. Environmental
Protection Agency, Environmental Monitoring Systems Laboratory,
Cincinnati, OH, EPA/600/4-88/039, December, 1988.
2. "The Determination of Halogenated Chemicals in Water by the Purge and Trap
Method", Method 502.1; U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, OH 45268, September, 1986.
8021B - 11 Revision 2
January 1995
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3. "Volatile Aromatic and Unsaturated Organic Compounds in Water by Purge and
Trap Gas Chromatography", Method 503.1; U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory: Cincinnati, OH,
September, 1986.
4. Glaser, J.A., Forest, D.L., McKee, G.D., Quave, S.A., Budde, W.L. "Trace
Analyses for Wastewaters", Environ. Sci. Technol., 1981, 15, 1426.
5. Bellar, T.A., Lichtenberg, J.J. "The Determination of Synthetic Organic
Compounds in Water by Purge and Sequential Trapping Capillary Column Gas
Chromatography", U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, OH, 45268.
8021B - 12 Revision 2
January 1995
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TABLE 1
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL) FOR
VOLATILE ORGANIC COMPOUNDS WITH PHOTOIONIZATION DETECTION (PID) AND
HALL ELECTROLYTIC CONDUCTIVITY DETECTOR (HECD) DETECTORS
Analyte
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
1,1-Dichloroethene
Methylene Chloride
trans- 1,2-Di chl oroethene
1,1-Dichloroethane
2 , 2-Di chl oropropane
cis- 1,2-Di chloroethane
Chloroform
Bromochl oromethane
1 , 1 , 1 -Tri chloroethane
1,1-Dichloropropene
Carbon Tetrachloride
Benzene
1,2-Dichloroethane
Trichloroethene
1,2-Dichloropropane
Bromodi chl oromethane
Dibromomethane
Toluene
1 , 1 , 2-Tri chl oroethane
Tetrachl oroethene
1, 3 -Di chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Chlorobenzene
Ethyl benzene
1,1,1 , 2-Tetrachl oroethane
m-Xylene
p-Xylene
o-Xylene
Styrene
Isopropyl benzene
Bromoform
1,1,2 , 2-Tetrachl oroethane
1,2, 3 -Tri chl oropropane
PID
Ret. Time"
minute
b
-
9.88
-
-
_
16.14
-
19.30
-
-
23.11
-
-
-
25.21
-
26.10
-
27.99
-
-
-
31.95
-
33.88
-
-
-
36.56
36.72
-
36.98
36.98
38.39
38.57
39.58
-
-
-
HECD
Ret. Time
minute
8.47
9.47
9.93
11.95
12.37
13.49
16.18
18.39
19.33
20.99
22.88
23.14
23.64
24.16
24.77
25.24
25.47
-
26.27
28.02
28.66
29.43
29.59
-
33.21
33.90
34.00
34.73
35.34
36.59
-
36.80
-
-
-
-
-
39.75
40.35
40.81
PID
MDL
M9/L
0.02
NDC
0.05
0.02
0.02
0.009
0.02
0.01
0.05
0.003
0.005
0.01
0.01
0.02
0.01
0.05
HECD
MDL
M9/L
0.05
0.03
0.04
1.1
0.1
0.03
0.07
0.02
0.06
0.07
0.05
0.01
0.02
0.01
0.03
0.02
0.01
0.03
0.01
0.006
0.02
2.2
ND
0.04
0.03
0.03
0.8
0.01
0.005
1.6
0.01
0.4
8021B - 13
Revision 2
January 1995
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Analyte
TABLE 1
(Continued)
PID
Ret. Time8
minute
HECD
Ret. Time
minute
PID
MDL
HECD
MDL
n-Propylbenzene 40.87
Bromobenzene 40.99
1,3,5-Trimethylbenzene 41.41
2-Chlorotoluene 41.41
4-Chlorotoluene 41.60
tert-Butyl benzene 42.92
1,2,4-Trimethylbenzene 42.71
sec-Butyl benzene 43.31
p-Isopropyltoluene 43.81
1,3-Dichlorobenzene 44.08
1,4-Dichlorobenzene 44.43
n-Butylbenzene 45.20
1,2-Dichlorobenzene 45.71
1,2-Di bromo-3-Chloropropane
1,2,4-Trichlorobenzene 51.43
Hexachlorobutadiene 51.92
Naphthalene 52.38
1,2,3-Trichlorobenzene 53.34
Internal Standards
Fluorobenzene 26.84
2-Bromo-l-chloropropane
41.03
41.45
41.63
44.11
44.47
45.74
48.57
51.46
51.96
53.37
33.08
0.004
0.006
0.004
ND
0.02
0.06
0.05
0.02
0.01
0.02
0.007
0.02
0.05
0.02
0.06
0.06
ND
0.03
0.01
0.01
0.02
0.01
0.02
3.0
0.03
0.02
0.03
Retention times determined on 60 m x 0.75 mm ID VOCOL capillary column.
Program: Hold at 10'C for 8 minutes, then program at 4°C/min to 180°C, and
hold until all expected compounds have eluted.
b Dash (-) indicates detector does not respond.
0 ND = Not determined.
8021B - 14
Revision 2
January 1995
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TABLE 2
SINGLE LABORATORY ACCURACY AND PRECISION DATA
FOR VOLATILE ORGANIC COMPOUNDS IN WATERd
Photoionization
Detector
Hall Electrolytic
Conductivity Detector
Benzene
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
l,2-Dibromo-3-chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1,2 Dichloroethene
trans- 1 , 2-Di chl oroethene
1,2-Dichloropropane
1,3-Dichloropropane
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Isopropyl benzene
p-Isopropyl toluene
99
99
-
-
-
-
100
97
98
-
100
-
-
-
ND°
101
-
-
-
-
102
104
103
-
-
-
100
ND
93
-
-
-
103
101
99
98
98
1.2
1.7
-
-
-
-
4.4
2.6
2.3
-
1.0
-
-
-
ND
1.0
-
-
-
-
2.1
1.7
2.2
-
-
-
2.4
ND
3.7
-
-
-
3.6
1.4
9.5
0.9
2.4
_b
97
96
97
106
97
-
-
-
92
103
96
98
96
97
97
86
102
97
109
100
106
98
89
100
100
103
105
99
103
100
105
103
-
98
-
-
.
2.7
3.0
2.9
5.5
3.7
-
-
-
3.3
3.7
3.8
2.5
8.9
2.6
3.1
9.9
3.3
2.7
7.4
1.5
4.3
2.3
5.9
5.7
3.8
2.9
3.5
3.7
3.8
3.4
3.6
3.4
-
8.3
-
-
8021B - 15
Revision 2
January 1995
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TABLE 2
(Continued)
Analyte
Photoionization
Detector
Recovery,"
%
Standard
Deviation
of Recovery
Hall Electrolytic
Conductivity Detector
Standard
Recovery,8 Deviation
% of Recovery
Methyl ene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichl oroethane
1 , 1 , 2-Tri chl oroethane
Trichl oroethene
Tr i chl orof 1 uoromethane
1,2,3-Trichloropropane
1 , 2 , 4-Tri methyl benzene
1 , 3 , 5-Tri methyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
-
102
103
104
-
-
101
99
106
104
-
-
100
-
-
99
101
109
99
100
99
-
6.3
2.0
1.4
-
-
1.8
0.8
1.9
2.2
-
-
0.78
-
-
1.2
1.4
5.4
0.8
1.4
0.9
97
-
-
-
99
99
97
-
98
102
104
109
96
96
99
-
-
95
-
-
-
2.8
-
-
-
2.3
6.8
2.4
-
3.1
2.1
3.4
6.2
3.5
3.4
2.3
-
-
5.6
-
-
" Recoveries and standard deviations were determined from seven samples and spiked at
10 M9/L of each analyte. Recoveries were determined by internal standard method using a
purge-and-trap. Internal standards were: Fluorobenzene for PID, 2-Bromo-l-chloropropane
for HECD.
b Detector does not respond.
c NO = Not determined.
d This method was tested in a single laboratory using water spiked at 10 ^g/L (see
Reference 8).
8021B - 16
Revision 2
January 1995
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TABLE 3
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES"
Matrix Factor6
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
Sample EQLs are highly matrix dependent. The EQLs listed herein
are provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)].
For non-aqueous samples, the factor is on a wet-weight basis.
8021B - 17 Revision 2
January 1995
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FIGURE 1
GAS CHROMATOGRAM OF VOLATILE ORGANICS
1120.ET
QUUP2
M2CMCL
8021B - 18
Revision 2
January 1995
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METHOD 80218
HALOGENATED VOLATILES BY GAS CHROMATOGRAPHY USING PHOTOIONIZATION
AND ELECTROLYTIC CONDUCTIVITY DETECTORS IN SERIES:
CAPILLARY COLUMN TECHNIQUE
7.2 Set
chromatographic
condition*.
7.3 Refer to
Method 8000 for
calibration technique*.
7.4.1 Introduce
•ample into QC uemg
direct infection or
purge-and-trap.
7.4.4 Record
•ample volume
introduced into QC
end peak eizee.
7.4.5 Refer
to Method 8000 for
calculation*.
7.4.6 Are
analytical
interference*
•u*p*cted?
Reanalyze cample
ueing eecond QC
column.
Dilute end reonalyze
•econd aliquot of
•ample.
8021B - 19
Revision 2
January 1995
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METHOD 8032A
ACRYLAMIDE BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8032 is used to determine trace amounts of acrylamide monomer
(CAS No. 79-06-1) in aqueous matrices. This method may be applicable to other
matrices and to other similar analytes.
1.2 The method detection limit (MDL) in an aqueous matrix is 0.032 M9/L-
1.3 This method is restricted for use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8032 is based on bromination of the acrylamide double bond.
The reaction product (2,3-dibromopropionamide) is extracted from the reaction
mixture with ethyl acetate, after salting out with sodium sulfate. The extract
is cleaned up using a Florisil column, and analyzed by gas chromatography with
electron capture detection (GC/ECD).
2.2 Compound identification should be supported by at least one additional
qualitative technique. Analysis using a second gas chromatographic column or gas
chromatography/mass spectrometry may be used for compound confirmation.
3.0 INTERFERENCES
3.1 No interference is observed from seawater or in the presence of 8.0%
of ammonium ions derived from ammonium bromide.
3.2 Impurities from potassium bromide are removed by the Florisil cleanup
procedure.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatographic system
4.1.1 Gas chromatograph suitable for on-column injections with all
required accessories, including detector, analytical columns, recorder,
gases, and syringes. A data system for measuring peak heights and/or peak
areas is recommended.
4.1.2 GC Column - 2 m x 3 mm glass column, 5% FFAP (free fatty acid
polyester) on 60-80 mesh acid washed Chromosorb W, or equivalent.
4.1.3 Detector - electron capture detector.
8032A - 1 Revision 1
January 1995
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4.2 Separatory funnel - 150-mL.
4.3 Volumetric flask (Class A) - 100-mL, with ground-glass stopper; 25-mL,
amber, with ground-glass stopper.
4.4 Syringe - 5-mL.
4.5 Microsyringes - 5-^L, 100-juL.
4.6 Pipets (Class A).
4.7 Glass chromatography column (30 cm x 2 cm).
4.8 Mechanical shaker.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents - All solvent must be pesticide quality, or equivalent.
5.3.1 Ethyl acetate, C2H5C02C2H5
5.3.2 Diethyl ether, C2H5OC2H5. Must be free of peroxides as
indicated by test strips (EM Quant, or equivalent). Procedures for removal
of peroxides are provided with the test strips. After cleanup, 20 ml of
ethyl alcohol preservative must be added to each liter of ether.
5.3.3 Methanol, CH3OH
5.3.4 Benzene, C6He
5.3.5 Acetone, CH3COCH3
5.4 Saturated bromine water - Prepare by shaking organic-free reagent
water with bromine and allowing to stand for 1 hour, in the dark, at 4°C. Use
the aqueous phase.
5.5 Sodium sulfate (anhydrous, granular), Na2S04. Purify this reagent by
heating at 4008C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there is
no interference from the sodium sulfate.
8032A - 2 Revision 1
January 1995
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5.6 Sodium thiosulfate, Na2S203, 1 M aqueous solution.
5.7 Potassium bromide, KBr, prepared for infrared analysis.
5.8 Concentrated hydrobromic acid, HBr, specific gravity 1.48.
5.9 Acrylamide monomer, H2C:CHCONH2, electrophoresis reagent grade, minimum
95% purity.
5.10 Dimethyl phthalate, C6H4(COOCH3)2, 99.0% purity.
5.11 Florisil (60/100 mesh): Prepare Florisil by activating at 130°C for
at least 16 hours. Alternatively, store Florisil in an oven at 130°C. Before
use, cool the Florisil in a desiccator. Pack 5 g of the Florisil, suspended in
benzene, in a glass column (Sec. 4.8).
5.12 Stock standard solution
Prepare a stock standard solution of acrylamide monomer as described
below. 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 standards can be used at any concentration
if they are certified by the manufacturer or by an independent source.
Dissolve 105.3 mg of acrylamide monomer in organic-free reagent water
in a 100-mL volumetric flask, and dilute to the mark with organic-free
reagent water. Dilute the solution of acrylamide monomer so as to obtain
standard solutions containing 0.1 - 10 mg/L of acrylamide monomer.
5.13 Calibration standards
Dilute the acrylamide stock solution with organic-free reagent water
to produce standard solutions containing 0.1-5 mg/L of acrylamide. Prior
to injection the calibration standards are reacted and extracted in the
same manner as environmental samples (Sec. 7.0).
5.14 Internal standards
The suggested internal standard is dimethyl phthalate. Prepare a
solution containing 100 mg/L of dimethyl phthalate in ethyl acetate. The
concentration of dimethyl phthalate in the sample extracts and calibration
standards should be 4 mg/L.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
8032A - 3 Revision 1
January 1995
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7.0 PROCEDURE
7.1 Bromination
7.1.1 Pipet 50 ml of sample into a 100-mL glass-stoppered flask.
Dissolve 7.5 g of potassium bromide into the sample, with stirring.
7.1.2 Adjust the pH of the solution with concentrated hydrobromic
acid until the pH is between 1 and 3.
7.1.3 Wrap the flask with aluminum foil in order to exclude light.
Add 2.5 ml of saturated bromine water, with stirring. Store the flask and
contents in the dark, at 0°C, for at least 1 hour.
7.1.4 After reacting the solution for at least 1 hour, decompose the
excess of bromine by adding 1 M sodium thiosulfate solution, drop by drop,
until the solution becomes colorless.
7.1.5 Add 15 g of sodium sulfate and stir vigorously using a
magnetic stirrer.
7.2 Extraction
7.2.1 Transfer the solution into a 150-mL separatory funnel. Rinse
the reaction flask three times with 1-mL aliquots of organic-free reagent
water. Transfer the rinsings into the separatory funnel.
7.2.2 Extract the aqueous solution twice with 10-mL portions of
ethyl acetate for 2 min each extraction, using a mechanical shaker at
approximately 240 strokes per minute. Dry the organic phase with 1 g of
sodium sulfate.
7.2.3 Transfer the organic phase into a 25-mL amber volumetric
flask. Rinse the sodium sulfate with three 1.5-mL portions of ethyl
acetate and combine the rinsings with the organic phase.
7.2.4 Add exactly 100 p,g of dimethyl phthalate to the flask and make
the solution up to the 25 ml mark with ethyl acetate. Inject a 5-/uL
aliquot of this solution into the gas chromatograph.
7.3 Florisil cleanup - Whenever interferences are observed, the samples
should be cleaned up as follows.
7.3.1 Transfer the dried extract into an evaporation vessel with
15 ml of benzene. Evaporate the solvent at 70°C under reduced pressure,
and concentrate the solution to about 3 mL.
7.3.2 Add 50 mL of benzene and subject the solution to Florisil
column chromatography at a flow rate of 3 mL/min. Elute the column first
with 50 ml of diethyl ether/benzene (1:4) at a flow rate of 5 mL/min, and
then with 25 mL of acetone/benzene (2:1) at a flow rate of 2 mL/min.
Discard all of the first eluate and the initial 9 mL portion of the second
eluate, and use the remainder for the determination, using dimethyl
phthalate (4 mg/L) as an internal standard.
8032A - 4 Revision 1
January 1995
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NOTE: Benzene is toxic, and should be only be used in a ventilated laboratory
hood.
7.4 Gas chromatographic conditions
Nitrogen carrier gas flow rate: 40 mL/min
Column temperature: 165°C.
Injector temperature: 180°C
Detector temperature: 185°C.
Injection volume: 5 ij.1
7.5 Calibration
7.5.1 Inject 5 p.1 of a method blank (organic-free reagent water
carried through all sample storage, handling, bromination and extraction
procedures) into the GC.
7.5.2 Prepare a minimum of five standard solutions of acrylamide as
described in Sec. 5.13.1. One standard should be near the detection limit
of the method. The remaining four standards should bracket the expected
sample concentrations and cover the linear working range of the instrument.
Brominate and extract each standard solution as described in Sees. 7.1 and
7.2.
7.5.3 Inject 5 nl of each of the brominated and extracted standards,
and record the instrument response.
7.5.4 Calculate the response factor relative to the internal
standard for each calibration standard according to the guidance in Sec.
7.0 of Method 8000.
7.5.5 Calculate the mean, standard deviation, and relative standard
deviation of the response factors from the five calibration standards,
using the equations found in Sec. 7.0 of Method 8000.
7.5.6 If the RSD of the response factors is less than or equal to
20%, then the calibration can be assumed to be linear, and an average
response factor may be used to calculate sample results. If the RSD is
greater than 20%, see Method 8000 for alternative approaches to
calibration.
7.6 Sample analysis
7.6.1 Inject 5-juL portions of each sample extract (containing 4
mg/L internal standard) into the gas chromatograph. An example GC/ECD
chromatogram is shown in Figure 1.
7.6.2 The concentration of acrylamide monomer in the sample is
calculated according to the following equation.
f\ _1_ _!_• / /l\
Concentration (ug/L) =
(AX)(CJ(D)(V.)
(A.J(RF)(VJ(1000)
8032A - 5
Revision 1
January 1995
-------
where:
Ax = Area (or height) of the peak for the analyte in the sample.
Ajs = Area (or height) of the peak for the internal standard.
Cis = Concentration of the internal standard in the concentrated
sample extract (/ug/L).
D = Dilution factor, if the sample or extract was diluted prior
to analysis. If no dilution was made, D = 1. The dilution
factor is always dimensionless.
Vj = Volume of the extract injected (juL). The injection volume
_ for samples and calibration standards must be the same.
RF = Mean response factor from the initial calibration.
V. = Volume of the aqueous sample extracted or purged (ml). If
units of liters are used for this term, multiple the results
by 1000.
The 1000 in the denominator represents the number of /zL in 1 ml. If the
injection (Vi) is expressed in ml, then the 1000 may be omitted.
Using the units specified here for these terms will result in a
concentration in units of ng/mL, which is equivalent to M9/L-
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
8032A - 6 Revision 1
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prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The following performance data have been generated under the
conditions described in this method:
9.1.1 The calibration curve for Method 8032 has been found to be
linear over the range 0-5 |ug/L of acrylamide monomer.
9.1.2 In previous analyses, the limit of detection for an aqueous
matrix has been found to be 0.032 ptg/L.
9.1.3 The yields of the brominated compound have been found to be
85.2 ± 3.3% and 83.3 ± 0.9%, at fortification concentrations of 1.0 and 5.0
jug/L, respectively.
9.2 Table 1 provides the recoveries of acrylamide monomer from river
water, sewage effluent, and sea water.
9.3 The recovery of the bromination product as a function of the amount
of potassium bromide and hydrobromic acid added to the sample is shown in
Figure 2.
9.4 The effect of the reaction time on the recovery of the bromination
product is shown in Figure 3. The yield was constant when the reaction time was
more than 1 hour.
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9.5 Figure 4 shows the recovery of the bromination product as a function
of the initial pH from 1 to 7.35. The yield was constant within this pH range.
The use of conventional buffer solutions, such as sodium acetate - acetic acid
solution or phosphate solution, caused a significant decrease in yield.
10.0 REFERENCES
1. Hashimoto, A., "Improved Method for the Determination of Acrylamide Monomer
in Water by Means of Gas-Liquid Chromatography with an Electron-Capture
Detector", Analyst, 101:932-938, 1976.
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TABLE 1
RECOVERY OF ACRYLAMIDE FROM WATER SAMPLES AS 2,3-DIBROMOPROPIONAMIDE
Acryl amide
Sample Monomer
Matrix Spiked//zg
Standard
River Water
Sewage
Effluent
Seawater
0.05
0.20
0.25
0.20
0.20
0.20
Amount of Bromi nation Recovery of
2,3-DBPA"/]Lig Recovery Acryl amide
Calculated Foundb %b Monomer (%)b
0.162
0.649
0.812
0.649
0.649
0.649
0.138
0.535
0.677
0.531
0.542
0.524
85.2
82.4
83.3
81.8 99.4
83.5 101.3
80.7 98.8
RSD (%)
3.3
1.0
0.9
2.5
3.0
3.5
" 2,3-Dibromopropionamide
b Mean of five replicate determinations
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FIGURE 1
02 4 f I 10 12 14 16
Typical gas chromatograms of the bromination product obtained from aqueous
acrylamide monomer solution:
A. Untreated
B. With Florisil cleanup
BL. Chromatogram of blank, concentrated
chromatographic analysis.
Peaks:
1. 2,3-Dibromopropionamide
2. Dimethyl phthalate
4-7. Impurities from potassium bromide
Sample size = 100 ml; acrylamide monomer = 0.1 /ig
five-fold before gas
8032A - 10
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FIGURE
*
•
<•
o
u
0 5 10 15 20 23
Amount of KBr/g ptr 50 ml
i i i i t L
0 24 6 8 10
Amount of H8r/ml ptr 50 ml
Effect of (A) potassium bromide and (B) hydrobromic acid on the
yield of bromination.
Sample size = 50 ml;
Acrylamide monomer = 0.25 /Ltg
8032A - 11
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FIGURE 3
Effect of reaction time on the bromination. Reaction conditions;
50 mi of sample;
0.25 iiq of acrylamide monomer;
7.5 g of potassium bromide;
2.5 ml of saturated bromine water
Extraction conditions:
15 g of sodium sulfate;
extraction at pH 2;
solvent = 10 mL of ethyl acetate (X2)
8032A - 12
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FIGURE 4
100
| so
o
I
i i t
0 1 J 3 4 S • 7 •
pH
Effect of initial pH on the bromination. Reaction and extraction
conditions as in Figure 3. The pH was adjusted to below 3 with
concentrated hydrobromic acid, and to 4 - 5 with dilute hydrobromic
acid. Reaction at pH 6 was in distilled water. A pH of 7.35 was
achieved by careful addition of dilute sodium hydroxide solution.
The broken line shows the result obtained by the use of sodium
acetate - acetic acid buffer solution.
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METHOD 8032A
ACRYLAMIDE BY GAS CHROMATOGRAPHY
Start
f
71
BrominaOon
7 1 1 Dissolve 7 5 g KBr into
50 ml sample in flask.
7 1 .2 Adjust soln. pH wrth
concentrated HBr to between
1 and 3
7.1.3 Wrap sola (ask with
aluminum. Add 2.5 ml satd.
bromine water, stir, store at
0 C for 1 hr.
7 1 4 Add 1 M sodium
thiosulfate dropwne to flask to
decompose excess bromine.
7 1 5 Add 15 g sodium
suttate, and stir.
7.2 Extractor)
7.2.1 Transfer flask sotn. to
sep funnel along with rinses
*
7.2.2 Extract som. twice w/ethyl
acetate. Dry organic phase
using sodium suttate
I
7.2.3 Transfer organic phase
and rinses into amber
glass flask.
7.2.4 Add 100 ug dimethyl
phthalate to flask. dKute to
mark. Inject 5 ul into GC
I
7 3 Rorisil Cleanup
7.3.1 Transfer dned extract to
K-D assembly w/benzene.
Concentrate to 3 mL at 70 C
under reduced pressure.
7.3.2 Add 50 mL benzene to
solution. Pass sola through
Rortsil column. Bute with
diethyl etwr/benzene, then
acetone/benzene. Collect
the second elubon tram (less
initial 9 ml) for analysis.
8032A - 14
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METHOD 8032A
continued
©
7 4 GC Conditions
7.5 Calibration
7 5 1 Infect 5 ul. sample blank.
7.5.2 Brominate and extract std.
sotns similar to the samples.
1 Infect 5 ul -i each of (he
minimum •; stris.
.2 Plot peak are vs. [ ].
.3 Calculate response factor
(RF) for each [ ].
7.5.3 Calculate mean RF from
eqn.2.
7.6 QC Analysis
7.6.1 Infect 5 uL sample containing
internal std. Into QC.
7.6.2 Calculate acrylamkte monomer
conoentratton in sample USWIQ
eqn.3.
8032A - 15
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METHOD 8033
ACETONITRILE BY GAS CHROMATOGRAPHY WITH NITROGEN-PHOSPHORUS DETECTION
1.0 SCOPE AND APPLICATION
1.1 Method 8033 may be used to determine the concentration of acetonitrile
(CAS No. 75-05-8) in aqueous matrices.
1.2 The method detection limit in water is approximately 1.7 to 2.8
picograms (1.7 to 2.8 jig/L with a 1 /iL injection) at 40 and 20 volts offset,
respectively. The upper limit of the range is approximately 5000 jug/L and is
limited by the non-linearity of the nitrogen-phosphorus detector (NPD). The
range may be extended by diluting the sample. This procedure may be applicable
to other matrices where the target compound can be extracted into water.
1.3 This method is also applicable to aqueous condensate collected from
the VOST or semi-VOST sampling trains.
1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
Water samples containing acetonitrile are introduced into a GC/NPD using
cool on-column direct aqueous injection. The GC/NPD system is externally
calibrated using a series of acetonitrile standards diluted in organic-free
reagent water. A series of blank organic-free reagent water samples are directly
injected until no peaks are observed in the chromatogram. The acetonitrile
concentration is quantitated using the external calibration method.
3.0 INTERFERENCES
3.1 There are no known chromatographic interferants when samples are
analyzed using the optimum operating conditions. However, at column temperatures
greater than 60°C, water becomes an interferant, as it coelutes with acetonitrile
and produces a negative disruption of the baseline, thereby preventing accurate
quantitations. This problem is eliminated by operating the column at 35°C. At
this temperature, water will eventually elute and produce a positive disruption
of the baseline. However, this effect can be minimized by heating the column to
120°C for 1 hour after a day's analysis.
3.2 Glassware containing traces of chromium should not be used because
chromium will form a complex with acetonitrile, thereby reducing the amount of
acetonitrile available for quantitation.
3.3 Reagents, glassware, and other sample processing hardware may yield
discrete artifacts and/or elevated baselines causing misinterpretation of gas
8033 - 1 Revision 0
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chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for cool on-column injections and all required
accessories, including detector, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Column: DB-WAX, 15 m X 0.53 mm, 1.0 /xm capillary column (J&W
Scientific or equivalent).
4.1.3 Detector: Nitrogen-Phosphorus (NPD).
4.2 Bubble flow-meter: 0-10 and 0-100 cm3/m\n, for measuring GC gas
flows.
4.3 Syringes: 10 p,L capacity, equipped with fused silica needles. Use
of a syringe with a stainless steel needle has not been evaluated.
4.4 Volumetric flasks: Class A, sizes as appropriate.
4.5 Pipets: Class A, assorted sizes.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Gases
5.3.1 Helium gas - 99.9999% purity, for carrier and make-up gas.
Helium is preferred as the make-up gas over nitrogen because helium
produces a more stable baseline with less noise.
5.3.2 Air - zero grade.
5.3.3 Nitrogen - zero grade.
5.4 Acetonitrile, CH3CN - 99.9% purity.
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5.5 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.6 Stock standard solutions - Prepare from pure standard materials or
purchase as certified solutions.
5.6.1 Prepare stock standard solutions by accurately weighing the
appropriate amount of acetonitrile and dissolving it in organic-free
reagent water to achieve a final concentration of at least 1000 mg/L.
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.
5.6.2 Transfer the stock standard solutions into bottles with
Teflon®-!ined screw-caps. The stock standard solution is stable at 20-
25°C for about 3 months. However, stock standard solutions should be
checked frequently for signs of degradation or evaporation, especially
when using them to prepare calibration standards.
5.8 Working standard solutions - Prepare working standards fresh daily by
diluting the stock standard solutions with organic-free reagent water. Prepare
at least five different acetonitrile concentrations to bracket the expected
concentration range of the samples.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1. Acetonitrile has a boiling point of approximately 81.6°C and should
therefore be handled as a volatile compound.
6.2 Calibration standards are stable for at least one day. Traces of
chromium in the glassware used to prepare the calibration standards may adversely
affect longer term stability.
7.0 PROCEDURE
7.1 This procedure utilizes direct aqueous injection (DAI) as the sample
introduction technique. Samples of condensate from a VOST or semi-VOST sampling
trains may need to be diluted to an appropriate concentration prior to analysis.
7.2 Chromatographic Conditions (Recommended):
Collector distance above the jet: 1 mm.
Column temperature: 35'C.
Carrier gas (He) flow rate: 7 cm3/min.
Make-up gas (He) flow rate: 26 cm3/min.
H2 flow rate: 3.5 cm3/min.
Air flow rate: 80 cm3/min.
Injection temperature: ambient temperature
Injection volume: 1 jiL
Detector temperature: 300°C.
8033 - 3 Revision 0
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Detector: Nitrogen-Phosphorus (NPD) operated at the conditions
specified by the manufacturer.
7.2.1 Under these chromatographic conditions the retention time of
acetonitrile is approximately 4.4 minutes. The offset voltage is very
sensitive to H2 flow rate.
7.2.2 Maximum control of the H2 flow rate is obtained by using a
pressure regulator at the instrument. The stability of the offset voltage
during the day's analysis will be enhanced by allowing the NPD to
equilibrate overnight, prior to carrying out an analysis.
7.3 Calibration - External standard calibration should be utilized for
this single analyte procedure. Refer to Method 8000 for guidance on
implementation of an external standard initial and continuing calibration
procedure.
7.4 Calibration standard and sample analysis
7.4.1 Allow the baseline to stabilize at the desired voltage.
Inject blank samples of water (a 1.0 juL volume is recommended) until no
peaks are observed in the chromatogram. An occasional positive baseline
disruption (caused by the water) may occur.
NOTE: The acetonitrile concentration of any standard or sample will decrease
with time if left exposed to the atmosphere. Use appropriate precautions
or use fresh sample for each injection to minimize this effect.
7.4.2 Inject the 5 calibration standards in order of increasing
concentration. It may be necessary to condition the syringe with organic-
free reagent water by rinsing it a minimum of 15 times between injections
to achieve acceptable method performance. Calculate the calibration
factors (CF) for each standard to determine the linearity of the
calibration.
7.4.3 Inject the sample into the GC. If the response for the
sample exceeds that of the initial calibration range, the sample must be
diluted and reanalyzed. Due to the relatively short retention time of the
target compound and potential instability of the detector, each sample or
sample dilution may need to be analyzed in duplicate. To achieve the
precision described in Section 9.0, it may be necessary to condition the
syringe with organic-free reagent water by rinsing it a minimum of 15
times between injections.
7.4.4 Calibration check samples should be analyzed after every 10
samples.
7.5 Refer to Method 8000 for guidance on GC peak identification, peak
confirmation, and sample concentration calculations.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicate a potential problem due to the sample matrix itself, the
LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
8033 - 5 Revision 0
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9.0 METHOD PERFORMANCE
Using laboratory produced samples containing 0.031 ng//iL and 0.33 ng//uL
acetonitrile in H20 and a 1.0 /zL injection volume, the relative standard
deviation was 2.0% and 1.4%, respectively. Analysis of an unknown (QC sample)
showed that calibration with external standards was accurate within ± 4% of the
true value. See Figure 1 for sample chromatogram of acetonitrile.
10.0 REFERENCES
1. "Draft Method for the Analysis of Acetonitrile in a Water Matrix", U.S.
Environmental Protection Agency, AREAL/RTP, April 3, 1990.
2. Margeson, J., Memorandum: Evaluation of Method 8033 (Acetonitrile by
GC/NPD), U.S. Environmental Protection Agency, AREAL/Source Methods
Research Branch, RTP, NC, August 14, 1991.
8033 - 6 Revision 0
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FIGURE 1
CHROMATOGRAM OF ACETONITRILE STANDARD
(Retention Time = 4.33 minutes)
o
cy
8033 - 7
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METHOD 8033
ACETQNITRILE BY GAS CHROMATOGRAPHY
WITH NITROGEN-PHOSPHORUS DETECTION
1
r
7.2 Establish
appropnata
chromatographic
conditions.
7.3 - 7.4
2 Calibrate
(refer to Method
8000 for
1
guidance).
r
7.4.3 Inject the
sample
Method
(refer to
8000 for
guidance on peak
identification, peak
confirmation, con-
centration calculations.
7.4.3
Does
sample response
exceed calibration
range?
7.4.3 Dilute
sample.
No
7.5 Perform
calculations.
>
1
8033 - 8
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METHOD 8041
PHENOLS BY GAS CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8041 describes open-tubular, capillary column gas
chromatography procedures for the analysis of phenols, using both single-column
and dual-column/dual-detector approaches. The following RCRA target analytes can
be determined by this method:
Compound Name
CAS No.1
Appropriate Technique
3510 3520 3540 3550 3580
4 - Chi oro- 3 -methyl phenol
2-Chlorophenol
2-Cyclohexyl-4,6-dinitrophenol
2,4-Dichlorophenol
2,6-Dichlorophenol
2,4-Ditnethylphenol
Dinoseb (DNBP)
2,4-Dinitrophenol
2 -Methyl -4,6-dinitrophenol
2-Methylphenol (o-Cresol)
3-Methylphenol (m-Cresol)
4-Methylphenol (p-Cresol )
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,3,4,5-Tetrachlorophenol
2,3,4,6-Tetrachlorophenol
2,3,5,6-Tetrachlorophenol
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
59-50-7
95-57-8
131-89-5
120-83-2
87-65-0
105-67-9
88-85-7
51-28-5
534-52-1
95-48-7
108-39-4
106-44-5
88-75-5
100-02-7
87-86-5
108-95-2
4901-51-3
58-90-2
935-95-5
95-95-4
88-06-2
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
DC(28)
ND
X
ND
X
X
X
X
ND
X
ND
X
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
X
X
X
ND
X
ND
X
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
ND
X
X
X
ND
X
ND
X
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
a Chemical Abstract Services Registry Number.
DC = Unfavorable distribution
recovery) .
LR = Low response.
ND = Not determined.
X = Greater than 70 percent
coefficient (number i
recovery by thi
s techni
n parenthesis
que.
is
percent
1.2 The single-column approach involves the use of a wide-bore fused-
silica open tubular column for analysis. The fused-silica, open-tubular wide-
bore column offers improved resolution, better selectivity, increased
sensitivity, and faster analysis than packed columns.
8041 - 1
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1.3 The dual-column/dual-detector approach involves the use of two wide-
bore fused-silica open-tubular columns of different polarities. The columns are
connected to an injection tee and two identical detectors.
1.4 Phenols may be analyzed underivatized by FID, although the sensitivity
of the method may not be suitable for all needs.
1.5 This method also includes a procedure for the derivatization of the
phenols and identification of the target phenols as methylated phenols (anisoles)
and as pentafluorobenzy] ether derivatives (PFBBr). Three phenols failed to
derivatize under the PFBBr method protocol: 2,4-dinitrophenol, 2-methyl-4,6-
dinitrophenol, and Dinoseb.
1.6 The following non-RCRA analytes may also be analyzed by this method:
Compound CAS No.
2-Chloro-5-methylphenol 615-74-7
4-Chloro-2-methyl phenol 1570-64-5
4-Chloro-3-methylphenol 59-50-7
3-Chlorophenol 108-43-0
4-Chlorophenol 106-48-9
2,3-Dichlorophenol 576-24-9
2,5-Dichlorophenol 583-78-8
3,4-Dichlorophenol 95-77-2
3,5-Dichlorophenol 591-35-5
2,3-Dimethylphenol 526-75-0
2,5-Dimethylphenol 95-87-4
2,6-Dimethylphenol 576-26-1
3,4-Dimethylphenol 95-65-8
2,5-Dinitrophenol 329-71-5
3-Nitrophenol 554-84-7
2,3,4-Trichlorophenol 15950-66-0
2,3,5-Trichlorophenol 933-78-8
2,3,6-Trichlorophenol 933-75-5
1.7 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.
1.8 Only experienced analysts should be allowed to work with diazomethane
due to the potential hazards associated with its use (explosive, carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Samples are extracted using an appropriate sample preparation method.
Prior to analysis, the extracts are cleaned up, as necessary, and the solvent
exchanged to 2-propanol.
2.2 Underivatized phenols may be analyzed by FID, using either the single-
column or dual-column approach.
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2.3 The target phenols also may be derivatized with diazomethane or
pentafluorobenzyl bromide (PFBBr) and analyzed by gas chromatography.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
syringe used for injection must be thoroughly rinsed between samples with
solvent. Whenever a highly concentrated sample is encountered, it should be
followed by the analysis of a solvent blank to check for cross-contamination.
Column blanks should be analyzed whenever the analysis of a solvent blank
indicates cross-contamination.
3.2 In certain cases some compounds coelute on either one or both columns.
In these cases the compounds must be reported as coeluting. The mixture can be
reanalyzed by GC/MS techniques, see Section 8.0 and Method 8270.
3.3 Non-specific interferences may occur in the analysis of the
underivatized phenols, reducing the sensitivity of the method.
3.4 The phenols listed in Sections 1.1 and 1.6 were derivatized with a-
Bromo-2,3,4,5,6-pentafluorotoluene (PFBBr) according to the method by Lee, et al.
(Ref. 1). Five compound pairs coeluted on the DB-5 column and three compound
pairs coeluted on the DB-1701 column.
DB-5: 2,6-dimethyl phenol/2,5-dimethyl phenol
2,4-dimethyl phenol/2-chlorophenol
2,6-dichlorophenol/4-chloro-2-methylphenol
2,4,5-trichlorophenol/2,3,5-trichlorophenol
2,3,4,5-tetrachlorophenol/2,5-dinitrophenol
DB-1701: 3-chlorophenol/3,4-dimethyl phenol
2,4-dichlorophenol/3,5-dichlorophenol
2,4,5-trichlorophenol/2,3,5-trichlorophenol
In addition, 3-methylphenol is only partially resolved from 4-methylphenol
on the two columns, and 2-chlorophenol is only partially resolved from 2,3-
dimethylphenol on the DB-1701 column.
3.5 Sample extracts should be dry prior to methylation or else poor
recoveries will be obtained.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - An analytical system complete with a gas
chromatograph suitable for on-column injection, and all required accessories,
including syringes, analytical columns, gases, flame ionization detector (FID),
electron capture detector (ECD), and a data system.
4.2 GC columns - This method describes procedures for both single-column
and dual-column analyses. The single-column approach involves one analysis to
determine that a compound is present, followed by a second analysis to confirm
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the identity of the compound (Sec. 8.0 describes how GC/MS confirmation
techniques may be employed). Both the single-column approach and the dual-column
approaches employ wide-bore (0.53 mm ID) columns. The dual-column approach
involves a single injection that is split between two columns that are mounted
in a single gas chromatograph.
The columns listed in this section were the columns used to develop the
method performance data. Listing these columns in this method is not intended
to exclude the use of other columns that may be developed. Laboratories may use
other capillary columns provided that they document method performance data
(e.g., chromatographic resolution, analyte breakdown, and MDLs) that demonstrate
performance appropriate for the intended application.
4.2.1 Column 1 - 30 m x 0.53 mm ID fused-silica open- tubular
column, cross-linked and chemically bonded with 95 percent dimethyl and 5
percent diphenyl-polysiloxane (DB-5, RTx-5, SPB-5, or equivalent), 0.83 ^m
or 1.5 urn film thickness.
4.2.2 Column 2 - 30 m x 0.53 mm ID fused-silica open-tubular column
cross-linked and chemically bonded with 14 percent cyanopropylphenyl and
86 percent dimethyl-polysiloxane (DB-1701, RTX-1701, or equivalent), 1.0
//m film thickness.
4.3 Splitter - When the dual-column approach is employed, the two columns
must be connected with a splitter such as those listed below (or equivalent).
4.3.1 J&W Scientific press-fit Y-shaped glass 3-way union splitter
(J&W Scientific, Catalog no. 705-0733).
4.3.2 Supelco 8-in glass injection tee, deactivated (Supelco,
Catalog no. 2-3665M).
4.3.3 Restek Y-shaped fused-silica connector (Restek, Catalog no.
20405).
4.4 Column rinsing kit - Bonded-phase column rinse kit (J&W Scientific,
Catalog no. 430-3000 or equivalent).
4.5 Diazomethane generators - Refer to Sec. 7.3 to determine which method
of diazomethane generation should be used for a particular generation.
4.5.2 As an alternative, assemble from two 20 mm x 150 mm test
tubes, two Neoprene rubber stoppers, and a source of nitrogen. Use
Neoprene rubber stoppers with holes drilled in them to accommodate glass
delivery tubes. The exit tube must be drawn to a point to bubble
diazomethane through the sample extract. The generator assembly is shown
in Figure 6.
4.5.1 Diazald kit - Recommended for the generation of diazomethane
(Aldrich Chemical Co., Catalog No. 210,025-0, or equivalent).
4.6 PFBBr Derivatization equipment - 10-mL graduated concentrator tubes
with screw caps, disposable pipets, beakers, and water bath.
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5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the chemicals are of sufficiently high purity to permit
their use without affecting the accuracy of the determinations.
5.2 Store the standard solutions (stock, composite, calibration, internal,
and surrogate) at 4°C in Teflon-sealed containers in the dark. All standard
solutions must be replaced after six months or sooner if routine QC (Section 8.0)
indicates a problem.
5.3 Solvents - all solvents must be pesticide quality or equivalent.
5.3.1 Hexane, C6H14
5.3.2 Acetone, CH3COCH3
5.3.3 Isooctane, (CH3)3CCH2CH(CH3)2
5.4 Stock standard solutions (1000 mg/L) - May be prepared from pure
standard materials or may be purchased as certified solutions.
5.4.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10 ml volumetric flask. If compound purity is
96 percent or greater, the weight may be used without correction to
calculate the concentration of the stock standard solution.
5.4.2 Transfer the stock standard solutions into bottles with
teflon lined screw-caps or crimp tops. Store at 4°C and protect from
light. Stock standards must be replaced after one year or sooner if
comparison with check standards indicate a problem. Stock standards
should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.4.3 Commercially-prepared stock standard solutions may be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.5 Composite stock standard - May be prepared from individual stock
solutions. For composite stock standards containing less than 25 components,
transfer exactly 1 ml of each individual stock solution at 1000 mg/L, add
solvent, mix the solutions and bring to volume in an appropriate volumetric
flask. This composite solution may be further diluted to obtain the desired
concentrations.
5.6 Calibration standards - These should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with hexane or other
appropriate solvent. The solvent or solvents used to dilute the standards should
be the same as the final solvent mixture in the sample extracts to be analyzed.
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The standard concentrations should correspond to the expected range of
concentrations present in the field samples and should bracket the linear range
of the detector. Concentrations of the target analytes at 5, 25, 50, 100, and
200 mg/L (except for 2,4- and 2,5-dinitrophenol and 2-methyl-4,6-dinitrophenol
at about 2x the given values) have been used as calibration solutions in soil
recovery studies. All standards should be prepared from the target phenols.
When derivatization is employed, the phenol standards should be prepared, and
then derivatized in the same fashion as the sample extracts, prior to
calibration.
5.7 Internal standard - When internal standard calibration is used,
prepare a solution of 1000 mg/L of 2,5-dibromotoluene and 2,2',5,5'-
tetrabromobiphenyl. For spiking, dilute this solution to 50 ng/juL. Use a
spiking volume of 10 juL/ml_ of extract. The spiking concentration of the
internal standards should be kept constant for all samples and calibration
standards.
5.8 Surrogate standard - The performance of the method should be monitored
using surrogate compounds. Surrogate standards are added to all samples, method
blanks, matrix spikes, and calibration standards. Prepare a solution of
1000 mg/L of 2,4-dibromophenol and dilute it to 1.6 ng/^L. Use a spiking volume
of 100 /it for a 1 L aqueous sample. Other appropriate surrogates are listed in
Sec. 1.6.
5.9 Reagents for derivatization
NOTE: Other derivatization techniques may be employed, provided that the analyst
can demonstrate acceptable precision and accuracy for the target compounds
(see Sec. 8.0) and for the particular application.
5.9.1 Diazomethane Derivatization
5.9.1.1 N-methyl-N-nitroso-p-tol uenesul fonamide (Diazald).
High purity (Aldrich Chemical Co., or equivalent).
5.9.1.2 Diethyl Ether stabilized with BHT, C2H5OC2H5. Must
be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with
the test strips. If ethanol stabilized diethyl ether is used, the
methylation reaction may not proceed efficiently.
5.9.1.3 Silicic acid, H2Si05. 100 mesh powder, store at
130°C.
5.9.1.4 HPLC-grade hexane.
5.9.2 PFBBr Derivatization
5.9.2.1 Standards for the target phenols are purchased as
phenols and derivatized prior to calibration.
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5.9.2.2 a-Bromo-2,3,4,5,6-pentafluorotoluene (PFBBr reagent)
- Dissolve 0.500 g of PFBBr in 9.5 ml acetone. Store in the dark at
4°C. Prepare fresh reagent biweekly.
5.9.2.3 Potassium carbonate solution (10 percent) - Dissolve
1 g anhydrous potassium carbonate in water and adjust volume to 10
ml.
5.9.2.4 HPLC-grade acetone.
5.9.2.5 HPLC-grade hexane.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
6.2. It is recommended that extracts to be methylated undergo
derivatization within 48 hours after extraction and methylated extracts be
analyzed immediately after derivatization to minimize other reactions that may
occur.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two and Method 3500 for guidance on choosing
the appropriate extraction procedure.
7.1.1.1 Water samples are extracted at a pH of less than or
equal to 2 with methylene chloride, using Method 3510 or 3520.
7.1.1.2 Solid samples are extracted using either Method 3540
or 3550, and non-aqueous sample using Method 3580. Acid-Base
Partition Cleanup using Method 3650 is suggested for extracts
obtained from application of either Method 3540 or 3550.
7.1.1.3 Other aqueous liquid or solid 3500 series extraction
techniques in this manual may be appropriate for this method.
7.1.2 If the phenols are to be determined without derivatization,
proceed to Sec. 7.2.
to be determined by derivatization,
concentrated down to 1 mL using
the
an
7.1.3 If the phenols are
extraction solvent should be
appropriate concentration technique. If the sample is to be analyzed by
GC/ECD the extraction solvent (methylene chloride) will need to be
exchanged to hexane or some other nonhalogenated compatable solvent. If
being performed, the sample should be
4 mL with diethyl ether. If PFB
methylation
diluted to
with diazomethane is
a final volume of
derivatization
volume of 4 mL
is being performed, the sample should be diluted to a final
with acetone.
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NOTE: It is very critical to ensure that the sample is dry when preparing it for
methylation. Any moisture remaining in the extract will result in low
methylated phenol recoveries. It may be appropriate to add approximately
10 g of acidified anhydrous sodium sulfate to the extract prior to
concentration and, periodically, vigorously shake the extract and drying
agent. The amount of sodium sulfate is adequate if some free flowing
crystals are visible when swirling the flask. If all of the sodium
sulfate solidifies in a cake, add a few additional grams of acidified
sodium sulfate and again test by swirling. The 2 hour drying time is a
minimum, however, the extracts may be held in contact with the sodium
sulfate overnight.
7.1.3.1 If the phenols are to be determined by methylation
derivatization, proceed to Sec. 7.3.
7.1.3.2 If the phenols are to be determined by PFBBr
derivatization, proceed to Sec. 7.4.
7.2 If the phenols are to be determined without derivatization then, prior
to gas chromatographic analysis, the extraction solvent must be exchanged to 2-
propanol. The exchange is performed as follows:
7.2.1 Concentrate the extract to 1 ml using the macro-Snyder column
and allow the apparatus to cool and drain for at least ten minutes.
7.2.2 Remove the micro-Snyder column and rinse its lower joint into
the concentrator tube with a small amount of 2-propanol. Adjust the
extract volume to 1.0 ml.
7.2.3 Stopper the concentrator tube and store refrigerated at 4°C
if further processing will not be performed immediately. If the extract
will be stored longer than two days, it should be transferred to a vial
with a Teflon-lined screw-cap or crimp top.
7.2.4 If the phenols are to be determined without derivatization,
proceed with gas chromatographic calibration and analysis (Sections 7.5,
7.6, and 7.7).
NOTE: Other derivatization techniques may be employed, provided that the analyst
can demonstrate acceptable precision and accuracy for the target compounds
(see Sec. 8.0).
7.3 Methylation derivatization procedures
7.3.1 Diazomethane derivatization - Two methods may be used for the
generation of diazomethane: the bubbler method, Sec. 7.3.1, and the
Diazald kit method, Sec. 7.3.2. The methylation of phenolic compounds for
this analysis procedure has been documented for the Diazald kit only
(Tables 3 and 4). However, the bubbler method should also be applicable.
CAUTION: Diazomethane is a carcinogen and can explode under certain conditions.
The bubbler method is suggested when small batches of samples (10 -
15) require methylation. The bubbler method works well with samples that
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have low concentrations of phenols (e.g., aqueous samples) and is safer to
use than the Diazald kit procedure. The Diazald kit method is good for
large quantities of samples needing methylation. The Diazald kit method
is more effective than the bubbler method for soils or samples that may
contain high concentrations of phenols (e.g., samples such as soils that
may result in yellow extracts following hydrolysis may be difficult to
handle by the bubbler method).
The diazomethane derivatization procedures described below will
react efficiently with all of the phenols described in this method and
should be used only by experienced analysts, due to the potential hazards
associated with its use.
The following precautions should be taken:
• Use a safety screen.
• Use mechanical pipetting aides.
• Do not heat above 90°C - EXPLOSION may result.
• Avoid grinding surfaces, ground-glass joints, sleeve bearings, and
glass stirrers - EXPLOSION may result,
• Store away from alkali metals - EXPLOSION may result.
• Solutions of diazomethane decompose rapidly in the presence of solid
materials such as copper powder, calcium chloride, and boiling
chips.
7.3.2 Bubbler method - Assemble the diazomethane bubbler (see
Figure 1).
7.3.2.1 Add 5 mL of diethyl ether to the first test tube.
Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5 mL of 37% KOH, and
0.1 - 0.2 g of Diazald to the second test tube. Immediately place
the exit tube into the concentrator tube containing the sample
extract. Apply nitrogen flow (10 mL/min) to bubble diazomethane
through the extract for 10 minutes or until the yellow color of
diazomethane persists. The amount of Diazald used is sufficient for
methylation of approximately three sample extracts. An additional
0.1 - 0.2 g of Diazald may be added (after the initial Diazald is
consumed) to extend the generation of the diazomethane. There is
sufficient KOH present in the original solution to perform a maximum
of approximately 20 minutes of total methylation.
7.3.2.2 Remove the concentrator tube and seal it with a
Neoprene or Teflon® stopper. Store at room temperature in a hood
for 20 minutes.
7.3.2.3 Destroy any unreacted diazomethane by adding 0.1 -
0.2 g of silicic acid to the concentrator tube. Allow to stand
until the evolution of nitrogen gas has stopped. Adjust the sample
volume to 10.0 mL with hexane. Stopper the concentrator tube or
transfer 1 mL of sample to a GC vial, and store refrigerated if
further processing will not be performed immediately. Analyze by
gas chromatography.
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7.3.2.4 Extracts should be stored at 4°C away from light. It
is recommended that the methylated extracts be analyzed immediately
after derivatization to minimize other reactions that may occur.
7.3.3 Diazald kit method - Instructions for preparing diazomethane
are provided with the generator kit.
7.3.3.1 Add 2 ml of diazomethane solution and let the sample
stand for 10 minutes with occasional swirling. The yellow color of
diazomethane should be evident and should persist for this period.
7.3.3.2 Rinse the inside wall of the ampule with 700 yl of
diethyl ether. Reduce the sample volume to approximately 2 ml to
remove excess diazomethane by allowing the solvent to evaporate
spontaneously at room temperature. Alternatively, 10 mg of silicic
acid can be added to destroy the excess diazomethane.
7.3.3.3 Dilute the sample to 10.0 ml with hexane. Analyze by
gas chromatography. It is recommended that the methylated extracts
be analyzed immediately to minimize other reactions that may occur.
Proceed to Sec. 7.5.
7.4 PFBBr derivatization procedure - Calibration standards and sample
extracts should be derivatized using the same procedures.
7.4.1 Using the individual phenol stock solutions at 1000 mg/L make
a composite solution and dilute with hexane or other appropriate solvent
to the appropriate concentrations for the calibration range of the
analysis.
7.4.2 Sample extracts should be in hexane and diluted to 4 mL with
acetone according to the procedure in Sec. 7.1.3.
WARNING; PFBBr is a lachrymator.
7.4.3 Add 100 p.1 of calibration standards and sample extracts to
8 ml acetone in a 10 ml graduated concentrator tube with screw caps. Add
100 nL of 5% PFBBr reagent and 100 /uL of K2C03 solution to the composite
standard.
7.4.4 Cap the tubes tightly and gently shake the contents. Heat
the tube in a water bath at 60°C for one hour.
7.4.5 After the reaction is complete, cool the solution and
concentrate it to 0.5 ml, using nitrogen blowdown.
7.4.6 Add 3 ml of hexane and concentrate the solution to a final
volume of 0.5 ml. If cleanup is not to be performed, proceed to Sec. 7.5
for the analysis of samples by GC/ECD.
7.4.7 Cleanup
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7.4.7.1 Refer to Method 3630 (Silica Gel Cleanup) for
specific instructions on the cleanup of derivatized phenols.
7.4.7.2 Following column clea
analysis of the samples using GC/ECD.
cleanup, proceed to Sec. 7.5 for
'rrn
7.5 GC Conditions - This method allows the analyst to choose between a
single-column or a dual-column configuration in the injector port. Either wide-
or narrow-bore columns may be used. Identifications based on retention times
from a single-column need to be confirmed on a second column or with an
alternative qualitative technique. The recommended gas chromatographic columns
and operating conditions for the instrument are provided in Figures 1, 2, 3, and
4 and Table 5.
7.6 Calibration
7.6.1 Prepare the calibration standards according to the guidance
in Sec. 5.6. Concentrations of the target analytes at 5, 25, 50, 100, and
200 jug/mL (except for 2,4- and 2,5-dinitrophenol and 2-methyl-4,6-dinitro-
phenol at about 2x the given values) have been used in the past as
calibration solutions in soil recovery studies. Calibration standards and
sample extracts should be derivatized using the same procedures. External
or internal calibration may be used for this procedure. Refer to Sec. 7.0
Method 8000 for guidance on either external and internal calibration
techniques.
7.6.2 Establish the GC operating conditions appropriate for the
single-column or dual column approach (see Sec. 7.7 and Figure 5).
Optimize the instrumental conditions for resolution of the target analytes
and sensitivity.
NOTE: Once established, the same operating conditions must be used for both
calibrations and sample analyses.
7.6.3 A 2 /ul injection volume of each calibration standard is
recommended. Other injection volumes may be employed, provided that the
analyst can demonstrate adequate sensitivity for the compounds of
interest.
7.6.4 Calibration factors - Refer to Sec. 7.0 of Method 8000 for
guidance on calculating calibration factors when external calibration is
used or on calculating response factors when internal calibration is used.
7.6.5 Retention time windows - Refer to Section 7.0 of Method 8000
for guidance on the establishment of retention time windows.
7.6.6 Initial calibration acceptance criteria - Refer to Section
7.0 of Method 8000 for guidance on initial calibration linearity and
acceptance criteria.
7.7 Gas chromatographic analysis of sample extracts
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7.7.1 Inject a 2 /uL aliquot of the concentrated sample extract.
Record the volume injected to the nearest 0.05 /j,L and the resulting peak
size in area units. The same GC operating conditions used for the initial
calibration must be employed for samples analyses.
NOTE: When using internal standard calibration, add 10 /uL of the internal
standard solution to the sample extract prior to injection.
7.7.2 Calibration verification - Verify calibration by injecting
calibration verification a standard prior to conducting any sample
analyses. Sample injections may continue for as long as the calibration
verification standards and standards interspersed with the samples meet
instrument QC requirements. It is recommended that standards be analyzed
after every 10 samples (required after every 20 samples and at the end of
a set) to minimize the number of samples that must be re-inject when the
standards fail the QC limits. The sequence ends when the set of samples
has been injected or when qualitative and/or quantitative QC criteria are
exceeded. Each sample analysis must be bracketed with an acceptable
initial calibration or calibration verification standards interspersed
between the sample analyses. When a calibration verification standard
fails to meet the QC criteria, all samples that were injected after the
last standard that last met the QC criteria must be re-injected.
7.7.2.1 The calibration factor for each analyte to be
quantitated must not exceed a ±15 percent difference when compared
to the initial calibration curve. Refer to Section 7.0 of Method
8000 for guidance on the proper calculation of percent difference
using either calibration factors or response factors.
7.7.2.2 If this criterion is exceeded, inspect the gas
chromatographic system to determine the cause and perform whatever
maintenance is necessary before verifying calibration and proceeding
with sample analysis.
7.7.2.3 If routine maintenance does not return the instrument
performance to meet the QC requirements (Sec. 7.9) based on the last
initial calibration, then a new initial calibration must be
performed.
7.7.3 Compare the retention time of each analyte in the calibration
standard with the absolute retention time windows established in Sec.
7.6.5. As described in Method 8000, the center of the absolute retention
time window for each analyte is its retention time in the mid-
concentration standard analyzed during the initial calibration. Each
analyte in each standard must fall within its respective retention time
window. If not, the gas chromatographic system must either be adjusted so
that a second analysis of the standard does result in all analytes falling
within their retention time windows, or a new initial calibration must be
performed and new retention time windows established.
7.7.4 Tentative identification of an analyte occurs when a peak
from a sample extract falls within the absolute retention time window.
Each tentative identification must be confirmed using either a second GC
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column of dissimilar stationary phase or using another technique such as
GC/MS (see Sec. 8.6). When using the dual-column technique, additional
confirmation is not required, provided that the analyte meets the
identification criteria in both columns.
7.7.5 Refer to Section 7.0 of Method 8000 for calculation of
results from either external or internal calibration. Both external and
internal standard quantitation can be applied to the analysis of either
the underivatized or derivatized phenols, provided that the initial
calibration is performed on the same type of standards.
7.7.5.1 Proper quantitation requires the appropriate
selection of a baseline from which the peak area or height can be
determined.
7.7.5.2 If the responses exceed the calibration range of the
system, dilute the extract and reanalyze. Peak height measurements
are recommended over peak area integration when overlapping peaks
cause errors in area integration.
7.7.5.3 If partially overlapping or coeluting peaks are
found, change columns or try GC/MS quantitation, see Sec. 8 and
Method 8270.
7.8 Confirmation
7.8.1 When the single-column approach is employed, all target
phenols must have their identities confirmed. Confirmatory techniques
such as gas chromatography with a dissimilar column, specific element
detector, or mass spectrometer should be used. Refer to Method 8000 for
further information on confirmation.
7.8.2 When the dual-column approach is employed, the target phenols
are identified and confirmed when they meet the identification criteria on
both columns.
7.9 Suggested chromatograph maintenance - Corrective measures may require
one or more of the following remedial actions.
7.9.1 Splitter connections - For dual columns which are connected
using a press-fit Y-shaped glass splitter or a Y-shaped fused-silica
connector (J&W Scientific, Restek, Supelco, or equivalent), clean and
deactivate the splitter port insert or replace with a cleaned and
deactivated splitter. Break off a few inches (up to one foot) of the
injection port side of the column. Remove the columns and solvent
backflush according to the manufacturer's instructions. If these
procedures fail to eliminate the degradation problem, it may be necessary
to deactivate the metal injector body and/or replace the columns.
7.9.2 Metal injector body - Turn off the oven and remove the
analytical columns when the oven has cooled. Remove the glass injection
port insert (instruments with on-column injection). Reduce the injection
port temperature to room temperature. Inspect the injection port and
remove any visible foreign material.
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7.9.2.1 Place a beaker beneath the injector port inside the
oven. Using a wash bottle, serially rinse the entire inside of the
injector port with acetone and then toluene.
7.9.2.2 Prepare a solution of a deactivating agent (Sylon-CT
or equivalent) following manufacturer's directions. After all metal
surfaces inside the injector body have been thoroughly coated with
the deactivation solution, serially rinse the injector body with
toluene, methanol, acetone, and hexane. Reassemble the injector and
replace the columns.
7.9.3 Column rinsing - The column should be rinsed with several
column volumes of an appropriate solvent. Both polar and nonpolar
solvents are recommended. Depending on the nature of the sample residues
expected, the first rinse might be water, followed by methanol and
acetone; methylene chloride is a satisfactory final rinse and in some
cases may be the only solvent required. The column should then be filled
with methylene chloride and allowed to remain flooded overnight to allow
materials within the stationary phase to migrate into the solvent. The
column is then flushed with fresh methylene chloride, drained, and dried
at room temperature with a stream of ultrapure nitrogen passing through
the column.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. QC to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method. Each laboratory should
maintain a formal quality assurance program. The laboratory should also maintain
records to document the quality of the data generated.
8.2 Quality control necessary to evaluate the GC system operation is found
in Method 8000, Sec. 7.0 under Retention Time Windows, Calibration Verification,
and Chromatographic Analysis of Samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. If appropriate, it is suggested that the quality
control (QC) reference sample concentrate contain each analyte of interest at 20
mg/L. See Method 8000, Sec. 8.0 for information on how to accomplish this
demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes a method blank, matrix spike, a duplicate, a laboratory control sample
(LCS), and the use of surrogate spikes in each analytical batch.
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8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, the laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 In-house method performance criteria should be developed
using the guidance found in Sec. 8.0 of Method 8000.
8.4.3 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicates a potential problem due to the sample matrix itself,
the LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control for preparation and analysis.
8.5 Surrogate recoveries: The laboratory should evaluate surrogate
recovery data from individual samples versus the surrogate control limits
developed by the laboratory. See Method 8000, Sec. 8.0 for information on
evaluating surrogate data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Tables 1 and 2 list the retention times and recovery data for the
underivatized analytes from sandy, loam soil that may be determined by this
method. Figures 1 and 2 provide chromatograms and GC operating conditions of
those analyses.
9.1 Tables 3 and 4 list the retention times for some of the methylated
analytes that may be determined by this method. Figures 3 and 4 provide
chromatograms and GC operating conditions of those analyses.
9.2 Table 5 lists the retention times for the PFB derivatives of the
analytes that may be determined by this method. Figure 5 provides a chromatogram
of the analytes under the GC conditions listed in Table 6.
8041 - 15 Revision 0
January 1995
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10.0 REFERENCES
1. Lee, H. B.; Weng, L. D.; Chau, A. S. Y. J. Assoc. Off. Anal. Chem. 1984,
67, 6, 1086-1090.
2. Lopez-Avila, V.; Baldin, E.; Benedicto, J; Milanes, J.; Beckert, W. F.
"Application of Open-Tubular Columns to SW-846 GC Methods"; final report
to the U.S. Environmental Protection Agency on Contract 68-03-3511; Mid-
Pacific Environmental Laboratory, Mountain View, CA, 1990.
3. Tsang, S.; Marsden, P.; Chau, N. "Performance Data for Methods 8041, 8091,
8111, and 8121A"; draft report to U.S. Environmental Protection Agency on
Contract 68-W9-0011; Science Applications International Corp., San Diego,
CA, 1992.
8041 - 16 Revision 0
January 1995
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TABLE 1
RETENTION TIMES AND RECOVERIES3 OF UNDERIVATIZED PHENOLS
(MIX 1)
Analyte
Phenol
2-Methylphenol
3-Methylphenol
2,4-Dimethylphenol
2,6-Dimethylphenol
2,3-Dimethylphenol
3-Chlorophenol
4 - Chi oro- 3 -methyl phenol
2,3,5-Trichlorophenol
2,4,5-Trichlorophenol
2,5-Dinitrophenol
2,4-Dinitrophenol
2,3,5,6-Tetrachlorophenol
2 -methyl -4,6-dinitrophenol
Dinoseb
Rt, min
6.37
8.17
8.65
9.63
10.54
11.32
11.68
14.07
15.47
16.05
18.37
19.29
20.42
21.72
25.71
Spiking Cone.
(M9/9)
20
20
20
20
20
20
20
20
20
20
40
40
20
40
20
Recovery
IV \
(/o)
93
95
98
93
101
106
116
128
136
139
177
157
236
201
210
% RSD
16.9
13.6
10.3
11.5
8.1
7.1
6.7
3.8
4.1
3.0
5.1
7.3
3.5
3.8
4.9
Five 5 g aliquots of clean, sandy loam soil were spiked separately and
extracted using Method 3540 (Soxhlet) with methylene chloride as a
solvent.
8041 - 17
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January 1995
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TABLE 2
RETENTION TIMES AND RECOVERIES3 OF UNDERIVATIZED PHENOLS
(MIX 2)
Analyte
2-Chlorophenol
4-Methylphenol
2,5-Dimethylphenol
2-Nitrophenol
2,4-Dichlorophenol
2,6-Dichlorophenol
2,4,6-Trichlorophenol
2,3,6-Trichlorophenol
3-Nitrophenol
4-Nitrophenol
2,3,4,6-Tetrachlorophenol
Pentachlorophenol
Rt, min
6.91
8.64
10.42
10.58
11.29
12.18
15.91
16.68
18.37
19.61
20.60
24.85
Spiking Cone.
(M9/9)
20
20
20
20
20
20
20
20
20
20
20
20
Recovery
i°/\
(/o)
93
96
101
99
102
104
122
125
124
123
146
168
% RSD
11.6
3.4
2.6
2.8
2.5
2.8
2.7
2.6
4.0
5.6
3.3
5.0
Five 5 g aliquots of clean, sandy loam soil were spiked separately and
extracted using Method 3540 (Soxhlet) with methylene chloride as a
sol vent.
8041 - 18
Revision 0
January 1995
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TABLE 3
RETENTION TIMES OF METHYLATED PHENOLS
(MIX 1)
Analyte (derivatized)
2,3,5-Trichlorophenol
2,4,5-Trichlorophenol
2,3,5,6-Tetrachlorophenol
2,5-Dinitrophenol
2 -methyl -4,6-dinitrophenol
2,4-Dinitrophenol
Dinoseb
Rt, min
15.873
15.873
17.50
20.07
20.92
22.15
23.87
a Co-eluting analytes.
TABLE 4
RETENTION TIMES OF METHYLATED PHENOLS
(MIX 2)
Analyte (derivatized)
2,6-Dichlorophenol
2,4-Di chlorophenol
2,4,6-Trichlorophenol
2-Nitrophenol
3-Ni trophenol
2,3,6-Trichlorophenol
4-Nitrophenol
2,3,4,6-Tetrachlorophenol
Pentachlorophenol
Rt, min
10.02
12.07
13.12
13.48a
13.483
14.15
14.64
17.56
21.54
Co-eluting analytes
8041 - 19
Revision 0
January 1995
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TABLE 5
RETENTION TIMES OF PFB DERIVATIVES OF PHENOLS"
Compound
Phenol
2-Methylphenol
3-Methyl phenol
4-Methyl phenol
2,6-Dimethylphenol
2,5-Dimethylphenol
2,4-Dimethylphenol
2,3-Dimethylphenol
2-Chlorophenol
3-Chlorophenol
3,4-Dimethylphenol
4-Chlorophenol
2 - Chi oro- 5 -methyl phenol
2,6-Dichlorophenol
4- Chi oro- 2 -methyl phenol
4 -Chi oro -3 -methyl phenol
2,5-Dichlorophenol
3,5-Dichlorophenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
2,3-Dichlorophenol
3,4-Dichlorophenol
2,3,6-Trichlorophenol
2-Nitrophenol
2,4,5-Trichlorophenol
2,3,5-Trichlorophenol
3-Nitrophenol
2,3,5,6-Tetrachlorophenol
2,3,4,6-Tetrachlorophenol
2,3,4-Trichlorophenol
4-Nitrophenol
2,3,4,5-Tetrachlorophenol
Pentachlorophenol
2, 5-Dini trophenol
2,5-Dibromotoluene (IS)
2,2' ,5,5'-Tetrabromobiphenyl (IS)
2,4-Dibromophenol (Surr.)
CAS No.
108-95-2
5-48-7
108-39-4
106-44-5
576-26-1
95-87-4
105-67-9
526-75-0
95-57-8
108-43-0
95-65-8
106-48-9
615-74-7
87-65-0
1570-64-5
59-50-7
583-78-8
591-35-5
120-83-2
88-06-2
576-24-9
95-77-2
933-75-5
88-75-5
95-95-4
933-78-8
554-84-7
935-95-5
58-90-2
15950-66-0
100-02-7
4901-51-3
87-86-5
329-71-5
615-59-8
59080-37-4
615-58-7
DB-5
RT (min)
4.69
5.68
6.05
6.21
7.08
7.08
7.34
7.96
7.34
7.86
8.46
8.19
9.12
9.73
9.73
10.18
10.71
11.02
11.02
12.85
12.01
12.51
13.93
12.51
15.02
15.02
13.69
17.71
17.96
16.81
15.69
20.51
22.96
20.51
3.16
25.16
16.02
DB-1701
RT (min)
6.36
7.44
7.99
8.13
8.83
9.02
9.27
10.11
10.24
10.78
10.78
11.31
12.25
12.52
12.89
13.31
14.37
14.75
14.75
15.76
16.22
16.67
17.36
19.19
19.35
19.35
20.06
21.18
21.49
21.76
22.93
25.52
26.81
30.15
3.18
28.68
20.56
a See Table 6 for GC operating conditions.
IS = Internal Standard
Surr. = Surrogate
8041 - 20
Revision 0
January 1995
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TABLE 6
OPERATING CONDITIONS FOR PFB DERIVATIVES'OF PHENOLS
DUAL-COLUMN APPROACH
Column 1:
Column 2:
Carrier gas:
Flow rate:
Makeup gas:
Flow rate:
Temperature program:
Injector temperature
Detector temperature
Injection volume:
Sol vent:
Type of injector:
Detector type:
Type of splitter:
DB-5 (J&W Scientific)
Dimensions: 30 m x 0.53 mm ID
Film Thickness (jum): 0.83
DB-1701 (J&W Scientific)
Dimensions: 30 m x 0.53 mm ID
Film Thickness (/urn): 1.0
Helium
6 mL/min
Nitrogen
20 mL/min
1 min hold
150°C to 275°C at 3°C/min
2 min hold
:250°C
:320°C
2 u,L
Hexane
Flash vaporization
Dual ECD
Supelco 8-in injection tee
8041 - 21
Revision 0
January 1995
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FIGURE 1
ANALYSIS OF UNDERIVATIZED PHENOLS FROM SOXHLET EXTRACTION - MIX 1
(See Table 1 for peak assignments.)
i . o
re?
a:1
ru
O.
o.O
3.
1 . O
r-
cd
co
C\2
1111
OS
jULJlj
CVZ
O
1 O
C3O
Operating Conditions:
Column: DB-5 30 m x 0.53 mm id
Injector: Packed, megabore liner, 200°C
Carrier gas: Nitrogen, 6 mL/min
Hydrogen: 30 mL/min
Total Nitrogen: 30 mL/min (carrier and makeup)
Detector: FID, 300'C
Temperature program: 80'C held for 1.5 minutes
6'C/min to 230°C
10°C/min to 275°C and held for 4.5 minutes
8041 - 22
Revision 0
January 1995
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FIGURE 2
ANALYSIS OF UNDERIVATIZED PHENOLS BY SOXHLET EXTRACTION - MIX 2
(See Table 2 for peak assignments.)
1 . O e 5-
3?
O2
O2
4.Oe4^
<35
CO
cvz
CO
=O
CD
CO
CO
r-
LO
^.?9r-
co
LTXO CO -—'
3
1 O
20
30
Operating Conditions:
Column: DB-5 30 m x 0.53 mm id
Injector: Packed, megabore liner, 200°C
Carrier gas: Nitrogen, 6 mL/min
Hydrogen: 30 mL/min
Total Nitrogen: 30 mL/min (carrier and makeup)
Detector: FID, 300°C
Temperature program: 80"C held for 1.5 minutes
6'C/min to 230°C
10°C/min to 275'C and held for 4.5 minutes
8041 - 23
Revision 0
January 1995
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FIGURE 3
ANALYSIS OF METHYLATED PHENOLS BY SOXHLET EXTRACTION - MIX 1
(See Table 3 for peak assignments.)
9 . Oe-4-
r-
C£5
O
C3
C\2
75
O
0
ri
^^
i
i
1
I
i
o
t~
c.
rv
j
1 i
\ J
-2
rs
D
J
tf
•T
c\
c\
I
i
I
Vj
r~
^
r
j
i
L j
o
1 O
20
C3O
Operating Conditions:
Column: DB-5 30 m x 0.53 mm id
Injector: Packed, megabore liner, 200°C
Carrier gas: Nitrogen, 6 mL/min
Hydrogen: 30 mL/min
Total Nitrogen: 30 mL/min (carrier and makeup)
Detector: FID, 300'C
Temperature program: 80"C held for 1.5 minutes
6°C/min to 230°C
10°C/min to 275°C and held for 4.5 minutes
8041 - 24
Revision 0
January 1995
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FIGURE 4
ANALYSIS OF METHYLATED PHENOLS BY SOXHLET EXTRACTION - MIX 2
(See Table 4 for peak assignments.)
o <
00
. O
CO
CV2
3 . O
-------
FIGURE 5
is
\
8 11
IS
14
DB-5
p,^
26
192.0 ' 2 •
21
22
I
i
24
\
IS
DB-1701
GC/ECD chromatogram of the PFB derivatives of phenolic compounds analyzed on a
DB-5/DB-1701 fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (0.83 /urn film thickness) and 30 m x
0.53 mm ID DB-1701 (1.0 p.m film thickness) connected to an 8" injection tee
(Supelco Inc.). Temperature program: 150°C (1 min hold) to 275°C (2 min hold) at
30°C/min.
8041 - 26
Revision 0
January 1995
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FIGURE 6
DIAZOMETHANE GENERATOR
nitrogen
rubber stopper
«•
o
•
gloss tubing
tub* 1
tube 2
8041 - 27
Revision 0
January 1995
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METHOD 8041
PHENOLS BY GAS CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
7.1 Choose approp-
riate extraction method
(refer to Chapter 2
and Method 3500).
No
7.2 Exchange
extraction solvent
to 2-propanol.
7.5 Set gas
chromatography
conditions (refer
to Table 2.)
derivatization
of phenols
required?
Exchange to
appropriate
solvent.
7.3 - 7.4 Proceed
with derivatization.
7.6 Perform
calibration (calculate
calibration factors
and establish retention
time windows.)
7.7.1 GC analysis of
sample extracts.
I
7.7.2 Calibration
verification.
7.7.3 - 7.7.4 Compound
identification.
7.7.5 Calculation
of concentrations found
in samples.
7.8 GC peak
confirmation.
8041 - 28
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January 1995
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METHOD 8061A
PHTHALATE ESTERS BY CAPILLARY GAS CHROMATOGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
1.0 SCOPE AND APPLICATION
1.1 Method 8061 is used to determine the identities and concentrations of
various phthalate esters in aqueous and solid matrices including groundwater,
leachate, soil, sludge and sediment. The following compounds can be determined
by this method:
Compound Name CAS No.'
Benzyl benzoate (Int. Std.) 120-51-4
Bis(2-ethylhexyl) phthalate 117-81-7
Butyl benzyl phthalate 85-68-7
Di-n-butyl phthalate 84-74-2
Diethyl phthalate 84-66-2
Dimethyl phthalate 131-11-3
Di-n-octyl phthalate 117-84-0
8 Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limits (MDLs) for the target
analytes in a water matrix. The MDLs for the components of a specific sample may
differ from those listed in Table 1 because MDLs depend on the nature of
interferences in the sample matrix. Table 2 lists the estimated quantitation
limits (EQLs) for other matrices.
1.3 The following compounds may also be analyzed by this procedure or may
be used as surrogates:
Compound Name CAS No.a
Bis(2-n-butoxyethyl) phthalate 117-83-9
Bis(2-ethoxyethyl) phthalate 605-54-9
Bis(2-methoxyethyl) phthalate 117-82-8
Bis(4-methyl-2-pentyl) phthalate 146-50-9
Diamyl phthalate 131-18-0
Dicyclohexyl phthalate 84-61-7
Dihexyl phthalate 84-75-3
Diisobutyl phthalate 84-69-5
Dinonyl phthalate 84-76-4
Hexyl 2-ethylhexyl phthalate 75673-16-4
8061A - 1 Revision 1
January 1995
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1.4 When this method is used to analyze for any or all of the target
analytes, compound identification should be supported by at least one additional
qualitative technique. This method describes conditions for parallel column,
dual electron capture detector analysis which fulfills the above requirement.
Retention time information obtained on two megabore fused-silica open tubular
columns is given in Table 1. Alternatively, gas chromatography/mass spectrometry
could be used for compound confirmation.
1.5 Phthalate esters will hydrolyze below pH 5 and above pH 7. The amount
of hydrolysis will increase with increasing or decreasing pH and with longer
contact times.
1.6 This method is restricted for use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 liter for
liquids, 10 to 30 grams for solids and sludges) is extracted using an appropriate
3500 series method.
2.1.1 Aqueous samples are extracted at a pH of 5 to 7, with
methylene chloride, in a separatory funnel (Method 3510). Alternatively,
particulate-free aqueous samples can be filtered through membrane disks
that contain C18-bonded silica (Method 3535). The phthalate esters are
retained by the bonded silica, eluted with acetonitrile, then exchanged to
hexane. Using either method, aqueous samples should not be adjusted to
basic pH, as phthalate esters will hydrolyze. Method 3520 is not
recommended for the extraction of aqueous samples containing phthalates
because the longer chain esters (dihexyl phthalate, bis(2-ethylhexyl)
phthalate, di-n-octyl phthalate, and dinonyl phthalate) tend to adsorb to
the glassware and consequently, their extraction recoveries are less than
40 percent.
2.1.2 Solid samples are extracted with hexane/acetone (1:1) or
methylene chloride/acetone (1:1) using an appropriate 3500 series method.
After cleanup, the extract is analyzed by gas chromatography with electron
capture detection (GC/ECD).
2.2 The sensitivity of Method 8061 usually depends on the level of
interferences rather than on instrumental limitations. If interferences prevent
detection of the analytes, cleanup of the sample extracts is necessary. Either
Method 3610 or 3620 alone or followed by Method 3660, Sulfur Cleanup, may be used
to eliminate interferences in the analysis. Method 3640, Gel Permeation Cleanup,
is applicable for samples that contain high amounts of lipids and waxes.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
8061A - 2 Revision 1
January 1995
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3.2 Interferences coextracted from the samples will vary considerably from
waste to waste. While general cleanup techniques are referenced or provided as
part of this method, unique samples may require additional cleanup approaches to
achieve desired sensitivities for the target analytes.
3.3 Glassware must be scrupulously clean. All glassware requires
treatment in a muffle furnace at 400°C for 2 to 4 hrs, or thorough rinsing with
pesticide-grade solvent, prior to use. Refer to Chapter Four, Sec. 4.1.4, for
further details regarding the cleaning of glassware. Volumetric glassware should
not be heated in a muffle furnace.
NOTE: If Soxhlet extractors are baked in the muffle furnace, care must be taken
to ensure that they are dry (breakage may result if any water is left in
the side-arm). Thorough rinsing with hot tap water, followed by deionized
water and acetone, is not an adequate decontamination procedure. Even
after a Soxhlet extractor was refluxed with acetone for three days, with
daily solvent changes, the concentration of bis(2-ethylhexyl) phthalate
was as high as 500 ng per washing. Storage of glassware in the laboratory
introduces contamination, even if the glassware is wrapped in aluminum
foil. Therefore, any glassware used in Method 8061 should be cleaned
immediately prior to use.
3.4 Florisil and alumina may be contaminated with phthalate esters and,
therefore, use of these materials in sample cleanup should be employed
cautiously. If these materials are used, they must be obtained packaged in glass
(plastic packaging will contribute to contamination with phthalate esters).
Washing of these materials prior to use with the solvent(s) used for elution
during extract cleanup was found helpful, however, heating at 320°C for Florisil
and 210eC for alumina is recommended. Phthalate esters were detected in Florisil
cartridge method blanks at concentrations ranging from 10 to 460 ng, with
5 phthalate esters in the 105 to 460 ng range. Complete removal of the phthalate
esters from Florisil cartridges does not seem possible, and it is therefore
desirable to keep the steps involved in sample preparation to a minimum.
3.5 Paper thimbles and filter paper must be exhaustively washed with the
solvent that will be used in the sample extraction. Soxhlet extraction of paper
thimbles and filter paper for 12 hrs with fresh solvent should be repeated a
minimum of three times. Method blanks should be obtained before any of the
precleaned thimbles or filter papers are used. Storage of precleaned thimbles
and filter paper in precleaned glass jars covered with aluminum foil is
recommended.
3.6 Glass wool used in any step of sample preparation should be a
specially treated Pyrex® wool, pesticide grade, and must be baked at 400°C for
4 hrs. immediately prior to use.
3.7 Sodium sulfate must be obtained packaged in glass (plastic packaging
will contribute to contamination with phthalate esters), and must be purified by
heating at 400°C for 4 hrs. in a shallow tray, or by precleaning with methylene
chloride. To avoid recontamination, the precleaned material must be stored in
glass-stoppered glass bottles, or glass bottles covered with precleaned aluminum
foil. The storage period should not exceed two weeks. To minimize
contamination, extracts should be dried directly in the glassware in which they
8061A - 3 Revision 1
January 1995
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are collected by adding small amounts of precleaned sodium sulfate until an
excess of free-flowing material is noted.
3.8 The presence of elemental sulfur will result in large peaks which
often mask the region of the compounds eluting before dicyclohexyl phthalate
(Compound No. 14) in the gas chromatograms shown in Figure 1. Method 3660 is
suggested for removal of sulfur.
3.9 Waxes and lipids can be removed by Gel Permeation Chromatography
(Method 3640). Extracts containing high concentrations of lipids are viscous,
and may even solidify at room temperature. Phthalates elute just after corn oil
in the GPC program.
4.0 APPARATUS AND MATERIALS
4.1 Gas Chromatography.
4.1.1 Gas chromatograph - analytical system complete with gas
chromatograph suitable for on-column and split/splitless injections and
all required accessories, including detector, analytical columns,
recorder, gases, and syringes. A data system for measuring peak heights
and/or peak areas is recommended.
4.1.1.1 Eight inch injection tee (Supelco, Inc., Catalog
No. 2-3665, or equivalent) or glass Y splitter for megabore columns
(J&W Scientific, "press-fit", Catalog No. 705-0733, or equivalent).
4.1.2 Columns.
4.1.2.1 Column 1 - 30 m x 0.53 mm ID, 5% phenyl/95% methyl
silicone fused-silica open tubular column (DB-5, J&W Scientific, or
equivalent), 1.5 p,m film thickness.
4.1.2.2 Column 2 - 30 m x 0.53 mm ID, 14% cyanopropyl phenyl
silicone fused-silica open tubular column (DB-1701, J&W Scientific,
or equivalent), 1.0 /zm film thickness.
4.1.3 Detector - Dual electron capture detector (ECD).
4.2 Glassware - see appropriate 3500 series method for specifications.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Phthalates are ubiquitous laboratory contaminants. Each lot of
reagents used for this method should be checked for phthalate contamination.
8061A - 4 Revision 1
January 1995
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Additional demonstration that reagents are free of contamination may be required
because reagents may become contaminated during storage in the laboratory
environment.
5.3 Hexane, C6H14 - Pesticide quality, or equivalent.
5.4 Stock standard solutions:
5.4.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in hexane
and diluting to volume in a 10 ml volumetric flask. When compound purity
is assayed to be 96 percent or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standard solutions can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.4.2 Transfer the stock standard solutions into glass vials with
Teflon®-!ined screw-caps or crimp tops. Store at 4°C and protect from
light. Stock standard solutions should be checked periodically by gas
chromatography for signs of degradation or evaporation, especially just
prior to preparation of calibration standards.
5.4.3 Stock standard solutions must be replaced after 6 months, or
sooner if comparison with check standards indicates a problem.
5.5 Calibration standards: Calibration standards are prepared at a minimum
of five concentrations for each parameter of interest through dilution of the
stock standard solutions with hexane. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples, or should define the working range of the GC. Calibration
solutions must be replaced after 1 to 2 months, or sooner if comparison with
calibration verification standards indicates a problem.
5.6 Internal standards (if internal standard calibration is used): To use
this approach, the analyst must select one or more internal standards that are
similar in analytical behavior to the compounds of interest. The analyst must
further demonstrate that the measurement of the internal standard is not affected
by method or matrix interferences. Benzyl benzoate has been tested and found
appropriate for Method 8061.
5.6.1 Prepare a spiking solution of benzyl benzoate in hexane at
5000 mg/L. Addition of 10 /xL of this solution to 1 ml of sample extract
is recommended. The spiking concentration of the internal standard should
be kept constant for all samples and calibration standards. Store the
internal standard spiking solution at 4°C in glass vials with Teflon®-
lined screw-caps or crimp tops. Standard solutions should be replaced
when ongoing QC (Sec. 8.0) indicates a problem,
5.7 Surrogate standards: The analyst should monitor the performance of the
extraction, cleanup (when used), analytical system, and the effectiveness of the
method in dealing with each sample matrix by spiking each sample, standard, and
8061A - 5 Revision 1
January 1995
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blank with surrogate compounds. Three surrogates are suggested for Method 8061:
diphenyl phthalate, diphenyl isophthalate, and dibenzyl phthalate.
5.7.1 Prepare a surrogate standard spiking solution in acetone
which contains 50 ng/juL of each compound. Addition of 500 /il_ of this
solution to 1 L of water or 30 g solid sample is equivalent to 25 jug/L of
water or 830 jitg/kg of solid sample. The spiking concentration of the
surrogate standards may be adjusted accordingly if the final volume of
extract is reduced below 2 ml for water samples or 10 ml for solid
samples. Store the surrogate spiking solution at 4°C in glass vials with
Teflon®-!ined screw-caps or crimp tops. The solution must be replaced
after 6 months, or sooner if ongoing QC (Sec. 8.0) indicates problems.
5.8 Matrix spike solution: Analysts should select phthalates of the
greatest interest as the matrix spike compounds. If no other guidance is
provided to the analyst, selected water samples should be spiked with 20-60 /ug/L
of butyl benzyl phthalate and diethylhexyl phthalate and selected solid samples
should be spiked with 1-3 mg/kg of butylbenzyl phthalate and diethylhexyl
phthalate. The matrix spiking solution should be prepared in acetone.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Extraction: Refer to Chapter Two and Method 3500 for guidance on
choosing the appropriate extraction procedure.
7.1.1 In general, water samples should be extracted at a pH of 5 to
7 with methylene chloride using an appropriate 3500 series method for
aqueous matrices (such as Methods 3510 or 3535). Using either method,
aqueous samples should not be adjusted to basic pH, as phthalate esters
will hydrolyze. Method 3520 is not recommended for the extraction of
aqueous samples because the longer chain esters (dihexyl phthalate, bis(2-
ethylhexyl) phthalate, di-n-octyl phthalate, and dinonyl phthalate) tend
to adsorb to the glassware and consequently, their extraction recoveries
are less than 40 percent.
7.1.2 Solid samples should be extracted with hexane/acetone (1:1)
or methylene chloride/acetone (1:1) using an appropriate 3500 series
method for solid matrices.
7.1.3 Immediately prior to extraction, spike 500 /zL of the
surrogate standard spiking solution (concentration = 50 ng//zL) into 1 L
of aqueous sample or 30 g solid sample.
7.2 Cleanup: Refer to Method 3600 for guidance on choosing an appropriate
cleanup procedure. Cleanup may not be necessary for extracts from a relatively
clean sample matrix.
8061A - 6 Revision 1
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7.2.1 Methods 3610 and 3620 describe procedures for sample cleanup
using alumina and Florisil Cartridges. With these methods,
bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and
bis(2-n-butoxyethyl) phthalate are recovered quantitatively.
NOTE: It is important to demonstrate through the analyses of standards that the
Florisil fractionation scheme is reproducible. When using the
fractionation schemes given in Methods 3610 or 3620, batch-to-batch
variations in the composition of the alumina or Florisil material may
cause variations in the recoveries of the phthalate esters.
7.2.2 Waxes and lipids can be removed by Gel Permeation
Chromatography (Method 3640). Phthalates elute just after corn oil in the
GPC program.
7.3 Gas chromatographic conditions (recommended):
7.3.1 Column 1 and Column 2 (Sec. 4.1.2):
Carrier gas (He) = 6 mL/min.
Makeup gas (N2) = 19 mL/min.
Injector temperature = 250°C.
Detector temperature = 320°C.
Injection volume = 2 yum
Column temperature:
Initial temperature = 150°C, hold for 0.5 min.
Temperature program = 150°C to 220"C at 5°C/min.,
followed by 220'C to 275°C at 3°C/min.
Final temperature = 275°C hold for 13 min.
7.3.2 Table 1 gives the retention times and MDLs that can be
achieved by this method for the 16 phthalate esters. An example of the
separation achieved with the DB-5 and DB-1701 fused-silica open tubular
columns is shown in Figure 1.
7.4 Calibration:
7.4.1 Refer to Method 8000 for proper calibration techniques. Use
Tables 1 and 2 for guidance on selecting the lowest point on the
calibration curve.
7.4.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for the description of each of these
procedures.
7.5 Gas chromatographic analysis:
7.5.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 ^L of internal standard solution to the sample
at 5000 mg/L prior to injection.
8061A - 7 Revision 1
January 1995
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7.5.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.5.3 Record the sample volume injected and the resulting peak
areas.
7.5.4 Using either the internal or the external calibration
procedure (Method 8000), determine the identity and the quantity of each
component peak in the sample chromatogram which corresponds to the
compounds used for calibration purposes.
7.5.5 At a minimum, a mid-concentration calibration standard should
be included after each group of 20 samples in the analysis sequence.
7.5.6 If the response of a peak exceeds the working range of the
system, dilute the extract and reanalyze.
7.5.7 Refer to Method 8000 for guidance on establishing retention
time windows and identifying target analytes.
7.6.8 GC/MS confirmation: Any compounds confirmed by two columns
may also be confirmed by GC/MS if the concentration is sufficient for
detection by GC/MS as determined by the laboratory-generated detection
limits.
7.6.8.1 The GC/MS would normally require a minimum
concentration of 10 ng//iL in the final extract for each
single-component compound.
7.6.8.2 The sample extract and associated blank should be
analyzed by GC/MS as per Sec. 7.0 of Method 8270. Normally,
analysis of a blank is not required for confirmation analysis,
however, analysis for phthalates is a special case because of the
possibility for sample contamination through septum punctures, etc.
7.6.8.3 A reference standard of the compound must also be
analyzed by GC/MS. The concentration of the reference standard must
be at a concentration that would demonstrate the ability to confirm
the phthalate esters identified by GC/ECD.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8061A - 8 Revision 1
January 1995
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8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicate a potential problem due to the sample matrix itself, the
LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDL concentrations listed in
Table 1 were obtained using organic-free reagent water. Details on how to
determine MDLs are given in Chapter One. The MDL actually achieved in a given
8061A - 9 Revision 1
January 1995
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analysis will vary, as it is dependent on instrument sensitivity and matrix
effects.
9.2 This method has been tested in a single laboratory by using different
types of aqueous samples and solid samples which were fortified with the test
compounds at two concentrations. Single-operator precision, overall precision,
and method accuracy were found to be related to the concentration of the
compounds and the type of matrix. Results of single-laboratory method evaluation
using extraction Methods 3510 and 3550 are presented in Tables 5 and 6.
9.3 Accuracy and precision data for extraction using C18-extraction disk
Method 3535 are presented in Table 4.
9.4 The accuracy and precision obtained is determined by the sample
matrix, sample preparation technique, cleanup techniques, and calibration
procedures used.
10.0 REFERENCES
1. Glazer, J.A., Foerst, G.D., McKee, G.D., Quave, S.A., and Budde, W.L.,
"Trace Analyses for Wastewaters", Environ. Sci. and Technol. 15: 1426,
1981.
2. Lopez-Avila, V., Baldin, E., Benedicto, J., Milanes, J., and Beckert,
W.F., "Application of Open-Tubular Columns to SW-846 GC Methods", U.S.
Environmental Protection Agency, EMSL-Las Vegas, NV, 1990.
3. Beckert, W.F. and Lopez-Avila, V., "Evaluation of SW-846 Method 8060 for
Phthalate Esters", Proceedings of Fifth Annual Waste Testing and Quality
Assurance Symposium, U.S. Environmental Protection Agency, 1989.
8061A - 10 Revision 1
January 1995
-------
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TABLE 2
ESTIMATED QUANTITATION LIMITS (EQL) FOR VARIOUS MATRICES8
Matrix Factor"
Groundwater 10
Low-concentration soil by ultrasonic extraction 670
with GPC cleanup
High-concentration soil and sludges by ultrasonic 10,000
extraction
Non-water miscible waste 100,000
" Sample EQLs are highly matrix dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
b EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For
nonaqueous samples, the factor is on a wet weight basis.
8061A - 13 Revision 1
January 1995
-------
TABLE 3
AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS USING METHODS 3610 AND 3620
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(Z-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Alumina
column3
64.5
62.5
77.0
76.5
89.5
70.5
75.0
67.0
90.5
73.0
87.0
62.5
91.0
84.5
108
71.0
Florisil
column3
40.0
57.0
80.0
85.0
84.5
0
81.5
0
105
74.5
90.0
0
82.0
83.5
115
72.5
Alumina
cartridge6
101
103
104
108
103
64. lc
103
111
101
108
103
108
97.6
97.5
112
97.3
Florisil
cartridged
89.4
97.3
91.8
102
105
78. 3e
94.5
93.6
96.0
96.8
98.6
91.5
97.5
90.5
97.1
105
3 2 determinations; alumina and Florisil chromatography performed according
to Methods 3610 and 3620, respectively.
b 2 determinations, using 1 g alumina cartridges; Fraction 1 was eluted with
5 ml of 20-percent acetone in hexane. 40 /zg of each component was spiked
per cartridge.
c 36.8 percent was recovered by elution with an additional 5 mL of
20-percent acetone in hexane.
d 2 determinations, using 1 g Florisil cartridges; Fraction 1 was eluted
with 5 mL of 10-percent acetone in hexane. 40 /ng of each component was
spiked per cartridge.
e 14.4 percent was recovered by elution with an additional 5 mL of
10-percent acetone in hexane.
8061A - 14 Revision 1
January 1995
-------
TABLE 4
ACCURACY AND PRECISION DATA FOR EXTRACTION USING
METHOD 3535 AND METHOD 8061
HPLC-qrade water
Groundwater
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(Z-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Average
recovery
(%)
88.6
92.3
87.6
90.3
87.2
107
93.6
108
93.9
98.4
97.3
94.8
91.3
106
84.9
96.9
Precision
(% RSD)
17.7
10.3
16.2
13.2
9.5
13.6
21.0
8.9
22.4
5.0
2.6
6.3
7.4
19.9
3.8
11.1
Average
recovery
(%)
86.6
92.6
89.3
95.0
86.7
113
78.9
102
83.4
97.7
66.0
98.7
96.3
108
90.1
95.2
Precision
(% RSD)
14.3
7.2
1.6
1.5
4.9
2.8
5.8
4.0
8.8
14.8
39.3
6.0
7.9
13.3
6.1
12.7
The number of determinations was 4.
100 /ng/L per component.
The spiking concentration was
8061A - 15
Revision 1
January 1995
-------
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FIGURE 1
DB-5
30 m x 0.53 mm ID
1.5-um Film
IS
6 8
5
11 12 SU-1 SU-2 SU-3
7 „ 1C
16
DB-1701
30 m x 0.53 mm ID
1.0-nmFilm
I-
O
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L
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40
GC/ECD chromatograms of a composite phthalate esters standard (concentration
10 ng/|iL per compound) analyzed on a DB-5 and a DB-1701 fused-silica open
tubular column. See Table 1 for peak assignments.
Temperature program: 150*C (0.5 min hold) to 220*C at 5*C/min,
220'C to 275*C (13 min hold) at 3*C/min.
8061A - 18
Revision 1
January 1995
-------
METHOD 8061A
PHTHALATE ESTERS BY CAPILLARY GAS CHROMATOGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
7.1 Extraction: Refer to Chapter 2
and Method 3500 for guidance on
choosing the appropriate extraction
procedure. Spike sample immediately
prior to extraction.
7.2 Cleanup: Refer to Method 3500
for guidance on choosing an
appropriate cleanup procedure.
7.3 Set gas chromatographic
conditions.
I
7.4 Calibration: Refer to Method
8000 for proper calibration
techniques using internal or
external calibration.
7.5 Gas chromatographic analysis:
Refer to Method 8000 for
instructions on analysis and
quantitation of component peaks.
8061A - 19
Revision 1
January 1995
-------
METHOD 8070A
NITROSAMINES BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8070 is a gas chromatographic (GC) method applicable to the
determination of nitrosamine in aqueous matrices such as groundwater and
municipal and industrial discharges. It is also applicable to solid matrices
such as soils, sediments, and sludges. Specifically, this method covers the
determination of the following compounds:
Appropriate Technique
Compound CAS No." 3510 3520 3540/1 3550 3580
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Ni trosodi -n-propyl ami ne
62-75-9
86-30-6
621-64-7
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8 Chemical Abstract Services Registry Number.
X Greater than 70 percent recovery by this preparation technique.
1.2 The method detection limit (MDL) for each analyte of interest 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. This
method has been tested for linearity of recovery from spiked organic-free reagent
water and has been demonstrated to be applicable for the concentration range from
4 x MDL to 1000 x MDL.
1.3 When this method is used to analyze samples from matrices that are not
well characterized, compound identifications should be confirmed by at least one
additional qualitative technique. Secondary confirmation can be performed using
a dissimilar GC column, specific element detector, or mass spectrometer (MS).
1.4 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 concentration by whatever means
available. The laboratory is responsible for maintaining a current awareness
file of OSHA regulations regarding the safe handling of the chemicals specified
in this method. A reference file of material safety data sheets should also be
made available to all personnel involved in the chemical analysis.
1.5 These nit^osamines are known carcinogens. Therefore, utmost care must
be exercised in the handling of these materials. Nitrosamine reference standards
and standard solutions should be handled and prepared in a ventilated glove box
within a properly ventilated room.
8070A - 1 Revision 1
January 1995
-------
1.6 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.
1.7 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample is solvent extracted with methylene
chloride using an appropriate sample preparation technique. The methylene
chloride extract is washed with dilute HC1 to remove free amines, dried, and
concentrated to a volume of 10 ml or less. Gas chromatographic conditions are
described which permit the separation and measurement of the compounds in the
extract after it has been exchanged to methanol.
2.2 Method 8070 provides gas chromatographic conditions for the detection
of ppb concentrations of nitrosamines. Prior to use of this method, appropriate
sample extraction techniques must be used. Both neat and diluted organic liquids
(Method 3580, Waste Dilution) may be analyzed by direct injection. A 2- to 5-jzL
aliquot of the extract is injected into a GC using the solvent flush technique,
and compounds in the GC effluent are detected by a nitrogen-phosphorus detector
(NPD), or a Thermal Energy Analyzer and the reductive Hall detector.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
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
(Methods 3610 or 3620) 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 Nitrosamines contaminate many types of products commonly found in the
laboratory. The analyst must demonstrate that no nitrosamine residues
contaminate the sample or solvent extract under the conditions of analysis.
Plastics, in particular, must be avoided because nitrosamines are commonly used
as plasticizers and are easily extracted from plastic materials. Serious
nitrosamine contamination may result at any time if consistent quality control
is not practiced.
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-mass spectrometric detectors.
8070A - 2 Revision 1
January 1995
-------
3.5 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences, under the conditions of the analysis, by analyzing reagent blanks.
Specific selection of reagents and purification of solvents by distillation in
all-glass systems may be required.
3.6 Interferences co-extracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - An analytical system complete with 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.
4.1.1 Column 1 - 1.8 m x 4 mm ID Pyrex® glass, packed with
Chromosorb W AW, (80/100 mesh) coated with 10% Carbowax 20 M/2% KOH or
equivalent. This column was used to develop the method performance
statements in Sec. 9.0. Guidelines for the use of alternate column
packings are provided in Sec. 7.3.2.
4.1.2 Column 2 - 1.8 m x 4 mm ID Pyrex® glass, packed with
Supelcoport (100/120 mesh) coated with 10% SP-2250, or equivalent.
4.1.3 Detector - Nitrogen-Phosphorus, reductive Hall, or Thermal
Energy Analyzer. These detectors have proven effective in the analysis of
wastewaters for the parameters listed in the scope. A nitrogen-phosphorus
detector was used to develop the method performance statements in Sec. 9.0.
Guidelines for the use of alternate detectors are provided in Sec. 7.3.2.
4.2 Boiling chips - Approximately 10/40 mesh. Heat to 400°C for
30 minutes or Soxhlet extract with methylene chloride.
4.3 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2"C). The bath should be used in a hood.
top.
4.4 Balance - Analytical, capable of accurately weighing 0.0001 g.
4.5 Vials - 10 to 15 ml, amber glass with Teflon®-! ined screw-cap or crimp
4.6 Volumetric flasks, Class A, Appropriate sizes with ground glass
stoppers.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
8070A - 3 Revision 1
January 1995
-------
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent.
5.4 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6 Stock standard solutions (1000 mg/L) - Stock standard solutions can
be prepared from pure standard materials or purchased as certified solutions.
5.6.1 Prepare stock standard solutions by accurately weighing
0.1000 ± 0.0010 g of pure material. Dissolve the material in pesticide
quality methanol and dilute to volume in a 100-mL volumetric flask. Larger
volumes can be used at the convenience of the analyst. If compound purity
is certified at 96% or greater, the weight can be used without correction
to calculate the concentration of the stock standard. Commercially-
prepared stock standards can be used at any concentration if they are
certified by the manufacturer or by an independent source.
5.6.2 Transfer the stock standard solutions into bottles with
Teflon®-!ined screw-caps or crimp tops. Store at 4°C and protect from
light. Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.6.3 Stock standard solutions must be replaced after six months,
or sooner if comparison with check standards indicates a problem.
5.7 Calibration standards - A minimum of five concentrations should be
prepared through dilution of the stock standards with isooctane. One of the
concentrations should be at a concentration near, but above, the method detection
limit. The remaining concentrations should correspond to the expected range of
concentrations found in real samples or should define the working range of the
GC. Calibration solutions must be replaced after six months, or sooner if
comparison with check standards indicates a problem.
5.8 Internal standards (if internal standard calibration is used) - To use
this approach, the analyst must select one or more internal standards that are
similar in analytical behavior to the compounds of interest. The analyst must
further demonstrate that the measurement of the internal standard is not affected
by method or matrix interferences. Because of these limitations, no internal
standard can be suggested that is applicable to all samples.
5.8.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest, as described in Sec. 5.7.
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5.8.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.8.3 Analyze each calibration standard according to Sec. 7.0.
5.9 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and reagent blank with one or two surrogates (e.g. nitrosamines that
are not expected to be in the sample) recommended to encompass the range of the
temperature program used in this method. Method 3500 details instructions on the
preparation of base/neutral surrogates. Deuterated analogs of analytes should
not be used as surrogates for gas chromatographic analysis due to coelution
problems.
5.10 Hydrochloric acid (HC1), 1 M.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
6.2 The nitrosamines validated for analysis by this procedure are
considered semivolatile organic compounds.
6.3 Extracts must be stored at 4°C and protected from light.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a neutral
pH, or as received, with methylene chloride, using an appropriate 3500
series method. Solid samples are extracted using a 3500 series method that
is appropriate for such matrices. Both neat and diluted organic liquids
(Method 3580, Waste Dilution) may be analyzed by direct injection.
7.1.2 In a separatory funnel, wash the methylene chloride extract
with 100 mL of 1 M HC1 to remove free amines.
7.1.3 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to methanol. The exchange is performed during the
extraction procedures listed in the appropriate 3500 series method.
7.1.4 N-nitrosodiphenylamine measured by gas chromatography
requires, the analyst must first use a cleanup column to eliminate
diphenylamine interference (Methods 3610 or 3620). If N-nitroso-
diphenylamine is of no interest, the analyst may proceed directly with gas
chromatographic analysis (Sec. 7.3).
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7.2 Cleanup
7.2.1 Cleanup procedures may not be necessary for a relatively clean
sample matrix. The cleanup procedure recommended in this method has been
used for the analysis of various clean waters and industrial effluents. If
particular circumstances demand the use of an alternative cleanup
procedure, the analyst must determine the elution profile and demonstrate
that the recovery of each compound of interest is no less than 85%.
Diphenylamine, if present in the original sample extract must be separate
from the nitrosamines if N-nitrosodiphenylamine is to be determined by this
method.
7.2.2 Proceed with either Method 3610 or 3620, using the 2-mL
methylene chloride extracts obtained from Sec. 7.1.2.5.
7.2.3 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
7.3 Gas Chromatography
7.3.1 GC Setup
7.3.1.1 N-nitrosodiphenylamine completely reacts to form
diphenylamine at the normal operating temperatures of a GC injection
port (200 - 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 (see Sec. 7.1.3).
7.3.1.2 Table 1 summarizes the recommended operating
conditions for the gas chromatograph. This table includes retention
times and MDLs that were obtained under these conditions. Examples
of the parameter separations achieved by these columns are shown in
Figures 1 and 2.
NOTE: Other columns, chromatographic conditions, or detectors may be used
if the requirements of Sec. 8.0 are met. Capillary (open-tubular)
columns may also be used if the relative standard deviations of
responses for replicate injections are demonstrated to be less than
6% and the requirements of Sec. 8.0 are met.
7.3.2 Calibration - Refer to Method 8000 for proper calibration
techniques.
7.3.2.1 The procedure for internal or external calibration
may be used. Refer to Method 8000 for a description of each of these
procedures.
7.3.2.2 If cleanup is performed on the samples, the analyst
should process a series of standards through the cleanup procedure and
then analyze the samples by GC. This will confirm elution patterns
and the absence of interferents from the reagents.
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7.3.3 GC Analysis
7.3.3.1 Refer to Method 8000. If the internal standard
calibration technique is used, add 10 /uL of internal standard to the
sample prior to injection.
7.3.3.2 Method 8000 provides instructions on the analysis
sequence, appropriate dilutions, establishing daily retention time
windows, and identification criteria. Include a mid-concentration
check standard after each group of 10 samples in the analysis
sequence.
7.3.3.3 Record the sample volume injected and the resulting
peak sizes (in area units or peak heights).
7.3.3.4 Using either the internal or external calibration
procedure (Method 8000), determine the identity and quantity of each
analyte peak in the sample chromatogram. See Method 8000 for
calculation equations.
7.3.3.5 If peak detection and identification are prevented
due to interferences, the hexane extract may undergo cleanup using
either Method 3610 or 3620.
7.3.3.6 Examples of GC/NPD chromatograms for nitrosamines
are shown in Figures 1 and 2.
NOTE: In order to confirm the presence of N-nitrosodiphenylamine an
appropriate cleanup procedure must be used.
7.3.4 Secondary confirmation - When this method is used to analyze
samples from matrices that are not well characterized, compound
identifications should be confirmed by at least one additional qualitative
technique. Secondary confirmation can be performed using one of the
following techniques:
7.3.4.1 Additional (or alternate) column listed in Sec. 4.1
may be used to document the retention time of the analytes of interest
on a dissimilar GC column.
7.3.4.2 Sec. 4.1 also lists three different GC detectors
with various compound selectivities that may be used to qualitatively
confirm peaks.
7.3.4.3 A GC/MS may also be utilized to confirm compounds
identified in the primary analysis. GC/MS Method 8270 is validated
for both the qualitative and quantitative confirmation of all the
target analytes in Method 8070.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
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various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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
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samples. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined 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. The MDL actually achieved in a given analysis will
vary depending on instrument sensitivity and matrix effects.
9.2 This method has been tested for linearity of recovery from spiked
organic-free reagent water and has been demonstrated to be applicable for the
concentration range from 4 x MDL to 1000 x MDL.
9.3 The average recoveries presented in Table 2 were obtained in a single
laboratory, using spiked wastewater samples. Each spiked sample was analyzed in
triplicate on three separate occasions. The standard deviation of the percent
recovery is also included in Table 2.
9.4 In a multi-laboratory study, 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 jug/L. Results
from these analyses have been used to generate accuracy and precision data in
Table 4 and the QC acceptance criteria in Table 3.
10.0 REFERENCES
1. "Determination of Nitrosamines in Industrial and Municipal Wastewaters",
EPA 600/4-82-016, U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, OH, 45268, May 1982.
2. Burgess, E.M., Lavanish, J.M., "Photochemical Decomposition of N-
nitrosamines", Tetrahedron Letters. 1964, 1221.
3. 40 Code of Federal Regulations (CFR): Protection of the Environment, Part
136 Appendix A, Method 607 - Nitrosamines.
4. "Method Detection Limit and Analytical Curve Studies EPA Methods 606, 607,
608", Special letter report for EPA Contract 68-03-2606, U.S.
Environmental Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, OH, 45268.
5. "EPA Method Validation Study 17, Method 607 (Nitrosamines)", Report for EPA
Contract 68-03-2606 (in preparation).
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TABLE 1
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS'1
Retention Time (minutes) Method
Detection Limit6
Compound Column 1 Column 2 (M9/L)
N-Nitrosodimethyl amine
N-Nitrosodi-n-propylamine
N-Nitrosodiphenyl amine8
4.1
12.1
12. 8b
0.88
4.2
6.4C
0.15
0.46
0.81
Column 1 conditions:
Carrier gas (He) flow rate: 40 mL/min
Column temperature: Isothermal, at IIO'C, except as otherwise indicated.
Column 2 conditions:
Carrier gas (He) flow rate: 40 mL/min
Column temperature: Isothermal, at 120°C, except as otherwise indicated.
a Measured as diphenylamine.
b Determined isothermally at 220°C.
c Determined isothermally at 210°C.
d Reference 3.
e MDLs were developed using reagent water.
TABLE 2
SINGLE OPERATOR ACCURACY AND PRECISION
Average Standard Spike Number
Percent Deviation Range of Matrix
Compound Recovery % (/-tg/L) Analyses Types
N-Nitrosodimethylamine 32 3.7 0.8 29 5
N-Nitrosodiphenylamine 79 7.1 1.2 29 5
N-Nitrosodi-n-propylamine 61 4.1 9.0 29 5
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TABLE 3
QC ACCEPTANCE CRITERIA
Analyte
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi -n-propyl ami ne
Test
Cone.
(M9/L)
20
20
20
Limit
for s
(M9/L)
3.4
6.1
5.7
Range
for X
(M9/L)
4.6-20.0
2.1-24.5
11.5-26.8
Recovery
Range
IO/\
(/o)
13-109
D-139
45-146
^ = Standard deviation for four recovery measurements, in M9/L.
X = Average recovery for four recovery measurements, in |ug/L.
D = Detected, result must be greater than zero.
a Reference 3.
TABLE 4
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Analyte
Accuracy, as
recovery, X'
(M9/L)
Single
analyst
precision,
V (M9/L)
Overall
precision,
S' (M9/D
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitroso-n-propylamine
0.37C + 0.06
0.64C + 0.52
0.96C - 0.07
0.25X - 0.04
0.36Y - 1.53
0.157 + 0.13
0.25X + 0.11
0.461 - 0.47
0.21X + 0.15
X' = Expected recovery for one or more measurements of a sample containing a
concentration of C, in M9/L.
C = True value for the concentration, in jug/L.
S' = Expected interlaboratory standard deviation of measurements at an
average concentration found of X, in M9/L-
s/ = Expected single analystjtandard deviation of measurements at an average
_ concentration found of X, in M9/L.
X = Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8 Reference 3.
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FIGURE i
GAS CHROMATOGRAM OF NITROSAMINES
Column: 10% Ctrtowu 20M + 3%
KOH on Chromoiorb W-AW
Otttetor Phosphorut/Nitrogtn
2 4 6 8 10 12 14
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FIGURE 2
GAS CHROMATOGRAM OF N-NITROSODIPHENYLAMINE AS DIPHENYLAMINE
Column: 10% Ctrbowtt 20M * 2% KOH on
Chromosorb W-AW
Tempirtturi: 220" C
Ofttctor Phosphorus/Nitrogtn
,»
:*.
c
2 4 S 8 10 12 14 16 It
Ruttntion time, rrunum
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METHOD 8070A
NITROSAMINES BY GAS CHROMATOGRAPHY
7.1.1 Choose
appropriate
extraction procedure.
7.1.2 Wash the
MeCl2extract with
HCI to remove
free amines.
7.1.3 Perform
solvent exchange
using methanol.
Adjust extract
volume, if necessary,
and proceed with
analysis.
7.2
Is cleanup
of the extract
required?
Perform
concentration
procedure using
methylene chloride.
7.2.2 Perform Method
3610 or 3620 using
methylene chloride.
7.2.3 Proceed with
GC Analysis.
.1.
Will
N-nitrosodi-
phenylamine be
measured?
7.1.4 Perform
column cleanup
using Method 3610
or 3620.
7.3.1.2 Refer to
Table 1 for
recommended
operating conditions
for the GC.
7.3.2 Refer to Method
8000 for proper
calibration techniques.
7.3.3.1 Refer to
Method 8000 for
guidance on GC
analysis.
7.3.3.3 - 7.3.3.4
Record sample
volume injected and
resulting peak size/
perform appropriate
calcualtions (refer
to Method 8000.)
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METHOD 8081A
ORGANOCHLORINE PESTICIDES BY CAPILLARY COLUMN GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8081 is used to determine the concentrations of various
organochlorine pesticides in extracts from solid and liquid matrices, using
fused-silica, open-tubular, capillary columns with electron capture detectors
(ECD). When compared to the packed columns, these columns offer improved
resolution, better selectivity, increased sensitivity, and faster analysis. The
compounds listed below may be determined by either a single- or dual-column
analysis system.
Compound
CAS Registry No.
Aldrin
a-BHC
0-BHC
7-BHC (Lindane)
5-BHC
Chiorobenzilate
a-Chlordane
7-Chlordane
DBCP
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dial late
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorocyclopentadi ene
Isodrin
Kepone
Methoxychlor
Toxaphene
309-00-2
319-84-6
319-85-7
58-89-9
319-86-8
510-15-6
5103-71-9
5103-74-2
96-12-8
72-54-8
72-55-9
50-29-3
2303-16-4
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
76-44-8
1024-57-3
118-74-1
77-47-4
465-73-6
143-50-0
72-43-5
8001-35-2
1.2 This revision of Method 8081 no longer includes the PCBs as Aroclors
in the list of target analytes. The analysis of PCBs should be undertaken using
Method 8082, which includes specific cleanup and quantitation procedures designed
for PCB analysis. This change was made to obtain PCB data of better quality and
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to eliminate the complications inherent in a combined organochlorine pesticide
and PCB method. Therefore, if the presence of PCBs is expected, use Method 8082
for PCB analyses, and this method (8081) for the organochlorine pesticides. If
there is no information of the likely presence of PCBs, either employ a PCB-
specific screening procedure such as an immunoassay (e.g., Method 4020), or split
the sample extract prior to any cleanup steps, and process part of the extract
for organochlorine pesticide analysis and the other portion for PCB analysis
using Method 8082.
1.3 The analyst must select columns, detectors and calibration procedures
most appropriate for the specific analytes of interest in a study. Matrix-
specific performance data must be established and the stability of the analytical
system and instrument calibration must be established for each analytical matrix
(e.g., hexane solutions from sample extractions, diluted oil samples, etc.).
1.4 Although performance data are presented for many of the target
analytes, it is unlikely that all of them could be determined in a single
analysis. The chemical and chromatographic behaviors of many of these chemicals
can result in co-elution of some target analytes. Several cleanup/fractionation
schemes are provided in this method and in Method 3600.
1.5 Several multi-component mixtures (i.e., Methoxychlor and Toxaphene)
are listed as target analytes. When samples contain more than one multi -
component analyte, a higher level of analyst expertise is required to attain
acceptable levels of qualitative and quantitative analysis. The same is true of
multi-component analytes that have been subjected to environmental degradation
or degradation by treatment technologies. These result in "weathered" multi-
component mixtures that may have significant differences in peak patterns than
those of standards.
1.6 Compound identification based on single-column analysis should be
confirmed on a second column, or should be supported by at least one other
qualitative technique. This method describes analytical conditions for a second
gas chromatographic column that can be used to confirm the measurements made with
the primary column. GC/MS Method 8270 is also recommended as a confirmation
technique, if sensitivity permits (Sec. 8.0).
1.7 This method includes a dual-column option. The option allows a
hardware configuration of two analytical columns joined to a single injection
port. The option allows one injection to be used for dual-column analysis.
Analysts are cautioned that the dual-column option may not be appropriate when
the instrument is subject to mechanical stress, many samples are to be run in a
short period, or when contaminated samples are analyzed.
1.8 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs (GC) and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.9 Extracts suitable for analysis by this method may also be analyzed for
organophosphorus pesticides (Method 8141). Some extracts may also be suitable
for triazine herbicide analysis, if low recoveries (normally samples taken for
triazine analysis must be preserved) are not a problem.
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1.10 The following compounds may also be determined using this method:
Compound
CAS Registry No.
Alachlor
Captafol
Chloroneb
Chloropropylate
Chlorothalonil
DC PA
Dichlone
Dicofol
Etridiazole
Halowax-1000
Halowax-1001
Halowax-1013
Halowax-1014
Halowax-1051
Halowax-1099
Mi rex
Nitrofen
PCNB
Perthane
Propachlor
Strobane
trans-Nonachlor
Permethrin
Trifluralin
15972-60-8
2425-06-1
2675-77-6
99516-95-7
1897-45-6
1861-32-1
117-80-6
115-32-2
2593-15-9
58718-66-4
58718-67-5
12616-35-2
12616-36-3
2234-13-1
39450-05-0
2385-85-5
1836-75-5
82-68-8
72-56-0
1918-16-17
8001-50-1
39765-80-5
51877-74-8
1582-09-8
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 L for liquids,
2 g to 30 g for solids) is extracted using the appropriate matrix-specific sample
extraction technique.
2.2 Liquid samples are extracted at neutral pH with methylene chloride
using either Method 3510 (separatory funnel), Method 3520 (continuous liquid-
liquid extractor), or other appropriate technique.
2.3 Solid samples are extracted with hexane-acetone (1:1) or methylene
chloride-acetone (1:1) using Method 3540 (Soxhlet), Method 3541 (automated
Soxhlet), Method 3545 (accelerated solvent extraction), Method 3550 (ultrasonic
extraction), or other appropriate technique.
2.4 A variety of cleanup steps may be applied to the extract, depending
on the nature of the matrix interferences and the target analytes. Suggested
cleanups include alumina (Method 3610), Florisil (Method 3620), silica gel
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(Method 3630), gel permeation chromatography (Method 3640), and sulfur (Method
3660).
2.5 After cleanup, the extract is analyzed by injecting a 1-juL sample into
a gas chromatograph with a narrow- or wide-bore fused silica capillary column and
electron capture detector (GC/ECD) or an electrolytic conductivity detector
(GC/ELCD).
3.0 INTERFERENCES
3.1 Refer to Methods 3500 (Sec. 3.0, in particular), 3600, and 8000, for
a discussion of interferences.
3.2 Sources of interference in this method can be grouped into three broad
categories.
3.2.1 Contaminated solvents, reagents, or sample processing
hardware.
3.2.2 Contaminated GC carrier gas, parts, column surfaces, or
detector surfaces.
3.2.3 Compounds extracted from the sample matrix to which the
detector will respond.
3.2.4 Interferences co-extracted from the samples will vary
considerably from waste to waste. While general cleanup techniques are
referenced or provided as part of this method, unique samples may require
additional cleanup approaches to achieve desired degrees of discrimination
and quantitation.
3.3 Interferences by phthalate esters introduced during sample preparation
can pose a major problem in pesticide determinations.
3.3.1 These materials may be removed prior to analysis using Method
3640 (Gel Permeation Cleanup - pesticide option) or Method 3630 (as
fraction III of the silica gel cleanup procedure).
3.3.2 Common flexible plastics contain varying amounts of phthalate
esters which are easily extracted or leached from such materials during
laboratory operations.
3.3.3 Cross-contamination of clean glassware routinely occurs when
plastics are handled during extraction steps, especially when solvent-
wetted surfaces are handled.
3.3.4 Interferences from phthalate esters can best be minimized by
avoiding contact with any plastic materials and checking all solvents and
reagents for phthalate contamination. Exhaustive cleanup of solvents,
reagents and glassware may be required to eliminate background phthalate
ester contamination.
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3.4 Glassware must be scrupulously cleaned. Clean all glassware as soon
as possible after use by rinsing with the last solvent used. This should be
followed by detergent washing with hot water, and rinses with tap water and
organic-free reagent water. Drain the glassware and dry it in an oven at 130°C
for several hours, or rinse with methanol and drain. Store dry glassware in a
clean environment.
3.5 The presence of elemental sulfur will result in broad peaks that
interfere with the detection of early-eluting organochlorine pesticides. Sulfur
contamination should be expected with sediment samples. Method 3660 is suggested
for removal of sulfur. Since the recovery of Endrin aldehyde (using the TBA
procedure) is drastically reduced, this compound must be determined prior to
sulfur cleanup.
3.6 Waxes, lipids, and other high molecular weight materials can be
removed by Method 3640 (gel-permeation cleanup).
3.7 Other halogenated pesticides or industrial chemicals may interfere
with the analysis of pesticides. Certain co-eluting organophosphorus pesticides
are eliminated by Method 3640 (gel permeation cleanup - pesticide option).
Co-eluting chlorophenols may be eliminated by using Method 3630 (silica gel),
Method 3620 (florisil), or Method 3610 (alumina).
3.8 Co-elution among the many target analytes in this method can cause
interference problems. The following target analytes coelute on the GC columns
listed, when using the single-column analysis scheme:
DB 608 Trifluralin/Dial late isomers
PCNP/Dichlone/Isodrin
DDD/Endosulfan II
DB 1701 Captafol/Mirex
DDD/Endosulfan II
Methoxychlor/Endosulfan sulfate
3.9 The following compounds coelute using the dual-column analysis scheme.
In general, the DB-5 column resolves fewer compounds that the DB-1701.
DB-5 Permethrin/Heptachlor epoxide
Endosulfan I/a-Chlordane
Perthane/Endrin
Endosulfan II/Chloropropylate/Chiorobenzi1 ate
4,4'-DDT/Endosulfan sulfate
Methoxychlor/Di cofol
DB-1701 Chlorothalonil/6-BHC
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph: an analytical system complete with gas
chromatograph suitable for on-column and split-splitless injection and all
required accessories including syringes, analytical columns, gases, electron
capture detectors (BCD), and recorder/integrator or data system.
4.2 GC columns
This method describes procedures for both single-column and dual-column
analyses. The single-column approach involves one analysis to determine that a
compound is present, followed by a second analysis to confirm the identity of the
compound (Sec. 8.4 describes how GC/MS confirmation techniques may be employed).
The single-column approach may employ either narrow-bore (<0.32 mm ID) columns
or wide-bore (0.53 mm ID) columns. The dual-column approach involves a single
injection that is split between two columns that are mounted in a single gas
chromatograph. The dual-column approach employs only wide-bore (0.53 mm ID)
columns.
The columns listed in this section were the columns used to develop the
method performance data. The mention of these columns in this method is not
intended to exclude the use of other columns that may be developed. Laboratories
may use other capillary columns provided that they document method performance
data (e.g., chromatographic resolution, analyte breakdown, and MDLs) that equals
or exceeds the performance described in this method, or as appropriate for the
intended application.
4.2.1 Narrow-bore columns for single-column analysis (use both
columns to confirm compound identifications unless another confirmation
technique such as GC/MS is employed).
4.2.1.1 30 m x 0.25 or 0.32 mm ID fused silica capillary
column chemically bonded with SE-54 (DB-5 or equivalent), 1 jum film
thickness.
4.2.1.2 30 m x 0.25 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608,
SPB-608, or equivalent), 2.5 urn coating thickness, 1 ^m film
thickness.
4.2.1.3 Narrow bore columns should be installed in
split/splitless (Grob-type) injectors.
4.2.2 Wide-bore columns for single-column analysis (use two of the
three columns listed to confirm compound identifications unless another
confirmation technique such as GC/MS is employed).
4.2.2.1 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608,
SPB-608, RTx-35, or equivalent), 0.5 /zm or 0.83 jum film thickness.
4.2.2.2 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 50 percent phenyl methylpolysiloxane (DB-1701,
or equivalent), 1.0 jum film thickness.
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4.2.2.3 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 95 percent dimethyl - 5 percent diphenyl
polysiloxane (DB-5, SPB-5, RTx-5, or equivalent), 1.5 jum film
thickness.
4.2.2.4 Wide-bore columns should be installed in 1/4 inch
injectors, with deactivated liners designed specifically for use with
these columns.
4.2.3 Wide-bore columns for dual-column analysis (choose one of the
two pairs of columns listed below).
4.2.3.1 Column pair 1
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5 /urn film
thickness.
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or
equivalent), 1.0 /im film thickness.
Column pair 1 is mounted in a press-fit Y-shaped glass 3-way
union splitter (J&W Scientific, Catalog No. 705-0733) or a Y-shaped
fused-silica connector (Restek, Catalog No. 20405), or equivalent.
4.2.3.2 Column pair 2
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 0.83 ^m film
thickness.
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or
equivalent), 1.0 jim film thickness.
Column pair 2 is mounted in an 8 in. deactivated glass
injection tee (Supelco, Catalog No. 2-3665M), or equivalent.
4.3 Column rinsing kit: Bonded-phase column rinse kit (J&W Scientific,
Catalog No. 430-3000), or equivalent.
4.4 Volumetric flasks, 10-mL and 25-mL, for preparation of standards.
5.0 REAGENTS
5.1 Reagent grade or pesticide grade chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
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NOTE: Store the standard solutions (stock, composite, calibration, internal, and
surrogate) at 4°C in Teflon®-sealed containers in the dark. When a lot of
standards is prepared, it is recommended that aliquots of that lot be
stored in individual small vials. All stock standard solutions must be
replaced after one year or sooner if routine QC tests (Sec. 8.0) indicate
a problem. All other standard solutions must be replaced after six months
or sooner if routine QC (Sec. 8.0) indicates a problem.
5.2 Solvents used in the extraction and cleanup procedures (appropriate
3500 and 3600 series methods) include n-hexane, diethyl ether, methylene
chloride, acetone, ethyl acetate, and isooctane (2,2,4-trimethylpentane) and must
be exchanged to n-hexane or isooctane prior to analysis.
Therefore, n-hexane and isooctane will be required in this procedure.
Acetone or toluene may be required for the preparation of some standard solutions
(see Sec. 5.4.2). All solvents should be pesticide quality or equivalent, and
each lot of solvent should be determined to be phthalate free.
5.3 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water as defined in Chapter One.
5.4 Stock standard solutions (1000 mg/L) - May be prepared from pure
standard materials or can be purchased as certified solutions.
5.4.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10-mL volumetric flask. If compound purity is
96 percent or greater, the weight can be used without correction to
calculate the concentration of the stock standard solution. Commercially
prepared stock standard solutions can be used at any concentration if they
are certified by the manufacturer or by an independent source.
5.4.2 IS-BHC, Dieldrin, and some other standards may not be
adequately soluble in isooctane. A small amount of acetone or toluene
should be used to dissolve these compounds during the preparation of the
stock standard solutions.
5.5 Composite stock standard - May be prepared from individual stock
solutions.
5.5.1 For composite stock standards containing less than 25
components, take exactly 1 ml of each individual stock solution at a
concentration of 1000 mg/L, add solvent, and mix the solutions in a 25-mL
volumetric flask. For example, for a composite containing 20 individual
standards, the resulting concentration of each component in the mixture,
after the volume is adjusted to 25 mL, will be 1 mg/25 mL. This composite
solution can be further diluted to obtain the desired concentrations.
5.5.2 For composite stock standards containing more than 25
components, use volumetric flasks of the appropriate volume (e.g., 50 mL,
100 mL), and follow the procedure described above.
5.6 Calibration standards should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with isooctane or
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hexane. The concentrations should correspond to the expected range of
concentrations found in real samples and should bracket the linear range of the
detector.
5.6.1 Although all single component analytes can be resolved on a
new 35 percent phenyl methyl silicone column (e.g., DB-608), two
calibration mixtures should be prepared for the single component analytes
of this method. This procedure is established to minimize potential
resolution and quantitation problems on confirmation columns or on older
35 percent phenyl methyl silicone (e.g. DB-608) columns and to allow
determination of Endrin and DDT breakdown for method QC (Sec. 8.0).
5.6.2 Separate calibration standards are required for each multi-
component target analyte (e.g., Toxaphene and Methoxychlor)
5.7 Internal standard (optional)
5.7.1 Pentachloronitrobenzene is suggested as an internal standard
for the single-column analysis, when it is not considered to be a target
analyte. l-bromo-2-nitrobenzene may also be used. Prepare a solution of
5000 mg/L (5000 ng//xL) of pentachloronitrobenzene or l-bromo-2-
nitrobenzene. Spike 10 /iL of this solution into each 1 ml sample extract.
5.7.2 l-bromo-2-nitrobenzene is suggested as an internal standard
for the dual-column analysis. Prepare a solution of 5000 mg/L (5000 ng/juL)
of l-bromo-2-nitrobenzene. Spike 10 ^L of this solution into each 1 ml of
sample extract.
5.8 Surrogate standards
The performance of the method should be monitored using surrogate
compounds. Surrogate standards are added to all samples, method blanks, matrix
spikes, and calibration standards.
5.8.1 For the single-column analysis, use decachlorobiphenyl as the
primary surrogate. However, if recovery is low, or late-eluting compounds
interfere with decachlorobiphenyl, then tetrachloro-m-xylene should be
evaluated as a surrogate. Method 3500, Sec. 5.0, describes the proper
procedure for preparing these surrogates.
5.8.2 For the dual-column analysis, prepare a solution of 500 mg/L
(500 ng/juL) of 4-chloro-3-nitrobenzotrifluoride. Use a spiking volume of
100 /LiL for a 1 L aqueous sample. Store the spiking solution at 4"C in
Teflon®-sealed containers in the dark.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See Chapter Four, Organic Analytes, Sec. 4.0, for sample collection
and preservation instructions.
6.2 Extracts must be stored under refrigeration in the dark and analyzed
within 40 days of extraction.
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7.0 PROCEDURE
7.1 Sample extraction
Refer to Chapter Two and Method 3500 for guidance in choosing the
appropriate extraction procedure. In general, water samples are extracted
at a neutral pH with methylene chloride using a separatory funnel (Method
3510) or a continuous liquid-liquid extractor (Method 3520). Solid samples
are extracted with hexane-acetone (1:1) or methylene chloride-acetone (1:1)
using one of the Soxhlet extraction (Method 3540 or 3541), accelerated
solvent extraction (Method 3545) or ultrasonic extraction (Method 3550)
procedures.
NOTE: Hexane-acetone (1:1) may be more effective as an extraction solvent for
organochlorine pesticides in some environmental and waste matrices than is
methylene chloride-acetone (1:1). Use of hexane-acetone generally reduces
the amount of interferences that are extracted and improves signal-to-
noise.
Spiked samples are used to verify the applicability of the chosen
extraction technique to each new sample type. Each sample type must be
spiked with the compounds of interest to determine the percent recovery and
the limit of detection for that sample (see Chapter One). See Method 8000
for guidance on demonstration of initial method proficiency as well as
guidance on matrix spikes for routine sample analysis.
7.2 Extract cleanup
Cleanup procedures may not be necessary for a relatively clean sample
matrix, but most extracts from environmental and waste samples will require
additional preparation before analysis. The specific cleanup procedure
used will depend on the nature of the sample to be analyzed and the data
quality objectives for the measurements. General guidance for sample
extract cleanup is provided in this section and in Method 3600.
7.2.1 If a sample is of biological origin, or contains high
molecular weight materials, the use of Method 3640 (GPC cleanup - pesticide
option) is recommended. Frequently, one of the adsorption chromatographic
cleanups (alumina, silica gel, or florisil) may also be required following
the GPC cleanup.
7.2.2 Method 3610 (alumina) may be used to remove phthalate esters.
7.2.3 Method 3620 (florisil) may be used to separate organochlorine
pesticides from aliphatic compounds, aromatics, and nitrogen-containing
compounds.
7.2.4 Method 3630 (silica gel) may be used to separate single
component organochlorine pesticides from some interferants.
7.2.5 Elemental sulfur, which may be present in certain sediments
and industrial wastes, interferes with the electron capture gas
chromatography of certain pesticides. Sulfur should be removed by the
technique described in Method 3660.
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7.3 GC conditions
This method allows the analyst to choose between a single-column or a dual-
column configuration in the injector port. Either wide- or narrow-bore columns
may be used. Identifications based on retention times from a single-column must
be confirmed on a second column or with an alternative qualitative technique.
7.3.1 Single-column analysis
This capillary GC/ECD method allows the analyst the option of using
0.25-0.32 mm ID capillary columns (narrow-bore) or 0.53 mm ID capillary
columns (wide-bore). Performance data are provided for both options.
Figures 1-6 provide example chromatograms.
7.3.1.1 The use of narrow-bore (<0.32 mm ID) columns is
recommended when the analyst requires greater chromatographic
resolution. Use of narrow-bore columns is suitable for relatively
clean samples or for extracts that have been prepared with one or more
of the clean-up options referenced in the method. Wide-bore columns
(0.53 mm ID) are suitable for more complex environmental and waste
matrices.
7.3.1.2 Table 1 lists average retention times and method
detection limits (MDLs) for the target analytes in water and soil
matrices, using wide-bore capillary columns. Table 2 lists average
retention times and method detection limits (MDLs) for the target
analytes in water and soil matrices, using narrow-bore capillary
columns. The MDLs for the components of a specific sample are
dependent upon the nature of interferences in the sample matrix and
may differ from those listed in Tables 1 and 2. Table 3 lists the
Estimated Quantitation Limits (EQLs) for other matrices.
7.3.1.3 Table 4 lists the GC operating conditions for the
single-column method of analysis.
7.3.2 Dual-column analysis
The dual-column/dual-detector approach involves the use of two 30 m
x 0.53 mm ID fused-silica open-tubular columns of different polarities,
thus, different selectivities towards the target analytes. The columns are
connected to an injection tee and separate electron capture detectors.
7.3.2.1 Retention times for the organochlorine analytes on
dual-columns are in Table 6. The GC operating conditions for the
compounds in Table 6 are given in Table 7.
7.3.2.2 Multi-component mixtures of Toxaphene and Strobane
were analyzed separately (Figures 6 and 7) using the GC operating
conditions found in Table 7.
7.3.2.3 Figure 8 is a sample chromatogram for a mixture of
organochlorine pesticides. The retention times of the individual
components detected in these mixtures are given in Tables 6 and 7.
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7.3.2.4 Operating conditions for a more heavily loaded DB-
5/DB-1701 pair are given in Table 8. This column pair was used for
the detection of multi-component organochlorine compounds.
7.3.2.5 Operating conditions for a DB-5/DB-1701 column pair
with thinner films, a different type of splitter, and a slower
temperature programming rate are provided in Table 7. These
conditions gave better peak shapes for Nitrofen and Dicofol. Table
6 lists the retention times for the compounds detected on this column
pair.
7.4 Calibration
7.4.1 Prepare calibration standards using the procedures in Sec.
5.0. Refer to Method 8000 (Sec. 7.0) for proper calibration techniques for
both initial calibration and calibration verification. The procedure for
either internal or external calibration may be used. In most cases,
external standard calibration is used with Method 8081 because of the
sensitivity of the electron capture detector and the probability of the
internal standard being affected by interferences. Because several of the
pesticides may co-elute on any single-column, analysts should use two
calibration mixtures (see Sec. 3.8). The specific mixture should be
selected to minimize the problem of peak overlap.
NOTE: Because of the sensitivity of the electron capture detector, the injection
port and column should always be cleaned prior to performing the initial
calibration.
7.4.1.1 A mid-point calibration standard of all multi-
component analytes must be included with the initial calibration (for
pattern recognition) so that the analyst is familiar with the patterns
and retention times on each column.
7.4.1.2 For calibration verification (each 12-hour shift)
all target analytes required in the project plan must be injected.
7.4.2 Establish the GC operating conditions appropriate for the
configuration (single-column or dual column, Sec. 7.3) using Tables 4, 5,
7, or 8 as guidance. Optimize the instrumental conditions for resolution
of the target analytes and sensitivity. An initial oven temperature <140 -
150°C is required to resolve the four BHC isomers. A final temperature of
240 - 270"C is required to elute decachlorobiphenyl. Use of injector
pressure programming will improve the chromatography of late eluting peaks.
NOTE: Once established, the same operating conditions must be used for both
calibrations and sample analyses.
7.4.3 A 2 juL injection volume of each calibration standard is
recommended. Other injection volumes may be employed, provided that the
analyst can demonstrate adequate sensitivity for the compounds of interest.
7.4.4 Because of the low concentration of pesticide standards
injected on a GC/ECD, column adsorption may be a problem when the GC has
not been used for a day or more. Therefore, the GC column should be primed
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(or deactivated) by injecting a pesticide standard mixture approximately
20 times more concentrated than the mid-concentration standard. Inject
this standard mixture prior to beginning the initial calibration or
calibration verification.
CAUTION: Several analytes, including Aldrin, may be observed in the injection
just following this system priming. Always run an acceptable blank
prior to running any standards or samples.
7.4.5 Calibration factors
When external standard calibration is employed, calculate the
calibration factor for each analyte at each concentration, the mean
calibration factor, and the relative standard deviation (RSD) of the
calibration factors, using the formulae below. If internal standard
calibration is employed, refer to Method 8000 for the calculation of
response factors.
7.4.5.1 Calculate the calibration factor for each analyte at
each concentration as:
CF =
Peak Area (or Height) of the Compound in the Standard
Mass of the Compound Injected (in nanograms)
7.4.5.2 Calculate the mean calibration factor for each
analyte as:
mean CF = CF = ±2
where n is the number of standards analyzed.
7.4.5.3 Calculate the standard deviation (SD) and the RSD of
the calibration factors for each analyte as:
SD =
E(CF,-CF):
n-1
RSD = — x 100
CT
If the RSD for each analyte is s 20%, then the response of the
instrument is considered linear and the mean calibration factor can
be used to quantitate sample results. If the RSD is greater than 20%,
then linearity through the origin cannot be assumed. The analyst must
use a calibration curve or a non-linear calibration model (e.g., a
polynomial equation) for quantitation. See Method 8000 for
information on non-linear calibrations.
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7.4.6 Retention time windows
7.4.6.1 Before establishing the retention time windows, make
sure the gas chromatographic system is within optimum operating
conditions. The width of the retention time window should be based
upon actual retention times of three standards analyzed over the
course of 72 hours. See Method 8000 for details.
7.4.6.2 The widths of the retention time windows are defined
as plus or minus three times the standard deviation of the absolute
retention times for each of the three standards. However, the
experience of the analyst should weigh heavily in the interpretation
of the chromatograms. Method 8000 provides guidance on establishing
absolute retention time windows.
7.4.6.3 Certain analytes, particularly Kepone, are subject
to changes in retention times. Dry Kepone standards prepared in
hexane or isooctane can produce Gaussian peaks. However, Kepone
extracted from samples or standards exposed to water or methanol may
produce peaks with broad tails that elute later than the standard by
up to 1 minute. This shift is presumably the result of the formation
of a hemi-acetal from the ketone functionality. As a result, the use
of Method 8270 is recommended for the analysis of Kepone.
7.5 Gas chromatographic analysis of sample extracts
7.5.1 The same GC operating conditions used for the initial
calibration must be employed for samples analyses.
7.5.2 Verify calibration each 12-hour shift by injecting calibration
verification standards prior to conducting any sample analyses. Analysts
should alternate the use of high and low concentration mixtures of single-
component analytes and multi-component analytes for calibration
verification. A calibration standard must also be injected at intervals
of not less than once every twenty samples (after every 10 samples is
recommended to minimize the number of samples requiring re-injection when
QC limits are exceeded) and at the end of the analysis sequence.
7.5.2.1 The calibration factor for each analyte to be
quantitated must not exceed a ± 15 percent difference when compared
to the initial calibration curve.
CF -CF
% Difference = v- x 100
CF
7.5.2.2 If this criterion is exceeded, inspect the gas
chromatographic system to determine the cause and perform whatever
maintenance is necessary before verifying calibration and proceeding
with sample analysis.
7.5.2.3 If routine maintenance does not return the
instrument performance to meet the QC requirements (Sec. 8.2) based
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on the last initial calibration, then a new initial calibration must
be performed.
7.5.3 Compare the retention time of each analyte in the calibration
standard with the absolute retention time windows established in Sec.
7.4.6. As described in Method 8000, the center of the absolute retention
time window for each analyte is its retention time in the mid-concentration
standard analyzed during the initial calibration. Each analyte in each
standard must fall within its respective retention time window. If not,
the gas chromatographic system must either be adjusted so that a second
analysis of the standard does result in all analytes falling within their
retention time windows, or a new initial calibration must be performed and
new retention time windows established.
7.5.4 Inject a 2-p.l aliquot of the concentrated sample extract.
Record the volume injected to the nearest 0.05 juL and the resulting peak
size in area units.
7.5.5 Tentative identification of an analyte occurs when a peak from
a sample extract falls within the absolute retention time window. Each
tentative identification must be confirmed using either a second 8C column
of dissimilar stationary phase or using another technique such as GC/MS
(see Sec. 8.4).
7.5.6 When using the external calibration procedure (Method 8000),
determine the quantity of each component peak in the sample chromatogram
which corresponds to the compounds used for calibration purposes, as
follows. Proper quantitation requires the appropriate selection of a
baseline from which the peak area or height can be determined.
7.5.6.1 For aqueous samples
(A)(Vt)(D)
Concentration (ug/L) = — - — - -
(CF)(V.)(Vs)
where:
Ax
Vt
D
Area (or height) of the peak for the analyte in the sample.
Total volume of the concentrated extract
Dilution factor, if the sample or extract was diluted prior to
analysis. If no dilution was made, D = 1. The dilution factor
is always dimensionless.
CF = Mean calibration factor from the initial calibration (area/ng).
Volume of the extract injected (/uL). The injection volume for
samples and calibration standards must be the same. For
purge-and-trap analysis, Vi is not applicable and therefore is
set at 1.
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V. = Volume of the aqueous sample extracted in mL. If units of liters
are used for this term, multiply the results by 1000.
Using the units specified here for these terms will result in a
concentration in units of ng/mL, which is equivalent to
7.5.6.2 For non-aqueous samples
(Ax)(Vt)(D)
Concentration (ug/kg) =
where Ax, Vt, D, CF, and Vf are the same as for aqueous samples, and
W. = Weight of sample extracted (g). The wet weight or dry weight may
be used, depending upon the specific application of the data.
If units of kilograms are used for this term, multiply the
results by 1000.
Using the units specified here for these terms will result in a
concentration in units of ng/g, which is equivalent to M9/kg.
7.5.6.3 See Method 8000 for the equation used for internal
standard quantitation.
7.5.6.4 If the responses exceed the calibration range of the
system, dilute the extract and reanalyze. Peak height measurements
are recommended over peak area integration when overlapping peaks
cause errors in area integration.
7.5.6.5 If partially overlapping or coeluting peaks are
found, change GC columns or try GC/MS quantitation (see Sec. 8.0 and
Method 8270).
7.5.7 Each sample analysis must be bracketed with an acceptable
initial calibration, calibration verification standard(s) (each 12-hour
analytical shift), or calibration standards interspersed within the
samples. When a calibration verification standard fails to meet the QC
criteria, all samples that were injected after the last standard that last
met the QC criteria must be re-injected.
Although analysis of a single mid-concentration standard (standard
mixture or multi-component analyte) will satisfy the minimum requirements,
analysts are urged to use different calibration verification standards
during organochlorine pesticide analyses. Also, multi-level standards
(mixtures or multi-component analytes) are highly recommended to ensure
that the detector response remains stable for all the analytes over the
calibration range.
7.5.8 Sample injections may continue for as long as the calibration
verification standards and standards interspersed with the samples meet
instrument QC requirements. It is recommended that standards be analyzed
after every 10 samples (required after every 20 samples and at the end of
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a set) to minimize the number of samples that must be re-injected when the
standards fail the QC limits. The sequence ends when the set of samples
has been injected or when qualitative and/or quantitative QC criteria are
exceeded.
7.5.9 If the peak response is less than 2.5 times the baseline noise
level, the validity of the quantitative result may be questionable. The
analyst should consult with the source of the sample to determine whether
further concentration of the sample is warranted.
7.5.10 Validation of GC system qualitative performance
7.5.10.1 Use the calibration standards analyzed during the
sequence to evaluate retention time stability. The retention time
windows are established using the absolute retention time of each
analyte in the mid-concentration standard during the initial
calibration as the mid-point of the window. The widths of the windows
are defined as the mid-point plus and minus three times the standard
deviations calculated in Sec. 7.4.6.
7.5.10.2 Each subsequent injection of a standard during the
12-hour analytical shift (i.e., those standards injected every 20
samples, or more frequently) must be checked against the retention
time windows. If any of these subsequent standards fall outside their
absolute retention time windows, the GC system is out of control.
Determine the cause of the problem and correct it. If the problem
cannot be corrected, a new initial calibration must be performed.
7.5.11 Identification of mixtures (i.e. Methoxychlor and Toxaphene)
is based on the characteristic "fingerprint" retention time and shape of
the indicator peak(s); and quantitation is based on the area under the
characteristic peaks as compared to the area under the corresponding
calibration peak(s) of the same retention time and shape generated using
either internal or external calibration procedures.
7.5.12 If compound identification or quantitation is precluded due
to interference (e.g., broad, rounded peaks or ill-defined baselines are
present) cleanup of the extract or replacement of the capillary column or
detector is warranted. Rerun the sample on another instrument to determine
if the problem results from analytical hardware or the sample matrix.
Refer to Method 3600 for the procedures to be followed in sample cleanup.
7.6 Quantitation of multi-component analytes - Multi-component analytes
present problems in measurement. Suggestions are offered in the following
sections for handing Toxaphene, Strobane, Chlordane, BHC, and DDT.
7.6.1 Toxaphene and Strobane - Toxaphene is manufactured by the
chlorination of camphenes, whereas Strobane results from the chlorination
of a mixture of camphenes and pinenes. Quantitation of Toxaphene or
Strobane is difficult, but reasonable accuracy can be obtained. To
calculate Toxaphene from GC/ECD results:
7.6.1.1. Adjust the sample size so that the major Toxaphene
peaks are 10-70% of full-scale deflection (FSD).
8081A - 17 Revision 1
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7.6.1.2 Inject a Toxaphene standard that is estimated to be
within ± 10 ng of the sample amount.
7.6.1.3 Quantitate using the five major peaks or the total
area of the Toxaphene pattern.
7.6.1.3.1 To measure total area, construct the
baseline of standard Toxaphene between its extremities and
construct the baseline under the sample peaks, using the
distances of the peak troughs to baseline on the standard as
a guide. This procedure is made difficult by the fact that
the relative heights and widths of the peaks in the sample
will probably not be identical to the standard.
7.6.1.3.2 A series of Toxaphene residues have been
calculated using the total peak area for comparison to the
standard and also using the area of the last four peaks only,
in both sample and standard. The agreement between the
results obtained by the two methods justifies the use of the
latter method for calculating Toxaphene in a sample where the
early eluting portion of the Toxaphene chromatogram shows
interferences from other substances such as DDT.
7.6.2 Chlordane - Chlordane is a technical mixture of at least 11
major components and 30 or more minor components. Trans- and c/s-Chlordane
(a and 7, respectively), are the two major components of technical
Chlordane. However, the exact percentage of each in the technical material
is not completely defined, and is not consistent from batch to batch.
7.6.2.1 The GC pattern of a Chlordane residue may differ
considerably from that of the technical standard. Depending on the
sample substrate and its history, residues of Chlordane can consist
of almost any combination of constituents from the technical
Chlordane, plant and/or animal metabolites, and products of
degradation caused by exposure to environmental factors such as water
and sunlight.
7.6.2.2 Whenever possible, when a Chlordane residue does not
resemble technical Chlordane, the analyst should quantitate the peaks
of a-Chlordane, y-Chlordane, and Heptachlor separately against the
appropriate reference materials, and report the individual residues.
7.6.2.3 When the GC pattern of the residue resembles that of
technical Chlordane, the analyst may quantitate Chlordane residues by
comparing the total area of the Chlordane chromatogram using the five
major peaks or the total area. If the Heptachlor epoxide peak is
relatively small, include it as part of the total Chlordane area for
calculation of the residue. If Heptachlor and/or Heptachlor epoxide
are much out of proportion, calculate these separately and subtract
their areas from the total area to give a corrected Chlordane area.
NOTE: Octachloro epoxide, a metabolite of Chlordane, can easily be mistaken for
Heptachlor epoxide on a nonpolar GC column.
8081A - 18 Revision 1
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7.6.2.4 To measure the total area of the Chlordane
chromatogram, inject an amount of a technical Chlordane standard which
will produce a chromatogram in which the major peaks are approximately
the same size as those in the sample chromatograms.
7.6.3 Hexachlorocyclohexane - Hexachlorocyclohexane is also known
as BHC, from the former name, benzene hexachloride. Technical grade BHC
is a cream-colored amorphous solid with a very characteristic musty odor.
It consists of a mixture of six chemically distinct isomers and one or more
heptachlorocyclohexanes and octachlorocyclohexanes. Commercial BHC
preparations may show a wide variance in the percentage of individual
isomers present. Quantitate each isomer (a, /3, 7, and s) separately
against a standard of the respective pure isomer.
7.6.4 DDT - Technical DDT consists primarily of a mixture of 4,4'-
DDT (approximately 75%) and 2, 4' -DDT (approximately 25%). As DDT weathers,
4,4'-DDE, 2,4'-DDE, 4,4'-DDD, and 2,4'-DDD are formed. Since the 4,4'-
isomers of DDT, DDE, and ODD predominate in the environment, these are the
isomers normally regulated by EPA. Therefore, sample extracts should be
quantitated against standards of the respective pure isomers of 4,4'-DDT,
4,4'-DDE, and 4,4'-DDD.
7.7 GC/MS confirmation may be used in conjunction with either single-
column or dual-column analysis if the concentration is sufficient for detection
by GC/MS.
7.7.1 Full -scan GC/MS will normally require a concentration of
approximately 10 ng//iL in the final extract for each single-component
compound. Ion trap or selected ion monitoring will normall-y require a
concentration of approximately 1
7.7.2 The GC/MS must be calibrated for the specific target
pesticides when it is used for quantitative analysis.
7.7.3 GC/MS may not be used for confirmation when concentrations are
below 1 ng/jLiL in the extract.
7.7.4 GC/MS confirmation should be accomplished by analyzing the
same extract that is used for GC/ECD analysis and the extract of the
associated method blank.
7.7.5 The base/neutral/acid extract and the associated blank may be
used for GC/MS confirmation if the surrogates and internal standards do not
interfere and if it is demonstrated that the analyte is stable during
acid/base partitioning. However, if the compounds are not detected in the
base/neutral/acid extract, then GC/MS analysis of the pesticide extract
should be performed.
7.7.6 A QC reference sample containing the compound must also be
analyzed by GC/MS. The concentration of the QC reference sample must
demonstrate that those pesticides identified by GC/ECD can be confirmed by
GC/MS.
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7.8 Suggested chromatographic system maintenance - When system performance
does not meet the established QC requirements, corrective action is required, and
may include one or more of the following.
7.8.1 Splitter connections - For dual-columns which are connected
using a press-fit Y-shaped glass splitter or a Y-shaped fused-silica
connector, clean and deactivate the splitter port insert or replace with
a cleaned and deactivated splitter. Break off the first few centimeters
(up to 30 cm) of the injection port side of the column. Remove the columns
and solvent backflush according to the manufacturer's instructions. If
these procedures fail to eliminate the degradation problem, it may be
necessary to deactivate the metal injector body and/or replace the columns.
7.8.2 GC injector ports can be of critical concern, especially in
the analysis of DDT and Endrin. Injectors that are contaminated,
chemically active, or too hot can cause the degradation ("breakdown") of
the analytes. Endrin and DDT breakdown to Endrin aldehyde, Endrin ketone,
ODD, or DDE. When such breakdown is observed, clean and deactivate the
injector port, break off at least 30 cm of the column and remount it.
Check the injector temperature and lower it to 205"C, if required. Endrin
and DDT breakdown are less of a problem when ambient on-column injectors
are used.
7.8.3 Metal injector body - Turn off the oven and remove the
analytical columns when the oven has cooled. Remove the glass injection
port insert (instruments with on-column injection). Lower the injection
port temperature to room temperature. Inspect the injection port and
remove any noticeable foreign material.
7.8.3.1 Place a beaker beneath the injector port inside the
oven. Using a wash bottle, serially rinse the entire inside of the
injector port with acetone and then toluene, catching the rinsate in
the beaker.
7.8.3.2 Prepare a solution of a deactivating agent (Sylon-CT
or equivalent) following manufacturer's directions. After all metal
surfaces inside the injector body have been thoroughly coated with the
deactivation solution, rinse the injector body with toluene, methanol,
acetone, then hexane. Reassemble the injector and replace the
columns.
7.8.4 Column rinsing - The column should be rinsed with several
column volumes of an appropriate solvent. Both polar and nonpolar solvents
are recommended. Depending on the nature of the sample residues expected,
the first rinse might be water, followed by methanol and acetone.
Methylene chloride is a good final rinse and in some cases may be the only
solvent required. The column should then be filled with methylene chloride
and allowed to stand flooded overnight to allow materials within the
stationary phase to migrate into the solvent. The column is then flushed
with fresh methylene chloride, drained, and dried at room temperature with
a stream of ultrapure nitrogen.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation techniques can be found in Method 3500. If an extract
cleanup procedure was performed, refer to Method 3600 for the appropriate quality
control procedures. Each laboratory should maintain a formal quality assurance
program. The laboratory should also maintain records to document the quality of
the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification, and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency
8.3.1 Each laboratory must demonstrate initial proficiency with each
sample preparation and determinative method combination it utilizes, by
generating data of acceptable accuracy and precision for target analytes
in a clean matrix. The laboratory must also repeat the following
operations whenever new staff are trained or significant changes in
instrumentation are made.
8.3.2 It is suggested that the quality control (QC) reference sample
concentrate (as discussed in Section 8.0 of Methods 8000 and 3500) contain
each analyte of interest at 10 mg/L. If this method is to be used for
analysis of Chlordane or Toxaphene only, the QC reference sample
concentrate should contain the most representative multi-component mixture
at a suggested concentration of 50 mg/L in acetone. See Method 8000, Sec.
8.0 for additional information on how to accomplish this demonstration.
8.3.3 If the recovery of any compound found in the QC reference
sample is less than 80 percent or greater than 120 percent of the certified
value, the laboratory performance is judged to be out of control, and the
problem must be corrected. A new set of calibration standards should be
prepared and analyzed.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, a laboratory control sample (LCS), and the addition of surrogates to
each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, the laboratories
should use a matrix spike and matrix spike duplicate pair.
8081A - 21 Revision 1
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8.4.2 Single laboratory accuracy and recovery data for this method
can be found in Tables 9, 10, and 11. In-house method performance criteria
should be developed using the guidance found in Sec. 8.0 of Method 8000.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicates a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.4 Include a calibration standard after each group of 20 samples
(it is recommended that a calibration standard be included after every 10
samples to minimize the number of repeat injections) in the analysis
sequence as a calibration check. The response factors for the calibration
should be within 15 percent of the initial calibration. When this
continuing calibration is out of this acceptance window, the laboratory
should stop analyses and take corrective action.
8.4.5 Whenever quantitation is accomplished using an internal
standard, internal standards must be evaluated for acceptance. The
measured area of the internal standard must be no more than 50 percent
different from the average area calculated during calibration. When the
internal standard peak area is outside the limit, all samples that fall
outside the QC criteria must be reanalyzed.
8.4.6 DDT and Endrin are easily degraded in the injection port.
Breakdown occurs when the injection port liner is contaminated high boiling
residue from sample injection or when the injector contains metal fittings.
Check for degradation problems by injecting a standard containing only
4,4'-DDT and Endrin. Presence of 4,4'-DDE, 4,4'-DDD, Endrin ketone or
Endrin indicates breakdown. If degradation of either DDT or Endrin exceeds
15%, take corrective action before proceeding with calibration.
8.4.6.1 Calculate percent breakdown as follows:
% breakdown of DDT - sum of degradation peak areas (ODD + DDE)xl()0
sum of all peak areas (DDT + DDE + ODD)
.. . . . c r . . sum of degradation peak areas (aldehyde + ketone) 1rtn
% breakdown of Endrin = - -xlOO
sum of all peak areas (Endrin + aldehyde + ketone)
8.4.6.2 The breakdown of DDT and Endrin should be measured
before samples are analyzed and at the beginning of each 12-hour
shift. Injector maintenance and recalibration should be completed if
the breakdown is greater than 15% for either compound (Sec. 7.8.2).
8.4.7 Whenever silica gel (Method 3630) or Florisil (Method 3620)
cleanups are used, the analyst must demonstrate that the fractionation
8081A - 22 Revision 1
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scheme is reproducible. Batch to batch variation in the composition of the
silica gel or Florisil or overloading the column may cause a change in the
distribution patterns of the organochlorine pesticides. When compounds are
found in two fractions, add the concentrations found in the fractions, and
correct for any additional dilution.
8.4.8 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries: The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined in Chapter One. The MDL
concentrations listed in Tables 1 and 2 were obtained using organic-free reagent
water and sandy loam soil.
9.2 The chromatographic separations in this method have been tested in a
single laboratory by using clean hexane and liquid and solid waste extracts that
were spiked with the test compounds at three concentrations. Single-operator
precision, overall precision, and method accuracy were found to be related to the
concentration of the compound and the type of matrix.
9.3 This method has been applied in a variety of commercial laboratories
for environmental and waste matrices. Performance data were obtained for a
limited number of target analytes spiked into sewage sludge and dichloroethene
stillbottoms at high concentrations. These data are provided in Tables 9 and 10.
9.4 The accuracy and precision obtainable with this method depend on the
sample matrix, sample preparation technique, optional cleanup techniques, and
calibration procedures used.
9.5 Single laboratory accuracy data were obtained for organochlorine
pesticides in a clay soil. The spiking concentration was 500 MQAg- The
spiking solution was mixed into the soil and then immediately transferred to the
extraction device and immersed in the extraction solvent. The spiked sample was
then extracted by Method 3541 (Automated Soxhlet). The data represent a single
determination. Analysis was by capillary column gas chromatography/electron
capture detector following Method 8081 for the organochlorine pesticides. These
data are listed in Table 11 and were taken from Reference 10.
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10.0 REFERENCES
1. Lopez-Avila, V., Baldin, E., Benedicto, J, Milanes, J., Beckert. W.F.,
"Application of Open-Tubular Columns to SW-846 GC Methods", final report
to the U.S. Environmental Protection Agency on Contract 68-03-3511; Mid-
Pacific Environmental Laboratory, Mountain View, CA, 1990.
2. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 10 - Pesticides and PCB Report for the
U.S. Environmental Protection Agency on Contract 68-03-2606.
3. Goerlitz, D.F., Law, L.M., "Removal of Elemental Sulfur Interferences from
Sediment Extracts for Pesticide Analysis", Bull. Environ. Contam. Toxicol.,
1971, 6, 9.
4. Jensen, S., Renberg, L., Reutergardth, L., "Residue Analysis of Sediment
and Sewage Sludge for Organochlorines in the Presence of Elemental Sulfur",
Anal. Chem., 1977, 49, 316-318.
5. Wise, R.H., Bishop, D.F., Williams, R.T., Austern, B.M., "Gel Permeation
Chromatography in the GC/MS Analysis of Organics in Sludges", U.S.
Environmental Protection Agency, Cincinnati, OH, 45268.
6. Pionke, H.B., Chesters, G., Armstrong, D.E., "Extraction of Chlorinated
Hydrocarbon Insecticides from Soil", Agron. J., 1968, 60, 289.
7. Burke, J.A., Mills, P.A., Bostwick, D.C., "Experiments with Evaporation of
Solutions of Chlorinated Pesticides", J. Assoc. Off. Anal. Chem., 1966, 49,
999.
8. Glazer, J.A., et al., "Trace Analyses for Wastewaters", Environ. Sci. and
Technol., 1981, 15, 1426.
9. Marsden, P.J., "Performance Data for SW-846 Methods 8270, 8081, and 8141",
U.S. Environmental Protection Agency, EMSL-las Vegas, EPA/600/4-90/015.
10. Lopez-Avila, V. (Beckert, W., Project Officer), "Development of a Soxtec
Extraction Procedure for Extracting Organic Compounds from Soils and
Sediments", EPA 600/X-91/140, US Environmental Protection Agency,
Environmental Monitoring Systems Laboratory, Las Vegas, NV, October 1991.
8081A - 24 Revision 1
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TABLE 1
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES
USING WIDE-BORE CAPILLARY COLUMNS
SINGLE-COLUMN METHOD OF ANALYSIS
Retention Time (min) MDLa Water MDL" Soil
Compound DB 608b DB 1701b (M9/L)
Aldrin
a-BHC
B-BHC
£-BHC
7-BHC (Lindane)
or-Chlordane
7-Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Toxaphene
11.84
8.14
9.86
11.20
9.52
15.24
14.63
18.43
16.34
19.48
16.41
15.25
18.45
20.21
17.80
19.72
10.66
13.97
22.80
MR
12.50
9.46
13.58
14.39
10.84
16.48
16.20
19.56
16.76
20.10
17.32
15.96
19.72
22.36
18.06
21.18
11.56
15.03
22.34
MR
0.034
0.035
0.023
0.024
0.025
0.008
0.037
0.050
0.058
0.081
0.044
0.030
0.040
0.035
0.039
0.050
0.040
0.032
0.086
NA
2.2
1.9
3.3
1.1
2.0
1.5
4.2
2.5
3.6
NA
2.1
2.4
3.6
3.6
1.6
2.0
2.1
5.7
NA
These MDLs are for organic-free reagent water or sandy loam soil.
NA = Data not available.
MR = Multiple response compound.
" MDL is the method detection limit. The MDL was determined from the
analysis of seven replicate aliquots of each matrix processed through the
entire analytical method (extraction, silica gel cleanup, and GC/ECD
analysis).
MDL = t(IV1 099I x SD, where t(tv1 099) is the Student's t value appropriate for a 99%
confidence interval and n-1 degrees of freedom, and SD is the standard
deviation of the seven replicate measurements.
b See Table 4 for GC operating conditions.
8081A - 25 Revision 1
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TABLE 2
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES
USING NARROW-BORE CAPILLARY COLUMNS
SINGLE-COLUMN METHOD OF ANALYSIS
Compound
Aldrin
a-BHC
6-BHC
-------
TABLE 3
FACTORS FOR DETERMINATION OF ESTIMATED QUANTITATION LIMITS8 (EQLs)
FOR VARIOUS MATRICES
Matrix Factor
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
EQL = [MDL for water (see Tables 1 and 2)] times [Factor in this table]
For nonaqueous samples, the factor is on a wet-weight basis. Sample
EQLs are highly matrix-dependent. EQLs determined using these factors
are provided as guidance and may not always be achievable.
8081A - 27 Revision 1
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TABLE 4
GC OPERATING CONDITIONS FOR ORGANOCHLORINE COMPOUNDS
SINGLE-COLUMN ANALYSIS USING NARROW-BORE COLUMNS
Column 1 - 30 m x 0.25 or 0.32 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5 or equivalent), 1 jitm film thickness.
Carrier gas
Carrier gas pressure
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
Helium
16 psi
225°C
300°C
100°C, hold 2 minutes
100'C to 160°C at 15eC/min, followed
by 160°C to 270°C at 5'C/min
270°C
Column 2 - 30 m x 0.25 mm ID fused silica capillary column chemically bonded
with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608, or equivalent), 25
/im coating thickness, 1 fj,m film thickness.
Carrier gas
Carrier gas pressure
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
Nitrogen
20 psi
225'C
300°C
160°C, hold 2 minutes
160°C to 290'C at 5°C/min
290°C, hold 1 min
8081A - 28
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TABLE 5
GC OPERATING CONDITIONS FOR ORGANOCHLORINE COMPOUNDS
SINGLE-COLUMN ANALYSIS USING WIDE-BORE COLUMNS
Column 1 - 30 m x 0.53 mm ID fused silica capillary column chemically bonded
with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608, RTx-35, or
equivalent), 0.5 jum or 0.83 jum film thickness.
Column 2 - 30 m x 0.53 mm ID fused silica capillary column chemically bonded
with 50 percent phenyl methylpolysiloxane (DB-1701, or equivalent), 1.0 /urn
film thickness.
Both Column 1 and Column 2 use the same GC operating conditions.
Carrier gas
Carrier gas flow rate
Makeup gas
Makeup gas flow rate
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
Helium
5-7 mL/minute
argon/methane (P-5 or P-10) or
nitrogen
30 mL/min
250°C
290°C
150°C, hold 0.5 minute
150°C to 270°C at 5°C/min
270°C, hold 10 min
Column 3 - 30 m x 0.53 mm ID fused silica capillary column chemically bonded
with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5 jum film thickness.
Carrier gas
Carrier gas flow rate
Makeup gas
Makeup gas flow rate
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
Helium
6 mL/minute
argon/methane (P-5 or P-10) or
nitrogen
30 mL/min
205°C
290°C
140°C, hold 2 min
140'C to 240°C at 10°C/min, hold 5
minutes at 240'C, 240°C to 265eC at
5°C/min
265'C, hold 18 min
8081A - 29
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TABLE 6
RETENTION TIMES OF THE ORGANOCHLORINE PESTICIDES8
DUAL-COLUMN METHOD OF ANALYSIS
Compound
DBCP
Hexachl orocycl opentadi ene
Etridiazole
Chloroneb
Hexachlorobenzene
Diallate
Propachlor
Trifluralin
a-BHC
PCNB
y-BHC
Heptachlor
Aldrin
Alachlor
Chlorothalonil
Alachlor
/3-BHC
Isodrin
DCPA
£-BHC
Heptachlor epoxide
Endosulfan-I
•y-Chlordane
a-Chlordane
tra/7s-Nonachlor
4,4'-DDE
Dieldrin
Perthane
Endrin
Chloropropylate
Chlorobenzilate
Nitrofen
4,4'-DDD
Endosulfan II
4,4'-DDT
Endrin aldehyde
Mi rex
Endosulfan sulfate
Methoxychlor
Captafol
DB-5 RT (min)
2.14
4.49
6.38
7.46
12.79
12.35
9.96
11.87
12.35
14.47
14.14
18.34
20.37
18.58
15.81
18.58
13.80
22.08
21.38
15.49
22.83
25.00
24.29
25.25
25.58
26.80
26.60
28.45
27.86
28.92
28.92
27.86
29.32
28.45
31.62
29.63
37.15
31.62
35.33
32.65
DB-1701 RT (min)
2.84
4.88
8.42
10.60
14.58
15.07
15.43
16.26
17.42
18.20
20.00
21.16
22.78
24.18
24.42
24.18
25.04
25.29
26.11
26.37
27.31
28.88
29.32
29.82
30.01
30.40
31.20
32.18
32.44
34.14
34.42
34.42
35.32
35.51
36.30
38.08
38.79
40.05
40.31
41.42
(continued)
8081A - 30
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TABLE 6 (continued)
RETENTION TIMES OF THE ORGANOCHLORINE PESTICIDES'
DUAL-COLUMN METHOD OF ANALYSIS
Compound DB-5 RT (min) DB-1701 RT (min)
Endrin ketone 33.79 42.26
Permethrin 41.50 45.81
Kepone 31.10 b
Dicofol 35.33 b
Dichlone 15.17 b
or,or'-Dibromo-m-xylene 9.17 11.51
2-Bromobiphenyl 8.54 12.49
8 See Table 7 for GC operating conditions.
b Not detected at 2 ng per injection.
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TABLE 7
GC OPERATING CONDITIONS FOR ORGANOCHLORINE PESTICIDES
FOR DUAL-COLUMN METHOD OF ANALYSIS
LOW TEMPERATURE, THIN FILM
Column 1:
Column 2;
DB-1701 or equivalent
30 m x 0.53 mm ID
1.0 juro film thickness
DB-5 or equivalent
30 m x 0.53 mm ID
0.83 jum film thickness
Carrier gas
Carrier gas flow rate
Makeup gas
Makeup gas flow rate
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
Helium
6 mL/minute
Nitrogen
20 mL/min
250'C
320eC
140°C, hold 2 minutes
140eC to 270°C at 2.8°C/min
270'C, hold 1 minute
8081A - 32
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TABLE 8
GC OPERATING CONDITIONS FOR ORGANOCHLORINE PESTICIDES
FOR THE DUAL COLUMN METHOD OF ANALYSIS
HIGH TEMPERATURE, THICK FILM
Column 1
Column 2:
DB-1701 (J&W) or equivalent
30 m x 0.53 mm ID
1.0 jum
DB-5 (J&W) or equivalent
30 m x 0.53 mm ID
1.5 jum
Carrier gas:
Carrier gas flowrate:
Makeup gas:
Makeup gas flowrate:
Injector temperature:
Detector temperature:
Initial temperature:
Temperature program:
Final temperature
Helium
6 mL/minute
Nitrogen
20 (mL/min)
250°C
320'C
150°C, hold 0.5 min
150°C to 190eC at 12°C/min, hold 2
min
190°C to 275°C at 4°C/min
275°C, hold 10 min
8081A - 33
Revision 1
January 1995
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TABLE 9
ANALYTE RECOVERY FROM SEWAGE SLUDGE
Compound
Sonication
%Recovery %RSD
Soxhlet
%Recovery %RSD
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
7-BHC
Heptachlor
Aldrin
/3-BHC
-------
TABLE 10
ANALYTE RECOVERY FROM DICHLOROETHANE STILLBOTTOMS
Compound
Sonication
%Recovery %RSD
Soxhlet
%Recovery %RSD
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
|8-BHC
Heptachlor
Aldrin
i8-BHC
5-BHC
Heptachlor epoxide
Endosulfan I
7-Chlordane
a-Chlordane
DDE
Dieldrin
Endrin
Endosulfan II
DDT
Endrin aldehyde
ODD
Tetrachloro-m-xylene
Decachlorobiphenyl
70
59
159
55
43
48
48
51
43
47
47
48
45
45
45
50
49
49
40
48
49
17
2
3
14
7
6
6
5
7
4
6
4
5
5
4
5
6
5
4
4
5
2
29
50
35
128
47
30
55
200
75
119
66
41
47
37
70
58
41
46
40
29
35
176
104
30
35
137
25
30
18
258
42
129
34
18
13
21
40
24
23
17
29
20
21
211
93
Concentration spiked in the sample: 500-1000 ng/g, three replicates analyses.
Soxhlet extraction by Method 3540 with methylene chloride.
Sonication extraction by Method 3550 with methylene chloride/acetone (1:1).
Cleanup by Method 3640.
GC column: DB-608, 30 m x 0.53 mm ID.
8081A - 35
Revision 1
January 1995
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TABLE 11
SINGLE LABORATORY ACCURACY DATA FOR THE EXTRACTION OF
ORGANOCHLORINE PESTICIDES FROM SPIKED CLAY SOIL BY METHOD 3541
(AUTOMATED SOXHLET)"
% Recovery
Compound Name DB-5 DB-1701
a-BHC
0-BHC
Heptachlor
Aldrin
Heptachlor epoxide
trans-Chlordane
Endosulfan I
Dieldrin
Endrin
Endosulfan II
4,4'-DDT
Mi rex
89
86
94
ND
97
94
92
ND
111
104
ND
108
94
ND
95
92
97
95
92
113
104
104
ND
102
* The operating conditions for the automated Soxhlet were:
Immersion time 45 min; extraction time 45 min; 10 g sample size; extraction
solvent, 1:1 acetone/hexane. No equilibration time following spiking.
ND = Not able to determine because of interference..
All compounds were spiked at 500
Data taken from Reference 10.
8081A - 36 Revision 1
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FIGURE 1.
GAS CHROMATOGRAM OF THE MIXED ORGANOCHLORINE PESTICIDE STANDARD
Start Time : 0.00 min
Scale Factor: 0
End Time : 33.00 mm
Plot Offset: 20 mV
Low Point : 20.00 mV High Pomt : «0.00 mV
Plot Scale: 400 nv
Response [rnV]
o-
u\—
o
'H
5
H
J
ro
o
04
O
-------
FIGURE 2
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX A
Scai« -Kl?
£"«2 T 'TW 53 30
' at :*»ser 20 nv
^3« eo'"t 20.30 *
»i3t Scan. 250 nv
Response [mV]
LH—
rt>
3
en
O
o
O
o
o
-5.13
-7.93
•12.33
-14.27
-17.08
•=S—
^2
20.22
20.77
22.68
-23.73
-28.52
-9.86
-10.98
-13.58
NO
Ln
O
-17.54
-18.47
-19.78
-19.24
-21.13
•1.95
-3. 5-;
-22.33
-30.05
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100'C (hold 2 minutes) to 160°C at 158C/min, then at
5*C/min to 270*C; carrier He at 16 psi.
8081A - 38
Revision 1
January 1995
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FIGURE 3
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX B
Start Time • 0 00 mm
Scale Factor: 0
End T ime . 33.00 mm
Plot Offset: 20 mv
Low Point : 20.00 mV High Point • 270.00 mV
Plot Scale: 250 mV
Response [mV]
o-
L.I—
0)
3
3
01
LJT
Ui
o
I I I I I I
o
o
I I I I I
(Ji O
O O
I I I I
to
r_n
O
I I I I I
--2.74
-6.97
4—
. 60
--10.71
-11.73
12:33
^-14.27
^-15.24
^Llfi,OR
-14.84
-16.23
—17.08
-17.63
W:
-18.31
19.54
-20.19
21.03
'.. 00
--22.68
30.04
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100'C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081A - 39
Revision 1
January 1995
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FIGURE 4.
GAS CHROMATOGRAM OF THE TOXAPHENE STANDARD
Start Time : 0.00 inin End Time : 53.00 urn Lou Point : 20.00 mv
Scile Factor: 0 Plat Offset: 20 «v Plot Sctlt: 60 *v
Point : 80.00 IHV
Response [mV]
r-O OJ
O
(D
-_
' N)
-
Li"
I I I I I I I I 11 I I
O O O
I I I I I I I 11 I I I I I I I I 11 I I I I I I I I II I I I I I I I I II I I I I ill I |_|] I II I I H I I
VSf
24. 32
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100'C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270*C; carrier He at 16 psi.
8081A - 40
Revision 1
January 1995
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FIGURE 5
GAS CHROMATOGRAM OF THE TECHNICAL CHLORDANE STANDARD
Start T"ne : 0.00 mm End Time : 33.00 mm Lou Point : 20.00 mV Higfe Point : 220.00
Scan Factor- 0 Plot Offset: 20 mV Plot Sctlf. 200 M
Response
o
O
NJ
O
o
m
niT
D
n"
I'D
' NO
J 0~
O—
-
-
Ul —
-
-
o~
-
(Ji~
•=&
-
•
~^t\Mt 1 1fj.59
•— 4. 33
-5.83
-i 8.87
4^*p-*>.
-Wil 1*3
%lh*i6
*T'?2
^1Jij°jj 13 60
W^"1*.,
*4EHv^ «n
Column:
Temperature program:
30 m x 0.25 mm ID DB-5 fused silica capillary.
100°C (hold 2 minutes) to 160'C at 15'C/min, then at
5°C/min to 270*C; carrier He at 16 psi.
8081A - 41
Revision 1
January 1995
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FIGURE 6
GAS CHROMATOGRAM OF TOXAPHENE
DB-1701
LJ
DB-5
Toxaphene analyzed on a DB-5/DB-1701 fused-silica open-tubular column pair. The
GC operating conditions were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jitm film
thickness) and 30 m x 0.53 mm ID DB-1701 (LO-^m film thickness) connected to
a J&W Scientific press-fit Y-shaped inlet splitter. Temperature program: 150"C
(0.5 min hold) to 190 °C (2 min hold) at 12°C/min then to 275°C (10 min hold) at
4'C/min.
8081A - 42
Revision 1
January 1995
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FIGURE 7
GAS CHROMATOGRAM OF STROBANE
CO
(M
DB-1701
r-
o
-------
FIGURE 8
GAS CHROMATOGRAM OF ORGANOCHLORINE PESTICIDES
IS SU
I U.o ..13
OB-5
t
21
12 3 4 SU IS 4
L
i .
1
DB-1701
10 11 12 IS
2S
14
uwuu
32
4 IS 11 4
20.
Organochlorine pesticides analyzed on a DB-5/DB-1701 fused-silica open-tubular
column pair. The GC operating conditions were as follows: 30 m x 0.53 mm ID DB-
5 (0.83-jum film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0-jiim film thickness)
connected to an 8 in injection tee (Supelco Inc.). Temperature program: 140°C
(2 min hold) to 270°C (1 min hold) at 2.8'C/min.
8081A - 44
Revision 1
January 1995
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METHOD 8081A
ORGANOCHLORINE PESTICIDES BY CAPILLARY COLUMN GAS CHROMATOGRAPHY
7.1 Choose
appropriate extraction
technique (see Chapter 2
and Method 3500).
7.1 Add specified
matrix spike to sample.
7.2 Routine cleanup/
fractionation.
7.3 Choose single-column
or dual-column GC
configuration.
7.4.1 Refer to Method 8000
for proper calibration
techniques.
7.4.2 Establish GC
operating conditions.
7.4.3 Inject each
calibration standard.
7.4.5 Calculate
calibration factors
for each analyte.
7.4.6 Calculate determination
time windows for
each analyte.
7.5.4 Inject in aliquot
of sample extract.
7.5.5 - 7.5.6 Identify
and quantify the peaks
observed in the
chromatogram.
7.5.7 Bracket the
sample analysis with
calibration standards
(every 10 samples).
7.5.9 Is
peak at least
2.5 times
noise?
7.6 Any
multicomponent
analytes
present?
7.5.8 Additional
cleanup or
concentration.
7.6.1 - 7.6.4 Calculate
concentration of
Toxaphene, Strobane,
Chlordane, BHC, or DOT.
8081A - 45
Revision 1
January 1995
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METHOD 8082
POLYCHLORINATED BIPHENYLS (PCBs)
BY CAPILLARY COLUMN GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8082 is used to determine the concentrations of polychlorinated
biphenyls (PCBs) as Aroclors or as individual PCB congeners in extracts from
solid and aqueous matrices. Open-tubular, capillary columns were employed with
electron capture detectors (ECD) or electrolytic conductivity detectors (ELCD).
When compared to packed columns, these fused-silica, open-tubular columns offer
improved resolution, better selectivity, increased sensitivity, and faster
analysis. The target compounds listed below may be determined by either a
single- or dual-column analysis system. The PCB congeners listed below have been
tested by this method, and the method may be appropriate for additional
congeners.
Compound
CAS Registry No.
IUPAC #
Aroclor 1016
Aroclor 1221
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1260
2-Chlorobiphenyl
2,3-Dichlorobiphenyl
2,2' ,5-Trichlorobiphenyl
2,4' ,5-Trichlorobiphenyl
2,2' ,3,5'-Tetrachlorobiphenyl
2 , 2 ' , 5 , 5 ' -Tetrachl orobi phenyl
2,3' ,4,4'-Tetrachlorobiphenyl
2,2' ,3,4,5'-Pentachlorobiphenyl
2,2' ,4,5,5'-Pentachlorobiphenyl
2,3,3' ,4' ,6-Pentachlorobiphenyl
2,2' ,3,4,4' ,5-Hexachl orobi phenyl
2,2',3,4,5,5'-Hexachlorobiphenyl
2,2',3,5,5',6-Hexachlorobiphenyl
2,2',4,4',5,5'-Hexachlorobiphenyl
2,2',3,3',4,4',5-Heptachlorobiphenyl
2,2',3,4,4',5,5'-Heptachlorobiphenyl
2,2' ,3,4,4',5',6-Heptachlorobiphenyl
2,2' ,3,4' ,5,5' ,6-Heptachlorobiphenyl
2,2',3,3',4,4',5,5',6-Nonachlorobiphenyl
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
2051-60-7
16605-91-7
37680-65-2
16606-02-3
41464-39-5
35693-99-3
33025-41-1
38380-02-8
37680-72-3
38380-03-9
35694-06-5
52712-04-6
52663-63-5
38380-01-7
35065-30-6
35065-29-3
52663-69-1
52663-68-0
40186-72-9
-
-
-
-
-
-
1
5
18
31
44
52
66
87
101
110
138
141
151
153
170
180
183
187
206
8082 - 1
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January 1995
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1.2 Aroclors are multi-component mixtures. When samples contain more than
one Aroclor, a higher level of analyst expertise is required to attain acceptable
levels of qualitative and quantitative analysis. The same is true of Aroclors
that have been subjected to environmental degradation ("weathering") or
degradation by treatment technologies. Such weathered multi-component mixtures
may have significant differences in peak patterns than those of Aroclor
standards.
1.3 Quantitation of PCBs as Aroclors is appropriate for many regulatory
compliance determinations, but is particularly difficult when the Aroclors have
been weathered by long exposure in the environment. Therefore, this method
provides procedures for the determination of selected individual PCB congeners.
The 19 PCB congeners listed above have been tested by this method.
1.4 The PCB congener approach potentially affords greater quantitative
accuracy when PCBs are known to be present. As a result, this method may be used
to determine Aroclors, some PCB congeners, or "total PCBs," depending on
regulatory requirements and project needs. The congener method is of particular
value in determining weathered Aroclors. However, analysts should use caution
when using the congener method when regulatory requirements are based on Aroclor
concentrations.
1.5 Compound identification based on single-column analysis should be
confirmed on a second column, or should be supported by at least one other
qualitative technique. This method describes analytical conditions for a second
gas chromatographic column that can be used to confirm the measurements made with
the primary column. GC/MS Method 8270 is also recommended as a confirmation
technique when sensitivity permits (Sec. 8.0).
1.6 This method also describes a dual-column option. The option allows
a hardware configuration of two analytical columns joined to a single injection
port. The option allows one injection to be used for dual-column analysis.
Analysts are cautioned that the dual-column option may not be appropriate when
the instrument is subject to mechanical stress, many samples are to be run in a
short period, or when highly contaminated samples are analyzed.
1.7 The analyst must select columns, detectors and calibration procedures
most appropriate for the specific analytes of interest in a study. Matrix-
specific performance data must be established and the stability of the analytical
system and instrument calibration must be established for each analytical matrix
(e.g., hexane solutions from sample extractions, diluted oil samples, etc.).
Example chromatograms and GC conditions are provided as guidance.
1.8 The MDLs for Aroclors vary in the range of 0.054 to 0.90 ^g/L in water
and 57 to 70 M9/kg in soils. Estimated quantisation limits may be determined
using the data in Table 1.
1.9 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs (GC) and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
8082 - 2 Revision 0
January 1995
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2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 L for liquids,
2 g to 30 g for solids) is extracted using the appropriate matrix-specific sample
extraction technique.
2.2 Aqueous samples are extracted at neutral pH with methylene chloride
using Method 3510 (separatory funnel), Method 3520 (continuous liquid-liquid
extractor), or other appropriate technique.
2.3 Solid samples are extracted with hexane-acetone (1:1) or methylene
chloride-acetone (1:1) using Method 3540 (Soxhlet), Method 3541 (automated
Soxhlet), or other appropriate technique.
2.4 Extracts for PCB analysis may be subjected to a sulfuric
acid/potassium permanganate cleanup (Method 3665) designed specifically for these
analytes. This cleanup technique will remove (destroy) many single component
organochlorine or organophosphorus pesticides. Therefore, Method 8082 is not
applicable to the analysis of those compounds. Instead, use Method 8081.
2.5 After cleanup, the extract is analyzed by injecting a 2-juL aliquot
into a gas chromatograph with a narrow- or wide-bore fused silica capillary
column and electron capture detector (GC/ECD).
2.6 The chromatographic data may be used to determine the seven Aroclors
in Sec. 1.1, individual PCB congeners, or total PCBs.
3.0 INTERFERENCES
3.1 Refer to Methods 3500 (Sec. 3.0, in particular), 3600, and 8000 for
a discussion of interferences.
3.2 Interferences co-extracted from the samples will vary considerably
from matrix to matrix. While general cleanup techniques are referenced or
provided as part of this method, unique samples may require additional cleanup
approaches to achieve desired degrees of discrimination and quantitation.
Sources of interference in this method can be grouped into three broad
categories.
3.2.1 Contaminated solvents, reagents, or sample processing
hardware.
3.2.2 Contaminated GC carrier gas, parts, column surfaces, or
detector surfaces.
3.2.3 Compounds extracted from the sample matrix to which the
detector will respond.
3.3 Interferences by phthalate esters introduced during sample preparation
can pose a major problem in PCB determinations.
3.3.1 Common flexible plastics contain varying amounts of phthalate
esters which are easily extracted or leached from such materials during
8082 - 3 Revision 0
January 1995
-------
laboratory operations. Interferences from phthalate esters can best be
minimized by avoiding contact with any plastic materials and checking all
solvents and reagents for phthalate contamination.
3.3.2 Exhaustive cleanup of solvents, reagents and glassware may be
required to eliminate background phthalate ester contamination.
3.3.3 These materials can be removed through the use of Method 3665
(sulfuric acid/permanganate cleanup).
3.4 Cross-contamination of clean glassware routinely occurs when plastics
are handled during extraction steps, especially when solvent-wetted surfaces are
handled. Glassware must be scrupulously cleaned.
Clean all glassware as soon as possible after use by rinsing with the last
solvent used. This should be followed by detergent washing with hot water, and
rinses with tap water and organic-free reagent water. Drain the glassware, and
dry it in an oven at 130"C for several hours, or rinse with methanol and drain.
Store dry glassware in a clean environment.
NOTE: Oven-drying of glassware used for PCB analysis can increase contamination
because PCBs are readily volatilized in the oven and spread to other
glassware. Therefore, exercise caution, and do not dry glassware from
samples containing high concentrations of PCBs with glassware that may be
used for trace analyses.
3.5 Elemental sulfur (S8) is readily extracted from soil samples and may
cause chromatographic interferences in the determination of PCBs. Sulfur can be
removed through the use of Method 3660.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - An analytical system complete with gas
chromatograph suitable for on-column and split-splitless injection and all
required accessories including syringes, analytical columns, gases, electron
capture detectors (ECD), and recorder/integrator or data system.
4.2 GC columns
This method describes procedures for both single-column and dual-column
analyses. The single-column approach involves one analysis to determine that a
compound is present, followed by a second analysis to confirm the identity of the
compound (Sec. 8.4 describes how GC/MS confirmation techniques may be employed).
The single-column approach may employ either narrow-bore (<0.32 mm ID) columns
or wide-bore (0.53 mm ID) columns. The dual-column approach involves a single
injection that is split between two columns that are mounted in a single gas
chromatograph. The dual-column approach employs only wide-bore (0.53 mm ID)
columns.
The columns listed in this section were the columns used to develop the
method performance data. Listing these columns in this method is not intended
to exclude the use of other columns that may be developed. Laboratories may use
8082 - 4 Revision 0
January 1995
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other capillary columns provided that they document method performance (e.g.,
chromatographic resolution, analyte breakdown, and MDLs) that equals or exceeds
the performance specified in this method.
4.2.1 Narrow-bore columns for single-column analysis (use both
columns to confirm compound identifications unless another confirmation
technique such as GC/MS is employed). Narrow bore columns should be
installed in split/splitless (Grob-type) injectors.
4.2.1.1 30 m x 0.25 or 0.32 mm ID fused silica capillary
column chemically bonded with SE-54 (DB-5 or equivalent), 1 jum film
thickness.
4.2.1.2 30 m x 0.25 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608,
SPB-608, or equivalent), 2.5 jum coating thickness, 1 jum film
thickness.
4.2.2 Wide-bore columns for single-column analysis (use two of the
three columns listed to confirm compound identifications unless another
confirmation technique such as GC/MS is employed). Wide-bore columns
should be installed in 1/4 inch injectors, with deactivated liners designed
specifically for use with these columns.
4.2.2.1 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608,
SPB-608, RTx-35, or equivalent), 0.5 jum or 0.83 jum film thickness.
4.2.2.2 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 50 percent phenyl methylpolysiloxane (DB-1701,
or equivalent), 1.0 ^m film thickness.
4.2.2.3 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5
jum film thickness.
4.2.3 Wide-bore columns for dual-column analysis (choose one of the
two pairs of columns listed below).
4.2.3.1 Column pair 1
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5 /*m film
thickness.
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or
equivalent), 1.0 jum film thickness.
Column pair 1 is mounted in a press-fit Y-shaped glass 3-way
union splitter (J&W Scientific, Catalog No. 705-0733) or a Y-shaped
fused-silica connector (Restek, Catalog No. 20405), or equivalent.
8082 - 5 Revision 0
January 1995
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4.2.3.2 Column pair 2
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 0.83 /xm film
thickness.
30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or
equivalent), 1.0 /urn film thickness.
Column pair 2 is mounted in an 8 in. deactivated glass
injection tee (Supelco, Catalog No. 2-3665M), or equivalent.
4.3 Column rinsing kit - Bonded-phase column rinse kit (J&W Scientific,
Catalog No. 430-3000), or equivalent.
4.4 Volumetric flasks - 10-mL and 25-mL, for preparation of standards.
5.0 REAGENTS
5.1 Reagent grade or pesticide grade chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
NOTE: Store the standard solutions (stock, composite, calibration,
internal, and surrogate standards) at 4°C in Teflon®-sealed
containers in the dark. When a lot of standards is prepared, it is
recommended that aliquots of that lot be stored in individual small
vials. All stock standard solutions must be replaced after one year
or sooner if routine QC (Sec. 8.0) indicates a problem. All other
standard solutions must be replaced after six months or sooner if
routine QC (Sec. 8.0) indicates a problem.
5.2 Sample extracts prepared by Methods 3510, 3520, 3540, 3541, 3545, or
3550 need to undergo a solvent exchange step prior to analysis. The following
solvents are necessary for dilution of sample extracts. All solvent lots should
be pesticide quality or equivalent and should be determined to be phthalate-free.
5.2.1 n-Hexane, C6H14
5.2.2 Isooctane, (CH3)3CCH2CH(CH3)2
5.3 The following solvents may be necessary for the preparation of
standards. All solvent lots must be pesticide quality or equivalent and should
be determined to be phthalate-free.
5.3.1 Acetone, (CH3)2CO
5.3.2 Toluene, C6H5CH3
8082 - 6 Revision 0
January 1995
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5.4 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water as defined in Chapter One.
5.5 Stock standard solutions (1000 mg/L) - May be prepared from pure
standard materials or can be purchased as certified solutions.
5.5.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10-mL volumetric flask. If compound purity is
96 percent or greater, the weight may be used without correction to
calculate the concentration of the stock standard solution.
5.5.2 Commercially-prepared stock standard solutions may be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.6 Calibration standards for Aroclors
5.6.1 A standard containing Aroclor 1016 and Aroclor 1260 will
include the major peaks represented in the other five Aroclor mixtures.
Therefore, GC calibration for Aroclors can be accomplished by the analysis
of five standards containing both Aroclor 1016 and Aroclor 1260. Prepare
a minimum of five calibration standards containing equal concentrations of
both Aroclor 1016 and Aroclor 1260 by dilution of the stock standard with
isooctane or hexane. The concentrations should correspond to the expected
range of concentrations found in real samples and should bracket the linear
range of the detector.
5.6.2 Standards of each of the other five Aroclors are required to
aid the analyst in pattern recognition. Prepare one standard for each of
the other Aroclors at a concentration in the mid-point of the calibration
range.
5.7 Calibration standards for PCB congeners
5.7.1 If results are to be determined for individual PCB congeners,
then standards for the pure congeners must be prepared. The table in Sec.
1.1 lists 19 PCB congeners that have been tested by this method along with
the IUPAC numbers designating these congeners. This procedure may be
appropriate for other congeners as well.
5.7.2 Stock standards may be prepared in a fashion similar to that
described for the Aroclor standards, or may be purchased as commercially-
prepared solutions. Stock standards should be used to prepare a minimum
of five concentrations by dilution of the stock standard with isooctane or
hexane. The concentrations should correspond to the expected range of
concentrations found in real samples and should bracket the linear range
of the detector.
5.8 Internal standard
5.8.1 When PCB congeners are to be determined, the use of an
internal standard is highly recommended. Decachlorobiphenyl may be used
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as an internal standard, added to each sample extract prior to analysis,
and included in each of the initial calibration standards.
5.8.2 When PCBs are to be determined as Aroclors, an internal
standard is not used, and decachlorobiphenyl is employed as a surrogate
(see Sec. 5.8).
5.9 Surrogate standards
5.9.1 When PCBs are to be determined as Aroclors, decachlorobiphenyl
is used as a surrogate, and is added to each sample prior to extraction.
Prepare a solution of decachlorobiphenyl at a concentration of 5 mg/L in
acetone.
5.9.2 When PCB congeners are to be determined, decachlorobiphenyl
is recommended for use as an internal standard, and therefore, cannot also
be used as a surrogate. Therefore, tetrachloro-meta-xylene may be used as
a surrogate for PCB congener analysis. Prepare a solution of tetrachloro-
meta-xylene at a concentration of 5 mg/L in acetone.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See Chapter Four, Organic Analytes for sample collection and
preservation instructions.
6.2 Extracts must be stored under refrigeration in the dark and analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Sample extraction
7.1.1 Refer to Chapter Two and Method 3500 for guidance in choosing
the appropriate extraction procedure. In general, water samples are
extracted at a neutral pH with methylene chloride using a separatory funnel
(Method 3510) or a continuous liquid-liquid extractor (Method 3520) or
other appropriate procedure. Solid samples are extracted with hexane-
acetone (1:1) or methylene chloride-acetone (1:1) using one of the Soxhlet
extraction (Method 3540 or 3541) procedures, ultrasonic extraction (Method
3550), or other appropriate procedure.
NOTE: Use of hexane-acetone generally reduces the amount of interferences
that are extracted and improves signal-to-noise.
7.1.2 Spiked samples are used to verify the applicability of the
chosen extraction technique to each new sample type. Each sample type must
be spiked with the compounds of interest to determine the percent recovery
and the limit of detection for that sample type (see Chapter One). See
Method 8000 for guidance on demonstration of initial method proficiency as
well as guidance on matrix spikes for routine sample analysis.
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7.2 Extract cleanup
Refer to Methods 3660 and 3665 for information on extract cleanup.
7.3 GC conditions
This method allows the analyst to choose between a single-column or a dual-
column configuration in the injector port. Either wide- or narrow-bore columns
may be used. Identifications based on retention times from a single-column must
be confirmed on a second column or with an alternative qualitative technique.
However, if true Aroclors are present, then identification can be based on
recognition of Aroclor patterns from a single-column analysis.
7.3.1 Single-column analysis
This capillary GC/ECD method allows the analyst the option of using
0.25-0.32 mm ID capillary columns (narrow-bore) or 0.53 mm ID capillary
columns (wide-bore). The use of narrow-bore (0.25-0.32 mm ID) columns is
recommended when the analyst requires greater chromatographic resolution.
Use of narrow-bore columns is suitable for relatively clean samples or for
extracts that have been prepared with one or more of the clean-up options
referenced in the method. Wide-bore columns (0.53 mm ID) are suitable for
more complex environmental and waste matrices.
7.3.2 Dual-column analysis
The dual-column/dual-detector approach involves the use of two 30 m
x 0.53 mm ID fused-silica open-tubular columns of different polarities,
thus different selectivities towards the target compounds. The columns are
connected to an injection tee and ECD detectors.
7.3.3 GC temperature programs and flow rates
7.3.3.1 Table 2 lists GC operating conditions for the
analysis of PCBs as Aroclors for single-column analysis, using either
narrow-bore or wide-bore capillary columns. Table 3 lists GC
operating conditions for the dual-column analysis. Use the conditions
in these tables as guidance and establish the GC temperature program
and flow rate necessary to separate the analytes of interest.
7.3.3.2 When determining PCBs as congeners, difficulties may
be encountered with coelution of congener 153 and other sample
components. When determining PCBs as Aroclors, chromatographic
conditions should be adjusted to give adequate separation of the
characteristic peaks in each Aroclor (see Sec. 7.4.5).
7.3.3.3 Tables 4 and 5 summarize the retention times of up
to 73 Aroclor peaks determined during dual-column analysis, using the
operating conditions in Table 2. Note that the peak numbers used in
these tables are not the IUPAC congener numbers, but represent the
elution order of the peaks on these GC columns.
7.3.3.4 Once established, the same operating conditions must
be used for the analysis of samples and standards.
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7.4 Calibration
7.4.1 Prepare calibration standards using the procedures in Sec.
5.0. Refer to Method 8000 (Sec. 7.0) for proper calibration techniques for
both initial calibration and calibration verification. When PCBs are to
be determined as congeners, the use of internal standard calibration is
highly recommended. Therefore, the calibration standards must contain the
internal standard (see Sec. 5.7) at the same concentration as the sample
extracts. When PCBs are to be determined as Aroclors, external standard
calibration is used.
NOTE: Because of the sensitivity of the electron capture detector, the
injection port and column should always be cleaned prior to performing
the initial calioration.
7.4.2 When PCBs are to be determined as congeners, an initial five-
point calibration must be performed that includes standards for all the
target analytes. When PCBs are to be determined as Aroclors, the initial
calibration includes the analysis of five standards containing a mixture
of Aroclor 1016 and Aroclor 1260 as well as the analysis of a single
standard of each of the other five Aroclors (for pattern recognition).
7.4.3 Establish the GC operating conditions appropriate for the
configuration (single-column or dual column, Sec. 7.3). Optimize the
instrumental conditions for resolution of the target compounds and
sensitivity. A final temperature of 240-270'C is required to elute
decachlorobiphenyl. Use of injector pressure programming will improve the
chromatography of late eluting peaks.
NOTE: Once established, the same operating conditions must be used for both
calibrations and sample analyses.
7.4.4 A 2 jiL injection of each calibration standard is recommended.
Other injection volumes may be employed, provided that the analyst can
demonstrate adequate sensitivity for the compounds of interest.
7.4.5. Record the peak area (or height) for each congener or each
characteristic Aroclor peak to be used for quantitation. A minimum of 3
peaks must be chosen for each Aroclor, and preferably 5 peaks. The peaks
must be characteristic of the Aroclor in question. A peak common to more
than one Aroclor may not be used for quantitation. Characteristic peaks
are defined as those peaks in the Aroclor standards that are at least 25%
of the height of the largest Aroclor peak. Late-eluting Aroclor peaks are
generally the most stable in the environment. Table 6 lists characteristic
peaks in each Aroclor, along with their retention times on two GC columns
suitable for single-column analysis. Table 7 lists 13 specific PCB
congeners found in Aroclor mixtures. Table 8 lists PCB congeners with
corresponding retention times on a DB-5 wide-bore GC column.
7.4.6 When determining PCB congeners by the internal standard
procedure, calculate the response factor (RF) for each congener in the
calibration standards relative to the internal standard,
decachlorobiphenyl, using the equation that follows.
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where:
Ai8 -
A x C.
RF = _! '1
A x C
Peak area (or height) of the analyte or surrogate.
Peak area (or height) of the internal standard.
Concentration of the analyte or surrogate, in M9/L.
Concentration of the internal standard, in M9/L-
7.4.7 When determining PCBs as Aroclors by the external standard
technique, calculate the calibration factor (CF) for each characteristic
Aroclor peak in each of the five initial calibration standards containing
the mixture of Aroclor 1016 and 1260, using the equation below. Do not
calculate calibration factors from the individual standards for the other
Aroclors, as these standards are used solely for pattern recognition.
CF =
Peak Area (or Height) in the Standard
Total Mass of the Standard Injected (in nanograms)
7.4.8 The response factors or calibration factors from the initial
calibration are used to evaluate the linearity of the initial calibration.
This involves the calculation of the mean response or calibration factor,
the standard deviation, and the relative standard deviation (RSD) for each
congener or Aroclor peak. See Method 8000 for the specifics of the
evaluation of the linearity of the calibration and guidance on performing
non-linear calibrations.
7.5 Retention time windows
Retention time windows are crucial to the identification of target
compounds. Absolute retention times are used for the identification of PCBs as
Aroclors. When PCBs are determined as congeners by an internal standard
technique, absolute retention times may be used in conjunction with relative
retention times (relative to the internal standard). Retention time windows are
established to compensate for minor shifts in absolute retention times as a
result of sample loadings and normal chromatographic variability. The width of
the retention time window should be carefully established to minimize the
occurrence of both false positive and false negative results. Tight retention
time windows may result in false negatives and/or may cause unnecessary
reanalysis of samples when surrogates or spiked compounds are erroneously not
identified. Overly wide retention time windows may result in false positive
results that cannot be confirmed upon further analysis.
7.5.1 Before establishing the retention time windows, make sure that
the gas chromatographic system is within optimum operating conditions. The
width of the retention time window should be based upon actual retention
times of three standards measured over the course of 72 hours. See Method
8000 for details.
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7.5.2 Retention time windows are defined as plus or minus three
times the standard deviation of the absolute retention times for each of
the three standards. However, the experience of the analyst should weigh
heavily in the interpretation of the chromatograms. Method 8000 provides
guidance on the establishment of absolute retention time windows.
7.6 Gas chromatographic analysis of sample extracts
7.6.1 The same GC operating conditions used for the initial
calibration must be employed for samples analyses.
7.6.2 Verify calibration each 12-hour shift by injecting calibration
verification standards prior to conducting any sample analyses. A
calibration standard must also be injected at intervals of not less than
once every twenty samples (after every 10 samples is recommended to
minimize the number of samples requiring re-injection when QC limits are
exceeded) and at the end of the analysis sequence. For Aroclor analyses,
the calibration verification standard should be a mixture of Aroclor 1016
and Aroclor 1260. The calibration verification process does not require
analysis of the other Aroclor standards used for pattern recognition, but
the analyst may wish to include a standard for one of these Aroclors after
the 1016/1260 mixture used for calibration verification throughout the
analytical sequence.
7.6.2.1 The calibration factor for each analyte calculated
from the calibration verification standard (CFJ must not exceed a
difference of more than ± 15 percent when compared to the mean
calibration factor from the initial calibration curve.
CF - CF
% Difference = x 100
CF
7.6.2.2 When internal standard calibration is used for PCB
congeners, the response factor calculated from the calibration
verification standard (RFJ must not exceed a ± 15 percent difference
when compared to the mean response factor from the initial calibration
RF - RF
% Difference = ——1 x 100
RF
7.6.2.3 If this criterion is exceeded for any calibration
factor or response factor, inspect the gas chromatographic system to
determine the cause and perform whatever maintenance is necessary
before verifying calibration and proceeding with sample analysis.
7.6.2.4 If routine maintenance does not return the
instrument performance to meet the QC requirements (Sec. 8.2) based
on the last initial calibration, then a new initial calibration must
be performed.
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7.6.3 Inject a 2-)uL aliquot of the concentrated sample extract.
Record the volume injected to the nearest 0.05 p.1 and the resulting peak
size in area (or peak height) units.
7.6.4 Tentative identification of an analyte occurs when a peak from
a sample extract falls within the daily retention time window. Each
tentative identification must be confirmed using either a second GC column
of dissimilar stationary phase (as in the dual-column analysis) or using
another technique such as GC/MS (see Sec. 8.4).
7.6.5 Using either the internal or the external calibration
procedure (Method 8000), determine the quantity of each component peak in
the sample chromatogram which corresponds to the compounds used for
calibration purposes (either Aroclors or congeners).
If the responses exceed the calibration range of the system, dilute
the extract and reanalyze. Peak height measurements are recommended over
peak area when overlapping peaks cause errors in area integration.
7.6.6 Each sample analysis must be bracketed with an acceptable
initial calibration, calibration verification standard(s) (each 12-hour
shift), or calibration standards interspersed within the samples. When a
calibration verification standard fails to meet the QC criteria, all
samples that were injected after the last standard that last met the QC
criteria must be re-injected.
Multi-level standards (mixtures or multi-component analytes) are
highly recommended to ensure that detector response remains stable for all
analytes over the calibration range.
7.6.7 Sample injections may continue for as long as the calibration
verification standards and standards interspersed with the samples meet
instrument QC requirements. It is recommended that standards be analyzed
after every 10 samples (required after every 20 samples and at the end of
a set) to minimize the number of samples that must be re-injected when the
standards fail the QC limits. The sequence ends when the set of samples
has been injected or when qualitative or quantitative QC criteria are
exceeded.
7.6.8 If the peak response is less than 2.5 times the baseline noise
level, the validity of the quantitative result may be questionable. The
analyst should consult with the source of the sample to determine whether
further concentration of the sample is warranted.
7.6.9 Use the calibration standards analyzed during the sequence to
evaluate retention time stability. If any of the standards fall outside
their daily retention time windows, the system is out of control.
Determine the cause of the problem and correct it.
7.6.10 If compound identification or quantitation is precluded due
to interference (e.g., broad, rounded peaks or ill-defined baselines are
present) cleanup of the extract or replacement of the capillary column or
detector is warranted. Rerun the sample on another instrument to determine
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if the problem results from analytical hardware or the sample matrix.
Refer to Method 3600 for the procedures to be followed in sample cleanup.
7.7 Quantitation of PCBs as congeners
7.7.1 The quantitation of PCB congeners is accomplished by the
comparison of the sample chromatogram to those of the PCB congener
standards, using the internal standard technique (see Method 8000).
Calculate the concentration of each congener.
7.7.2 Depending on project requirements, the PCB congener results
may be reported as congeners, or may be summed and reported as total PCBs.
The analyst should use caution when using the congener method for
quantitation when regulatory requirements are based on Aroclor
concentrations. See Sec. 9.3.
7.8 Quantitation of PCBs as Aroclors
The quantitation of PCB residues as Aroclors is accomplished by comparison
of the sample chromatogram to that of the most similar Aroclor standard. A
choice must be made as to which Aroclor is most similar to that of the residue
and whether that standard is truly representative of the PCBs in the sample.
7.8.1 Use the individual Aroclor standards (not the 1016/1260
mixtures) to determine the pattern of peaks on Aroclors 1221, 1232, 1242,
1248, and 1254. The patterns for Aroclors 1016 and 1260 will be evident
in the mixed calibration standards.
7.8.2 Once the Aroclor pattern has been identified, compare the
responses of 3 to 5 major peaks in the calibration standards (the 1016/1260
mixture) with the peaks observed in the sample extract, using the external
standard calibration technique (see Method 8000). The amount of Aroclor
is calculated using an individual calibration factor for each of the 3 -
5 characteristic peaks chosen in Sec. 7.4.4. The quantitative results for
those peaks are averaged to determine the concentration of that Aroclor.
7.8.3 When samples appear to contained weathered PCBs, treated PCBs,
or mixtures of Aroclors, the quantitation of PCBs as Aroclors is not
appropriate, and analyses should be repeated using the PCB congener
approach described in this method.
7.9 GC/MS confirmation may be used in conjunction with either single-or
dual-column analysis if the concentration is sufficient for detection by GC/MS.
7.9.1 Full-scan GC/MS will normally require a concentration of
approximately 10 ng/juL in the final extract for each single-component
compound. Ion trap or selected ion monitoring will normally require a
concentration of approximately 1 ng//iL.
7.9.2 The GC/MS must be calibrated for the specific target analytes
when it is used for quantitative analysis.
7.9.3 GC/MS may not be used for confirmation when concentrations are
below 1 ng/juL in extract.
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7.9.4 GC/MS confirmation should be accomplished by analyzing the
same extract used for GC/ECD analysis and the extract of the associated
blank.
7.9.5 The base/neutral/acid extract and the associated blank may be
used for GC/MS confirmation if the surrogates and internal standards do not
interfere. However, if the compounds are not detected in the
base/neutral/acid extract, then GC/MS analysis of the pesticide extract
should be performed.
7.9.6 A QC reference sample containing the compound must also be
analyzed by GC/MS. The concentration of the QC reference sample must
demonstrate that those PCBs identified by GC/ECD can be confirmed by GC/MS.
7.10 Chromatographic System Maintenance as Corrective Action
When system performance does not meet the established QC requirements,
corrective action is required, and may include one or more of the following.
7.10.1 Splitter connections
For dual columns which are connected using a press-fit Y-shaped glass
splitter or a Y-shaped fused-silica connector, clean and deactivate the
splitter port insert or replace with a cleaned and deactivated splitter.
Break off the first few inches (up to one foot) of the injection port side
of the column. Remove the columns and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate the
degradation problem, it may be necessary to deactivate the metal injector
body and/or replace the columns.
7.10.2 Metal injector body
Turn off the oven and remove the analytical columns when the oven has
cooled. Remove the glass injection port insert (instruments with on-column
injection). Lower the injection port temperature to room temperature.
Inspect the injection port and remove any noticeable foreign material.
7.10.2.1 Place a beaker beneath the injector port inside the
oven. Using a wash bottle, rinse the entire inside of the injector
port with acetone and then rinse it with toluene, catching the rinsate
in the beaker.
7.10.2.2 Prepare a solution of a deactivating agent (Sylon-CT
or equivalent) following manufacturer's directions. After all metal
surfaces inside the injector body have been thoroughly coated with the
deactivation solution, rinse the injector body with toluene, methanol,
acetone, and then hexane. Reassemble the injector and replace the
columns.
7.10.3 Column rinsing
The column should be rinsed with several column volumes of an
appropriate solvent. Both polar and nonpolar solvents are recommended.
Depending on the nature of the sample residues expected, the first rinse
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might be water, followed by methanol and acetone. Methyl ene chloride is
a good final rinse and in some cases may be the only solvent required. The
column should then be filled with methylene chloride and allowed to stand
flooded overnight to allow materials within the stationary phase to migrate
into the solvent. The column is then flushed with fresh methylene
chloride, drained, and dried at room temperature with a stream of ultrapure
nitrogen.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation techniques can be found in Method 3500. If an extract
cleanup procedure was performed, refer to Method 3600 for the appropriate quality
control procedures. Each laboratory should maintain a formal quality assurance
program. The laboratory should also maintain records to document the quality of
the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.3.1 The QC Reference Sample concentrate (Method 3500) should
contain PCBs as Aroclors at 10-50 mg/L for water samples, or PCBs as
congeners at the same concentrations. A 1-mL volume of this concentrate
spiked into 1 L of reagent water will result in a sample concentration of
10-50
8.3.1.1 The frequency of analysis of the QC reference sample
analysis is equivalent to a minimum of 1 per 20 samples or 1 per batch
if less than 20 samples.
8.3.1.2 If the recovery of any compound found in the QC
reference sample is less than 80 percent or greater than 120 percent
of the certified value, the laboratory performance is judged to be out
of control, and the problem must be corrected. A new set of
calibration standards should be prepared and analyzed.
8.3.2 Include a calibration standard after each group of 20 samples
(it is recommended that a calibration standard be included after every 10
samples to minimize the number of repeat injections) in the analysis
sequence as a calibration check. The response factors for the calibration
should be within 15 percent of the initial calibration. When this
continuing calibration is out of this acceptance window, the laboratory
should stop analyses and take corrective action.
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8.3.3 Whenever quantitation is accomplished using an internal
standard, internal standards must be evaluated for acceptance. The
measured area of the internal standard must be no more than 50 percent
different from the average area calculated during calibration. When the
internal standard peak area is outside the limit, all samples that fall
outside the QC criteria must be reanalyzed.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDLs for Aroclors vary in the
range of 0.054 to 0.90 jug/L in water and 57 to 70 M9Ag in soils, with the
higher MDLs for the more heavily chlorinated Aroclors. Estimated quantitation
limits may be determined using the data in Table 1.
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9.2 Estimated quantitation limits for PCBs as congeners vary by congener,
in the range of 5 - 25 ng/L in water and 160 - 800 ng/kg in soils, with the
higher values for the more heavily chlorinated congeners.
9.3 The accuracy and precision obtainable with this method depend on the
sample matrix, sample preparation technique, optional cleanup techniques, and
calibration procedures used. Table 9 provides single laboratory recovery data
for Aroclors spiked into clay and soil and extracted with automated Soxhlet.
Table 10 provides multiple laboratory data on the precision and accuracy for
Aroclors spiked into soil and extracted by automated Soxhlet.
9.4 During method performance studies, the concentrations determined as
Aroclors were larger than those obtained using the congener method. In certain
soils, interference prevented the measurement of congener 66. Recoveries of
congeners from soils spiked with Aroclor 1254 and Aroclor 1260 were between 80%
and 90%. Recoveries of congeners from environmental reference materials ranged
from 51 - 66% of the certified Aroclor values.
10.0 REFERENCES
1. Lopez-Avila, V., Baldin, E., Benedicto, J, Milanes, J., Beckert. W.F.,
Application of Open-Tubular Columns to SW-846 GC Methods", final report to
the U.S. Environmental Protection Agency on Contract 68-03-3511, Mid-
Pacific Environmental Laboratory, Mountain View, CA, 1990.
2. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 10 - Pesticides and PCB Report for the
U.S. Environmental Protection Agency on Contract 68-03-2606.
3. Ahnoff, M., Josefsson, B., "Cleanup Procedures for PCB Analysis on River
Water Extracts", Bull. Environ. Contam. Toxicol., 1975, 13, 159.
4. Marsden, P.J., "Performance Data for SW-846 Methods 8270, 8081, and 8141",
U.S. Environmental Protection Agency, EMSL-Las Vegas, EPA/600/4-90/015.
5. Marsden, P.J., "Analysis of PCBs", U.S. Environmental Protection Agency,
EMSL-Las Vegas, NV, EPA/600/8-90/004.
6. Erickson, M., Analytical Chemistry of PCBs, Butterworth Publishers, Ann
Arbor Science Book, (1986).
7. Stewart, J., "EPA Verification Experiment for Validation of the SOXTEC* PCB
Extraction Procedure", Oak Ridge National Laboratory, Oak Ridge, TN,
37831-6138, October 1988.
8. Lopez-Avila, V. (Beckert, W., Project Officer), "Development of a Soxtec
Extraction Procedure for Extracting Organic Compounds from Soils and
Sediments", EPA 600/X-91/140, U.S. Environmental Protection Agency,
Environmental Monitoring Systems Laboratory, Las Vegas, NV, October 1991.
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9. Stewart, J.H., Bayne, C.K., Holmes, R.L., Rogers, W.F., and Maskarinec,
M.P., "Evaluation of a Rapid Quantitative Organic Extraction System for
Determining the Concentration of PCB in Soils", Proceedings of the U.S. EPA
Symposium on Waste Testing and Quality Assurance, Oak Ridge National
Laboratory, Oak Ridge, TN, 37831, July 11-15, 1988.
10. Tsang, S.F., Marsden, P.O., and Lesnik, B., "Quantitation of
Polychlorinated Biphenyls Using 19 Specific Congeners", Proceedings of the
9th Annual Waste Testing and Quality Assurance Symposium, Office of Solid
Waste and Emergency Response, U.S. Environmental Protection Agency,
Washington, DC, July 1993.
8082 - 19 Revision 0
January 1995
-------
TABLE 1
FACTORS FOR DETERMINATION OF ESTIMATED QUANTITATION LIMITS" (EQLs)
FOR VARIOUS MATRICES
Matrix Factor
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
"EQL = [MDL for water (see Sec. 1.8)] times [Factor in this table]
For nonaqueous samples, the factor is on a wet-weight basis. Sample EQLs are
highly matrix-dependent. EQLs determined using these factors are provided as
guidance and may not always be achievable.
8082 - 20 Revision 0
January 1995
-------
TABLE 2
GC OPERATING CONDITIONS FOR PCBs AS AROCLORS
SINGLE COLUMN ANALYSIS
Narrow-bore columns
Narrow-bore Column 1 - 30 m x 0.25 or 0.32 mm ID fused silica capillary column
chemically bonded with SE-54 (DB-5 or equivalent), 1 p,m film thickness.
Carrier gas (He) 16 psi
Injector temperature 225'C
Detector temperature 300eC
Initial temperature 100'C, hold 2 minutes
Temperature program 100°C to 160°C at 15'C/min, followed
by 160°C to 270°C at 5°C/min
Final temperature 270°C
Narrow-bore Column 2 - 30 m x 0.25 mm ID fused silica capillary column chemically
bonded with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608, or equivalent)
25 /UN coating thickness, 1 jLtm film thickness
Carrier gas (N2)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
20 psi
225°C
300°C
160°C, hold 2 minutes
160°C to 290'C at 5°C/min
290'C, hold 1 min
Wide-bore columns
Wide-bore Column 1 - 30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608, RTx-35, or
equivalent), 0.5 /xm or 0.83 pm film thickness.
Wide-bore Column 2 - 30 m x 0.53 mm ID fused silica capillary column chemically
bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or equivalent), 1.0
/im film thickness.
Carrier gas (He) 5-7 mL/minute
Makeup gas (argon/methane
[P-5 or P-10] or N2) 30 mL/min
Injector temperature 250°C
Detector temperature 290"C
Initial temperature 150"C, hold 0.5 minute
Temperature program 150°C to 270°C at 5eC/min
Final temperature 270°C, hold 10 min
(continued)
8082 - 21 Revision 0
January 1995
-------
TABLE 2 (continued)
GC OPERATING CONDITIONS FOR PCBs AS AROCLORS
SINGLE COLUMN ANALYSIS
Wide-bore Columns (continued)
Wide-bore Column 3 - 30 m x 0.53 mm ID fused silica capillary column chemically
bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5 jum film thickness.
Carrier gas (He)
Makeup gas (argon/methane
[P-5 or P-10] or N2)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
6 mL/minute
30 mL/min
205°C
290°C
140'C, hold 2 min
140eC to 240°C at 10'C/min,
hold 5 minutes at 240'C,
240eC to 265eC at 5'C/min
265°C, hold 18 min
8082 - 22
Revision 0
January 1995
-------
TABLE 3
GC OPERATING CONDITIONS FOR PCBs AS AROCLORS
FOR THE DUAL COLUMN METHOD OF ANALYSIS
HIGH TEMPERATURE, THICK FILM
Column 1 - DB-1701 or equivalent, 30 m x 0.53 mm ID, 1.0 Aim film thickness.
Column 2 - DB-5 or equivalent, 30 m x 0.53 mm ID, 1.5 jum film thickness.
Carrier gas (He) flow rate
Makeup gas (N2) flow rate
Temperature program
Injector temperature
Detector temperature
Injection volume
Solvent
Type of injector
Detector type
Range
Attenuation
Type of splitter
6 mL/min
20 mL/min
0.5 min hold
150eC to 190°C, at 12°C/min, 2 min
hold
190eC to 275°C, at 4°C/min, 10 min
hold
250eC
320eC
2 ML
Hexane
Flash vaporization
Dual ECD
10
64 (DB-1701)/64 (DB-5)
J&W Scientific press-fit Y-shaped
inlet splitter
8082 - 23
Revision 0
January 1995
-------
TABLE 4
SUMMARY OF RETENTION TIMES OF AROCLORS
ON THE DB-5 COLUMN", DUAL COLUMN ANALYSIS
Peak
No.b
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
Aroclor
1016
8.41
8.77
8.98
9.71
10.49
10.58
10.90
11.23
11.88
11.99
12.27
12.66
12.98
13.18
13.61
13.80
13.96
14.48
14.63
14.99
15.35
16.01
16.27
Aroclor
1221
5.85
7.63
8.43
8.77
8.99
10.50
10.59
11.24
12.29
12.68
12.99
Aroclor
1232
5.85
7.64
8.43
8.78
9.00
10.50
10.59
10.91
11.24
11.90
12.00
12.29
12.69
13.00
13.19
13.63
13.82
13.97
14.50
14.64
15.02
15.36
16.14
16.29
17.04
17.22
17.46
18.41
18.58
18.83
Aroclor
1242
7.57
8.37
8.73
8.94
9.66
10.44
10.53
10.86
11.18
11.84
11.95
12.24
12.64
12.95
13.14
13.58
13.77
13.93
14.46
14.60
14.98
15.32
15.96
16.08
16.26
17.19
17.43
17.92
18.16
18.37
18.56
18.80
Aroclor
1248
8.95
10.45
10.85
11.18
11.85
12.24
12.64
12.95
13.15
13.58
13.77
13.93
14.45
14.60
14.97
15.31
16.08
16.24
16.99
17.19
17.43
17.69
17.91
18.14
18.36
18.55
18.78
Aroclor
1254
13.59
13.78
13.90
14.46
14.98
15.32
16.10
16.25
16.53
16.96
17.19
17.44
17.69
17.91
18.14
18.36
18.55
18.78
Aroclor
1260
13.59
16.26
16.97
17.21
18.37
18.68
18.79
(continued)
8GC operating conditions are given in Table 3. All retention times in minutes.
bPeaks are sequentially numbered in elution order and are not isomer numbers.
8082 - 24
Revision 0
January 1995
-------
TABLE 4 (continued)
SUMMARY OF RETENTION TIMES OF AROCLORS
ON THE DB-5 COLUMN", DUAL COLUMN ANALYSIS
Peak
No."
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
Aroclor Aroclor Aroclor Aroclor Aroclor Aroclor
1016 1221 1232 1242 1248 1254
19.33 19.30 19.29 19.29
19.48
19.81
20.03 19.97 19.92 19.92
20.28
20.46 20.45
20.57
20.85 20.83 20.83
21.18 21.14 21.12 20.98
21.36 21.38
21.78
22.08 22.05 22.04
22.38
22.74
22.96
23.23
23.75
23.99
24.27
24.61
24.93
26.22
Aroclor
1260
19.29
19.48
19.80
20.28
20.57
20.83
21.38
21.78
22.03
22.37
22.73
22.95
23.23
23.42
23.73
23.97
24.16
24.45
24.62
24.91
25.44
26.19
26.52
26.75
27.41
28.07
28.35
29.00
bPeaks are sequentially numbered in elution order and are not isomer numbers,
8082 - 25
Revision 0
January 1995
-------
TABLE 5
SUMMARY OF RETENTION TIMES OF AROCLORS
ON THE DB-1701 COLUMN8, DUAL COLUMN ANALYSIS
Peak Aroclor Aroclor
No.b 1016 1221
Aroclor Aroclor Aroclor Aroclor Aroclor
1232 1242 1248 1254 1260
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
4.45
5.38
5.78
5.86
6.33 6.34
6.78 6.78
6.96 6.96
7.64
8.23 8.23
8.62 8.63
8.88
9.05 9.06
9.46
9.77 9.79
10.27 10.29
10.64 10.65
11.01
11.09
11.98
12.39
12.92
12.99
13.14
13.49
13.58
4.45
5.86
6.34
6.79
6.96
8.23
8.63
8.89
9.06
9.47
9.78
10.29
10.66
11.02
11.10
11.99
12.39
12.77
13.00
13.16
13.49
13.61
14.08
14.30
14.49
15.38
15.65
6.28
6.72
6.90
7.59
8.15
8.57
8.83
8.99
9.40
9.71
10.21
10.59
10.96
11.02
11.94
12.33
12.71
12.94
13.09
13.44
13.54
13.67
14.03
14.26
14.46
15.33
15.62
6.91
8.16
8.83
8.99
9.41
9.71
10.21
10.59
10.95
11.03
11.93
12.33
12.69
12.93
13.09
13.44
13.54
14.03
14.24
14.39
14.46
15.10
15.32
15.62
10.95
11.93
12.33
13.10
13.24
13.51
13.68
14.03
14.24
14.36
14.56
15.10
15.32
15.61
13.52
14.02
14.25
14.56
16.61
(continued)
"GC operating conditions are given in Table 3. All retention times in minutes.
bPeaks are sequentially numbered in elution order and are not isomer numbers.
8082 - 26
Revision 0
January 1995
-------
TABLE 5 (continued)
SUMMARY OF RETENTION TIMES OF AROCLORS
ON THE DB-1701 COLUMN", DUAL COLUMN ANALYSIS
Peak
No.b
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
Aroclor Aroclor Aroclor Aroclor Aroclor
1016 1221 1232 1242 1248
15.78 15.74 15.74
16.13 16.10 16.10
16.77 16.73 16,74
17.13 17.09 17.07
17.46 17.44
17.69 17.69
18.19
18.48 18.49
19.13 19.13
20.57
Aroclor
1254
15.74
16.08
16.34
16.44
16.55
16.77
17.07
17.29
17.43
17.68
18.17
18.42
18.59
18.86
19.10
19.42
19.55
20.20
20.34
20.55
20.62
20.88
21.53
21.83
23.31
Aroclor
1260
15.79
16.19
16.34
16.45
16.77
17.08
17.31
17.43
17.68
18.18
18.40
18.86
19.09
19.43
19.59
20.21
20.43
20.66
20.87
21.03
21.53
21.81
23.27
23.85
24.11
24.46
24.59
24.87
25.85
27.05
27.72
bThese are sequentially numbered from elution order and are not isomer numbers
8082 - 27
Revision 0
January 1995
-------
TABLE 6
PEAKS DIAGNOSTIC OF PCBs OBSERVED ON 0.53 mm ID COLUMN
DURING SINGLE COLUMN ANALYSIS
Peak
No.'
I
II
III
IV
V
VI
VII
VIII
IX
X
XI
RT on
DB-608b
4.90
7.15
7.89
9.38
10.69
14.24
14.81
16.71
19.27
21.22
22.89
RT on
DB-1701b
4.66
6.96
7.65
9.00
10.54
14.12
14.77
16.38
18.95
21.23
22.46
Aroclor0
1221
1221, 1232, 1248
1061, 1221, 1232, 1242,
1016, 1232, 1242, 1248,
1016, 1232, 1242,
1248, 1254
1254
1254
1254, 1260
1260
1260
" Peaks are sequentially numbered in elution order and are not isomer numbers
b Temperature program: T( = 150°C, hold 30 seconds; 5°C/minutes to 275°C.
0 Underline indicates largest peak in the pattern for that Aroclor
8082 - 28 Revision 0
January 1995
-------
Congener
TABLE 7
SPECIFIC PCB CONGENERS IN AROCLORS
IUPAC number
Aroclor
1016 1221 1232 1242 1248 1254 1260
Biphenyl
2-CB
23-DCB
34-DCB
244' -TCB
22'35'-TCB
23'44'-TCB
233'4'6-PCB
23'44'5-PCB
22'44'55'-HCB
22'344'5'-HCB
22'344'55'-HpCB
22'33'44'5-HpCB
..
1
5
12
28*
44
66*
110
118*
153
138
180
170
X
X X X X
X X X X X
X XXX
X XXX
XXX
X
X
X
X
X
X
X
X
X
X
X
X
*Apparent co-elution of:
28 with 31 (2,4',5-trichlorobiphenyl)
66 with 95 (2,2',3,5',6-pentachlorobiphenyl)
118 with 149 (2,2',3,4',5',6-hexachlorobiphenyl)
8082 - 29
Revision 0
January 1995
-------
TABLE 8
RETENTION TIMES OF PCB CONGENERS
ON THE DB-5 WIDE-BORE COLUMN
IUPAC # Retention Time (min)
1
5
18
31
52
44
66
101
87
110
151
153
138
141
187
183
180
170
206
209
6.52
10.07
11.62
13.43
14.75
15.51
17.20
18.08
19.11
19.45
19.87
21.30
21.79
22.34
22.89
23.09
24.87
25.93
30.70
32.63 (internal standard)
8082 - 30 Revision 0
January 1995
-------
TABLE 9
SINGLE LABORATORY RECOVERY DATA FOR EXTRACTION OF
PCBs FROM CLAY AND SOIL BY METHOD 3541" (AUTOMATED SOXHLET)
Matrix
Clay
Clay
Clay
Clay
Aroclor Spike Level Trial
(ppm)
1254 5 1
2
3
4
5
6
1254 50 1
2
3
4
5
6
1260 5 1
2
3
4
5
6
1260 50 1
2
3
4
5
6
Percent
Recovery11
87.0
92.7
93.8
98.6
79.4
28.3
65.3
72.6
97.2
79.6
49.8
59.1
87.3
74.6
60.8
93.8
96.9
113.1
73.5
70.1
92.4
88.9
90.2
67.3
(continued)
8 The operating conditions for the automated Soxhlet
Immersion time: 60 min
Reflux time: 60 min
b Multiple results from two different extractors.
Data from Reference 9.
8082 - 31
Revision 0
January 1995
-------
TABLE 9 (continued)
SINGLE LABORATORY RECOVERY DATA FOR EXTRACTION OF
PCBs FROM CLAY AND SOIL BY METHOD 3541" (AUTOMATED SOXHLET)
Matrix
Soil
Soil
Soil
Soil
Aroclor Spike Level Trial
(ppm)
1254 5 1
2
3
4
5
1254 50 1
2
3
4
5
6
1260 5 1
2
3
4
5
6
7
1260 50 1
2
3
4
5
6
Percent
Recovery1"
69.7
89.1
91.8
83.2
62.5
84.0
77.5
91.8
66.5
82.3
61.6
83.9
82.8
81.6
96.2
93.7
93.8
97.5
76.9
69.4
92.6
81.6
83.1
76.0
The operating conditions for the automated Soxhlet
Immersion time: 60 min
Reflux time: 60 min
b Multiple results from two different extractors.
Data from Reference 9.
8082 - 32
Revision 0
January 1995
-------
TABLE 10
MULTIPLE LABORATORY PRECISION AND ACCURACY DATA
FOR THE EXTRACTION OF PCBs FROM SPIKED SOIL
BY METHOD 3541 (AUTOMATED SOXHLET)
Laboratory 1
Laboratory 2
Laboratory 3
Laboratory 4
Laboratory 5
Laboratory 6
Laboratory 7
Laboratory 8
All
Laboratories
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
Percent Recovery
Aroclor 1254
Spike Cone.
(MgAg)
5
3
101.2
34.9
3
72.8
10.8
6
112.6
18.2
2
140.9
4.3
3
100.1
17.9
3
65.0
16.0
20
98.8
28.7
50
3
74.0
41.8
6
56.5
7.0
3
63.3
8.3
6
144.3
30.4
3
97.1
8.7
3
127.7
15.5
3
123.4
14.6
3
38.3
21.9
30
92.5
42.9
500
6
66.9
15.4
3
80.1
5.1
9
71.3
14.1
Aroclor 1260
Spike Cone.
(M9/kg)
5
3
83.9
7.4
3
70.6
2.5
6
100.3
13.3
3
138.7
15.5
3
82.1
7.9
3
92.8
36.5
21
95.5
25.3
50
3
78.5
7.4
6
70.1
14.5
3
57.2
5.6
6
84.8
3.8
3
79.5
3.1
4
105.9
7.9
3
94.1
5.2
3
51.9
12.8
31
78.6
18.0
500
6
74.5
10.3
3
77.0
9.4
9
75.3
9.5
All
Levels
12
84.4
26.0
24
67.0
13.3
12
66.0
9.1
24
110.5
28.5
12
83.5
10.3
12
125.4
18.4
12
99.9
19.0
12
62.0
29.1
120
87.6
29.7
Data from Reference 7.
8082 - 33
Revision 0
January 1995
-------
0
0}
s.
CD
in
c
i
i
0-
1
<
<
i
(
t
i
Illull
OB-170
•O
0-
OB-5
FIGURE 1. GC/ECD chromatogram of Aroclor 1016 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/um film thickness) and
30 m x 0.53 mm ID DB-1701 (l.O-pm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150'C (0.5 min hold) to 1908C (2 min hold) at 12eC/min then to 275°C
(10 min hold) at 4'C/min.
8082 - 34
Revision 0
January 1995
-------
p.
o
1
0)
r-
p*o .
rx> ir.
DB-1701
s>
(r
r-o
•CD
U/v,
JL
DB-5
10
— t
FIGURE 2. GC/ECD chromatogram of Aroclor 1221 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150'C (0.5 min hold) to 190'C (2 min hold) at 12'C/min then to 275eC
(10 min hold) at 4'C/min.
8082 - 35
Revision 0
January 1995
-------
r
j
J
(M
m
ru
0.
r-
UJ
08-1701
r
OB-5
FIGURE 3. GC/ECD chromatogram of Aroclor 1232 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150'C (0.5 min hold) to 190'C (2 min hold) at 12'C/min then to 275eC
(10 min hold) at 4'C/min.
8082 - 36
Revision 0
January 1995
-------
o-
CD
r-
n
T -0
Jl
ill
T
(M
DB-1701
Ifl
in
bJU
OB-5
r-
r-
10
u
01
o
FIGURE 4. GC/ECD chromatogram of Aroclor 1242 analyzed on a D8-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/xm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150'C (0.5 min hold) to 190'C (2 min hold) at 12'C/min then to 275'C
(10 min hold) at 4'C/min.
8082 - 37
Revision 0
January 1995
-------
0
LU
10 Ifl
- 0
IT)
in
DB-1701
at ir.
ii K»
O
I-
DB-5
e T
O u>
FIGURE 5. GC/ECD chromatogram of Aroclor 1248 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a JiW
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150*C (0.5 min hold) to 190*C (2 min hold) at 12'C/min then to 275°C
(10 min hold) at 4'C/min.
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in
m
DB-1701
tfl
<>•
•o
DB-5
U G*li u b» *•»
n«* tov m
FIGURE 6. GC/ECD chromatogram of Aroclor 1254 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-Mtn film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150*C (0.5 min hold) to 190'C (2 min hold) at 12'C/min then to 275°C
(10 min hold) at 4'C/min.
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DB-1701
DB-5
FIGURE 7. GC/ECD chromatogram of Aroclor 1260 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (LS-^im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150*C (0.5 min hold) to 190'C (2 min hold) at 12'C/mln then to 275'C
(10 min hold) at 4eC/min.
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METHOD 8082
POLYCHLORINATED BIPHENYLS (PCBs) AS AROCLORS BY CAPILLARY COLUMN GAS
CHROMATOGRAPHY
7.1 Choose
appropriate extraction
technique.
7.1 Add specified
matrix spike to sample.
7.2 Perform
extract cleanup.
7.3 Set
chromatographic
conditions.
7.4 Perform
initial calibration.
7.5 Establish retention
time windows.
7.6 Perform GC
analysis of sample
extracts.
7.6.3 Inject sample
extract.
7.6.5
Does
response fall
within
calibration
range?
7.6.10
Any sample
peak inter-
ferences?
7.6.5 Dilute
extract.
7.6.10 Additional
cleanup (possible
that replacement
of capillary column or
detector is warranted.)
7.7 - 7.8 Choose
appropriate standard
and calculate sample
concentrations.
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METHOD 8091
NITROAROMATICS AND CYCLIC KETONES: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8091 is a gas chromatographic (GC) method used to determine the
concentration of nitroaromatics and cyclic ketones. It describes wide-bore,
open-tubular, capillary column gas chromatography procedures using either
electron capture (ECD) or nitrogen-phosphorous (NPD) detectors. The following
RCRA analytes can be determined by this method:
Compound
CAS No."
2,4-Dinitrotoluene
1,4-Dinitrobenzene
2,6-Dinitrotoluene
1,4-Naphthoquinone
Nitrobenzene
Pentachloronitrobenzene
121-14-2
100-25-4
606-20-2
130-15-4
98-95-3
82-68-8
* Chemical Abstract Services Registry Number.
1.2 The following non-RCRA analytes can also be determined by this method:
Compound
CAS No.'
Benefin
Butralin
1-Chioro-2,4-dinitrobenzene
1-Chioro-3,4-dinitrobenzene
1-Chioro-2-nitrobenzene
l-Chloro-4-nitrobenzene
2-Chloro-6-nitrotoluene
4-Chloro-2-nitrotoluene
4-Chloro-3-nitrotoluene
2,3-Dichloronitrobenzene
2,4-Dichloronitrobenzene
3,5-Dichloronitrobenzene
3,4-Dichloronitrobenzene
2,5-Dichloronitrobenzene
Dinitramine
1,3-Dinitrobenzene
1861-40-1
33629-47-9
97-00-7
610-40-2
88-73-3
100-00-5
83-42-1
89-59-8
89-60-1
3209-22-1
611-06-3
618-62-2
99-54-7
89-61-2
29091-05-2
99-65-0
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Compound CAS No."
1,2-Dinitrobenzene 528-29-0
Isopropalin 33820-53-0
1,2-Naphthoquinone 524-42-5
2-Nitrotoluene 88-72-2
3-Nitrotoluene 99-08-1
4-Nitrotoluene 99-99-0
Penoxalin (Pendimethalin) 40487-42-1
Profluralin 26399-36-0
2,3,5,6-Tetrachloronitrobenzene 117-18-0
2,3,4,5-Tetrachloronitrobenzene 879-39-0
l,2,3-Trichloro-4-nitrobenzene 17700-09-3
l,2,4-Trichloro-5-nitrobenzene 89-69-0
2,4,6-Trichloronitrobenzene 18708-70-8
Trifluralin 1582-09-8
1.3 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8091 provides gas chromatographic conditions for the detection
of ppb concentrations of nitroaromatics and cyclic ketones in water and soil or
ppm concentrations in waste samples. Prior to use of this method, appropriate
sample extraction techniques must be used for environmental samples (refer to
Chapter Two and Method 3500). Both neat and diluted organic liquids (Method
3580) may be analyzed by direct injection. Analysis is accomplished by gas
chromatography utilizing an instrument equipped with wide bore capillary columns
and one or more electron capture detectors or nitrogen-phosphorus detectors
(NPD).
3.0 INTERFERENCES
3.1 Refer to Method 3500, 3600, and 8000.
3.2 The electron capture detector responds to all electronegative
compounds. Therefore, interferences are possible from other halogenated
compounds, as well as phthalates and other oxygenated compounds such as
organonitrogen, organosulfur, and organophosphorus compounds. Second column
confirmation or GC/MS confirmation is necessary to ensure proper analyte
identification unless previous characterization of the sample source will ensure
proper identification.
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3.3 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
syringe used for injection must be thoroughly rinsed between samples with
solvent. Whenever a highly concentrated sample extract is encountered, it should
be followed by the analysis of a solvent blank to check for cross-contamination.
Additional solvent blanks interspersed with the sample extracts should be
considered whenever the analysis of a solvent blank indicates cross-contamination
problems.
3.4 In certain cases some compounds coelute on either one or both columns.
In these cases the compounds must be reported as coeluting. The mixture can be
reanalyzed by GC/MS techniques if concentration permits (see Method 8270).
3.4.1 DB-5 column:
2,4,6-trichloronitrobenzene/1,3-dinitrobenzene
l-chloro-2,4-dinitrobenzene/l-chloro-3,4-dinitrobenzene/
l,2,3-trichloro-4-nitrobenzene
3.4.2 DB-1701 column:
2,4-dichloronitrobenzene/4-chloro-3-nitrotoluene
2,4,6-trichloronitrobenzene/l,4-naphthoquinone
l-chloro-2,4-dinitrobenzene/
2,3,4,5-tetrachloronitrobenzene
3.4.3 In addition, on the DB-5 column, 2,5-dichloronitrobenzene is
not well resolved from 4-chloro-3-nitrotoluene. Also, Trifluralin is not
well resolved from Benefin. On the DB-1701 column, compound pairs that
are not well resolved include 4-nitrotoluene/l-chloro-3-nitrobenzene and
Trifluralin/Benefin.
3.5 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph: An analytical system complete with a gas
chromatograph suitable for on-column and split/splitless injection, and all
accessories, including syringes, analytical columns, gases, electron capture
detectors or nitrogen-phosphorus detectors. A GC equipped with a single GC
column and detector or other configurations of column and detector is also
acceptable. A data system for measuring peak areas and/or peak heights, and dual
display of chromatograms is recommended.
4.1.1 Suggested GC Columns: Alternative columns may be used to
provide the separation needed to resolve all target analytes listed in
Sec. 1.1 of this method. Refer to Chapter One for additional information
regarding column performance and QA requirements.
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4.1.1.1 Column 1 - 30 m x 0.53 mm ID fused-silica open-
tubular column, crosslinked and chemically bonded with 95 percent
dimethyl and 5 percent diphenyl-polysiloxane (DB-5, RTx-5, SPB-5, or
equivalent), 0.83 jim or 1.5 p.m film thickness.
4.1.1.2 Column 2 - 30 m x 0.53 mm ID fused-silica
open-tubular column crosslinked and chemically bonded with 14
percent cyanopropylphenyl and 86 percent dimethyl-polysiloxane
(DB-1701, RTX-1701, or equivalent), 1.0 /urn film thickness.
4.1.2 Splitter: If the splitter approach to dual column injection
is chosen, following are three suggested splitters. An equivalent
splitter is acceptable. See Sec. 7.5.1 for a caution on the use of
splitters.
4.1.2.1 Splitter 1 - J&W Scientific press-fit Y-shaped glass
3-way union splitter (J&W Scientific, Catalog No. 705-0733).
4.1.2.2 Splitter 2 - Supelco 8-in glass injection tee,
deactivated (Supelco, Catalog No. 2-3665M).
4.1.2.3 Splitter 3 - Restek Y-shaped fused-silica connector
(Restek, Catalog No. 20405).
4.1.3 Column rinsing kit (optional): Bonded-phase column rinse kit
(J&W Scientific, Catalog No. 430-3000 or equivalent).
4.2 Microsyringes - 100 juL, 50 nl, 10 ij.1 (Hamilton 701 N or equivalent),
and 50 jtL (Blunted, Hamilton 705SNR or equivalent).
4.3 Balances - Analytical, 0.0001 g, Top-loading, 0.01 g.
4.4 Volumetric flasks, Class A - 10 ml to 1000 ml.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, all reagents shall conform to the specifications of the Committee on
Analytical Reagents of the American Chemical Society, where such specifications
are available. Other grades may be used, provided it is first ascertained that
the chemicals are of sufficiently high purity to permit their use without
affecting the accuracy of the determinations.
5.2 Solvents
5.2.1 Hexane, CeH14 - Pesticide quality or equivalent.
5.2.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.2.3 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or
equivalent.
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5.3 Stock standard solutions (1000 mg/L): Can be prepared from pure
standard materials or can be purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10 mL volumetric flask. (Isooctane is preferred
because it is less volatile than hexane.) If compound purity is 96
percent or greater, the weight can be used without correction to calculate
the concentration of the stock standard solution. Commercially prepared
stock standard solutions can be used at any concentration if they are
certified by the manufacturer or by an independent source.
5.3.2 For those compounds which are not adequately soluble in
hexane or isooctane, dissolve the compound initially with a small volume
of toluene, ethyl acetate or acetone and dilute to volume with isooctane
or hexane.
5.4 Composite stock standard: Can be prepared from individual stock
solutions. For composite stock standards containing less than 25 components,
transfer exactly 1 mL of each individual stock solution at 1000 mg/L, add
solvent, mix the solutions, and bring to volume in a 25 mL volumetric flask. For
example, for a composite containing 20 individual standards, the resulting
concentration of each component in the mixture, after the volume is adjusted to
25 mL, will be 40 mg/L. This composite solution can be further diluted to obtain
the desired concentrations. For composite stock standards containing more than
25 components, use volumetric flasks of the appropriate volume (e.g., 50 mL,
100 mL).
5.5 Calibration standards: These should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with isooctane or
hexane. The standard concentrations should correspond to the expected range of
concentrations present in the field samples and should bracket the linear range
of the detector.
5.6 Recommended internal standard: Prepare a solution of 1000 mg/L of
hexachlorobenzene. For spiking, dilute this solution to 50 ng//iL. (This
concentration may need to be more dilute depending on the detector chosen and its
sensitivity. The internal standard response should be approximately 50 to 90%
of full scale.) Use a spiking volume of 10.0 /nL/mL of extract. The spiking
concentration of the internal standards should be kept constant for all samples
and calibration standards.
5.7 Recommended surrogate standard: Monitor the performance of the method
using surrogate compounds. Surrogate standards are added to all samples, method
blanks, matrix spikes, and calibration standards. Prepare a solution of
1000 mg/L of l-chloro-3-nitrobenzene and dilute it to 10 ng//iL. (This
concentration may need to be adjusted depending on the detector chosen and its
sensitivity. The surrogate standard response should be approximately 100% of
full scale.) Use a spiking volume of 100 /A for a 1 L aqueous sample.
5.8 Store the standard solutions (stock, composite, calibration, internal,
and surrogate) at 4"C or cooler in Teflon®-sealed containers in the dark. All
standard solutions must be replaced after six months or sooner if routine QC
(Sec. 8.0) indicates a problem.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
6.2 Extracts must be stored in the dark at or below 4"C and analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction and Cleanup:
7.1.1 Refer to Chapter Two and Method 3500 for guidance on choosing
the appropriate extraction procedure. In general, water samples are
extracted at a pH between 5 to 9 with methylene chloride, using either
Method 3510 or 3520. Solid samples are extracted using any of the
extraction methods for solids listed in Method 3500, as appropriate.
7.1.2 If necessary, the samples may be cleaned up using Method 3620
(Florisil) and/or Method 3640 (Gel Permeation Chromatography). See
Chapter Two, Sec. 2.0 and Method 3600 for general guidance on cleanup and
method selection. Method 3660 is used for sulfur removal.
7.1.3 Prior to gas chromatographic analysis, the extraction solvent
needs to be exchanged to hexane. The exchange is performed using the K-D
procedures listed in each of the extraction methods. Any methylene
chloride remaining in the extract will cause a very broad solvent peak.
7.2 Gas Chromatographic Conditions: Retention time information for each
of the analytes is presented in Tables 1 and 3. The recommended GC operating
conditions are provided in Tables 2 and 4. Figures 1, 2, and 3 illustrate
typical chromatography of the method analytes for both columns when operated at
the conditions specified.
7.3 Calibration:
7.3.1 Prepare calibration standards using the procedures in Sec.
5.0. Refer to Method 8000, Sec. 7.0 for proper calibration procedures.
The procedure for internal or external calibration may be used.
7.3.2 Refer to Method 8000, Sec. 7.0 for the establishment of
retention time windows.
7.4 Gas chromatographic analysis:
7.4.1 Method 8000, Sec. 7.0 provides instructions on calibration,
establishing retention time windows, the analysis sequence, appropriate
dilutions, and identification criteria.
7.4.2 Automatic injections of 1 juL are recommended. Hand
injections of no more than 2 jxL may be used if the analyst demonstrates
quantitation precision less than or equal to 10 percent relative standard
deviation. The solvent flush technique may be used if the amount of
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solvent is kept at a minimum. If the internal standard calibration
technique is used, add 10 /iL of the internal standard to each ml of sample
extract prior to injection.
7.4.3 Tentative identification of an analyte occurs when a peak
from a sample extract falls within the absolute retention time window.
Normally, confirmation is necessary. Confirmation techniques include
analysis on a second column with dissimilar stationary phase, by GC/MS
(full scan or SIM) or by using a different detector and getting comparable
data. See Sec. 7.0 of Method 8000 on "Compound Identification" for
further information.
7.4.3.1 If partially overlapping or coeluting peaks are
present, install columns with a dissimilar liquid phase or use a
GC/MS technique. Interferences that prevent analyte identification
and/or quantitation may possibly be removed by the cleanup
techniques mentioned above.
7.4.4 Record the volume injected to the nearest 0.05 ^L and the
resulting peak size in area units or peak height. Using either the
internal or the external calibration procedure (Method 8000), determine
the quantity of each component peak in the sample chromatogram which
corresponds to the compounds used for calibration purposes.
7.4.4.1 If the responses exceed the linear range of the
system, dilute the extract and reanalyze. Peak height measurements
are recommended, rather than peak area integration, when overlapping
peaks may cause errors in area integration.
7.4.4.2 If the peak response is less than 2.5 times the
baseline noise level, the validity of the quantitative result may be
questionable. The analyst should consult with the source of the
sample to determine whether further concentration of the sample is
warranted.
7.4.5 Determine the concentration of each identified analyte using
the calculation formulae in Sec. 7.0 of Method 8000.
7.5 Instrument Maintenance:
7.5.1 Injection of sample extracts from waste sites often leaves a
high boiling residue in: the injection port area, splitters when used, and
the injection port end of the chromatographic column. This residue
affects chromatography in many ways (i.e., peak tailing, retention time
shifts, analyte degradation, etc.) and, therefore, instrument maintenance
is very important. Residue buildup in a splitter may limit flow through
one leg and therefore change the split ratios. If this occurs during an
analytical run, the quantitative data may be incorrect. Proper cleanup
techniques will minimize the problem and instrument QC will indicate when
instrument maintenance is required.
7.5.2 Suggested chromatograph maintenance: Corrective measures may
require any one or more of the following remedial actions. Also see Sec.
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7.0 in Method 8000 for additional guidance on corrective action for
capillary columns and the injection port.
7.5.2.1 Splitter connections: For dual columns which are
connected using a press-fit Y-shaped glass splitter or a Y-shaped
fused-silica connector, clean and deactivate the splitter or replace
with a cleaned and deactivated splitter. Break off the first few
inches (up to one foot) of the injection port side of the column.
Remove the columns and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate
the degradation problem, it may be necessary to deactivate the metal
injector body and/or replace the columns.
7.5.2.2 Column rinsing: The column should be rinsed with
several column volumes of an appropriate solvent. Both polar and
nonpolar solvents are recommended. Depending on the nature of the
sample residues expected, the first rinse might be water, followed
by methanol and acetone; methylene chloride is a satisfactory final
rinse and in some cases may be the only solvent required. The
column should then be filled with methylene chloride and allowed to
remain flooded overnight to allow materials within the stationary
phase to migrate into the solvent. The column is then flushed with
fresh methylene chloride, drained, and dried at room temperature
with a stream of ultrapure nitrogen passing through the column.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
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8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories
should use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with
each analytical batch. The LCS consists of an aliquot of a clean
(control) matrix similar to the sample matrix and of the same weight or
volume. The LCS is spiked with the same analytes at the same
concentrations as the matrix spike. When the results of the matrix spike
analysis indicate a potential problem due to the sample matrix itself, the
LCS results are used to verify that the laboratory can perform the
analysis in a clean matrix.
8.4.3 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Table 1 lists the retention times of the target analytes. Figure 1
shows a chromatogram of the target analytes eluted from a pair of DB-5/DB-1701
columns and detected using electron capture detectors (ECD) under the GC
conditions listed in Table 2.
9.2 Table 3 provides the retention times and recovery data of the target
analytes. GC conditions used during the recovery study are listed in Table 4.
Chromatograms of the standard mixes used in the recovery study are provided in
Figures 2 and 3.
9.3 The laboratory should perform a Method Detection Limit (MDL) study and
generate its own performance data (precision and accuracy) for matrix spike and
surrogate compounds. Refer to Method 8000 for guidance.
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10.0 REFERENCES
1. Lopez-Avila, V., Baldin, E., Benedicto, J, Milanes, J., Beckert, W.F.,
"Application of Open-Tubular Columns to SW 846 GC Methods", final report
to the U.S. Environmental Protection Agency on Contract 68-03-3511,
Mid-Pacific Environmental Laboratory, Mountain View, CA, 1990.
2. Tsang, S., Marsden, P.O., Chau, N., "Performance Data for Methods 8041,
8091, Bill, and 8121A", draft report to U.S. Environmental Protection
Agency on Contract 68-W9-0011, Science Applications International Corp.,
San Diego, CA, 1992.
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TABLE 1
RETENTION TIMES8 OF THE NITROAROMATICS AND CYCLIC KETONES
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
DB-5 DB-1701
Compound
Nitrobenzene
2-Nitrotoluene
3-Nitrotoluene
4-Nitrotoluene
l-Chloro-3-nitrobenzene (Surr.)
1 -Chi oro-4- nitrobenzene
l-Chloro-2-nitrobenzene
2-Chloro-6-nitrotoluene
4-Chloro-2-nitrotoluene
3 , 5-Di chl oroni trobenzene
2,5-Dichloronitrobenzene
2, 4 -Di chl oroni trobenzene
4-Chloro-3-nitrotoluene
3, 4-Di chl oroni trobenzene
2, 3 -Di chl oroni trobenzene
2, 4, 6 -Tri chl oroni trobenzene
1,4-Naphthoquinone
1,2, 4 -Tri chl oro-5-ni trobenzene
1,4-Dinitrobenzene
2,6-Dinitrotoluene
1,3-Dinitrobenzene
1,2, 3 -Tri chl oro-4- nitrobenzene
2, 3, 5, 6-Tetrachl oroni trobenzene
1,2-Dinitrobenzene
2,4-Dinitrotoluene
1 -Chl oro-2,4-di nitrobenzene
2, 3, 4, 5 -Tetrachl oroni trobenzene
1 -Chl oro-3,4-di nitrobenzene
Trifluralin
Benefin
Pent achl oroni trobenzene
Profluralin
Dinitramine
Butralin
Isopropalin
CAS No. RT(min) RT(min)
98-95-3
88-72-2
99-08-1
99-99-0
121-73-3
100-00-5
88-73-3
83-42-1
89-59-8
618-62-2
89-61-2
611-06-3
89-60-1
99-54-7
3209-22-1
18708-70-8
130-15-4
89-69-0
100-25-4
606-20-2
99-65-0
17700-09-3
117-18-0
528-29-0
121-14-2
97-00-7
879-39-0
610-40-2
1582-09-8
1861-40-1
82-68-8
26399-36-0
29091-05-2
33629-47-9
33820-53-0
4.71
6.08
6.93
7.35
7.66
7.9
8.09
9.61
9.76
10.42
11.46
11.73
11.31
12.24
12.58
13.97
12.98
15.97
13.41
14.44
13.97
17.61
19.41
14.76
16.92
17.85
21.51
17.85
21.81
21.94
25.13
25.39
26.45
32.41
32.71
4.23
5.32
6.22
6.73
6.85
7.15
7.78
8.32
8.62
8.84
10.62
10.84
10.84
11.04
12.01
12.31
12.31
14.46
14.72
15.16
15.68
16.51
17.11
17.51
18.16
19.55
19.55
19.85
20.31
20.46
22.33
23.81
27.06
31.03
31.33
(continued)
8091 - 11
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TABLE 1 (continued)
Compound
No.
36
37
38
Int. Std.
Compound
Penoxalin (Pendimethal in)
1,2-Naphthoquinone
2-Chloro-4-nitrotoluene
Hexachlorobenzene
CAS No.
40487-42-1
524-42-5
121-86-8
118-74-1
DB-5 DB-1701
RT(min) RT(min)
33.05 31.67
c c
b b
23.18 18.72
a See Table 2 for operating conditions.
b Not available.
c Not detected at 1 ng per injection.
NOTE: These data are from Reference 1.
TABLE 2
DUAL COLUMN GC OPERATING CONDITIONS FOR NITROAROMATICS
GC Instrument: Varian 6000 with dual electron capture detectors
Column 1: Type: DB-5 (J&W Scientific)
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 1.5 ^m
Column 2: Type: DB-1701 (J&W Scientific)
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 1.0 /urn
Type of splitter: J&W Scientific press-fit Y-shaped inlet splitter
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Nitrogen)
Injector temperature: 250°C
Detector temperature: 320°C
Temperature program: 120°C (1.0 min hold) to 200°C (1 min hold) at 3°C/min
then to 250°C (4 min hold) at 8°C/min.
Injection volume: 2 juL
Type of injection: Flash vaporization
Solvent: Hexane
Range: 10
Attenuation: 64 (DB-1701)/64 (DB-5)
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TABLE 3
RETENTION TIMES AND RECOVERY OF NITROAROMATICS
Analyte
Rt, min
Spiking Cone.
(ng/g)
Recovery
(%)
% RSD
MIX 1
l,2:3,4-diepoxy butane
Nitrobenzene
2-Nitrotoluene
3-Nitrotoluene
4-Nitrotoluene
1 -Chi oro-3 -nitrobenzene8
2,3-Dichloronitrobenzene
1,4-Naphthoquinone
1,3-Dinitrobenzene
1,2-Dinitrobenzene
3-Nitroaniline
2,4-Dinitrotoluene
4-Nitroaniline
Trifluralin
Pentachloronitrobenzene
4-Nitroquinoline-l -oxide
3.23
11.51
14.13
15.52
16.22
16.64
22.48
23.29
24.25
24.69
25.44
26.95
28.91
30.25
32.26
36.05
5,000
5,000
5,000
5,000
5,000
100
100
200
400
200
10,000
200
5,000
200
100
5,000
22
85
80
83
97
103
102
35
80
99
54
75
53
127
129
6.7
18.1
6.9
5.4
6.8
6.2
6.2
7.3
23.1
13.1
17.0
17.8
13.9
29.6
4.4
5.8
18.5
MIX 2
1 -Chi oro -3 -nitrobenzene8
2-Nitroanil ine
1 , 4 -Di nitrobenzene
2,6-Dinitrotoluene
5-Nitro-o-toluidine
16.64
22.87
23.82
24.49
28.91
100
5,000
200
200
5,000
98
88
142
192
60
3.0
3.6
2.9
6.2
42
8 Recommended Surrogate
n = 5 samples
NOTE: This table is from Reference 2. See Table 4 for operating conditions used
in this table.
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TABLE 4
GC OPERATING CONDITIONS USED FOR RECOVERY DATA IN TABLE 3
Column: DB-5 30 m x 0.53 mm ID.
Carrier gas: Nitrogen at 6 mL/min with hydrogen at 30 mL/min.
Total nitrogen flow: 60 mL/min (carrier and makeup).
Injector: Packed, megabore liner at 200°C.
Detector: ECD at 300°C.
Temperature Program:
708C held for 1.5 minutes
4°C/min to 170°C
8°C/min to 275°C and held for 5.4 minutes
The total run time was 45 minutes.
NOTE: This table is from Reference 2.
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FIGURE 1
GC/ECD CHROMATOGRAM OF NITROAROMATICS ANALYZED ON A
DB-5/DB-1701 FUSED-SILICA, OPEN-TUBULAR COLUMN PAIR
See Table 2 for operating conditions.
JL,
32
33
DB-1701
35
34
36
9 10 12 15 fi
DB-5
28
26
\l V
33
34
35
36
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FIGURE 2
RECOVERY OF NITROAROMATICS - MIX 1
See Table 4 for operating conditions.
3 . Oe-4
2. O
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FIGURE 3
RECOVERY OF NITROAROMATICS - MIX 2
See Table 4 for operating conditions.
"3 . O
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METHOD 8091
NITROAROMATICS AND CYCLIC KETONES: CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose
appropriate
extraction procedure.
7.1.2
Is cleanup
required?
7.1.3 Choose
appropriate
cleanup method.
7.1.3 Exchange
extraction solvent
with hexane.
7.2 Set GC column
operating
conditions.
7.3 Calibrate (see
Method 8000.)
I
7.4 Perform GC
analysis (see
Method 8000.)
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METHOD 8111
HALOETHERS: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8111 is a gas chromatographic (GC) method used to determine the
concentration of haloethers. It describes wide-bore open-tubular, capillary
column gas chromatography procedures using the a dual-column/dual-detector
approach, however, a single column/single detector approach is acceptable. The
following RCRA analytes can be determined by this method:
Compound
CAS No.8
Bis(Z-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-Chlorophenyl phenyl ether
111-91-1
111-44-4
108-60-1
7005-72-3
aChemical Abstract Services Registry Number
1.2 The following non-RCRA analytes may also be analyzed by this method:
Compound
CAS No.
4-Bromophenyl phenyl ether
2-Chlorophenyl 4-nitrophenyl ether
3-Chlorophenyl 4-nitrophenyl ether
4-Chlorophenyl 4-nitrophenyl ether
2,4-Dibromophenyl 4-nitrophenyl ether
2,4-Dichlorophenyl 3-methyl-4-nitrophenyl ether
6-Dichlorophenyl 4-nitrophenyl ether
2.
3..
2,5-Dichlorophenyl 4-nitrophenyl
2,4-Dichlorophenyl 4-nitrophenyl
2,3-Dichlorophenyl
,5-Dichlorophenyl 4-nitrophenyl ether
ether
ether
4-nitrophenyl ether
3,4-Dichlorophenyl 4-nitrophenyl ether
4-Nitrophenyl phenyl ether
2,4,6-Trichlorophenyl 4-nitrophenyl ether
2,3,6-Trichlorophenyl 4-nitrophenyl ether
2,3,5-Trichlorophenyl
2,4,5-Trichlorophenyl
4-nitrophenyl ether
4-nitrophenyl ether
3,4,5-Trichlorophenyl 4-nitrophenyl ether
2,3,4-Trichlorophenyl 4-nitrophenyl ether
101-55-3
209-61-4
2303-23-3
1836-74-4
2671-93-4
42488-57-3
2093-28-9
NA
391-48-7
1836-75-5
82239-20-1
22532-80-5
620-88-2
1836-77-7
NA
NA
22532-68-9
NA
NA
NA = CAS numbers not assigned at this time.
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1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph and in the interpretation
of gas chromatograms.
2.0 SUMMARY OF METHOD
2.1 Method 8111 provides gas chromatographic conditions for the detection
of ppb concentrations of haloethers in water and soil or ppm concentrations in
waste samples.
2.2 Prior to use of this method, appropriate sample extraction techniques
must be used for environmental samples (refer to Chap. Two and Method 3500).
2.3 Both neat and diluted organic liquids (Method 3580) may be analyzed
by direct injection.
2.4 Analysis is accomplished by gas chromatography utilizing an instrument
equipped with a wide-bore capillary column and an electron capture detector.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 The electron capture detector responds to all electronegative
compounds. Therefore, interferences are possible by other halogenated compounds,
as well as phthalates and other oxygenated compounds such as organonitrogen,
organosulfur, and organophosphorus compounds. Second column confirmation or
GC/MS confirmation are necessary to ensure proper analyte identification unless
previous characterization of the sample source will ensure proper identification.
3.3 Dichlorobenzenes are known to coelute with haloethers under some gas
chromatographic conditions. If these materials are present in a sample, it may
be necessary to analyze the extract with two different column packings to
completely resolve all of the compounds.
3.4 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
syringe used for injection must be thoroughly rinsed between samples with
solvent. Whenever a highly concentrated sample is encountered, it should be
followed by the analysis of a solvent blank to check for cross-contamination.
Additional solvent blanks interspersed with the sample extracts should be
considered whenever the analysis of a solvent blank indicates cross-contamination
problems.
3.5 Some compounds coelute using this procedure. In these cases, the
compounds must be reported as coeluting. The mixture may be reanalyzed for peak
confirmation by GC/MS techniques if concentration permits (see Method 8270).
3.6 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - An analytical system complete with a gas
chromatograph suitable for on-column and split/splitless injection, and all
necessary accessories, including syringes, analytical columns, gases and two
electron capture detectors, A data system for measuring peak areas and/or peak
heights is recommended.
4.2 Suggested GC columns - Alternative columns may be used to provide the
separation required to resolve all target analytes listed in Sec. 1.1 of this
method. Refer to Chapter 1 and Method 8000 for additional information regarding
column performance and QC requirements.
4.2.1 Column 1 - 30 m x 0.53 mm ID fused-silica open-tubular column,
crosslinked and chemically bonded with 95 percent dimethyl and 5 percent
diphenyl-polysiloxane (DB-5, RTx-5, SPB-5, or equivalent), 0.83 urn or 1.5
/urn film thickness.
4.2.2 Column 2 - 30 m x 0.53 mm ID fused-silica open-tubular column
crosslinked and chemically bonded with 14 percent cyanopropylphenyl and 86
percent dimethyl-polysiloxane (DB-1701, RTX-1701, or equivalent), 1.0 jum
film thickness.
4.3 Column rinsing kit (optional) - Bonded-phase column rinse kit (J&W
Scientific, Catalog no. 430-3000 or equivalent).
4.4 Microsyringes - 100-^tL, 50-/A, 10-/xL (Hamilton 701 N or equivalent),
and 50-juL (Blunted, Hamilton 705SNR or equivalent).
4.5 Balances - Analytical, capable of accurately weighing 0.0001 g, and
top-loading, capable of accurately weighing 0.01 g.
4.6 Volumetric flasks, Class A - 10-mL to 1000-mL.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the chemicals are of sufficiently high purity to permit
their use without affecting the accuracy of the determinations.
5.2 Solvents - All solvents must be pesticide quality or equivalent.
5.2.1 Hexane, C6H14
5.2.2 Acetone, CH3COCH3
5.2.3 Isooctane, (CH3)3CCH2CH(CH3)2
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5.3 Stock standard solutions (1000 mg/L) - May be prepared from pure
standard materials or can be purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10-mL volumetric flask. If compound purity is
96 percent or greater, the weight can be used without correction to
calculate the concentration of the stock standard solution. Commercially-
prepared stock standard solutions may be used at any concentration if they
are certified by the manufacturer or by an independent source.
5.3.2 For those compounds which are not adequately soluble in hexane
or isooctane, dissolve the compound initially with a small volume of
toluene, ethyl acetate or acetone and dilute to volume with isooctane or
hexane.
5.4 Composite stock standard - Can be prepared from individual stock
solutions. For composite stock standards containing less than 25 components,
transfer exactly 1 ml of each individual stock solution at 1000 mg/L, add
solvent, mix the solutions, and bring to volume in a 25-mL volumetric flask. For
example, for a composite containing 20 individual standards, the resulting
concentration of each component in the mixture, after the volume is adjusted to
25 ml, will be 40 mg/L. This composite solution can be further diluted to obtain
the desired concentrations. *
5.5 Calibration standards - These should be prepared at a minimum of five
concentrations with dilution of the composite stock standard with isooctane or
hexane. The standard concentrations should correspond to the expected range of
concentrations present in the field samples and should bracket the linear range
of the detector.
5.6 Recommended internal standard - Prepare a solution of 1000 mg/L of
4,4'-dibromobiphenyl. For spiking, dilute this solution to 50 ng//^L. (This
concentration may need to be more dilute depending on the detector sensitivity.
The internal standard response should be approximately 50 to 90% of full scale.)
Use a spiking volume of 10 juL/ml of extract. The spiking concentration of the
internal standards should be kept constant for all samples and calibration
standards. Store the internal standard spiking solutions at 4°C in Teflon-sealed
containers in the dark.
5.7 Recommended surrogate standards - The performance of the method should
be monitored using surrogate compounds. Surrogates are added to all samples,
method blanks, matrix spikes, and calibration standards. Prepare a solution of
1000 mg/L each of 2,4-dichlorodiphenyl ether and 2,3,4-trichlorodiphenyl ether
and dilute them to 20 ng//zL. Use a spiking volume of 100 jLtL for a 1-L aqueous
sample. (This concentration may need to be adjusted depending on the detector
sensitivity. The surrogate standard response should be approximately 100% of
full scale.)
5.8 Store the standard solutions (stock, composite, calibration, internal,
and surrogate) at 4°C or cooler in Teflon-sealed containers in the dark. All
standard solutions must be replaced after six months or sooner if routine QC
(Sec. 8.0) indicates a problem.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 Extracts must stored in the dark at or below 4°C and be analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction and cleanup
7.1.1 Refer to Chapter Two and Method 3500 for guidance on choosing
the appropriate extraction procedure. In general, water samples are
extracted at a neutral, or as is, pH with methylene chloride, using either
Method 3510 or 3520. Solid samples are extracted using any of the
extraction methods for solids listed in Method 3500, as appropriate.
7.1.2 If necessary, the samples may be cleaned up using Method 3620
(Florisil) and/or Method 3640 (Gel Permeation Chromatography). See Chapter
Two, Sec. 2, and Method 3600 for general guidance on cleanup and method
selection. Method 3660 may be used for sulfur removal.
7.1.3 Prior to gas chromatographic analysis, the extraction solvent
should be exchanged to hexane. The exchange is performed during the K-D
procedures listed in each of the extraction methods. Any methylene
chloride remaining in the extract will cause a very broad solvent peak.
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.
7.2 Gas Chromatography (GC) Conditions - Retention time information for
each of the target analytes is presented in Table 1. Retention times of non-RCRA
analytes are presented in Table 2. GC operating conditions under which these
retention times were obtained are provided in the appropriate table. Figures 1
and 2 illustrate typical Chromatography of the haloethers.
7.3 Calibration
7.3.1 Prepare calibration standards using the procedures in Sec.
5.0. Refer to Method 8000 for proper calibration procedures. The
procedure for internal or external calibration may be used.
7.3.2 Refer to Method 8000, for procedures for establishing
retention time windows.
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7.4 Gas chromatographic analysis
7.4.1 Method 8000, provides instructions on calibration,
establishing retention time windows, the analysis sequence, appropriate
dilutions, and identification criteria.
7.4.2 Automatic injections of 1 jttL are recommended. Hand injections
of no more than 2 nl may be used if the analyst demonstrates quantitation
precision of < 10 percent relative standard deviation. The solvent flush
technique may be used if the amount of solvent is kept at a minimum. If
the internal standard calibration technique is used, add 10 //L of the
internal standard to each 1 ml of sample extract prior to injection.
7.4.3 Tentative identification of an analyte occurs when a peak from
a sample extract falls within the absolute retention time window. Normally
confirmation is required. Confirmation techniques include analysis on a
second column with dissimilar stationary phase, by GC/MS (full scan or SIM)
or by using a different detector and getting comparable data. See Method
8000 for further information.
7.4.3.1 If partially overlapping or coeluting peaks are
present, install columns with a dissimilar liquid phase or use a GC/MS
technique.
7.4.3.1 Interferences that prevent analyte identification
and/or quantitation may possibly be removed by the cleanup techniques
mentioned above.
7.4.4 Record the volume injected to the nearest 0.05 yuL and the
resulting peak size in area units or peak height. Using either the
internal or the external calibration procedure (Method 8000), determine the
quantity of each component peak in the sample chromatogram which
corresponds to the compounds used for calibration purposes.
7.4.4.1 If the responses exceed the linear range of the
system, dilute the extract and reanalyze. Peak height measurements
are recommended, rather than peak area integration, when overlapping
peaks may cause errors in area integration.
7.4.4.2 If the peak response is less than 2.5 times the
baseline noise level, the validity of the quantitative result may be
questionable. The analyst should consult with the source of the
sample to determine whether further concentration of the sample is
warranted.
7.4.5 Determine the concentration of each identified analyte using
the calculation formulas in Sec. 7.0 of Method 8000.
7.5 Instrument Maintenance
7.5.1 Injection of sample extracts from waste sites often leaves a
high boiling residue in: the injection port area, splitters (when used),
and the injection port end of the chromatographic column. This residue
effects chromatography in many ways (i.e., peak tailing, retention time
8111 - 6 Revision 0
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shifts, analyte degradation, etc.)- Therefore, instrument maintenance is
very important. Residue buildup in a splitter may limit flow through one
leg and therefore change the split ratios. If this occurs during an
analytical run, the quantitative data may be incorrect. Proper cleanup
techniques will minimize the problem and instrument QC will indicate when
instrument maintenance is required.
7.5.2 Suggested chromatograph maintenance - Corrective measures may
require remedial actions.
7.5.2.1 Column rinsing - The column should be rinsed with
several column volumes of an appropriate solvent. Both polar and
nonpolar solvents are recommended. Depending on the na ure of the
sample residues expected, the first rinse might be water, followed by
methanol and acetone-; methylene chloride is a satisfactory final rinse
and in some cases may be the only solvent required. The column should
then be filled with methylene chloride and allowed to remain flooded
overnight to allow materials within the stationary phase to migrate
into the solvent. The column is then flushed with fresh methylene
chloride, drained, and dried at room temperature with a stream of
ultrapure nitrogen passing through the column.
7.5.2.2 See Method 8000 for additional guidance on
corrective action for capillary columns and the injection port.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
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8.4.1 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time
a set of samples is analyzed or there is a change in reagents, a method
blank should be analyzed as a safeguard against chronic laboratory
contamination. The blanks should be carried through all stages of sample
preparation and measurement.
8.4.2 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Table 1 lists the retention times and recoveries of the target
analytes. The recoveries presented were obtained from the analysis of spiked
sandy loam soils. No recovery data is currently available on Bis(2-chloro-
ethoxy)methane and Bis(Z-chloroethyl) ether.
9.2 Table 2 lists the compounds that may be determined by this method and
their retention times. Figure 1 shows a chromatogram of the target analytes
eluted from a pair of DB-5/DB-1701 columns and detected with electron capture
detectors (ECD) under the GC conditions listed in Table 2.
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10.0 REFERENCES
1. Lopez-Avila, V.; Baldin, E.; Benedicto, J; Milanes, J.; Beckert, W. F.
"Application of Open-Tubular Columns to SW-846 GC Methods"; final report
to the US EPA on Contract 68-03-3511; Mid-Pacific Environmental Laboratory,
Mountain View, CA, 1990.
2. Tsang, S.; Marsden, P.; Chau, N. "Performance Data for Methods 8041, 8091,
8111, and 8121A"; draft report to US EPA on Contract 68-W9-0011; Science
Applications International Corp., San Diego, CA, 1992.
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TABLE 1
RETENTION TIMES AND RECOVERY OF TARGET HALOETHERS
Analyte
Bis(2-chloroisopropy1) ether
4-Chlorophenyl phenyl ether
4-Bromophenyl phenyl ether
RT
(min)
3.06
15.75
18.21
Spiking Cone.
(M9/9)
2.5
5.0
0.5
Recovery
(%)
112
91.5
97.0
RSD
(%)
4.3
3.5
2.1
Column:
Temperature program:
Injector:
Injector temperature:
Detector:
Detector temperature:
Nitrogen carrier gas:
Nitrogen makeup gas:
DB-5, 30 m x 0.53 mm id
125°C (1.0 min hold) to 135°C at 2°C/min.,
135°C to 200°C at 5°C/min.,
200°C to 275°C at 10°C/min., (3.5 min hold)
Packed, wide-bore liner
200°C
ECD
320°C
5 mL/min
55 mL/min
8111 - 10
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TABLE 2
RETENTION TIMES OF NON-TARGET HALOETHERS
r
Peak8 Compound
Retention Time (min)
DB-5 DB-1701
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
IS
SU-1
SU-2
4-Bromophenyl-phenyl ether
Phenyl 4-nitrophenyl ether
2-Chlorophenyl 4-nitrophenyl ether
3-Chlorophenyl 4-nitrophenyl ether
4-Chlorophenyl 4-nitrophenyl ether
2,6-Dichlorophenyl 4-nitrophenyl ether
3,5-Dichlorophenyl 4-nitrophenyl ether
2,5-Dichlorophenyl 4-nitrophenyl ether
2,4-Dichlorophenyl 4-nitrophenyl ether
2,3-Dichlorophenyl 4-nitrophenyl ether
3,4-Dichlorophenyl 4-nitrophenyl ether
2,4,6-Trichlorophenyl 4-nitrophenyl ether
2,3,6-Trichlorophenyl 4-nitrophenyl ether
2,3,5-Trichlorophenyl 4-nitrophenyl ether
2,4,5-Trichlorophenyl 4-nitrophenyl ether
2,4-Dibromophenyl 4-nitrophenyl ether
3,4,5-Trichlorophenyl 4-nitrophenyl ether
2,3,4-Trichlorophenyl 4-nitrophenyl ether
2,4-Dichlorophenyl 3-methyl-4-nitrophenyl ether
4,4'-Dibromobiphenyl
2,4-Dichlorodiphenyl ether
2,3,4-Trichlorodiphenyl ether
4.28
6.85
10.44
10.78
11.37
14.02
14.55
14.55
15.08
16.11
16.65
17.89
19.40
19.70
20.03
21.63
21.83
22.28
21.83
9.44
4.82
8.31
5.57
10.86
16.31
16.70
17.68
20.84
21.33
21.54
22.30
23.87
24.54
24.93
27.27
27.56
28.05
30.03
30.42
31.18
31.60
12.66
6.17
10.95
aPeak numbers refer to the chromatogram in Figure 2.
IS = Internal Standard
SU = Surrogate
The GC operating conditions for the above analysis were as follows:
Columns:
Temperature program:
Injector temperature:
Detector temperature:
Helium carrier gas:
Nitrogen makeup gas:
30 m x 0.53 mm ID DB-5 (0.83 p,m film thickness)
and 30 m x 0.53 mm ID DB-1701 (1.0 urn film
thickness) connected to an 8 in. injection tee
(Supelco, Inc.).
180°C (0.5 min hold) to 260°C (1.0 min hold) at
2°C/min.
250°C
320°C
6 mL/min
20 mL/min
8111 - 11
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FIGURE 1
GC/ECD CHROMATOGRAM OF TARGET ANALYTE HALOETHERS
ANALYZED ON A DB-5 CAPILLARY COLUMN FOR RECOVERY STUDIES
The GC operating conditions are listed in Table 1.
n H
-|J 0
0 0 0 0 C 0 0
0000000
oooocoo
0 (E
-------
FIGURE 2
GC/ECD CHROMATOGRAM OF HALOETHERS ANALYZED ON A
DB-5/DB-1701 FUSED-SILICA OPEN-TUBULAR COLUMN PAIR
The GC operating conditions are listed in Table 2.
so
DB-1701
su
IS
J
19
17
It
DB-5
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METHOD 8111
HALOETHERS: CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose
appropriate
extraction procedure.
7.1.2 Perform cleanup
using Method 3620
and/or Method 3640.
7.1.2
Is cleanup
of the extract
required?
7.2 Refer to Table 2
for recommended
operating condition*
for the GC.
7.3 Refer to Method
8000 for proper
calibration technique.
7.4 Refer to
Method SOOO for
guidance on GC
analyst*.
7.4.4 Record *ample
volume injected and
resulting peak size.
7.4.5 Perform
appropriate
calculations (refer
to Method 8000.)
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METHOD 8131
ANILINE AND SELECTED DERIVATIVES BY GC: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8131 is used to determine (by gas chromatography with a
specific detector) the concentration of aniline and certain derivatives of
aniline in extracts prepared from environmental samples and RCRA wastes. It has
been validated for aqueous matrices. Application to other matrices must be
validated by developing spike recovery data. The following compounds can be
determined by this method:
Compound CAS No.a
Aniline 62-53-3
4-Bromoaniline 106-40-1
2-Bromo-6-chloro-4-nitroaniline 99-29-6
2-Bromo-4,6-dinitroaniline 1817-73-8
2-Chloroaniline 95-51-2
3-Chloroaniline 108-42-9
4-Chloroaniline 106-47-8
2-Chloro-4,6-dinitroaniline 3531-19-9
2-Chloro-4-nitroaniline 121-87-9
4-Chloro-2-nitroaniline 89-63-4
2,6-Dibromo-4-nitroaniline 827-94-1
3,4-Dichloroaniline 95-76-1
2,6-Dichloro-4-nitroaniline 99-30-9
2,4-Dinitroaniline 97-02-9
2-Nitroaniline 88-74-4
3-Nitroaniline 99-09-2
4-Nitroaniline 100-01-6
2,4,6-Trichloroaniline 634-93-5
2,4,5-Trichloroaniline 636-30-6
aChemical Abstract Services Registry Number.
1.2 When this method is used to analyze unfamiliar samples for any or all
of the target analytes, 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. It is highly recommended that gas
chromatography/mass spectrometry be utilized for absolute analyte identification
when analyzing unfamiliar samples, if concentration permits. See Section 8.6 for
guidance. However, the use of the NPD minimizes the possibility of false
positives.
1.3 The method detection limit (MDL) for each target analyte is given in
Table 1. The MDL for a specific sample may differ from those listed, depending
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upon the nature of interferences in the sample matrix. Table 2 provides guidance
on the calculation of estimated quantisation limits (EQLs) for various matrices.
1.4 Aniline and many aniline derivatives often result in erratic
responses, thereby requiring frequent column maintenance and recalibration.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography and in the interpretation
of gas chromatograms. Each analyst must demonstrate the ability to generate
acceptable results with this method using the procedure described in Section 8.2.
2.0 SUMMARY OF METHOD
2.1 A measured volume of aqueous sample, approximately 1 liter, is solvent
extracted at basic pH according to Method 3510 (separatory funnel), Method 3520
(continuous liquid-liquid extraction), or other appropriate technique.
Extraction from solid matrices may be performed using Methods 3540, 3541, 3545,
or 3550, or other appropriate technique. Both neat and diluted organic liquids
may be prepared by Method 3580 (waste dilution) and analyzed by direct injection.
2.2 If interferences are present, the extract may be cleaned up according
to Method 3620, Florisil Column Cleanup. Gel Permeation Chromatography Cleanup
(Method 3640) has also been validated for aniline and certain derivatives to
remove high boiling material that causes chromatography problems.
2.3 The target analytes in the extract are determined by capillary gas
chromatography with a nitrogen phosphorus detector (GC/NPD).
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
3.2.1 Volumetric flasks and glassware used for making up calibration
standards must be scrupulously cleaned. Clean all glassware as soon as
possible after use by rinsing with the last solvent used in it. This
should be followed by detergent washing with hot water and rinses with tap
and distilled water. It should then be drained dry, and heated in a muffle
furnace at 400°C for 15 to 30 min.
3.2.2 Some thermally stable materials may not be eliminated by this
treatment. Solvent rinses with acetone and 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.
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3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from sample to sample. The judicious use of one or more cleanup
techniques as listed in Section 7.1.2 may be necessary to eliminate or minimize
matrix interferences. The use of the NPD will help to minimize many of the
interference problems.
3.4 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
syringe used for injection must be rinsed out between samples with solvent.
Whenever an unusually concentrated sample is encountered, it should be followed
by the analysis of a solvent blank to check for cross-contamination. Additional
solvent blanks interspersed with the sample extracts should be considered
whenever the analysis of a solvent blank indicates cross-contamination problems.
3.5 Retention time data for two capillary columns are found in Table 1.
The SE-54 fused silica capillary column does not adequately resolve the following
two pairs of compounds: 2-nitroaniline/2,4,6-trichloroaniline, and
4-nitroaniline/4-chloro-2-nitroaniline. Only partial resolution of
3-chloroaniline and 4-chloroaniline is achieved. Unless the purpose for the
analysis can be served by reporting the sum of an unresolved pair, the alternate
capillary column must be used for these compounds and to verify the absence of
either compound in a pair. The alternate fused silica capillary column (SE-30)
gives resolution of these compound pairs, but fails to resolve
2,6-dibromo-4-nitroaniline/2,4-dinitroaniline and gives only partial resolution
of 3-chloroaniline/4-chloroaniline. Guidelines for selecting alternate capillary
columns are given in Section 4.1.2.3.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - an analytical system complete with gas
chromatograph suitable for on-column splitless injections and all required
accessories, including detector, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.2 Suggested GC columns
4.2.1 Column 1 - 30 m x 0.25 mm fused silica capillary column coated
with SE-54 (J&W Scientific or equivalent). The outlet end of the fused
silica column should be threaded through the burner tip of the NPD to
within 2 to 4 mm from the flame jet in order to minimize losses of
anilines.
4.2.2 Column 2 - 30 m x 0.25 mm fused silica capillary column coated
with SE-30 (J&W Scientific or equivalent).
4.2.3 The fused silica capillary columns will not resolve certain
pairs of aniline compounds, as indicated in Section 1.3 and Table 1.
Alternate capillary columns may be used if the relative standard deviations
of responses for replicate injections meet the requirements of Section 8.2.
The analyst should be aware of chromatographic peak shape as an indicator
of method performance.
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4.3 Detector- Nitrogen/Phosphorus (NPD) or equivalent [i.e., Alkali-Flame
Detector (AFD) or Thermionic Specific Detector (TSD)].
4.4 Vials - sizes as appropriate, glass with Teflon®-lined screw-caps or
crimp tops.
4.5 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.6 Glassware - refer to the applicable 3500 and 3600 series methods.
5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the chemicals are of sufficiently high purity to permit
their use without affecting the accuracy of the determinations.
5.3 Reagents for sample preservation
5.3.1 Sodium hydroxide, NaOH - (1.0 M in organic-free reagent
water).
5.3.2 Sulfuric acid, H2S04 - concentrated, specific gravity 1.84.
5.4 Solvents - All solvents must be pesticide quality or equivalent.
5.4.1 Acetone, CH3COCH3
5.4.2 Toluene, C6H5CH3
5.4.3 Refer to the applicable 3500 and 3600 series methods.
5.5 Stock standard solutions (1000 mg/L) - Stock standard solutions can
be prepared from pure standard materials or purchased as certified solutions.
5.5.1 Prepare stock standard solutions by accurately weighing about
0.0100 grams of pure materials. Dissolve the material in pesticide quality
toluene and dilute to volume in a 10 ml volumetric flask. Larger volumes
can be used at the convenience of the analyst. If compound purity is
certified at 96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards can be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.5.2 Transfer the stock standard solutions into Teflon®-sealed
bottles. Store at 4°C and protect from light. Stock standard solutions
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should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.5.3 Stock standard solutions must be replaced after six months,
or sooner if comparison with check standards indicates a problem.
5.6 Working standard solutions - Prepare working standards weekly by
adding volumes of one or more stock standards to a volumetric flask and diluting
to volume with toluene. Prepare at least five different concentrations to cover
the expected concentration range of the samples. Aniline and derivatives are not
as stable as many of the common semivolatile organics, therefore, their responses
must be closely monitored.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 The samples must be iced or refrigerated at 4°C from the time of
collection until extraction. For chlorinated waste, immediately add 35 mg sodium
thiosulfate per part per million of free chlorine per liter.
6.3 Adjust the pH of the sample to 6 to 8 with sodium hydroxide or
sulfuric acid immediately after sampling.
7.0 PROCEDURE
7.1 Extraction and cleanup
7.1.1 Refer to Chapter Two and Method 3500 for guidance on choosing
the appropriate extraction procedure. In general, water samples are
extracted at a pH > 11 with methylene chloride, using either Methods 3510
or 3520. Solid samples are extracted using either Methods 3540, 3541,
3545, or 3550 with methylene chloride/acetone (1:1) as the extraction
solvent. Prepare waste liquids (non-aqueous) by Method 3580 (Waste
Dilution).
7.1.2 If necessary, the samples may be cleaned up using Method 3620
(Florisil) and/or Method 3640 (Gel Permeation Chromatography). See Chapter
Two, Sec. 2.3 and Method 3600 for general guidance on cleanup and method
selection. Method 3660 is used for sulfur removal.
7.1.3 Prior to gas chromatographic analysis by NPD, the extraction
solvent must exchanged into toluene by adding 3 - 4 mL of toluene to the
vial just prior to the final concentration by nitrogen blowdown.
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7.2 Chromatographic conditions (recommended)
Column 1:
Carrier gas:
Carrier gas flow rate:
Temperature program:
Column 2:
Carrier gas:
Flow rate:
Temperature program:
SE-54 Fused Silica 30 m x 0.25 mm
Helium
28.5 cm/sec at room temperature
Initial temperature 80°C for 4 min
80°C to 230°C at 4°C/min
Hold at 230°C for 4 min
SE-30 Fused Silica 30 m x 0.25 mm
Helium
30 cm/sec at room temperature
Initial temperature 80°C for 4 min
80°C to 230°C at 4°C/min
Hold at 230°C for 4 min
Chromatographic conditions should be optimized to give separation
equivalent to that shown in Table 1.
7.3 Calibration
7.3.1 Prepare calibration standards using the procedures in Section
5.0. Refer to Method 8000 for proper calibration procedures. The procedure
for internal or external calibration may be used. Aniline and many aniline
derivatives often result in erratic responses, thereby requiring frequent
column maintenance and recal ibration.
7.3.2
windows.
Refer to Method 8000 for the establishment of retention time
7.4 Gas Chromatographic analysis of samples
7.4.1 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.4.2 Automatic injections of 1 fj,L are recommended. Manual
injections of no more than 2 jzL may be used if the analyst demonstrates
quantitation precision of < 10 percent relative standard deviation. The
solvent flush technique may be used if the amount of solvent is kept at a
minimum. If the internal standard calibration technique is used, add 10
Hi of the internal standard to each 1 ml of sample extract prior to
injection.
7.4.3 Tentative identification of an analyte occurs when a peak from
a sample extract falls within the daily retention time window.
7.4.4 Record the volume injected to the nearest 0.05 p,L and the
resulting peak size in peak height or area units. Using either the
internal or the external calibration procedure (Method 8000), determine the
identity and the quantity of each component peak in the sample chromatogram
which corresponds to the compounds used for calibration purposes. See
Method 8000 for calculation equations.
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7.4.5 If the responses exceed the linear range of the system, dilute
the extract and reanalyze. Peak height measurements are recommended over
peak area integration when overlapping peaks cause errors in area
integration.
7.4.6 If partially overlapping or coeluting peaks are found, change
columns or employ a GC/MS technique (see Section 8.6 and Method 8270).
Interferences that prevent analyte identification and/or quantitation may
be removed by the cleanup techniques mentioned above.
7.4.7 If the peak response is less than 2.5 times the baseline noise
level, the validity of the quantitative result may be questionable. The
analyst should consult with the source of the sample to determine whether
further concentration of the sample is warranted.
7.5 Instrument maintenance
7.5.1 Injection of sample extracts from waste sites often leaves a
high boiling residue in the injection port area, splitters, when used, and
the injection port end of the chromatographic column. This residue affects
chromatography in many ways (i.e., peak tailing, retention time shifts,
analyte degradation, etc.) and, therefore, instrument maintenance is very
important. Residue buildup in a splitter may limit flow through one leg
and therefore change the split ratios. If this occurs during an analytical
run, the quantitative data may be incorrect. Proper cleanup techniques
will minimize the problem and instrument QC will indicate when instrument
maintenance is required.
7.5.2 Suggested chromatograph maintenance - See Section 7.0 of
Method 8000 for guidance on corrective action for capillary columns and the
injection port.
7.6 GC/MS confirmation
7.6.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Follow the GC/MS
operating requirements specified in Method 8270. Ensure that there is
sufficient concentration of the analyte(s) to be confirmed, in the extract
for GC/MS analysis.
7.6.2 When available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process.
7.6.3 To confirm an identification of a compound, the background
corrected mass spectrum of the compound must be obtained from the sample
extract and must be compared with a mass spectrum from a stock or
calibration standard analyzed under the same chromatographic conditions.
At least 20 ng of material should be injected into the GC/MS. The
identification criteria specified in Method 8270 must be met for
qualitative confirmation.
7.6.4 Should the MS procedure fail to provide satisfactory results,
additional steps may be taken before reanalysis. These steps may include
the use of alternate GC columns or additional sample cleanup.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time
a set of samples is analyzed or there is a change in reagents, a method
blank should be analyzed as a safeguard against chronic laboratory
contamination. The blanks should be carried through all stages of sample
preparation and measurement.
8.4.2 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
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8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
8.7 Data from systems that automatically identify target analytes on the
basis of retention time or*retention time indices should be reviewed by an
experienced analyst before they are reported.
9.0 METHOD PERFORMANCE
9.1 Table 1 provides MDLs calculated from the analysis of spiked water
samples. Table 2 presents EQLs for various matrices.
9.2 The average recoveries presented in Table 3 were obtained in a single
laboratory, using spiked wastewater samples. The standard deviation of the
percent recovery is also included in Table 3.
9.3 This method has also been tested for linearity of recovery from spiked
organic-free reagent water and has been demonstrated to be applicable over the
concentration range from 3 x MDL to 300 x MDL with the following exceptions:
4-chloroaniline recovery was linear over the range 40-400 /jg/L (40-400 x MDL).
Aniline recovery was linear over the range 40-800 /jg/L (16-320 x MDL).
10.0 REFERENCES
1. U.S. EPA Method 650, Aniline and Selected Substituted Derivatives.
2. Analytical Procedures for Aniline and Selected Derivatives in Wastewater
and Sludge. Report for U.S. Environmental Protection Agency, Contract
Number 68-03-2952.
3. Interagency Testing Committee: Receipt of Fourth Report and Request for
Comments, Federal Register, June 1, 1979, V. 44, p. 31866-31889.
4. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis: Some
Practical Aspects", Journal of the Association of Official Analytical
Chemists, 48, 1037 (1965).
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TABLE 1
RETENTION TIMES AND METHOD DETECTION LIMITS
Retention Time (min)
Method8
Detection
Analyte
Anil ine
2-Chloroaniline
3-Chloroaniline
4-Chloroanil ine
4-Chloroaniline
2-Nitroanil ine
2,4,6-Trichloroaniline
3,4-Dichloroaniline
3-Nitroanil ine
2,4,5-Trichloroanil ine
4-Nitroanil ine
4-Chloro-2-nitroani1 ine
2-Chloro-4-nitroanil ine
2,6-Dichloro-4-nitroaniline
2-Bromo-6-chloro-4-nitroanil ine
2-Chloro-4,6-dinitroaniline
2,6-Dibromo-4-nitroanil ine
2,4-Dinitroaniline
2-Bromo-4,6-dinitroanil ine
Column 1
7.5
12.1
14.6
14.7
18.0
21.9
21.9
22.7
24.5
26.3
28.3
28.3
31.2
31.9
34.8
37.1
37.6
38.4
39.8
Column 2
6.3
7.1
9.0
9.1
12.1
15.6
16.3
16.6
18.0
20.4
21.7
22.0
24.8
26.0
28.8
30.1
31.6
31.6
33.4
Limit (M9/L)
2.3
1.4
1.8
0.66
4.6
1.0
5.8
3.2
3.3
3.0
11.0
2.7
3.2
2.9
3.4
3.6
3.8
8.9
3.7
MDL based upon seven replicate determinations in organic-free reagent water.
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TABLE 2
ESTIMATED QUANTITATION LIMITS (EQL) FOR VARIOUS MATRICES3
Matrix Factorb
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
a Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
b EQL = [MDL for water (Table 1)] times [Factor (Table 3)].
For nonaqueous samples, the factor is on a wet-weight basis.
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TABLE 3
SINGLE OPERATOR ACCURACY AND PRECISION
Average
"I
/a
Compounds Recovery
Aniline
2-Chloroanil ine
3-Chloroaniline
4-Chloroanil ine
4-Bromoaniline
3,4-Dichloroanil ine
2,4,6-Trichloroaniline
2,4,5-Trichloroaniline
2-Nitroanil ine
3-Nitroanil ine
4-Nitroanil ine
2,4-Dinitroaniline
4-Chloro-2-nitroani1ine
2-Chloro-4-nitroanil ine
2,6-Dichloro-4-nitroaniline
2,6-Dibromo-4-nitroanil ine
2-Bromo-6-chloro-4-nitroanil ine
2-Chloro-4,6-dinitroanil ine
2-Bromo-4,6-dinitroanil ine
70
88
75
64
78
79
93
85
92
80
94
93
94
96
92
89
110
92
81
Standard
Deviation
%
8.7
9.1
10
7.5
10
13
15
12
14
11
21
20
16
16
12
13
16
17
14
Spike
Range
(M9/L)
8.0
4.0
4.0
4.0
16.0
8.0
8.0
8.0
4.0
8.0
8.0
16.0
8.0
8.0
8.0
8.0
8.0
8.0
8.0
- 100
- 100
- 100
- 100
- 160
- 100
- 100
- 100
- 100
- 100
- 100
- 160
- 100
- 100
- 100
- 100
- 100
- 100
- 100
A total of forty-eight (48) samples comprising four (4) different matrix types
were used for this study.
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METHOD 8131
ANILINE AND SELECTED DERIVATIVES BY GC: CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose appropriate
extraction technique and
perform extraction.
I
7.1.2 If necessary, choose
appropriate cleanup technique
and perform cleanup.
7.1.3 Solvent exchange
to toluene (only if using
NPD).
7.2 Set chromatographic
conditions to allow
adequate separation
(See Table 1).
7.3 Prepare calibration
standards and perform
calibration. Establish
retention time windows.
7.4 Record volume injected
to 0.05_pL and the resulting
peak size (height or area
units).
7.4 Determine identity and
quantity of component
peaks using calibration.
Does
response
exceed the linear
range of the
system?
Dilute and reanalyze.
Are
overlapping or
co-eluting peak
present?
Change columns or
try a GC/MS technique.
Is peak
response
less than 2.5
times the base
line noise
level?
The validity of the
quantitative result may be
questionable. Take
appropriate action.
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METHOD 8151A
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATION
PERIVATIZATION: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8151 is a capillary gas chromatographic (GC) method for
determining certain chlorinated acid herbicides and related compounds in aqueous,
soil and waste matrices. Specifically, Method 8151 may be used to determine the
following compounds:
Compound CAS No.'
2,4-D 94-75-7
2,4-DB 94-82-6
2,4,5-TP (Silvex) 93-72-1
2,4,5-T 93-76-5
Dalapon 75-99-0
Dicamba 1918-00-9
Dichloroprop 120-36-5
Dinoseb 88-85-7
MCPA 94-74-6
MCPP 93-65-2
4-Nitrophenol 100-02-1
Pentachlorophenol 87-86-5
a Chemical Abstract Services Registry Number
1.2 Because these compounds are produced and used in various forms (i.e.,
acid, salt, ester, etc.), Method 8151 describes a hydrolysis step that can be
used to convert herbicide esters into the acid form prior to analysis. Herbicide
esters generally have a half-life of less than one week in soil.
1.3 When Method 8151 is used to analyze unfamiliar samples, compound
identifications should be supported by at least one additional qualitative
technique. Sec. 8.4 provides gas chromatograph/mass spectrometer (GC/MS)
criteria appropriate for the qualitative confirmation of compound
identifications.
1.4 The estimated detection limits for each of the compounds in aqueous
and soil matrices are listed in Table 1. The detection limits for a specific
waste sample may differ from those listed, depending upon the nature of the
interferences and the sample matrix.
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1.5 The following compounds may also be determined using this method:
Compound CAS No.'
Acifluorfen 50594-66-6
Bentazon 25057-89-0
Chloramben 133-90-4
DCPA diacidb 2136-79-0
3,5-Dichlorobenzoic acid 51-36-5
5-Hydroxydicamba 7600-50-2
Picloram 1918-02-1
a Chemical Abstract Services Registry Number
b DCPA monoacid and diacid metabolites included in method scope; DCPA
diacid metabolite used for validation studies. DCPA is a dimethyl ester.
1.6 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.7 Only experienced analysts should be allowed to work with diazomethane
due to the potential hazards associated with its use (explosive, carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Method 8151 provides extraction, derivatization, and gas
chromatographic conditions for the analysis of chlorinated acid herbicides in
water, soil, and waste samples. An option for the hydrolysis of esters is also
described.
2.2 Water samples are extracted with diethyl ether and then esterified
with either diazomethane or pentafluorobenzyl bromide. The derivatives are
determined by gas chromatography with an electron capture detector (GC/ECD). The
results are reported as acid equivalents.
2.3 Soil and waste samples are extracted and esterified with either
diazomethane or pentafluorobenzyl bromide. The derivatives are determined by gas
chromatography with an electron capture detector (GC/ECD). The results are
reported as acid equivalents.
2.4 If herbicide esters are to be determined using this method, hydrolysis
conditions for the esters in water and soil extracts are described.
2.5 The sensitivity of Method 8151 depends on the level of interferences
in addition to instrumental limitations. Table 1 lists the GC/ECD and GC/MS
detection limits that can be obtained in aqueous and soil matrices in the absence
of interferences. Detection limits for a typical waste sample should be higher.
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3.0 INTERFERENCES
3.1 Refer to Method 8000.
3.2 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis, by analyzing reagent blanks.
3.2.1 Glassware must be scrupulously cleaned. Clean each piece of
glassware as soon as possible after use by rinsing it with the last solvent
used in it. This should be followed by detergent washing with hot water
and rinses with tap water, then with organic-free reagent water. Glassware
should be solvent-rinsed with acetone and pesticide-quality hexane. After
rinsing and drying, glassware should be sealed and stored in a clean
environment to prevent any accumulation of dust or other contaminants.
Store glassware inverted or capped with aluminum foil. Immediately prior
to use, glassware should be rinsed with the next solvent to be used.
3.2.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all-
glass systems may be required.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from waste to waste, depending upon the nature and diversity of the
waste being sampled.
3.4 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination by methylation. Phenols, including
chlorophenols, may also interfere with this procedure. The determination using
pentafluorobenzylation is more sensitive, and more prone to interferences from
the presence of organic acids or phenols than by methylation.
3.5 Alkaline hydrolysis and subsequent extraction of the basic solution
removes many chlorinated hydrocarbons and phthalate esters that might otherwise
interfere with the electron capture analysis. However, hydrolysis may result in
the loss of dinoseb and the formation of aldol condensation products if any
residual acetone remains from the extraction of solids.
3.6 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Therefore, glassware must
be acid-rinsed and then rinsed to constant pH with organic-free reagent water.
Sodium sulfate must be acidified.
3.7 Sample extracts should be dry prior to methylation or else poor
recoveries will be obtained.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - Analytical system complete with gas chromatograph
suitable for Grob-type injection using capillary columns, and all required
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accessories including detector, capillary analytical columns, recorder, gases,
and syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.2 GC columns
The analyst may chose either narrow-bore or wide-bore GC columns. Narrow-
bore column la is useful for GC/MS confirmation of these analytes. When using
an electron capture detector, analyses two columns are necessary to provide
confirmation of identifications.
Although not specifically evaluated under the chromatographic conditions
described in this procedure, the analyst may opt to configure the GC for
simultaneous dual-column operation using commercially-available Y-adapters to
connect both columns to a single injector port and employing a separate electron
capture detector for each column.
4.2.1 Narrow-bore columns
4.1.2.1 Primary column 1 - 30 m x 0.25 mm, 5% phenyl/95%
methyl silicone (DB-5, J&W Scientific, or equivalent), 0.25 p.m film
thickness.
4.1.2.2 Primary column la (GC/MS) - 30 m x 0.32 mm, 5%
phenyl/95% methyl silicone, (DB-5, J&W Scientific, or equivalent), 1
/iin film thickness.
4.1.2.3 Column 2 - 30 m x 0.25 mm, 35% phenyl
methylpolysiloxane (DB-608, J&W Scientific or equivalent), a 0.25 urn
film thickness.
4.1.2.4 Confirmation column - 30 m x 0.25 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or equivalent),
0.25 /urn film thickness.
4.2.2 Wide-bore columns
4.2.2.1 Primary Column - 30 m x 0.53 mm DB-608 (J&W
Scientific or equivalent) with 0.83 /zm film thickness.
4.2.2.2 Confirmation Column - 30 m x 0.53 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or equivalent),
1.0 urn film thickness.
4.3 Electron capture detector (ECD).
4.4 Kuderna-Danish (K-D) apparatus.
4.4.1 Concentrator tube - 10-mL graduated (Kontes K-570050-1025 or
equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
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4.4.2 Evaporation flask - 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.4.3 Snyder column - Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.4.4 Snyder column - Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.4.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
NOTE: The following glassware is recommended for the purpose of solvent
recovery during the concentration procedures requiring the use of
Kuderna-Danish evaporative concentrators. Incorporation of this
apparatus may be required by State or local municipality regulations that
govern air emissions of volatile organics. EPA recommends the
incorporation of this type of reclamation system as a method to implement
an emissions reduction program. Solvent recovery is a means to conform
with waste minimization and pollution prevention initiatives.
4.4.6 Solvent vapor recovery system (Kontes K-545000-1006 or K-
547300-0000, Ace Glass 6614-30, or equivalent).
4.5 Diazomethane generator - Refer to Sec. 7.5 to determine which method
of diazomethane generation should be used for a particular generation.
4.5.1 Diazald kit - Recommended for the generation of diazomethane
(Aldrich Chemical Co., Catalog No. 210,025-0, or equivalent).
4.5.2 As an alternative, assemble from two 20 mm x 150 mm test
tubes, two Neoprene rubber stoppers, and a source of nitrogen. Use
Neoprene rubber stoppers with holes drilled in them to accommodate glass
delivery tubes. The exit tube must be drawn to a point to bubble
diazomethane through the sample extract. The generator assembly is shown
in Figure 1. The procedure for use of this type of generator is given in
Sec. 7.5.
4.6 Beaker - 400-mL, thick-walled.
4.7 Funnel - 75 mm diameter.
4.8 Separatory funnel - 500-mL, with Teflon® stopcock.
4.9 Centrifuge bottle - 500-mL, Pyrex® 1260 or equivalent.
4.10 Erlenmeyer flasks - 250-mL and 500-mL, with a ground-glass joint at
the neck.
4.11 Pipet - Pasteur, glass, disposable (140 mm x 5 mm ID).
4.12 Vials - 10-mL, glass, with Teflon®-lined screw-caps.
4.13 Volumetric flasks, Class A - 10-mL to 1000-mL.
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4.14 Filter paper - 15 cm diameter (Whatman No. 1 or equivalent).
4.15 Glass wool - Pyrex®, acid washed.
4.16 Boiling chips - Solvent-extracted with methylene chloride,
approximately 10/40 mesh (silicon carbide or equivalent).
4.17 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
4.18 Balance - Analytical, capable of accurately weighing to 0.0001 g.
4.19 Centrifuge.
4.20 Ultrasonic extraction system - A horn-type device equipped with a
titanium tip, or a device that will give equivalent performance, should be used.
The disrupter must have a minimum power wattage of 300 watts, with pulsing
capability. A device designed to reduce the cavitation sound is recommended.
Follow the manufacturer's instructions for preparing the disrupter for extraction
of samples. Use a 3/4" horn for most samples.
4.21 Sonabox - Recommended with above disrupters for decreasing cavitation
sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.22 pH paper - wide range
4.23 Silica gel cleanup column (Bond Elut™ - Analytichem, Harbor City, CA
or equivalent).
4.24 Microsyringe - 10-//L.
4.25 Wrist shaker - Burrell Model 75 or equivalent.
4.26 Drying column - 400 mm x 20 mm ID Pyrex® chromatographic column with
Pyrex® glass wool at bottom and a Teflon® stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex® glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed by
50 ml of elution solvent prior to packing the column with adsorbent.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free water, as defined in Chapter One.
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5.3 Sodium hydroxide solution (0.1 N), NaOH - Dissolve 4 g of NaOH in
organic-free reagent water and dilute to 1.0 L.
5.4 Potassium hydroxide solution (37% aqueous solution (w/v)), KOH -
Dissolve 37 g of potassium hydroxide pellets in organic-free reagent water and
dilute to 100 ml.
5.5 Phosphate buffer (0.1 M), pH = 2.5 - Dissolve 12 g sodium phosphate
(NaH2P04) in organic-free reagent water and dilute to 1.0 L. Add phosphoric acid
to adjust the pH to 2.5.
5.6 N-methyl-N-nitroso-p-toluenesulfonamide (Diazald) - High purity
(Aldrich Chemical Co., or equivalent).
5.7 Silicic acid, H2Si05 - 100-mesh powder, store at 130°C.
5.8 Potassium carbonate, K2C03.
5.9 2,3,4,5,6-Pentafluorobenzyl bromide (PFBBr), C6F5CH2Br - Pesticide
quality or equivalent.
5.10 Sodium sulfate (granular, acidified, anhydrous), Na2S04 - Purify by
heating at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there is
no interference from the sodium sulfate. Acidify by slurrying 100 g sodium
sulfate with enough diethyl ether to just cover the solid; then add 0.1 ml of
concentrated sulfuric acid and mix thoroughly. Remove the ether under vacuum.
Mix 1 g of the resulting solid with 5 ml of organic-free reagent water and
measure the pH of the mixture. It must be below a pH of 4. Store the remaining
solid at 130'C.
5.11 Solvents - All solvents should be pesticide quality or equivalent.
5.11.1 Methylene chloride, CH2C12.
5.11.2 Acetone, CH3COCH3.
5.11.3 Methanol, CH3OH.
5.11.4 Toluene, C6H5CH3.
5.11.5 Diethyl Ether, C2H5OC2H5. Must be free of peroxides as
indicated by test strips (EM Quant, or equivalent). Procedures for removal
of peroxides are provided with the test strips.
NOTE: Diethyl ether used for this procedure should be stabilized with BHT, not
with ethanol, as when ethanol-stabilized ether is used, the methylation
reaction may not proceed efficiently, leading to low recoveries of target
analytes.
5.11.6 Isooctane, (CH3)3CH2CH(CH3)2.
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5.11.7 Hexane, C6H14.
5.11.8 Carbitol (diethylene glycol monoethyl ether),
C2H5OCH2CH2OCH2CH20 - optional, for producing alcohol-free diazomethane.
5.12 Stock standard solutions (1000 mg/L) - May be prepared from pure
standard materials or purchased as certified solutions.
5.12.1 Prepare stock standard solutions by accurately weighing about
0.010 g of pure acid. Dissolve the material in pesticide quality acetone
and dilute to volume in a 10-mL volumetric flask. Stocks prepared from
pure methyl esters are dissolved in 10% acetone/isooctane (v/v). Larger
volumes may be used at the convenience of the analyst. If compound purity
is certified at 96% or greater, the weight may be used without correction
to calculate the concentration of the stock standard.
5.12.2 Transfer the stock standard solutions to vials with Teflon®-
lined screw-caps. Store at 4°C, protected from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially immediately prior to preparing calibration
standards from them.
5.12.3 Stock standard solutions of the derivatized acids must be
replaced after 1 year, or sooner, if comparison with check standards
indicates a problem. Stock standard solutions of the free acids degrade
more quickly and should be replaced after two months, or sooner if
comparison with check standards indicates a problem.
5.13 Internal Standard Spiking Solution (if internal standard calibration
is used) - To use this approach, the analyst must select one or more internal
standards that are similar in analytical behavior to the compounds of interest.
The analyst must further demonstrate that the measurement of the internal
standard is not affected by method or matrix interferences.
5.13.1 The compound 4,4'-dibromooctafluorobiphenyl (DBOB) has been
shown to be an effective internal standard, but other compounds, such as
1,4-dichlorobenzene, may be used if there is a DBOB interference.
5.13.2 Prepare an internal standard spiking solution by accurately
weighing approximately 0.0025 g of pure DBOB. Dissolve the DBOB in acetone
and dilute to volume in a 10 ml volumetric flask. Transfer the internal
standard spiking solution to a vial with a Teflon®-lined screw-cap, and
store at room temperature. Addition of 10 /uL of the internal standard
spiking solution to 10 mL of sample extract results in a final internal
standard concentration of 0.25 M9/L. The solution should be replaced if
there is a change in internal standard response greater than 20 percent of
the original response recorded.
5.14 Calibration standards - Prepare a minimum of five concentrations for
each parameter of interest, through dilution of the stock standards with diethyl
ether or hexane. One of the standards should be at a concentration near, but
above, the method detection limit. The remaining standards should correspond to
the expected range of concentrations found in real samples or should define the
8151A - 8 Revision 1
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working range of the GC. Calibration solutions must be replaced after six
months, or sooner if comparison with check standards indicates a problem.
5.14.1 Derivatize each calibration standard prepared from free acids
in a 10-mL K-D concentrator tube, according to the procedures beginning at
Sec. 7.5.
5.14.2 Add a known, constant, amount of one or more internal
standards to each derivatized calibration standard, and dilute to volume
with the solvent indicated in the derivative option used.
5.15 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and determinative step, and the
effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two herbicide surrogates (e.g.,
herbicides that are not expected to be present in the sample) recommended to
encompass the range of the temperature program used in this method. Deuterated
analogs of analytes should not be used as surrogates in gas chromatographic
analysis due to coelution problems.
5.15.1 The recommended surrogate is 2,4-Dichlorophenylacetic acid
(DCAA).
5.15.2 Prepare a surrogate spiking solution by accurately weighing
approximately 0.001 g of pure DCAA. Dissolve the DCAA in acetone, and
dilute to volume in a 10-mL volumetric flask. Transfer the surrogate
spiking solution to a vial with a Teflon®-lined screw-cap, and store at
room temperature. Addition of 50 /uL of the surrogate spiking solution to
1 L of sample, prior to extraction, results in a final concentration in the
extract of 0.5 mg/L.
5.16 pH Adjustment Solutions
5.16.1 Sodium hydroxide, NaOH, 6 N.
5.16.2 Sulfuric acid, H2S04, 12 N.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1. 1-L samples should be collected.
6.2 Extracts must be stored under refrigeration (4°C) and protected from
light.
7.0 PROCEDURE
7.1 Extraction and hydrolysis of high concentration waste samples
7.1.1 Follow Method 3580, Waste Dilution, with the following
exceptions:
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7.1.1.1 Use diethyl ether as the dilution solvent.
7.1.1.2 Use acidified anhydrous sodium sulfate and acidified
glass wool.
7.1.1.3 Spike the sample with surrogate(s) according to Sec.
5.15.
7.1.2 If the sample is to be analyzed for both herbicide esters and
acids, then the sample extract must be hydrolyzed. In this case, transfer
1.0 ml (a smaller volume or a dilution may be required if herbicide
concentrations are large) to a 250-mL Erlenmeyer flask with a ground-glass
joint at the neck. Proceed to Sec. 7.2.3. If the analysis is for acid
herbicides only, proceed to Sec. 7.5 for derivatization by diazomethane (if
PFB derivatization is selected, reduce the volume of diethyl ether to 0.1 -
0.5 ml as per Sec. 7.2 and then dilute to 4 ml with acetone).
7.2 Extraction and hydrolysis of soil, sediment, and other solid samples
Two extraction procedures are applicable to solid samples: ultrasonic
extraction and shaker extraction. The same hydrolysis procedures (Sec. 7.2.3)
apply to both types of extracts.
7.2.1 Ultrasonic extraction
7.2.1.1 Add 30 g (dry weight) of the well-mixed solid sample
to a 400-mL thick-wall beaker. Adjust the pH to 2 with concentrated
hydrochloric acid or acidify solids in the beaker with 85 ml of 0.1
M phosphate buffer (pH = 2.5) and thoroughly mix the contents with a
glass stirring rod. Spike the sample with surrogate(s) (Sec. 5.15).
7.2.1.2 The ultrasonic extraction of solids must be
optimized for each type of sample. In order for the ultrasonic
extractor to efficiently extract solid samples, the sample must be
free flowing when the solvent is added. Acidified anhydrous sodium
sulfate should be added to clay type soils (normally 1:1), or any
other solid that is not a free flowing sandy mixture, until a free
flowing mixture is obtained.
7.2.1.3 Add 100 ml of methylene chloride/acetone (1:1 v/v)
to the beaker. Perform ultrasonic extraction for 3 minutes, with
output control knob set at 10 (full power) and with mode switch on
Pulse (pulsing energy rather than continuous energy) and percent-duty
cycle knob set at 50% (energy on 50% of time and off 50% of time).
Allow the solids to settle. Transfer the organic layer into a 500-mL
centrifuge bottle.
7.2.1.4 Ultrasonically extract the sample twice more using
100 mL of methylene chloride and the same ultrasonic conditions.
7.2.1.5 Combine the three organic extracts from the sample
in the centrifuge bottle and centrifuge 10 minutes to settle the fine
particles. Filter the combined extract through filter paper
(Whatman #1, or equivalent) containing 7-10 g of acidified sodium
8151A - 10 Revision 1
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sulfate into a 500-mL 24/40 Erlenmeyer flask. Add 10 g of acidified
anhydrous sodium sulfate. Periodically, vigorously shake the extract
and drytfig agent and allow the drying agent to remain in contact with
the extract for a minimum of 2 hours. See NOTE in Sec. 7.3.6 that
emphasizes the need for a dry extract prior to esterification.
7.2.1.6 Quantitatively transfer the contents of the flask to
a 500-mL K-D flask with a 10-mL concentrator tube attached. Add
boiling chips and attach the macro Snyder column. Evaporate the
extract on the water bath to a volume of approximately 5 ml. Remove
the flasks from the water bath and allow them to cool.
7.2.1.7 If hydrolysis or additional cleanup is not required
and the sample is dry, proceed to Sec. 7.4.4. Otherwise, proceed to
Sec. 7.2.3 for hydrolysis or Sec. 7.2.4 for cleanup.
7.2.2 Shaker extraction
7.2.2.1 Add 50 g (dry weight) of the well mixed, moist solid
sample to a 500-mL wide-mouth Erlenmeyer flask. Adjust the pH to 2
with concentrated HC1 and monitor the pH for 15 minutes with
occasional stirring. If necessary, add additional HC1 until the pH
remains at 2. Spike the sample with surrogate(s) (Sec. 5.15).
7.2.2.2 Add 20 mL of acetone to the flask and mix the
contents with the wrist shaker for 20 minutes. Add 80 mL diethyl
ether to the same flask and shake again for 20 minutes. Decant the
extract and measure the volume of solvent recovered.
7.2.2.3 Extract the sample twice more using 20 mL of acetone
followed by 80 mL of diethyl ether. After addition of each solvent,
the mixture should be shaken with the wrist shaker for 10 minutes and
the acetone-ether extract decanted.
7.2.2.4 After the third extraction, the volume of extract
recovered should be at least 75% of the volume of added solvent. If
this is not the case, additional extractions may be necessary.
Combine the extracts in a 2-L separatory funnel containing 250 mL of
reagent water. If an emulsion forms, slowly add 5 g of acidified
sodium sulfate (anhydrous) until the solvent-water mixture separates.
A quantity of acidified sodium sulfate equal to the weight of the
sample may be added, if necessary.
7.2.2.5 Check the pH of the extract. If it is not at or
below pH 2, add more concentrated HC1 until stabilized at the desired
pH. Gently mix the contents of the separatory funnel for 1 minute and
allow the layers to separate. Collect the aqueous phase in a clean
beaker and the extract phase (top layer) in a 500-mL ground glass-
stoppered Erlenmeyer flask. Place the aqueous phase back into the
separatory funnel and re-extract using 25 mL of diethyl ether. Allow
the layers to separate and discard the aqueous layer. Combine the
ether extracts in a 500-mL K-D flask.
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7.2.2.6 If hydrolysis or additional cleanup is not required
and the sample is dry, proceed to Sec. 7.4.4. Otherwise , proceed to
Sec. 7.2.3 for hydrolysis or Sec. 7.2.4 for extract cleanup.
7.2.3 Hydrolysis of soil, sediment, or other solid sample extracts
Use this step only if herbicide esters in addition to herbicide acids
are to be determined.
7.2.3.1 Add 5 ml of 37% aqueous potassium hydroxide and 30
mL of water to the extract. Add additional boiling chips to the K-D
flask. Reflux the mixture on a water bath at 60 - 65°C until the
hydrolysis step is completed (usually 1-2 hours). Remove the flasks
from the water bath and cool to room temperature.
CAUTION: The presence of residual acetone will result in the formation of aldol
condensation products which will cause GC interference.
7.2.3.2 Transfer the hydrolyzed aqueous solution to a 500-mL
separatory funnel and extract the solution three times with 100-mL
portions of methylene chloride. Discard the methylene chloride phase.
At this point, the basic (aqueous) solution contains the herbicide
salts.
7.2.3.3 Adjust the pH of the solution to <2 with cold (4°C)
sulfuric acid (1:3) and extract once with 40 ml of diethyl ether and
twice with 20-mL portions of ether. Combine the extracts and pour
them through a pre-rinsed drying column containing 7 to 10 cm of
acidified anhydrous sodium sulfate. Collect the dried extracts in a
500-mL Erlenmeyer flask (with a 24/40 joint) containing 10 g of
acidified anhydrous sodium sulfate. Periodically, vigorously shake
the extract and drying agent and allow the drying agent to remain in
contact with the extract for a minimum of 2 hours. See NOTE in Sec.
7.3.6 that emphasizes the need for a dry extract prior to
esterification. Quantitatively transfer the contents of the flask to
a 500-mL Kuderna-Danish flask with a 10-mL concentrator tube attached
when the extract is known to be dry.
7.2.3.4 Proceed to Sec. 7.4 for extract concentration. If
additional cleanup is required, proceed to Sec. 7.2.4.
7.2.4 Cleanup of non-hydrolyzed herbicides
Use this step if additional cleanup of the non-hydrolyzed herbicides
is required.
7.2.4.1 Partition the herbicides by extracting the methylene
chloride from 7.2.1.7 (or diethyl ether from 7.2.3.4) three times with
15-mL portions of aqueous base prepared by carefully mixing 30 mL of
reagent water into 15 mL of 37% aqueous potassium hydroxide. Discard
the methylene chloride or ether phase. At this point the basic
(aqueous) solution contains the herbicide salts.
8151A - 12 Revision 1
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7.2.4.2 Adjust the pH of the solution to <2 with cold (4°C)
sulfuric acid (1:3) and extract once with 40 ml of diethyl ether and
twice with 20-mL portions of ether. Combine the extracts and pour
them through a pre-rinsed drying column containing 7-10 cm of
acidified anhydrous sodium sulfate. Collect the dried extracts in a
500-mL Erlenmeyer flask (with a 24/40 joint) containing 10 g of
acidified anhydrous sodium sulfate. Periodically, vigorously shake
the extract and drying agent and allow the drying agent to remain in
contact with the extract for a minimum of 2 hours. See NOTE in Sec.
7.3.6 that emphasizes the need for a dry extract prior to
esterification. Quantitatively transfer the contents of the flask to
a 500-mL Kuderna-Danish flask with a 10-mL concentrator tube attached
when the extract is known to be dry.
7.2.4.3 Proceed to Section 7.4 for extract concentration.
7.3 Preparation of aqueous samples
7.3.1 Using a graduated cylinder, transfer a 1-L sample aliquot to
2-L separatory funnel. Spike the sample with surrogate compound(s)
"•• L Sec. 5.15.
a 6-L beyar di.ui jr i uinie
according to Sec. 5.15.
7.3.2 Add 250 g of NaCl to the sample, seal, and shake to dissolve
the salt.
7.3.3 Use this step only if herbicide esters, in addition to
herbicide acids, are to be determined
7.3.3.1 Add 17 mL of 6 N NaOH to the sample, seal, and
shake. Check the pH of the sample with pH paper. If the sample does
not have a pH greater than or equal to 12, adjust the pH by adding
more 6 N NaOH. Let the sample sit at room temperature until the
hydrolysis step is completed (usually 1-2 hours), shaking the
separatory funnel and contents periodically.
7.3.3.2 Add 60 mL of methylene chloride to the sample bottle
and rinse both the bottle and the graduated cylinder. Transfer the
methylene chloride to the separatory funnel and extract the sample by
vigorously shaking the funnel for 2 minutes, with periodic venting to
release excess pressure. Allow the organic layer to separate from the
water phase for a minimum of 10 minutes. If the emulsion interface
between the layers is more than one-third the volume of the solvent
layer, the analyst must employ mechanical techniques to complete the
phase separation. The optimum technique depends upon the sample, but
may include stirring, filtration through glass wool, centrifugation,
or other physical methods. Discard the methylene chloride phase.
7.3.3.3 Add a second 60-mL volume of methylene chloride to
the separatory funnel and repeat the extraction procedure a second
time, discarding the methylene chloride layer. Perform a third
extraction in the same manner.
7.3.4 Add 17 mL of cold (4°C) 12 N sulfuric acid to the sample (or
hydrolyzed sample), seal, and shake to mix. Check the pH of the sample
8151A - 13 Revision 1
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with pH paper. If the sample does not have a pH less than or equal to 2,
adjust the pH by adding more acid.
7.3.5 Add 120 mL diethyl ether to the sample, seal, and extract the
sample by vigorously shaking the funnel for 2 min with periodic venting to
release excess pressure. Allow the organic layer to separate from the
water phase for a minimum of 10 min. If the emulsion interface between
layers is more than one third the volume of the solvent layer, the analyst
must employ mechanical techniques to complete the phase separation. The
optimum techniques to complete the phase separation depends upon the
sample, but may include stirring, filtration through glass wool,
centrifugation, or other physical methods. Remove the aqueous phase to a
2-L Erlenmeyer flask and collect the ether phase in a 500-mL Erlenmeyer
flask containing approximately 10 g of acidified anhydrous sodium sulfate.
Periodically, vigorously shake the extract and drying agent.
7.3.6 Return the aqueous phase to the separatory funnel, add 60 mL
of diethyl ether to the sample, and repeat the extraction procedure a
second time, combining the extracts in the 500-mL Erlenmeyer flask.
Perform a third extraction with 60 ml diethyl ether in the same manner.
Allow the extract to remain in contact with the sodium sulfate for
approximately 2 hours.
NOTE: The drying step is very critical to ensuring complete esterification. Any
moisture remaining in the ether will result in low herbicide recoveries.
The amount of sodium sulfate is adequate if some free flowing crystals are
visible when swirling the flask. If all of the sodium sulfate solidifies
in a cake, add a few additional grams of acidified sodium sulfate and
again test by swirling. The 2 hour drying time is a minimum, however, the
extracts may be held in contact with the sodium sulfate overnight.
7.3.7 Pour the dried extract through a funnel plugged with acid
washed glass wool, and collect the extract in the K-D concentrator. Use
a glass rod to crush any caked sodium sulfate during the transfer. Rinse
the Erlenmeyer flask and funnel with 20 to 30 ml of diethyl ether to
complete the quantitative transfer. Proceed to Sec. 7.4 for extract
concentration.
7.4 Extract concentration
7.4.1 Add one or two clean boiling chips to the flask and attach a
three-ball Snyder column. Prewet the Snyder column by adding about 1 mL
of diethyl ether to the top of the column. Attach the solvent vapor
recovery glassware (condenser and collection device) (Sec. 4.4.6) to the
Snyder column of the K-D apparatus following manufacturer's instructions.
Place the K-D apparatus on a hot water bath (15 - 20°C above the boiling
point of the solvent) so that the concentrator tube is partially immersed
in the hot water and the entire lower rounded surface of the flask is
bathed with hot vapor. Adjust the vertical position of the apparatus and
the water temperature, as required, to complete the concentration in 10 -
20 minutes. At the proper rate of distillation the balls of the column
will actively chatter, but the chambers will not flood. When the apparent
volume of liquid reaches 1 mL, remove the K-D apparatus from the water bath
and allow it to drain and cool for at least 10 minutes.
8151A - 14 Revision 1
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7.4.2 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1 - 2 ml of diethyl ether. The
extract may be further concentrated by using either the micro Snyder column
technique (Sec. 7.4.3) or nitrogen blowdown technique (Sec. 7.4.4).
7.4.3 Micro Snyder column technique
7.4.3.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two-ball micro Snyder column. Prewet
the column by adding about 0.5 mL of diethyl ether to the top of the
column. Place the K-D apparatus in a hot water bath so that the
concentrator tube is partially immersed in the hot water. Adjust the
vertical position of the apparatus and the water temperature, as
required, to complete the concentration in 5-10 minutes. At the
proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 mL, remove the K-D apparatus from the water bath
and allow it to drain and cool for at least 10 minutes. Remove the
Snyder column, rinse the flask and its lower joints with about 0.2 mL
of diethyl ether and add to the concentrator tube. Proceed to Sec.
7.4.5.
7.4.4 Nitrogen blowdown
7.4.4.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the required
level using a gentle stream of clean, dry nitrogen (filtered through
a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon trap and the sample.
7.4.4.2 The internal wall of the tube must be rinsed down
several times with diethyl ether during the operation. During
evaporation, the solvent level in the tube must be positioned to
prevent water from condensing into the sample (i.e., the solvent level
should be below the level of the water bath). Under normal operating
conditions, the extract should not be allowed to become dry. Proceed
to Sec. 7.4.5.
7.4.5 Dilute the extract with 1 mL of isooctane and 0.5 mL of
methanol. Dilute to a final volume of 4 mL with diethyl ether. The sample
is now ready for methylation with diazomethane. If PFB derivation is being
performed, dilute to 4 mL with acetone.
7.5 Esterification - For diazomethane derivatization proceed with Sec.
7.5.1. For PFB derivatization proceed with Sec. 7.5.2.
7.5.1 Diazomethane derivatization - Two methods may be used for the
generation of diazomethane: the bubbler method (see Figure 1), Sec.
7.5.1.1, and the Diazald kit method, Sec. 7.5.1.2.
CAUTION: Diazomethane is a carcinogen and can explode under certain conditions.
8151A - 15 Revision 1
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The bubbler method is suggested when small batches of samples (10 -
15) require esterification. The bubbler method works well with samples
that have low concentrations of herbicides (e.g., aqueous samples) and is
safer to use than the Diazald kit procedure. The Diazald kit method is
good for large quantities of samples needing esterification. The Diazald
kit method is more effective than the bubbler method for soils or samples
that may contain high concentrations of herbicides (e.g., samples such as
soils that may result in yellow extracts following hydrolysis may be
difficult to handle by the bubbler method).
The diazomethane derivatization procedures described below will react
efficiently with all of the chlorinated herbicides described in this method
and should be used only by experienced analysts, due to the potential
hazards associated with its use.
The following precautions should be taken:
• Use a safety screen.
• Use mechanical pipetting aides.
• Do not heat above 90°C - EXPLOSION may result.
• Avoid grinding surfaces, ground-glass joints, sleeve bearings, and
glass stirrers - EXPLOSION may result.
• Store away from alkali metals - EXPLOSION may result.
• Solutions of diazomethane decompose rapidly in the presence of
solid materials such as copper powder, calcium chloride, and
boil ing chips.
7.5.1.1 Bubbler method - Assemble the diazomethane bubbler
(see Figure 1).
7.5.1.1.1 Add 5 mL of diethyl ether to the first
test tube. Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5
mL of 37% KOH, and 0.1 - 0.2 g of Diazald to the second test
tube. Immediately place the exit tube into the concentrator
tube containing the sample extract. Apply nitrogen flow
(10 mL/min) to bubble diazomethane through the extract for
10 minutes or until the yellow color of diazomethane
persists. The amount of Diazald used is sufficient for
esterification of approximately three sample extracts. An
additional 0.1 - 0.2 g of Diazald may be added (after the
initial Diazald is consumed) to extend the generation of the
diazomethane. There is sufficient KOH present in the
original solution to perform a maximum of approximately 20
minutes of total esterification.
7.5.1.1.2 Remove the concentrator tube and seal
it with a Neoprene or Teflon® stopper. Store at room
temperature in a hood for 20 minutes.
7.5.1.1.3 Destroy any unreacted diazomethane by
adding 0.1 - 0.2 g of silicic acid to the concentrator tube.
Allow to stand until the evolution of nitrogen gas has
stopped. Adjust the sample volume to 10.0 mL with hexane.
Stopper the concentrator tube or transfer 1 mL of sample to
8151A - 16 Revision 1
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a GC vial, and store refrigerated if further processing will
not be performed immediately. Analyze by gas chromatography.
7.5.1.1.4 Extracts should be stored at 4°C away
from light. Preservation study results indicate that most
analytes are stable for 28 days; however, it is recommended
that the methylated extracts be analyzed immediately to
minimize the trans-esterification and other potential
reactions that may occur.
7.5.1.2 Diazald kit method - Instructions for preparing
diazomethane are provided with the generator kit.
7.5.1.2.1 Add 2 mL of diazomethane solution and
let the sample stand for 10 minutes with occasional swirling.
The yellow color of diazomethane should be evident and should
persist for this period.
7.5.1.2.2 Rinse the inside wall of the ampule with
700 /iL of diethyl ether. Reduce the sample volume to
approximately 2 mL to remove excess diazomethane by allowing
the solvent to evaporate spontaneously at room temperature.
Alternatively, 10 mg of silicic acid can be added to destroy
the excess diazomethane.
7.5.1.2.3 Dilute the sample to 10.0 ml with
hexane. Analyze by gas chromatography. It is recommended
that the methylated extracts be analyzed immediately to
minimize the trans-esterification and other potential
reactions that may occur.
7.5.2 PFB derivatization
7.5.2.1 Add 30 ML of 10% K2C03 and 200 /zL of 3% PFBBr in
acetone. Close the tube with a glass stopper and mix on a vortex
mixer. Heat the tube at 60°C for 3 hours.
7.5.2.2 Evaporate the solution to 0.5 mL with a gentle
stream of nitrogen. Add 2 mL of hexane and repeat evaporation just
to dryness at ambient temperature.
7.5.2.3 Redissolve the residue in 2 mL of toluene:hexane
(1:6) for column cleanup.
7.5.2.4 Top a silica column (Bond Elut™ or equivalent) with
0.5 cm of anhydrous sodium sulfate. Prewet the column with 5 mL
hexane and let the solvent drain to the top of the adsorbent.
Quantitatively transfer the reaction residue to the column with
several rinsings of the toluenerhexane solution (total 2-3 mL).
7.5.2.5 Elute the column with sufficient toluene:hexane to
collect 8 mL of eluent. Discard this fraction, which contains excess
reagent.
8151A - 17 Revision 1
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7.5.2.6 Elute the column with toluene:hexane (9:1) to
collect 8 ml of eluent containing PFB derivatives in a 10-mL
volumetric flask. Dilute to 10 mL with hexane. Analyze by GC/ECD.
7.6 Gas chromatographic conditions (recommended)
7.6.1 Narrow-bore columns
Temperature program: 60"C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /nl_, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.2 Wide-bore columns
Temperature program: 0.5 minute at 150°C, 150°C to 270°C,
at 5°C/min
Helium carrier flow: 7 mL/min
Injection volume: 1 jiL
Injector temperature: 250°C
Detector temperature: 320°C
7.7 Calibration
The procedure for internal or external calibration may be used. Refer
to Method 8000 for a description of each of these procedures. Use Table
1 for guidance on selecting the lowest point on the calibration curve.
7.8 Gas chromatographic analysis of samples
7.8.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 juL of internal standard to the sample prior to
injection.
7.8.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.8.3 An example of a chromatogram for a methylated chlorophenoxy
herbicide is shown in Figure 2. Tables 2 and 3 present retention times for
the target analytes after esterification, using the diazomethane
derivatization procedure and the PFB derivatization procedure,
respectively.
7.8.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.8.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
8151A - 18 Revision 1
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7.8.6 If calibration standards have been analyzed in the same manner
as the samples (e.g. have undergone hydrolysis and esterification), then
the calculation of concentration given in Method 8000 should be used.
However, if calibration is performed using standards made from methyl ester
compounds (compounds not esterified by application of this method), then
the calculation of concentration must include a correction for the
molecular weight of the methyl ester versus the acid herbicide.
7.8.7 If peak detection and identification are prevented due to
interferences, further cleanup is required. Before using any cleanup
procedure, the analyst must process a series of standards through the
procedure to validate elution patterns and the absence of interferences
from reagents.
7.9 GC/MS confirmation
7.9.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Refer to Method 8270
for the appropriate GC/MS operating conditions and analysis procedures.
7.9.2 When available, chemical ionization mass spectra may be
employed to aid the qualitative identification process.
7.9.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps may
include the use of alternate GC columns or additional cleanup.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Method 3500. Each laboratory should maintain a formal quality assurance program.
The laboratory should also maintain records to document the quality of the data
generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
8151A - 19 Revision 1
January 1995
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duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time
a set of samples is analyzed or there is a change in reagents, a method
blank should be analyzed as a safeguard against chronic laboratory
contamination. The blanks should be carried through all stages of sample
preparation and measurement.
8.4.2 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 In single laboratory studies using organic-free reagent water and
clay/still bottom samples, the mean recoveries presented in Tables 4 and 5 were
obtained for diazomethane derivatization. The standard deviations of the percent
recoveries of these measurements are also in Tables 4 and 5.
9.2 Table 6 presents relative recoveries of the target analytes obtained
using the PFB derivatization procedure with spiked water samples.
8151A - 20 Revision 1
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10.0 REFERENCES
1. Goerlitz, D.G., Lamar, W.L., "Determination of Phenoxy Acid Herbicides in
Water by Electron Capture and Microcoulometric Gas Chromatography", U.S.
Geol. Survey Water Supply Paper 1967, 1817-C.
2. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis: Some
Practical Aspects", J. Assoc. Off. Anal. Chem. (JAOAC), 1965, 48, 1037.
3. "Extraction and Cleanup Procedures for the Determination of Phenoxy Acid
Herbicides in Sediment", U.S. Environmental Protection Agency, EPA Toxicant
and Analysis Center, Bay St. Louis, MS, 1972.
4. Shore, F.L., Amick, E.N., Pan, S.T., "Single Laboratory Validation of EPA
Method 8151 for the Analysis of Chlorinated Herbicides in Hazardous Waste",
U.S. Environmental Protection Agency, Environmental Monitoring Systems
Laboratory, Office of Research and Development, Las Vegas, NV, 1985;
EPA-60014-85-060.
5. Method 515.1, "Determination of Chlorinated Acids in Water by Gas
Chromatography with an Electron Capture Detector", Revision 4.0, U.S.
Environmental Protection Agency, Office of Research and Development,
Environmental Monitoring Systems Laboratory, Cincinnati, OH.
6. Gurka, D.F, Shore, F.L., Pan, S.T., "Single Laboratory Validation of EPA
Method 8150 for Determination of Chlorinated Herbicides in Hazardous
Waste", JAOAC, 69, 970, 1986.
8151A - 21 Revision 1
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TABLE 1
ESTIMATED METHOD DETECTION LIMITS FOR
DIAZOMETHANE DERIVATIZATION
Compound
Aqueous Samples
GC/ECD
Estimated
Detection
Limit8 (/ig/L)
Soil
GC/ECD
Estimated
Detection
Limit" (/jg/kg)
Samples
GC/MS
Estimated
Identification
Limit0 (ng)
Acifluorfen 0.096
Bentazon 0.2
Chloramben 0.093
2,4-D 0.2
Dalapon 1.3
2,4-DB 0.8
DCPA diacid6 0.02
Dicamba 0.081
3,5-Dichlorobenzoic acid 0.061
Dichloroprop 0.26
Dinoseb 0.19
5-Hydroxydicamba 0.04
MCPP 0.09d
MCPA 0.056d
4-Nitrophenol 0.13
Pentachlorophenol 0.076
Picloram 0.14
2,4,5-T 0.08
2,4,5-TP 0.075
4.0
0.11
0.12
0.38
66
43
0.34
0.16
0.28
1.7
1.25
0.5
0.65
0.43
0.3
0.44
1.3
4.5
a EDL = estimated detection limit; defined as either the MDL, or a
concentration of analyte in a sample yielding a peak in the final extract
with signal-to-noise ratio of approximately 5, whichever value is higher.
b Detection limits determined from standard solutions corrected back to 50-g
samples, extracted and concentrated to 10 mL, with 5 jj.1 injected.
Chromatography using narrow-bore
5% phenyl/95% methyl silicone.
capillary column, 0.25 jum film,
The minimum amount of analyte to give a Finnigan INCOS FIT value of 800 as
the methyl derivative vs. the spectrum obtained from 50 ng of the respective
free acid herbicide.
40 CFR Part 136, Appendix B (49 FR 43234).
capillary column.
Chromatography using wide-bore
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151A - 22
Revision 1
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TABLE 2
RETENTION TIMES (MINUTES) OF METHYL DERIVATIVES OF CHLORINATED HERBICIDES
Narrow Bore Columns
Hide-bore Columns
Analyte
Dalapon
3,5-Dichlorobenzoic acid
4-Nitrophenol
DCAA (surrogate)
Dicamba
Dichloroprop
2,4-D
DBOB (internal std.)
Pentachlorophenol
Chloramben
2,4,5-TP
5-Hydroxydicamba
2,4,5-T
2,4-DB
Dinoseb
Bentazon
Picloram
DCPA diacidc
Acifluorfen
MCPP
MCPA
Primary8 Confirmation8
Column Column
3.4
18.6
18.6
22.0
22.1
25.0
25.5
27.5
28.3
29.7
29.7
30.0
30.5
32.2
32.4
33.3
34.4
35.8
41.5
-
_
4.7
17.7
20.5
14.9
22.6
25.6
27.0
27.6
27.0
32.8
29.5
30.7
30.9
32.2
34.1
34.6
37.5
37.8
42.8
-
"
Primary13
Column
.
-
-
-
4.39
5.15
5.85
-
-
-
6.97
-
7.92
8.74
-
-
_
-
-
4.24
4.74
Confirmation15
Column
_
-
-
-
4.39
5.46
6.05
-
-
*
7.37
-
8.20
9.02
-
-
.
-
-
4.55
4.94
Primary Column:
Confirmation Column:
Temperature program:
Helium carrier flow:
Injection volume:
Injector temperature:
Detector temperature:
Primary Column:
Confirmatory Column:
Temperature program:
Helium carrier flow:
Injection volume:
5% phenyl/95% methyl silicone
14% cyanopropyl phenyl silicone
60°C to 300°C, at 4°C/min
30 cm/sec
2 juL, splitless, 45 sec delay
250°C
320°C
DB-608
14% cyanopropyl phenyl silicone
0.5 minute at 150°C,
150°C to 270°C, at 5°C/min
7 mL/min
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151A - 23
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TABLE 3
RETENTION TIMES (MINUTES) OF PFB DERIVATIVES OF CHLORINATED HERBICIDES
Gas Chromatoqraphic Column
Compound
Thin-film DB-5a
SP-2250b
Thick-film DB-5C
Dalapon
MCPP
Dicamba
MCPA
Dichloroprop
2,4-D
Silvex
2,4,5-T
Dinoseb
2,4-DB
10.41
18.22
18.73
18.88
19.10
19.84
21.00
22.03
22.11
23.85
12.94
22.30
23.57
23.95
24.10
26.33
27.90
31.45
28.93
35.61
13.54
22.98
23.94
24.18
24.70
26.20
29.02
31.36
31.57
35.97
a DB-5 capillary column, 0.25 /urn film thickness, 0.25 mm ID x 30 m long. Column
programmed: 70°C for 1 minute, program 10°C/min. to 240°C, hold for 17
minutes.
b SP-2550 capillary column, 0.25 /xm film thickness, 0.25 mm ID x 30 m long.
Column programmed: 70°C for 1 minute, program 10°C/min. to 240°C, hold for
10 minutes.
0 DB-5 capillary column, 1.0 jum film thickness, 0.32 mm ID x 30 m long. Column
programmed: 70°C for 1 minute, program 10°C/min. to 240°C, hold for
10 minutes.
8151A - 24
Revision 1
January 1995
-------
TABLE 4
ACCURACY AND PRECISION FOR DIAZOMETHANE DERIVATIZATION
ORGANIC-FREE REAGENT WATER MATRIX
Compound
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacidb
Dicamba
3,5-Dichlorobenzoic acid
Dichloroprop
Dinoseb
5-Hydroxydicamba
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-TP
2,4,5-T
Spike
Concentration
(M9/L)
0.2
1
0.4
1
10
4
0.2
0.4
0.6
2
0.4
0.2
1
0.04
0.6
0.4
0.2
Mean3 Standard
Percent Deviation of
Recovery Percent Recovery
121
120
111
131
100
87
74
135
102
107
42
103
131
130
91
117
134
15.7
16.8
14.4
27.5
20.0
13.1
9.7
32.4
16.3
20.3
14.3
16.5
23.6
31.2
15.5
16.4
30.8
Mean percent recovery calculated from 7-8 determinations of spiked organic-
free reagent water.
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151A - 25
Revision 1
January 1995
-------
TABLE 5
ACCURACY AND PRECISION FOR DIAZOMETHANE DERIVATIZATION
CLAY MATRIX
Linear Percent
Concentration Relative
Mean Range6 Standard Deviation0
Compound Percent Recovery8 (ng/g) (n=20)
Dicamba
MCPP
MCPA
Dichloroprop
2,4-D
2,4,5-TP
2,4,5-T
2,4-DB
Dinoseb
95.7
98.3
96.9
97.3
84.3
94.5
83.1
90.7
93.7
0.52
620
620
1.5
1.2
0.42
0.42
4.0
0.82
104
- 61,800
- 61,200
- 3,000
- 2,440
828
828
- 8,060
- 1,620
7.5
3.4
5.3
5.0
5.3
5.7
7.3
7.6
8.7
a Mean percent recovery calculated from 10 determinations of spiked clay and
clay/still bottom samples over the linear concentration range.
b Linear concentration range was determined using standard solutions and
corrected to 50 g solid samples.
c Percent relative standard deviation was calculated using standard solutions,
10 samples high in the linear concentration range, and 10 samples low in the
range.
8151A - 26 Revision 1
January 1995
-------
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FIGURE 1
DIAZOMETHANE GENERATOR
gloss tubing
nitrogen
rubber stepper
-\ F
tub* 1
tube 2
8151A - 28
Revision 1
January 1995
-------
FIGURE 2
CHROMATOGRAM OF METHYL ESTERS OF CHLOROPHENOXYACIDS
lOOO-i
J
1O46
RIC-
C
•13 E
•66
A
271
\J
393
443
ZOO
3:20
317
479 S43
•33 693
4OO
• :4O
•00
10:00
G
•64
H
I
•00
13.20
1OOO
16:40
A - Dalapon. methyl ester
B = Dicamba, methyl ••t«r
C MCPP. methyl estw
D : MCPA. methyl ester
E - Oichlorprop. methyl ester
f -• 2.4. D methyl ester
G Silvek. nieihyl ester
H 2.4.b T. methyl ester
I 2.4 OB. methyl ester
J Dinoseb, methyl ether
12OO
20:00
Scan Time
8151A - 29
Revision 1
January 1995
-------
METHOD 8151A
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATION
DERIVATIZATION: CAPILLARY COLUMN TECHNIQUE
Extraction/Hydrolysis of Waste and Soil Samples
NO
1
1
Concentrate and/or
dilute based on
whether denvaua&on
s by diazomethane
or PFB
7000*8
sample con
am a high
cone of
waste?
7211 Wwgn sample
and add to backer.
add aod and spike.
mix we*.
72181 Add KOH and
water Reflux for 2 hrs
Allow to cool.
72182 Transtor the
hydrolyzed solution ID a
sep funnel and extract 3
Wneewrth MeCI.
Discard •xracts
72.1 8.3 Aodrfyand
•xtract 3 DmM wnh
diMhylctw Cornbm*
and dry lh* «xtracti 2 hrs
7 2.1 8 4 ProoMd to
Sactxxi 7 4 to oono»rrtr»*»
extract.
72.1 91 Extract 3 tmM
with KOH Dwcardtrw
72.1 92 Acxtttyand
extract 3 nrnce wnh
dwtnylctwr Combme
and dry tfw extracS 2 hn.
72.1 2 Opdrmz*
urtraiontc aoHd extrac
oon tar Men matnx
72.1.3
aoMon* to sampt* &
extract 3 mn«
A decant extract
72.1 445 Ultra-
soncaVy •xtract samptoj
2 rnore am** with MeCI
72.1.5 Comttn* orgaracj
extract, centnfuga. and
W«»r extract Dry lor
2 hrs
7216 Cooceorate
extract to about 5 mL
wrth Snyd*r column
YES
If hydrolysis a not
required, proceed to Section
744 NrtogenSlowdown.
8151A - 30
Revision 1
January 1995
-------
METHOD 8151A
(continued)
Extraction/Hydrolysis of Aqueous Samples and Extract Concentration
7.3.1 Measure 1L of
sample and transfer to
a 2 L sep. funnel.
I
7.3.1 Add 250 g NaCI
to sample and shake
to dissolve.
A
/ 7.3 3\
/ D°" \
Mo
\^ herbicide /
\esters? /
7.2.2 1 Add 12N HCI
and shake. Add until
pH <2.
i
7.2.2.2 Add diethyl
ether to sample and
extract. Save both
phases.
A
/ Does \
/ difficult \Ves ^
Nv form? /
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centrifugation, or
other physical methods.) Save
both phases.
No]
Yes
7.3.3 1 Add 6N NaOH
to sample and shake
Add until pH > 1 2. Let
stand 1 hour
7.2.2 3 • 7 2.2.4 Return aqueous
phase to separatory funnel and
repeat extraction 2 more times.
combine extracts, and allow
extract to remain in contact with
sodium sulfate for 2 hrs.
Pour extract
through glass wool
and proceed to
Section 7.4.1.
7.3.3.2 Add MeCI
and extract by
shaking for 2 mm.
Discard MeCI
7.4.1 Place K-0
apparatus in water
bath, concentrate
and cool.
nulsi
> 1/3
Dlu
No
oes\^
ficult\ Yes
on from\___w
solvent /
urne' /
L.
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centrifugation. or
other physical methods I
Discard MeCI.
I
7.4.2 - 7.4.4
Complete con-
centration with micro-
Snyder column or
nitrogen blowdown.
7.3.1 3 3 Repeat
extraction twice
more Discard MeCI
7.4.6 Dilute extract
with 1 ml isoctane
and 0.5 ml methanol.
8151A - 31
Revision 1
January 1995
-------
METHOD 8151A
(continued)
Extract Derivatization
7.4.5 Dilute extract
to 4 ml with acetone.
7.5.2 1 Add
potassium carbonate
and PFBBr. Close
tube, mix & heat.
7.5.2.2 Evaporate
with nitrogen to
0.5 mL. Add 2 ml
hexane »nd repeat.
7.5.2.3 Redissolve
the residue in 2 ml
toluene: hexane (1:6).
7.5.2.4 Load sodium
sulfate/silica cleanup
column with residue.
7 5.2.5 Elute column
with enough toluene:
hexane to collect
8 mL eluant.
7.5.2.6 Discard 1st
fraction and continue
elution with enough
toluene:hexane (9:11
to collect 8 mL more eluant
Transfer to a 10 mL
volumetric flask and dilute
to the mark with hexana.
L
7.4.5 Dilute extract
to 4 mL with diethyl
ether.
7.5 1.1 Assemble the
diazomethane bubbler
(Figure 11.
7.5.1.1 1 Add 5 mL to 1st
test tube. Add 1 mL diethyl
ether. 1 mL carbitol, 1.5 mL
of 37% KOH and 0.1-0.2 g
Diazald to the 2nd tube.
Bubble with nitrogen for
10 min. or until yellow persists
7.5.1.1.2 Remove
concentrator tube
and seal it. Store at
room temp.
7.5 1.1.3 Add silicic acid
to concentrator tuba and let
stand until nitrogen evolution
has stopped. Adjust sample
volume to 10 mL with hexane.
Stopper Immediate analysis
is recommended.
7.5.1.1.5 If
necessary store
at 4 C in the dark
for a max of 26 days
7.6.1 81 7.6.2 Set
GC conditions
8151A - 32
7.5.1.2.1 Add 2 mL
diazomethane solution.
Let stand for 10 min.
and twirl.
7.5.1.2.2 Rinse ampule
with diethyl ether and
evaporate to 2 mL to
remove diazomethane.
Alternatively, silicic acid
may be added.
7.5.1.2.3 Dilute
sample to 10 mL
with hexane.
Revision 1
January 1995
-------
METHOD 8151A
(continued)
Analysis by Gas Chromatography
7.7 Internal or
external calibra-
tion may be used
(See method 8000.1
7.B.1 Add 10 uL
internal standard
to the sample prior
to injection.
7.8.2 See Method 8000 for
analysis sequence, appropriate
dilutions, establishing daily
retention time windows, and
identification criteria Check
stds. every 10 samples.
7.8 4 Record volume
injected and the
resulting peak sizes.
7 8.5 Determine the
identity and quantity
component peaks
Calculate the correction
for molecular weight
of methyl ether vs.
herbicide.
7 8.6 Calculate con-
centration using
procedure in
Method 8000
7.8.7 Perform further
cleanup if necessary.
8151A - 33
Revision 1
January 1995
-------
4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.2 GAS CHROMATOGRAPHIC/MASS SPECTROMETRIC METHODS
The following methods are included in this section:
Method 8260B:
Volatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS):
Capillary Column Technique
Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS):
Capillary Column Technique
The Analysis of Polychlorinated Dibenzo-p-Dioxins
and Polychlorinated Dibenzofurans by High
Resolution Gas Chromatography/Low Resolution Mass
Spectrometry (HRGC/LRMS)
Signal-to-Noise Determination Methods
Recommended Safety and Handling Procedures
for PCDDs/PCDFs
Polychlorinated Dibenzodioxins (PCDDs) and
Polychlorinated Dibenzofurans (PCDFs) by High-
Resolution Gas Chromatography/High-Resolution
Mass Spectrometry (HRGC/HRMS)
Attachment A: Procedures for the Collection, Handling,
Analysis, and Reporting of Wipe Tests
Performed within the Laboratory
Method 8270C:
Method 8280A:
Appendix A:
Appendix B:
Method 8290:
FOUR - 11
Revision 3
January 1995
-------
METHOD 8260B
VOLATILE ORGANIC COMPOUNDS BY GAS CHROHATOGRAPHY/
MASS SPECTROMETRY (GC/MS): CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8260 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including various air sampling trapping
media, ground and surface water, aqueous sludges, caustic liquors, acid liquors,
waste solvents, oily wastes, mousses, tars, fibrous wastes, polymeric emulsions,
filter cakes, spent carbons, spent catalysts, soils, and sediments. The
following compounds can be determined by this method:
Compound
CAS No.'
Appropriate Technique3
5030/ Direct
5035 5031 5032 5021 5041 Inject.
Acetone
Acetonitrile
Acrolein (Propenal)
Acrylonitrile
Allyl alcohol
Allyl chloride
Benzene
Benzyl chloride
Bromoacetone
Bromochloromethane
Bromodichloromethane
4-Bromofluorobenzene (surr)
Bromoform
Bromomethane
n-Butanol
2-Butanone (MEK)
t-Butyl alcohol
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chlorobenzene
Chlorobenzene-d5 (IS)
Chlorodibromomethane
Chloroethane
2-Chloroethanol
Bis-(2-chloroethyl)sulfide
2-Chloroethyl vinyl ether
Chloroform
Chloromethane
Chloroprene
3-Chloropropionitrile
67-64-1
75-05-8
107-02-8
107-13-1
107-18-6
107-05-1
71-43-2
100-44-7
598-31-2
74-97-5
75-27-4
75-25-2
74-83-9
71-36-3
78-93-3
75-65-0
75-15-0
56-23-5
302-17-0
108-90-7
124-48-1
75-00-3
107-07-3
505-60-2
110-75-8
67-66-3
74-87-3
126-99-8
542-76-7
PP
PP
PP
PP
ht
c
c
c
PP
c
c
c
c
c
ht
PP
PP
PP
c
PP
c
c
c
c
PP
PP
c
c
c
c
i
c
c
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
nd
c
c
nd
nd
c
nd
nd
c
c
c
c
c
nd
c
nd
c
c
nd
c
c
c
c
nd
nd
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
c
nd
nd
c
c
c
c
c
nd
nd
nd
nd
c
nd
c
c
nd
c
nd
nd
nd
c
c
nd
nd
c
nd
nd
c
nd
nd
c
nd
nd
c
c
c
c
c
nd
nd
nd
c
c
nd
c
c
c
c
nd
nd
nd
c
c
nd
nd
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
pc
8260B - 1
Revision 2
January 1995
-------
Appropriate Technique3
Compound
Crotonaldehyde
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Dibromomethane
1 ,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (IS)
cis-l,4-Dichloro-2-butene
trans -l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surr)
1, 1-Dichloroethene
trans-l,2-Dichloroethene
1,2-Dichloropropane
l,3-Dichloro-2-propanol
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1,2,3,4-Diepoxybutane
Diethyl ether
1,4-Difluorobenzene (IS)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethylene oxide
Ethyl methacrylate
Fluorobenzene (IS)
Hexachlorobutadiene
Hexachloroethane
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Isopropylbenzene
Malononitri le
Methacrylonitrile
Methanol
Methylene chloride
Methyl methacrylate
4-Methyl-2-pentanone (MIBK)
Naphthalene
Nitrobenzene
2-Nitropropane
CAS No.b
123-73-9
96-12-8
106-93-4
74-95-3
95-50-1
541-73-1
106-46-7
1476-11-5
110-57-6
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
96-23-1
10061-01-5
10061-02-6
1464-53-5
60-29-7
123-91-1
106-89-8
64-17-5
141-78-6
100-41-4
75-21-8
97-63-2
87-68-3
67-72-1
591-78-6
78-97-7
74-88-4
78-83-1
98-82-8
109-77-3
126-98-7
67-56-1
75-09-2
80-62-6
108-10-1
91-20-3
98-95-3
79-46-9
5030/
5035
PP
PP
c
c
c
c
c
c
c
PP
c
c
c
c
c
c
c
PP
c
c
c
c
nd
PP
i
i
i
c
PP
c
c
c
i
PP
i
c
PP
c
PP
PP
i
c
c
PP
c
c
c
5031
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
nd
c
c
nd
c
nd
nd
nd
nd
nd
nd
nd
c
nd
nd
i
c
nd
nd
c
nd
nd
nd
5032
nd
nd
nd
c
nd
nd
nd
nd
c
c
c
c
c
c
c
c
c
nd
c
c
nd
nd
nd
c
nd
c
nd
c
nd
c
nd
nd
nd
c
nd
c
nd
nd
nd
nd
nd
c
nd
c
nd
nd
nd
5021
nd
c
c
c
c
c
c
c
nd
nd
c
c
c
c
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
nd
nd
nd
c
nd
nd
nd
nd
nd
c
nd
nd
nd
c
nd
nd
c
nd
nd
5041
nd
nd
nd
c
nd
nd
nd
nd
nd
nd
nd
c
c
c
c
c
c
nd
c
c
nd
nd
c
nd
nd
nd
nd
c
nd
nd
nd
nd
nd
nd
nd
c
nd
nd
nd
nd
nd
c
nd
nd
nd
nd
nd
Direct
Inject.
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
nd
c
c
c
c
c
c
c
nd
c
c
c
pc
c
c
c
c
c
c
c
c
c
c
c
c
8260B - 2
Revision 2
January 1995
-------
Compound
CAS No.
Appropriate Technique3
5030/ Direct
5035 5031 5032 5021 5041 Inject.
N-Nitroso-di -n -butyl amine
Paraldehyde
Pentachloroethane
2-Pentanone
2-Picoline
1-Propanol
2-Propanol
Propargyl alcohol
/3-Propiolactone
Propionitrile (ethyl cyanide)
n-Propyl amine
Pyridine
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
Toluene-d8 (surr)
o-Toluidine
1 , 2 , 4-Tri chl orobenzene
1 , 1 , 1-Tri chl oroethane
1, 1, 2 -Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
Vinyl acetate
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
924-16-3
123-63-7
76-01-7
107-87-9
109-06-8
71-23-8
67-63-0
107-19-7
57-57-8
107-12-0
107-10-8
110-86-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
95-53-4
120-82-1
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
108-05-4
75-01-4
95-47-6
108-38-3
106-42-3
PP
PP
i
PP
PP
PP
PP
PP
PP
ht
c
i
c
c
c
c
c
c
PP
c
c
c
c
c
c
c
c
c
c
c
c
c
nd
c
c
c
c
i
nd
c
nd
c
nd
nd
nd
nd
nd
nd
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
nd
c
c
c
c
nd
nd
c
c
c
c
c
c
c
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
c
c
c
c
c
nd
c
c
c
c
c
c
nd
c
c
c
c
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
nd
c
c
c
c
c
c
nd
nd
c
c
c
c
c
nd
c
c
c
c
c
c
c
c
c
c
c
c
c
pc
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
c
a See Sec. 1.2 for other appropriate sample preparation techniques
b Chemical Abstract Services Registry Number
c = Adequate response by this technique
ht = Method analyte only when purged at 80°C
nd = Not determined
i = Inappropriate technique for this analyte
pc = Poor chromatographic behavior
pp = Poor purging efficiency resulting in high Estimated Quantitation Limits
surr = Surrogate
IS = Internal Standard
8260B - 3
Revision 2
January 1995
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1.2 There are various techniques by which these compounds may be
introduced into the GC/MS system. The more common techniques are listed in the
table above. Purge-and-trap, by Methods 5030 (aqueous samples) and 5035 (solid
samples and waste oil samples), is the most commonly used technique for volatile
organic analytes. However, other techniques are also acceptable. These include
direct injection following dilution with hexadecane (Method 3585) for waste oil
samples; automated static headspace by Method 5021 for solid samples; direct
injection of an aqueous sample (concentration permitting) or injection of a
sample concentrated by azeotropic distillation (Method 5031); and closed system
vacuum distillation (Method 5032) for aqueous, solid and tissue samples. For air
samples, Method 5041 provides methodology for desorbing volatile organics from
trapping media (Methods 0010, 0030, and 0031). In addition, direct analysis
utilizing a sample loop is used for sub-sampling from Tedlar® bags (Method 0040).
Method 5000 provides more general information on the selection of the appropriate
introduction method.
1.3 Method 8260 can be used to quantitate most volatile organic compounds
that have boiling points below 200°C. Volatile, water soluble compounds can be
included in this analytical technique by the use of azeotropic distillation or
closed system vacuum distillation. Such compounds include low molecular weight
halogenated hydrocarbons, aromatics, ketones, nitriles, acetates, acrylates,
ethers, and sulfides. See Tables 1 and 2 for analytes and retention times that
have been evaluated on a purge and trap GC/MS system. Also, the method detection
limits for 25-mL sample volumes are presented. The following compounds are also
amenable to analysis by Method 8260:
Bromobenzene 1,3-Dichloropropane
n-Butyl benzene 2,2-Dichloropropane
sec-Butyl benzene 1,1-Dichloropropene
tert-Butylbenzene p-Isopropyltoluene
Chloroacetonitrile Methyl acrylate
1-Chlorobutane Methyl-t-butyl ether
1-Chlorohexane Pentafluorobenzene
2-Chlorotoluene n-Propylbenzene
4-Chlorotoluene 1,2,3-Trichlorobenzene
Dibromofluoromethane 1,2,4-Trimethyl benzene
cis-l,2-Dichloroethene 1,3,5-Trimethyl benzene
1.4 The estimated quantitation limit (EQL) of Method 8260 for an
individual compound is somewhat instrument dependent and also dependent on the
choice of sample preparation/introduction method. Using standard quadrapole
instrumentation and the purge-and-trap technique, limits should be approximately
5 M9Ag (wet weight) for soil/sediment samples, 0.5 mg/kg (wet weight) for
wastes, and 5 fj,g/l for ground water (see Table 3). Somewhat lower limits may
be achieved using an ion trap mass spectrometer or other instrumentation of
improved design. No matter which instrument is used, EQLs will be
proportionately higher for sample extracts and samples that require dilution or
when a reduced sample size is used to avoid saturation of the detector.
1.5 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatograph/mass spectrometers, and
skilled in the interpretation of mass spectra and their use as a quantitative
tool.
8260B - 4 Revision 2
January 1995
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2.0 SUMMARY OF METHOD
2.1 The volatile compounds are introduced into the gas chromatograph by
the purge-and-trap method or by other methods (see Sec. 1.2). The analytes are
introduced directly to a large-bore capillary column or cryofocussed on a
capillary pre-column before being flash evaporated to a narrow-bore capillary for
analysis. The column is temperature-programmed to separate the analytes, which
are then detected with a mass spectrometer (MS) interfaced to the gas
chromatograph (GC).
2.2 Analytes eluted from the capillary column are introduced into the mass
spectrometer via a jet separator or a direct connection. (Wide-bore capillary
columns normally require a jet separator, whereas narrow-bore capillary columns
may be directly interfaced to the ion source). Identification of target analytes
is accomplished by comparing their mass spectra with the electron impact (or
electron impact-like) spectra of authentic standards. Quantitation is
accomplished by comparing the response of a major (quantitation) ion relative to
an internal standard with a five-point calibration curve.
2.3 The method includes specific calibration and quality control steps
that supersede the general requirements provided in Method 8000.
3.0 INTERFERENCES
3.1 Major contaminant sources are volatile materials in the laboratory and
impurities in the inert purging gas and in the sorbent trap. The use of
non-polytetrafluoroethylene (PTFE) thread sealants, plastic tubing, or flow
controllers with rubber components should be avoided, since such materials
out-gas organic compounds which will be concentrated in the trap during the purge
operation. Analyses of calibration and reagent blanks provide information about
the presence of contaminants. When potential interfering peaks are noted in
blanks, the analyst should change the purge gas source and regenerate the
molecular sieve purge gas filter. Subtracting blank values from sample results
is not permitted. If reporting values without correcting for the blank results
in what the laboratory feels is a false positive result for a sample, the
laboratory should fully explained this in text accompanying the uncorrected
data.
3.2 Contamination may occur when a sample containing low concentrations
of volatile organic compounds is analyzed immediately after a sample containing
high concentrations of volatile organic compounds. A technique to prevent this
problem is to rinse the purging apparatus and sample syringes with two portions
of organic-free reagent water between samples. After the analysis of a sample
containing high concentrations of volatile organic compounds, one or more blanks
should be analyzed to check for cross-contamination.
3.3 For samples containing large amounts of water-soluble materials,
suspended solids, high boiling compounds, or high concentrations of compounds
being determined, it may be necessary to wash the purging device with a soap
solution, rinse it with organic-free reagent water, and then dry the purging
device in an oven at 105°C. In extreme situations, the entire purge-and-trap
device may require dismantling and cleaning. Screening of the samples prior to
purge-and-trap GC/MS analysis is highly recommended to prevent contamination of
8260B - 5 Revision 2
January 1995
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the system. This is especially true for soil and waste samples. Screening may
be accomplished with an automated headspace technique (Method 5021) or by Method
3820 (Hexadecane Extraction and Screening of Purgeable Organics).
3.4 Many analytes exhibit low purging efficiencies from a 25-mL sample.
This often results in significant amounts of these analytes remaining in the
sample purge vessel after analysis. After removal of the sample aliquot that was
purged, and rinsing the purge vessel three times with organic-free water, the
empty vessel should be subjected to a heated purge cycle prior to the analysis
of another sample in the same purge vessel. This will reduce sample-to-sample
carryover.
3.5 Special precautions must be taken to analyze for methylene chloride.
The analytical and sample storage area should be isolated from all atmospheric
sources of methylene chloride. Otherwise, random background levels will result.
Since methylene chloride will permeate through PTFE tubing, all gas
chromatography carrier gas lines and purge gas plumbing should be constructed
from stainless steel or copper tubing. Laboratory clothing worn by the analyst
should be clean, since clothing previously exposed to methylene chloride fumes
during liquid/liquid extraction procedures can contribute to sample
contamination.
3.6 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal of
the sample container into the sample during shipment and storage. A trip blank
prepared from organic-free reagent water and carried through the sampling,
handling, and storage protocols can serve as a check on such contamination.
3.7 Use of sensitive mass spectrometers to achieve lower detection level
will increase the potential to detect laboratory contaminants as interferences.
3.8 Direct injection - Some contamination may be eliminated by baking out
the column between analyses. Changing the injector liner will reduce the
potential for cross-contamination. A portion of the analytical column may need
to be removed in the case of extreme contamination. The use of direct injection
will result in the need for more frequent instrument maintenance.
3.9 If hexadecane is added to waste samples or petroleum samples that are
analyzed, some chromatographic peaks will elute after the target analytes. The
oven temperature program must include a post-analysis bake out period to ensure
that semivolatile hydrocarbons are volatilized.
4.0 APPARATUS AND MATERIALS
4.1 Purge-and-trap device for aqueous samples - Described in Method 5030.
4.2 Purge-and-trap device for solid samples - Described in Method 5035.
4.3 Automated static headspace device for solid samples - Described in
Method 5021.
4.4 Azeotropic distillation apparatus for aqueous and solid samples -
Described in Method 5031.
8260B - 6 Revision 2
January 1995
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4.5 Vacuum distillation apparatus for aqueous, solid and tissue samples -
Described in Method 5032.
4.6 Desorption device for air trapping media for air samples - Described
in Method 5041.
4.7 Air sampling loop for sampling from Tedlar® bags for air samples -
Described in Method 0040.
4.8 Injection port liners (HP Catalog #18740-80200, or equivalent) -
modified for direct injection analysis by placing a 1-cm plug of Pyrex® wool
approximately 50-60 mm down the length of the injection port towards the oven
(see illustration below). A 0.53 mm ID column is mounted 1 cm into the liner
from the oven side of the injection port, according to manufacturer's
specifications.
Sesptxim SO — GO Oven
4.9 Gas chromatography/mass spectrometer/data system
4.9.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless injection
with appropriate interface for sample introduction device. The system
includes all required accessories, including syringes, analytical columns,
and gases.
4.9.1.1 The GC should be equipped with variable constant
differential flow controllers so that the column flow rate will remain
constant throughout desorption and temperature program operation.
4.9.1.2 For some column configurations, the column oven must
be cooled to less than 30°C, therefore, a subambient oven controller
may be necessary.
4.9.1.3 The capillary column is either directly coupled to
the source or interfaced through a jet separator, depending on the
size of the capillary and the requirements of the GC/MS system.
4.9.1.4 Capillary pre-column interface - This device is the
interface between the sample introduction device and the capillary gas
chromatograph, and is, necessary when using cryogenic cooling. The
interface condenses the desorbed sample components and focuses them
into a narrow band on an uncoated fused-silica capillary pre-column.
When the interface is flash heated, the sample is transferred to the
analytical capillary column.
4.9.1.5 During the cryofocussing step, the temperature of
the fused-silica in the interface is maintained at -150°C under a
stream of liquid nitrogen. After the desorption period, the interface
must be capable of rapid heating to 250°C in 15 seconds or less to
complete the transfer of analytes.
8260B - 7 Revision 2
January 1995
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4.9.2 Gas chromatographic columns
4.9.2.1 Column 1 - 60 m x 0.75 mm ID capillary column coated
with VOCOL (Supelco), 1.5 /xm film thickness, or equivalent.
4.9.2.2 Column 2 - 30 - 75 m x 0.53 mm ID capillary column
coated with DB-624 (J&W Scientific), Rtx-502.2 (RESTEK), or VOCOL
(Supelco), 3 /urn film thickness, or equivalent.
4.9.2.3 Column 3 - 30 m x 0.25 - 0.32 mm ID capillary column
coated with 95% dimethyl - 5% diphenyl polysiloxane (DB-5, Rtx-5,
SPB-5, or equivalent), 1 ^m film thickness.
4.9.2.4 Column 4 - 60 m x 0.32 mm ID capillary column coated
with DB-624 (J&W Scientific), 1.8 urn film thickness, or equivalent.
4.9.3 Mass spectrometer - Capable of scanning from 35 to 300 amu
every 2 sec or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable of
producing a mass spectrum for 4-Bromofluorobenzene (BFB) which meets all
of the criteria in Table 4 when 5-50 ng of the GC/MS tuning standard (BFB)
are injected through the GC. To ensure sufficient precision of mass
spectral data, the desirable MS scan rate allows acquisition of at least
five spectra while a sample component elutes from the GC.
4.9.3.1 An ion trap mass spectrometer may be used if it is
capable of axial modulation to reduce ion-molecule reactions and can
produce electron impact-like spectra that match those in the EPA/NIST
Library. Because ion-molecule reactions with water and methanol in
an ion trap mass spectrometer may produce interferences that coelute
with chloromethane and chloroethane, the base peak for both of these
analytes will be at m/z 49. This ion should be used as the
quantitation ion in this case. The mass spectrometer must be capable
of producing a mass spectrum for BFB which meets all of the criteria
in Table 3 when 5 or 50 ng are introduced.
4.9.4 GC/MS interface - Two alternatives may be used to interface
the GC to the mass spectrometer.
4.9.4.1 Direct coupling, by inserting the column into the
mass spectrometer, is generally used for 0.25 - 0.32 mm ID columns.
4.9.4.2 A jet separator, including an all-glass transfer
line and glass enrichment device or split interface, is used with a
0.53 mm column.
4.9.4.3 Any enrichment device or transfer line may be used,
if all of the performance specifications described in Sec. 8.0
(including acceptable calibration at 50 ng or less) can be achieved.
GC/MS interfaces constructed entirely of glass or of glass-lined
materials are recommended. Glass may be deactivated by silanizing
with dichlorodimethylsilane.
8260B - 8 Revision 2
January 1995
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4.9.5 Data system - A computer system that allows the continuous
acquisition and storage on machine-readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows searching any GC/MS data file for ions of a specified mass and
plotting such ion abundances versus time or scan number. This type of plot
is defined as an Extracted Ion Current Profile (EICP). Software must also
be available that allows integrating the abundances in any EICP between
specified time or scan-number limits. The most recent version of the
EPA/NIST Mass Spectral Library should also be available.
4.10 Microsyringes - 10-, 25-, 100-, 250-, 500-, and l,000-/xL.
4.11 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.12 Syringes - 5-, 10-, or 25-mL, gas-tight with shutoff valve.
4.13 Balance - Analytical, capable of weighing 0.0001 g, and top-loading,
capable of weighing 0.1 g.
4.14 Glass scintillation vials - 20-mL, with Teflon®-lined screw-caps or
glass culture tubes with Teflon®-lined screw-caps.
4.15 Vials - 2-mL, for GC autosampler.
4.16 Disposable pipets - Pasteur.
4.17 Volumetric flasks, Class A - 10-mL and 100-mL, with ground-glass
stoppers.
4.18 Spatula - Stainless steel.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to be
free of analytes. Store apart from other solvents.
5.4 Reagent Hexadecane - Reagent hexadecane is defined as hexadecane in
which interference is not observed at the method detection limit of compounds of
interest. Hexadecane quality is demonstrated through the analysis of a solvent
blank injected directly into the GC/MS. The results of such a blank analysis
8260B - 9 Revision 2
January 1995
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must demonstrate that all interfering volatiles have been removed from the
hexadecane.
5.5 Polyethylene glycol, H(OCH2CH2)nOH - Free of interferences at the
detection limit of the target analytes.
5.6 Hydrochloric acid (1:1 v/v), HC1 - Carefully add a measured volume of
concentrated HC1 to an equal volume of organic-free reagent water.
5.7 Stock solutions - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standard solutions
in methanol, using assayed liquids or gases, as appropriate.
5.7.1 Place about 9.8 ml of methanol in a 10-mL tared
ground-glass-stoppered volumetric flask. Allow the flask to stand,
unstoppered, for about 10 minutes or until all alcohol-wetted surfaces have
dried. Weigh the flask to the nearest 0.0001 g.
5.7.2 Add the assayed reference material, as described below.
5.7.2.1 Liquids - Using a 100-jiiL syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.7.2.2 Gases - To prepare standards for any compounds that
boil below 30°C (e.g. bromomethane, chloroethane, chloromethane, or
vinyl chloride), fill a 5-mL valved gas-tight syringe with the
reference standard to the 5.0 ml mark. Lower the needle to 5 mm above
the methanol meniscus. Slowly introduce the reference standard above
the surface of the liquid. The heavy gas will rapidly dissolve in the
methanol. Standards may also be prepared by using a lecture bottle
equipped with a septum. Attach Teflon® tubing to the side arm relief
valve and direct a gentle stream of gas into the methanol meniscus.
5.7.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially-prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.7.4 Transfer the stock standard solution into a bottle with a
Teflon®-!ined screw-cap. Store, with minimal headspace, at -10°C or less
and protect from light.
5.7.5 Prepare fresh standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months, or
sooner, if comparison with check standards indicates a problem. Both gas
and liquid standards must be monitored closely by comparison to the initial
8260B - 10 Revision 2
January 1995
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calibration curve and by comparison to QC check standards. It may be
necessary to replace the standards more frequently if either check exceeds
a 20% drift.
5.7.6 Preparation of Calibration Standards From a Gas Mixture
An optional calibration procedure involves using a certified gaseous
mixture daily, utilizing a commercially-available gaseous analyte mixture
of bromomethane, chloromethane, chloroethane, vinyl chloride, dichloro-
difluoromethane and trichlorofluoromethane in nitrogen. Mixtures of
documented quality are stable for as long as six months without
refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.7.6.1 Before removing the cylinder shipping cap, be sure
the valve is completely closed (turn clockwise). The contents are
under pressure and should be used in a well-ventilated area.
5.7.6.2 Wrap the pipe thread end of the Luer fitting with
PTFE tape. Remove the shipping cap from the cylinder and replace it
with the Luer fitting.
5.7.6.3 Transfer half the working standard containing other
analytes, internal standards, and surrogates to the purge apparatus.
5.7.6.4 Purge the Luer fitting and stem on the gas cylinder
prior to sample removal using the following sequence:
a) Connect either the 100-juL or 500-/xL Luer syringe
to the inlet fitting of the cylinder.
b) Make sure the on/off valve on the syringe is in
the open position.
c) Slowly open the valve on the cylinder and
withdraw a full syringe volume.
d) Be sure to close the valve on the cylinder
before you withdraw the syringe from the Luer
fitting.
e) Expel the gas from the syringe into a
well-ventilated area.
f) Repeat steps a through e one more time to fully
purge the fitting.
5.7.6.5 Once the fitting and stem have been purged, quickly
withdraw the volume of gas you require using steps 5.6.6.1.4(a)
through (d). Be sure to close the valve on the cylinder and syringe
before you withdraw the syringe from the Luer fitting.
8260B - 11 Revision 2
January 1995
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5.7.6.6 Open the syringe on/off valve for 5 seconds to
reduce the syringe pressure to atmospheric pressure. The pressure in
the cylinder is -30 psi.
5.7.6.7 The gas mixture should be quickly transferred into
the reagent water through the female Luer fitting located above the
purging vessel.
NOTE: Make sure the arrow on the 4-way valve is pointing toward the female Luer
fitting when transferring the sample from the syringe. Be sure to switch
the 4-way valve back to the closed position before removing the syringe
from the Luer fitting.
5.7.6.8 Transfer the remaining half of the working standard
into the purging vessel. This procedure insures that the total volume
of gas mix is flushed into the purging vessel, with none remaining in
the valve or lines.
5.7.6.9 The concentration of each compound in the cylinder
is typically 0.0025 M9/ML.
5.7.6.10 The following are the recommended gas volumes spiked
into 5 mL of water to produce a typical 5-point calibration:
Gas Volume Calibration Concentration
40 ML 20 jug/L
100 ML 50 Mg/L
200 ML 100
300 ML 150
400 ML 200 jug/L
5.7.6.11 The following are the recommended gas volumes spiked
into 25 mL of water to produce a typical 5-point calibration:
Gas Volume Cal i brat ion Concentration
10 //L
20 ML
50 ML
100 ML
250 ML
1 M9/L
2 M9/L
5 Mg/L
10 M9/L
25 M9/L
5.8 Secondary dilution standards - Using stock standard solutions, prepare
secondary dilution standards in methanol containing the compounds of interest,
either singly or mixed together. Secondary dilution standards must be stored
with minimal headspace and should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards from
them. Store in a vial with no headspace. Replace after one week.
5.9 Surrogate standards - The recommended surrogates are toluene-da,
4-bromofluorobenzene, l,2-dichloroethane-d4, and dibromofluoromethane. Other
compounds may be used as surrogates, depending upon the analysis requirements.
8260B - 12 Revision 2
January 1995
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A stock surrogate solution in methanol should be prepared as described above, and
a surrogate standard spiking solution should be prepared from the stock at a
concentration of 50-250 /ng/10 mL, in methanol. Each sample undergoing GC/MS
analysis must be spiked with 10 /xL of the surrogate spiking solution prior to
analysis. If a more sensitive mass spectrometer is employed to achieve lower
detection levels, then more dilute surrogate solutions may be required.
5.10 Internal standards - The recommended internal standards are
fluorobenzene, chlorobenzene-d5, and l,4-dichlorobenzene-d4. Other compounds may
be used as internal standards as long as they have retention times similar to the
compounds being detected by GC/MS. Prepare internal standard stock and secondary
dilution standards in methanol using the procedures described in Sees. 5.7 and
5.8. It is recommended that the secondary dilution standard be prepared at a
concentration of 25 mg/L of each internal standard compound. Addition of 10 /zL
of this standard to 5.0 ml of sample or calibration standard would be the
equivalent of 50 /ng/L. If a more sensitive mass spectrometer is employed to
achieve lower detection levels, then more dilute internal standard solutions may
be required. Area counts of the internal standard peaks should be between
50-200% of the areas of the target analytes in the mid-point calibration
analysis.
5.11 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng//iL of BFB in methanol should be prepared. If a more sensitive mass
spectrometer is employed to achieve lower detection levels, then a more dilute
BFB standard solution may be required.
5.12 Calibration standards - Calibration standards should be prepared at
a minimum of five concentrations from the secondary dilution of stock standards
(see Sees. 5.7 and 5.8). Prepare these solutions in organic-free reagent water.
One of the standards should be at a concentration near, but above, the method
detection limit. The remaining standards should correspond to the range of
concentrations found in real samples but should not exceed the working range of
the GC/MS system. Each standard should contain each analyte for detection by
this method.
5.12.1 It is the intent of EPA that all target analytes for a
particular analysis be included in the calibration standard(s). These
target analytes may not include the entire list of analytes (Sec. 1.1) for
which the method has been demonstrated. However, the laboratory shall not
report a quantitative result for a target analyte that was not included in
the calibration standard(s).
5.12.2 The calibration standards must also contain the internal
standards chosen for the analysis. Calibration standards must be prepared
fresh daily.
5.13 Matrix spiking and laboratory control sample (LCS) standards - Matrix
spiking standards should be prepared from volatile organic compounds which are
representative of the compounds being investigated. At a minimum, the matrix
spike should include 1,1-dichloroethene, trichloroethene, chlorobenzene, toluene,
and benzene. The matrix spiking solution should contain compounds that are
expected to be found in the types of samples to be analyzed.
8260B - 13 Revision 2
January 1995
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5.13.1 Some permits may require the spiking of specific compounds of
interest, especially if polar compounds are a concern, since the spiking
compounds listed above would not be representative of such compounds. The
standard should be prepared in methanol, with each compound present at a
concentration of 250 /jg/10.0 mL.
5.13.2 The spiking solutions should not be prepared from the same
standards as the calibration standards. However, the same spiking standard
prepared for the matrix spike may be used for the LCS.
5.13.3 If a more sensitive mass spectrometer is employed to achieve
lower detection levels, more dilute matrix spiking solutions may be
required.
5.14 Great care must be taken to maintain the integrity of all standard
solutions. It is recommended all standards in methanol be stored at -10°C or
less, in amber bottles with Teflon®-!ined screw-caps.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See the introductory material to this chapter, Organic Analytes, Sec. 4.1.
7.0 PROCEDURE
7.1 Various alternative methods are provided for sample introduction. All
internal standards, surrogates, and matrix spiking compounds (when applicable)
must be added to the samples before introduction into the GC/MS system. Consult
the sample introduction method for the procedures by which to add such standards.
7.1.1 Direct injection - this includes: injection of an aqueous
sample containing a very high concentration of analytes; injection of
aqueous concentrates from Method 5031 (azeotropic distillation); and
injection of a waste oil diluted 1:1 with hexadecane (Method 3585).
Direct injection of aqueous samples (non-concentrated) has very limited
applications. It is only used for the determination of volatiles at the
toxicity characteristic (TC) regulatory limits or at concentrations in
excess of 10,000 jug/L. It may also be used in conjunction with the test
for ignitability in aqueous samples (along with Methods 1010 and 1020), to
determine if alcohol is present at greater than 24%.
7.1.2 Purge-and-trap - this includes purge-and-trap for aqueous
samples (Method 5030) and purge-and-trap for solid samples (Method 5035).
Method 5035 also provides techniques for extraction of solid and oily waste
samples by methanol (and other water miscible solvents) with subsequent
purge-and-trap from an aqueous matrix using Method 5030. Normally purge-
and-trap of aqueous samples is performed at ambient temperature, while
purging of soil/solid samples is performed at 40°C, to improve purging
efficiency. Occasionally, there may be a need to perform a heated purge
for aqueous samples to lower detection limits, however, a 25-mL sample will
provide the needed sensitivity in most situations.
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7.1.3 Vacuum distillation - this technique may be used for the
introduction of volatile organics from aqueous, solid, or tissue samples
(Method 5032) into the GC/MS system.
7.1.4 Automated static headspace - this technique may be used for
the introduction of volatile organics from solid samples (Method 5021) into
the GC/MS system.
7.1.5 Cartridge desorption - this technique may be for the
introduction of volatile organics from sorbent cartridges (Method 5041)
used in the sampling of air. The sorbent cartridges are from the volatile
organics sampling train (VOST) or SMVOC (Method 0031).
7.2 Recommended chromatographic conditions
7.2.1 General conditions
cooling (example
Injector temperature: 200 - 225°C
Transfer line temperature: 250 - 300°C
7.2.2 Column 1 and Column 2 with cryogenic
chromatograms are presented in Figures 1 and 2)
Carrier gas (He) flow rate: 15 mL/min
Initial temperature: 10°C, hold for 5 minutes
Temperature program: 6°C/min to 160 °C
Final temperature: 160°C, hold until all expected
compounds have eluted.
7.2.3 Column 2, non-cryogenic cooling (an example chromatogram is
presented in Figure 3).
7.2.3.1 Carrier gas flow and split and make-up gas flows
should be set using the performance of standards as guidance. Set the
carrier gas head pressure to -10 psi and the split to ~30 mL/min.
Optimize the make-up gas flow for the separator (approximately 30
mL/min) by injecting BFB, and determining the optimum response when
varying the make-up gas. This will require several injections of BFB.
7.2.3.2 After completing Sec. 7.2.3.1, make several
injections of the volatile working standard with all analytes of
interest. Adjust the carrier and split to provide optimum
chromatography and response. This is an especially critical
adjustment for the volatile gas analytes. The head pressure should
optimize between 8-12 psi and the split between 20 - 60 mL/min. The
use of the splitter is important to minimize the effect of water on
analyte response, to allow the use of a larger volume of helium during
trap desorption, and to slow column flow.
Initial temperature: 45°C, hold for 2 minutes
Temperature program: 8°C/min to 200°C
Final temperature: 200°C, hold for 6 minutes.
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A trap preheated to 150°C prior to trap desorption is required to
provide adequate chromatography of the gas analytes.
7.2.4 Column 3 (A sample chromatogram is presented in Figure 4)
Carrier gas (He) flow rate: 4 mL/min
Initial temperature: 10°C, hold for 5 minutes
Temperature program: 6°C/min to 70°C, then
15°C/min to 145°C
Final temperature: 145°C, hold until all expected
compounds have eluted.
7.2.5 Direct injection - Column 2
Carrier gas (He) flow rate: 4 mL/min
Column: J&W DB-624, 70m x 0.53 mm
Initial temperature: 40°C, hold for 3 minutes
Temperature program: 8°C/min
Final temperature: 260°C, hold until all expected
compounds have eluted.
Column Bake out: 75 minutes
Injector temperature: 200-225°C
Transfer line temperature: 250-300°C
7.2.6 Direct split interface - Column 4
Carrier gas (He) flow rate: 1.5 mL/min
Initial temperature: 35°C, hold for 2 minutes
Temperature program: 4°C/min to 50°C
10°C/min to 220°C
Final temperature: 220°C, hold until all expected
compounds have eluted
Split ratio: 100:1
Injector temperature: 125°C
7.3 Initial calibration
Establish the GC/MS operating conditions, using the following
recommendations as guidance.
Mass range: 35 - 260 amu
Scan time: 0.6 - 2 sec/scan
Source temperature: According to manufacturer's specifications
Ion trap only: Set axial modulation, manifold temperature,
and emission current to manufacturer's
recommendations
7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 4 for a 5-50 ng injection or purging of 4-bromofluorobenzene (2-/iL
injection of the BFB standard). Analyses must not begin until these
criteria are met.
7.3.2 Set up the sample introduction system as outlined in the
method of choice (see Sec. 7.1). A different calibration curve is
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necessary for each method because of the differences in conditions and
equipment. A set of at least five calibration standards containing the
method analytes is necessary. One calibration standard should contain each
analyte at a concentration near, but greater than, the method detection
limit (Table 1) for that compound; the other calibration standards should
contain analytes at concentrations that define the range of the method.
Calibration must be performed using the sample introduction technique that
will be used for samples. For Method 5030, the purging efficiency for 5
mL of water is greater than for 25 mL. Therefore, develop the standard
curve with whichever volume of sample that will be analyzed.
7.3.2.1 To prepare a calibration standard, add an
appropriate volume of a secondary dilution standard solution to an
aliquot of organic-free reagent water in a volumetric flask. Use a
microsyringe and rapidly inject the alcoholic standard into the
expanded area of the filled volumetric flask. Remove the needle as
quickly as possible after injection. Mix by inverting the flask three
times only. Discard the contents contained in the neck of the flask.
Aqueous standards are not stable and should be prepared daily.
Transfer 5.0 ml (or 25 ml if lower detection limits are required) of
each standard to a gas tight syringe along with 10 p,l of internal
standard. Then transfer the contents to the appropriate device or
syringe. Some of the introduction methods may have specific guidance
on the volume of calibration standard and the way the standards are
transferred to the device.
7.3.2.2 The internal standards selected in Sec. 5.4 should
permit most of the components of interest in a chromatogram to have
retention times of 0.80 - 1.20, relative to one of the internal
standards. Use the base peak ion from the specific internal standard
as the primary ion for quantitation (see Table 1). If interferences
are noted, use the next most intense ion as the quantitation ion.
7.3.2.3 To prepare a calibration standard for direct
injection analysis of waste oil, dilute standards in hexadecane.
7.3.3 Proceed with the analysis of the calibration standards
following the procedure in the introduction method of choice. For direct
injection, inject 1 - 2 juL into the GC/MS system. The injection volume
will depend upon the chromatographic column chosen and the tolerance of the
specific GC/MS system to water.
7.3.4 Tabulate the area response of the characteristic ions (see
Table 5) against concentration for each compound and each internal
standard. Calculate response factors (RF) for each compound relative to
one of the internal standards. The internal standard selected for the
calculation of the RF for a compound should be the internal standard that
has a retention time closest to the compound being measured (Sec. 7.6.2).
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The RF is calculated as follows:
A x C.
RF =
where: is 8
As = Peak area (or height) of the analyte or surrogate.
Ais = Peak area (or height) of the internal standard.
Cs = Concentration of the analyte or surrogate.
Cjs = Concentration of the internal standard.
7.3.5 Calculate the mean RF for each compound using the five RF
values calculated from the initial (5-point) calibration curve. A system
performance check should be made before this calibration curve is used.
Five compounds (the System Performance Check Compounds, or SPCCs) are
checked for a minimum average response factor. These compounds are
Chloromethane; 1,1-dichloroethane; bromoform; chlorobenzene; and
1,1,2,2-tetrachloroethane. These compounds are used to check compound
instability and to check for degradation caused by contaminated lines or
active sites in the system. Example problems include:
7.3.5.1 Chloromethane is the most likely compound to be lost
if the purge flow is too fast.
7.3.5.2 Bromoform is one of the compounds most likely to be
purged very poorly if the purge flow is too slow. Cold spots and/or
active sites in the transfer lines may adversely affect response.
Response of the quantitation ion (m/z 173) is directly affected by the
tuning of BFB at ions m/z 174/176. Increasing the m/z 174/176 ratio
relative to m/z 95 may improve bromoform response.
7.3.5.3 Tetrachloroethane and 1,1-dichloroethane are
degraded by contaminated transfer lines in purge-and-trap systems
and/or active sites in trapping materials.
7.3.5.4 The minimum mean response factors for the volatile
SPCCs are as follows:
Chloromethane 0.10
1,1-Dichloroethane 0.10
Bromoform 0.10
Chlorobenzene 0.30
1,1,2,2-Tetrachloroethane 0.30
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7.3.6 Calculate the standard deviation (SD) and relative standard
deviation (RSD) of the response factors for all compounds from the initial
calibration, as follows:
SD =
1=1
RSD = IE x 100
RF
n-1
where:
RFi = RF for each of the calibration standards
RF = mean RF for each compound from the initial calibration
n = Number of calibration standards, e.g., 5
7.3.6.1 The RSD should be less than 15% for each compound.
However, the RSD for each individual Calibration Check Compound (CCC)
must be less than 30%. The CCCs are:
1,1-Dichloroethene
Chloroform
1,2-Dichloropropane
Toluene
Ethyl benzene
Vinyl chloride
NOTE:
7.3.6.2 If an RSD of greater than 30% is measured for any
CCC, then corrective action to eliminate a system leak and/or column
reactive sites is necessary before reattempting calibration.
7.3.6.3 The relative retention times of each compound in
each calibration standard should agree within 0.06 relative retention
time units. Late-eluting compounds usually have much better
agreement.
7.3.7 Linearity
7.3.7.1 If the RSD of any compound is 15% or less, then the
response factor is assumed to be constant over the calibration range,
and the average response factor may be used for quantitation (Sec.
7.7.2).
7.3.7.2 If the RSD of any compound is greater than 15%, see
Sec. 7.0 in Method 8000 for options on dealing with non-linear
calibrations. One of the options must be applied to GC/MS calibration
in this situation, or a new initial calibration must be performed.
Method 8000 specifies a linearity criterion of 20% RSD. That criterion
pertains to GC and HPLC methods other than GC/MS. Method 8260 requires
15% RSD as evidence of sufficient linearity to employ an average response
factor.
7.3.7.3 When the RSD exceeds 15%, the plotting and visual
inspection of a calibration curve can be a useful diagnostic tool.
The inspection may indicate analytical problems, including errors in
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standard preparation, the presence of active sites in the
chromatographic system, analytes that exhibit poor chromatographic
behavior, etc.
NOTE: The 20% RSD criteria in Method 8000 pertains to GC and HPLC methods other
than GC/MS. Method 8260 requires 15% RSD.
7.4 GC/MS calibration verification
7.4.1 Prior to the analysis of samples, inject or introduce 5-50 ng
of the 4-bromofluorobenzene standard into the GC/MS system following the
sample introduction method of choice. The resultant mass spectra for the
BFB must meet the criteria given in Table 4 before sample analysis begins.
These criteria must be demonstrated each 12-hour shift during which samples
are analyzed.
7.4.2 The initial calibration curve (Sec. 7.3) for each compound of
interest must be verified once every 12 hours during analysis, using the
introduction technique used for samples. This is accomplished by analyzing
a calibration standard that is at a concentration near the midpoint
concentration for the working range of the GC/MS by checking the SPCC and
CCC.
NOTE: A method blank should be analyzed prior to the calibration standard to
ensure that the total system (introduction device, transfer lines and
GC/MS system) is free of contaminants.
7.4.3 System Performance Check Compounds (SPCCs)
A system performance check must be made during every 12-hour
analytical shift. Each SPCC compound in the calibration verification
standard must meet its minimum response factor (see Sec. 7.3.5.4). This
is the same check that is applied during the initial calibration. If the
minimum response factors are not met, the system must be evaluated, and
corrective action must be taken before sample analysis begins. Possible
problems include standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column, and
active sites in the column or chromatographic system. This check must be
met before sample analysis begins.
7.4.4 Calibration Check Compounds (CCCs)
7.4.4.1 After the system performance check is met, the CCCs
listed in Sec. 7.3.6 are used to check the validity of the initial
calibration. Calculate the percent difference using the following
equation:
RF - RF
% Difference = v x 100
RF
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where:
W = Mean RF from the initial calibration
RFW = RF from the calibration verification standard
V
7.4.4.2 If the percent difference for each CCC is less than
20%, the initial calibration is assumed to be valid. If the criterion
is not met (i.e., greater than 20% difference), for any one CCC, then
corrective action must be taken prior to the analysis of samples.
7.4.4.3 Problems similar to those listed under SPCCs could
affect the CCCs. If the problem cannot be corrected by other
measures, a new five-point initial calibration must be generated. The
CCC criteria must be met before sample analysis begins. If the CCCs
are not included in the list of analytes for a project, and therefore
not included in the calibration standards, then all analytes must meet
the 20% difference criterion.
7.4.5 The internal standard responses and retention times in the
calibration verification standard must be evaluated immediately after or
during data acquisition. If the retention time for any internal standard
changes by more than 30 seconds from the last calibration verification (12
hours), the chromatographic system must be inspected for malfunctions and
corrections must be made, as required. If the EICP area for any of the
internal standards changes by a factor of two (-50% to +100%) from the
previous calibration verification standard, the mass spectrometer must be
inspected for malfunctions and corrections must be made, as appropriate.
When corrections are made, reanalysis of samples analyzed while the system
was malfunctioning is necessary.
7.5 GC/MS analysis of samples
7.5.1 It is highly recommended that the sample be screened to
minimize contamination of the GC/MS system from unexpectedly high
concentrations of organic compounds. Some of the screening options
available utilizing SW-846 methods are automated headspace-GC/FID (Methods
5021/8015), automated headspace-GC/PID/ELCD (Methods 5021/8021), or waste
dilution-GC/PID/ELCD (Methods 3585/8021) using the same type of capillary
column. When used only for screening purposes, the quality control
requirements in the methods above may be reduced as appropriate. Sample
screening is particularly important when Method 8260 is used to achieve low
detection levels.
7.5.2 BFB tuning criteria and GC/MS calibration verification
criteria must be met before analyzing samples.
7.5.3 All samples and standard solutions must be allowed to warm to
ambient temperature before analysis. Set up the introduction device as
outlined in the method of choice.
7.5.4 The process of taking an aliquot destroys the validity of
remaining volume of an aqueous sample for future analysis. Therefore, if
only one VOA vial is provided to the laboratory, the analyst should prepare
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two aliquots for analysis at this time, to protect against possible loss
of sample integrity. This second sample is maintained only until such time
when the analyst has determined that the first sample has been analyzed
properly. For aqueous samples, one 20-mL syringe could be used to hold two
5-mL aliquots. If the second aliquot is to be taken from the syringe, it
must be analyzed within 24 hours. Care must be taken to prevent air from
leaking into the syringe.
7.5.5 Remove the plunger from a 5-mL syringe and attach a closed
syringe valve. Open the sample or standard bottle, which has been allowed
to come to ambient temperature, and carefully pour the sample into the
syringe barrel to just short of overflowing. Replace the syringe plunger
and compress the sample. Open the syringe valve and vent any residual air
while adjusting the sample volume to 5.0 ml. If lower detection limits are
required, use a 25-mL syringe, and adjust the final volume to 25.0 mL.
7.5.6 The following procedure may be used to dilute aqueous samples
for analysis of volatiles. All steps must be performed without delays,
until the diluted sample is in a gas-tight syringe.
7.5.6.1 Dilutions may be made in volumetric flasks (10 to
100 mL). Select the volumetric flask that will allow for the
necessary dilution. Intermediate dilution steps may be necessary for
extremely large dilutions.
7.5.6.2 Calculate the approximate volume of organic-free
reagent water to be added to the volumetric flask, and add slightly
less than this quantity of organic-free reagent water to the flask.
7.5.6.3 Inject the appropriate volume of the original sample
from the syringe into the flask. Aliquots of less than 1 mL are not
recommended. Dilute the sample to the mark with organic-free reagent
water. Cap the flask, invert, and shake three times. Repeat above
procedure for additional dilutions.
7.5.6.4 Fill a 5-mL syringe with the diluted sample, as
described in Sec. 7.5.5.
7.5.7 Compositing aqueous samples prior to GC/MS analysis
7.5.7.1 Add 5 mL of each sample (up to 5 samples are
allowed) to a 25-mL glass syringe. Special precautions must be made
to maintain zero headspace in the syringe. Larger volumes of a
smaller number of samples may be used, provided that equal volumes of
each sample are composited.
7.5.7.2 The samples must be cooled to 4°C or less during
this step to minimize volatilization losses. Sample vials may be
placed in a tray of ice during the processing.
7.5.7.3 Mix each vial well and draw out a 5-mL aliquot with
the 25-mL syringe.
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7.5.7.4 Once all the aliquots have been combined on the
syringe, invert the syringe several times to mix the aliquots.
Introduce the composited sample into the instrument, using the method
of choice (see Sec. 7.1).
7.5.7.5 If less than five samples are used for compositing,
a proportionately smaller syringe may be used, unless a 25-mL sample
is to be purged.
7.5.8 Add 10.0 juL of the surrogate spiking solution and 10.0 p,l of
the internal standard spiking solution to each sample. The surrogate and
internal standards may be mixed and added as a single spiking solution.
The addition of 10 /zL of the surrogate spiking solution to 5 ml of aqueous
sample will yield a concentration of 50 /Ltg/L of each surrogate standard.
The addition of 10 juL of the surrogate spiking solution to 5 g of a non-
aqueous sample will yield a concentration of 50 fj,g/kg of each standard.
7.5.8.1 If a more sensitive mass spectrometer is employed to
achieve lower detection levels, more dilute surrogate and internal
standard solutions may be required.
7.5.9 Add 10 juL of the matrix spike solution (Sec. 5.13) to a 5-mL
aliquot of the sample chosen for spiking. Disregarding any dilutions, this
is equivalent to a concentration of 50 /ug/L of each matrix spike standard.
7.5.9.1 Follow the same procedure in preparing the
laboratory control sample (LCS), except the spike is added to a clean
matrix. See Sec. 8.4 and Method 5000 for more guidance on the
selection and preparation of the matrix spike and the LCS.
7.5.9.2 If a more sensitive mass spectrometer is employed to
achieve lower detection levels, more dilute matrix spiking and LCS
solutions may be required.
7.5.10 Analyze the sample following the procedure in the introduction
method of choice.
7.5.10.1 For direct injection, inject 1 to 2 /xL into the
GC/MS system. The volume limitation will depend upon the
chromatographic column chosen and the tolerance of the specific GC/MS
system to water (if an aqueous sample is being analyzed).
7.5.10.2 The concentration of the internal standards,
surrogates, and matrix spiking standards (if any) added to the
injection aliquot must be adjusted to provide the same concentration
in the 1-2 fj,L injection as would be introduced into the GC/MS by
purging a 5-mL aliquot.
7.5.11 If the initial analysis of the sample or a dilution of the
sample has a concentration of any analyte that exceeds the initial
calibration range, the sample must be reanalyzed at a higher dilution.
Secondary ion quantitation is allowed only when there are sample
interferences with the primary ion.
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7.5.11.1 When ions from a compound in the sample saturate the
detector, this analysis must be followed by the analysis of an
organic-free reagent water blank. If the blank analysis is not free
of interferences, then the system must be decontaminated. Sample
analysis may not resume until the blank analysis is demonstrated to
be free of interferences.
7.5.11.2 All dilutions should keep the response of the major
constituents (previously saturated peaks) in the upper half of the
linear range of the curve.
7.5.12 The use of selected ion monitoring (SIM) is acceptable in
situations requiring detection limits below the normal range of full El
spectra. However, SIM may provide a lesser degree of confidence in the
compound identification unless multiple ions are monitored for each
compound.
7.6 Qualitative analysis
7.6.1 The qualitative identification of compounds determined by this
method is based on retention time, and on comparison of the sample mass
spectrum, after background correction, with characteristic ions in a
reference mass spectrum. The reference mass spectrum must be generated by
the laboratory using the conditions of this method. The characteristic
ions from the reference mass spectrum are defined to be the three ions of
greatest relative intensity, or any ions over 30% relative intensity if
less than three such ions occur in the reference spectrum. Compounds are
identified as present when the following criteria are met.
7.6.1.1 The intensities of the characteristic ions of a
compound maximize in the same scan or within one scan of each other.
Selection of a peak by a data system target compound search routine
where the search is based on the presence of a target chromatographic
peak containing ions specific for the target compound at a
compound-specific retention time will be accepted as meeting this
criterion.
7.6.1.2 The relative retention time (RRT) of the sample
component is within ± 0.06 RRT units of the RRT of the standard
component.
7.6.1.3 The relative intensities of the characteristic ions
agree within 30% of the relative intensities of these ions in the
reference spectrum. (Example: For an ion with an abundance of 50%
in the reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.6.1.4 Structural isomers that produce very similar mass
spectra should be identified as individual isomers if they have
sufficiently different GC retention times. Sufficient GC resolution
is achieved if the height of the valley between two isomer peaks is
less than 25% of the sum of the two peak heights. Otherwise,
structural isomers are identified as isomeric pairs.
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7.6.1.5 Identification is hampered when sample components
are not resolved chromatographically and produce mass spectra
containing ions contributed by more than one analyte. When gas
chromatographic peaks obviously represent more than one sample
component (i.e., a broadened peak with shoulder(s) or a valley between
two or more maxima), appropriate selection of analyte spectra and
background spectra is important.
7.6.1.6 Examination of extracted ion current profiles of
appropriate ions can aid in the selection of spectra, and in
qualitative identification of compounds. When analytes coelute (i.e.,
only one chromatographic peak is apparent), the identification
criteria may be met, but each analyte spectrum will contain extraneous
ions contributed by the coeluting compound.
7.6.2 For samples containing components not associated with the
calibration standards, a library search may be made for the purpose of
tentative identification. The necessity to perform this type of
identification will be determined by the purpose of the analyses being
conducted. Data system library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other.
For example, the RCRA permit or waste delisting requirements may
require the reporting of non-target analytes. Only after visual comparison
of sample spectra with the nearest library searches may the analyst assign
a tentative identification. Use the following guidelines for making
tentative identifications:
(1) Relative intensities of major ions in the reference spectrum
(ions greater than 10% of the most abundant ion) should be
present in the sample spectrum.
(2) The relative intensities of the major ions should agree within ±
20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%).
(3) Molecular ions present in the reference spectrum should be
present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination
or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes
create these discrepancies.
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7.7 Quantitative analysis
7.7.1 Once a compound has been identified, the quantitation of that
compound will be based on the integrated abundance from the EICP of the
primary characteristic ion. The internal standard used shall be the one
nearest the retention time of that of a given analyte.
7.7.2 If the RSD of a compound's response factors is 15% or less,
then the concentration in the extract may be determined using the average
response factor (RF) from initial calibration data (7.3.6). See Method
8000, Sec. 7.0, for the equations describing internal standard calibration
and either linear or non-linear calibrations.
7.7.3 Where appl icable, the concentration of any non-target analytes
identified in the sample (Sec. 7.6.2) should be estimated. The same
formulae should be used with the following modifications: The areas Ax and
Ais should be from the total ion chromatograms, and the RF for the compound
should be assumed to be 1.
7.7.4 The resulting concentration should be reported indicating:
(1) that the value is an estimate, and (2) which internal standard was used
to determine concentration. Use the nearest internal standard free of
interferences.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Methods 3500 and 5000. Each laboratory should maintain a formal quality
assurance program. The laboratory should also maintain records to document the
quality of the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
In addition, instrument QC requirements may be found in the following sections
of Method 8260:
8.2.1 The GC/MS system must be tuned to meet the BFB specifications
in Sees. 7.3.1 and 7.4.1.
8.2.2 There must be an initial calibration of the GC/MS system as
described in Sec. 7.3.
8.2.3 The GC/MS system must meet the SPCC criteria described in Sec.
7.4.3 and the CCC criteria in Sec. 7.4.4, each 12 hours.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
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the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time
a set of samples is analyzed or there is a change in reagents, a method
blank should be analyzed as a safeguard against chronic laboratory
contamination. The blanks should be carried through all stages of sample
preparation and measurement.
8.4.2 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
8.6 The experience of the analyst performing GC/MS analyses is invaluable
to the success of the methods. Each day that analysis is performed, the
calibration verification standard should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal? Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still performing acceptably, the
injector is leaking, the injector septum needs replacing, etc. If any changes
8260B - 27 Revision 2
January 1995
-------
are made to the system (e.g., the column changed), recalibration of the system
must take place.
8.7 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL actually achieved in a given
analysis will vary depending on instrument sensitivity and matrix effects.
9.2 This method has been tested using purge-and-trap (Method 5030) in a
single laboratory using spiked water. Using a wide-bore capillary column, water
was spiked at concentrations between 0.5 and 10 /zg/L. Single laboratory
accuracy and precision data are presented for the method analytes in Table 6.
Calculated MDLs are presented in Table 1.
9.3 The method was tested using purge-and-trap (Method 5030) with water
spiked at 0.1 to 0.5 /j.g/1 and analyzed on a cryofocussed narrow-bore column.
The accuracy and precision data for these compounds are presented in Table 7.
MDL values were also calculated from these data and are presented in Table 2.
9.4 Direct injection (Method 3585) has been used for the analysis of waste
motor oil samples using a wide-bore column. Single laboratory precision and
accuracy data are presented in Tables 10 and 11 for TCLP volatiles in oil. The
performance data were developed by spiking and analyzing seven replicates each
of new and used oil. The oils were spiked at the TCLP regulatory concentrations
for most analytes, except for the alcohols, ketones, ethyl acetate and
chlorobenzene which are spiked at 5 ppm, well below the regulatory
concentrations. Prior to spiking, the new oil (an SAE 30-weight motor oil) was
heated at 80°C overnight to remove volatiles. The used oil (a mixture of used
oil drained from passenger automobiles) was not heated and was contaminated with
20 - 300 ppm of BTEX compounds and isobutanol. These contaminants contributed
to the extremely high recoveries of the BTEX compounds in the used oil.
Therefore, the data from the deuterated analogs of these analytes represent more
typical recovery values.
9.5 Single laboratory accuracy and precision data were obtained for the
Method 5035 analytes in three soil matrices: sand; a soil collected 10 feet below
the surface of a hazardous landfill, called C-Horizon; and a surface garden soil.
Sample preparation was by Method 5035. Each sample was fortified with the
analytes at a concentration of 4 jug/kg. These data are listed in Tables 17, 18,
and 19. All data were calculated using fluorobenzene as the internal standard
added to the soil sample prior to extraction. This causes some of the results
to be greater than 100% recovery because the precision of results is sometimes
as great as 28%.
8260B - 28 Revision 2
January 1995
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9.5.1 In general, the recoveries of the analytes from the sand
matrix are the highest, the C-Horizon soil results are somewhat less, and
the surface garden soil recoveries are the lowest. This is due to the
greater adsorptive capacity of the garden soil. This illustrates the
necessity of analyzing matrix spike samples to assess the degree of matrix
effects.
9.5.2 The recoveries of some of the gases, or very volatile
compounds, such as vinyl chloride, trichlorofluoromethane, and
1,1-dichloroethene, are somewhat greater than 100%. This is due to the
difficulty encountered in fortifying the soil with these compounds,
allowing an equilibration period, then extracting them with a high degree
of precision. Also, the garden soil results in Table 19 include some
extraordinarily high recoveries for some aromatic compounds, such as
toluene, xylenes, and trimethylbenzenes. This is due to contamination of
the soil prior to sample collection, and to the fact that no background was
subtracted.
9.6 Performance data for non-purgeable volatiles using azeotropic
distillation (Method 5031) are included in Tables 12 to 16.
9.7 Performance data for volatiles prepared using vacuum distillation
(Method 5032) in soil, water, oil and fish tissue matrices are included in Tables
20 to 27.
9.8 Single laboratory accuracy and precision data were obtained for the
Method 5021 analytes in two soil matrices: sand and a surface garden soil.
Replicate samples were fortified with the analytes at concentrations of 10
Mg/kg. These data are listed in Table 30. All data were calculated using the
internal standards listed for each analyte in Table 28. The recommended internal
standards were selected because they generated the best accuracy and precision
data for the analyte in both types of soil.
9.8.1 If a detector other than an MS is used for analysis,
consideration must be given to the choice of internal standards and
surrogates. They must not coelute with any other analyte and must have
similar properties to the analytes. The recoveries of the analytes are 50%
or higher for each matrix studied. The recoveries of the gases or very
volatile compounds are greater than 100% in some cases. Also, results
include high recoveries of some aromatic compounds, such as toluene,
xylenes, and trimethylbenzenes. This is due to contamination of the soil
prior to sample collection.
9.8.2 The method detection limits using Method 5021 listed in Table
29 were calculated from results of seven replicate analyses of the sand
matrix. Sand was chosen because it demonstrated the least degree of matrix
effect of the soils studied. These MDLs were calculated utilizing the
procedure described in Chapter One and are intended to be a general
indication of the capabilities of the method.
9.9 The MDL concentrations listed in Table 31 were determined using Method
5041 in conjunction with Method 8260. They were obtained using cleaned blank
VOST tubes and reagent water. Similar results have been achieved with field
samples. The MDL actually achieved in a given analysis will vary depending upon
8260B - 29 Revision 2
January 1995
-------
instrument sensitivity and the effects of the matrix. Preliminary spiking
studies indicate that under the test conditions, the MDLs for spiked compounds
in extremely complex matrices may be larger by a factor of 500 - 1000.
9.10 The EQL of sample taken by Method 0040 and analyzed by Method 8260 is
estimated to be in the range of 0.03 to 0.9 ppm (See Table 33). Matrix effects
may cause the individual compound detection limits to be higher.
10.0 REFERENCES
1. Methods for the Determination of Organic Compounds in Finished Drinking
Water and Raw Source Water Method 524.2, U.S. Environmental Protection
Agency, Office of Research Development, Environmental Monitoring and
Support Laboratory, Cincinnati, OH, 1986.
2. Bellar, T.A., Lichtenberg, J.J, J. Amer. Water Works Assoc.. 1974, 66(12),
739-744.
3. Bellar, T.A., Lichtenberg, J.J., "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds"; in Van Hall, Ed.; Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp 108-129, 1979.
4. Budde, W.L., Eichelberger, J.W., "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass Spectrometry Equipment and
Laboratories"; U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, OH, April 1980;
EPA-600/4-79-020.
5. Eichelberger, J.W., Harris, L.E., Budde, W.L., "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems"; Analytical Chemistry 1975, 47, 995-1000.
6. Olynyk, P., Budde, W.L., Eichelberger, J.W., "Method Detection Limit for
Methods 624 and 625"; Unpublished report, October 1980.
7. Non Cryogenic Temperatures Program and Chromatogram, Private
Communications; M. Stephenson and F. Allen, EPA Region IV Laboratory,
Athens, GA.
8. Marsden, P.J., Helms, C.L., Colby, B.N., "Analysis of Volatiles in Waste
Oil"; Report for B. Lesnik, OSW/EPA under EPA contract 68-W9-001, 6/92.
9. Methods for the Determination of Organic Compounds in Drinking Water,
Supplement II Method 524.2; U.S. Environmental Protection Agency, Office
of Research and Development, Environmental Monitoring Systems Laboratory,
Cincinnati, OH, 1992.
10. Flores, P., Bellar, T., "Determination of Volatile Organic Compounds in
Soils Using Equilibrium Headspace Analysis and Capillary Column Gas
Chromatography/Mass Spectrometry", U.S. Environmental Protection Agency,
Office of Research and Development, Environmental Monitoring Systems
Laboratory, Cincinnati, OH, December, 1992.
8260B - 30 Revision 2
January 1995
-------
11. Bruce, M.L., Lee, R.P., Stephens, M.W., "Concentration of Water Soluble
Volatile Organic Compounds from Aqueous Samples by Azeotropic
Microdistillation", Environmental Science and Technology 1992, 26, 160-163.
12. Cramer, P.M., Wilner, J., Stanley, J.S., "Final Report: Method for Polar,
Water Soluble, Nonpurgeable Volatile Organics (VOCs)", For U.S.
Environmental Protection Agency, Environmental Monitoring Support
Laboratory, EPA Contract No. 68-C8-0041.
13. Hiatt, M.H., "Analysis of Fish and Sediment for Volatile Priority
Pollutants", Analytical Chemistry 1981, 53, 1541.
14. Validation of the Volatile Organic Sampling Train (VOST) Protocol. Volumes
I and II. EPA/600/4-86-014A, January, 1986.
8260B - 31 Revision 2
January 1995
-------
TABLE 1
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON WIDE-BORE CAPILLARY COLUMNS
Compound
Di chl orodi fl uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Trichlorofluorometh~
-------
Compound
1 , 1 , 2-Tri chl oroethane
Ethyl methacrylate
2-Hexanone
Tetrachloroethene
1,3-Dichloropropane
Di bromochl oromethane
1,2-Dibromoethane
1-Chlorohexane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
o-Xylene
Styrene
Bromoform
Isopropyl benzene (Cumene)
cis-l,4-Dichloro-2-butene
1,1,2 , 2-Tetrachl oroethane
Bromobenzene
1,2,3-Trichloropropane
n-Propyl benzene
2-Chlorotoluene
trans-l,4-Dichloro-2-butene
1,3, 5-Tri methyl benzene
4-Chlorotoluene
Pentachl oroethane
1, 2, 4-Tri methyl benzene
sec-Butyl benzene
tert-Butyl benzene
p- Isopropyl toluene
1, 3 -Di chlorobenzene
1, 4 -Di chlorobenzene
Benzyl chloride
n-Butyl benzene
1 , 2-Di chl orobenzene
l,2-Dibromo-3-chloropropane
1,2, 4-Tri chl orobenzene
Hexachlorobutadiene
Naphthalene
1,2, 3 -Tri chl orobenzene
TABLE 1
(continued)
Retention Time
Column la
19.59
20.01
20.30
20.26
20.51
21.19
21.52
--
23.17
23.36
23.38
23.54
23.54
25.16
25.30
26.23
26.37
27.12
27.29
27.46
27.55
27.58
28.19
28.26
28.31
28.33
29.41
29.47
30.25
30.59
30.59
30.56
31.22
32.00
32.23
32.31
35.30
38.19
38.57
39.05
40.01
Column
11.05
11.15
11.31
11.85
11.83
13.29
13.01
13.33
13.39
13.69
13.68
14.52
14.60
14.88
15.46
16.35
15.86
16.23
16.41
16.42
16.90
16.72
17.70
18.09
17.57
18.52
18.14
18.39
19.49
19.17
21.08
23.08
23.68
23.52
24.18
(minutes)
2b Column 2/c
18.30
18.60
18.70
19.20
19.40
--
20.67
20.87
21.00
21.30
21.37
22.27
22.40
22.77
23.30
24.07
24.00
24.13
24.33
24.53
24.83
24.77
31.50
26.13
26.60
26.50
26.37
26.60
27.32
27.43
--
31.50
32.07
32.20
32.97
MDLd
(M9/L)
0.10
0.14
0.04
0.05
0.06
0.05
0.04
0.05
0.06
0.13
0.05
0.11
0.04
0.12
0.15
0.04
0.03
0.32
0.04
0.04
0.05
0.06
0.13
0.13
0.14
0.12
0.12
0.03
0.11
0.03
0.26
0.04
0.11
0.04
0.03
8260B - 33
Revision 2
January 1995
-------
TABLE 1
(continued)
Compound
Retention Time (minutes)
Column laColumn 2bColumn 2/c
MDLd
(M9/L)
INTERNAL STANDARDS/SURROGATES
1,4-Di f1uorobenzene
Chlorobenzene-d5
l,4-Dichlorobenzene-d4
4-Bromofluorobenzene
l,2-Dichlorobenzene-d4
Dichloroethane-d4
Di bromof1uoromethane
Toluene-d8
Pentafluorobenzene
Fluorobenzene
13.26
23.10
31.16
27.83
32.30
12.08
18.27
13.00
15.71
19.08
23.63
27.25
6.27
14.06
Column 1 - 60 meter x 0.75 mm ID VOCOL capillary.
8 minutes, then program to 180°C at 4°C/min.
Hold at 10°C for
b Column 2-30 meter x 0.53 mm ID DB-624 wide-bore capillary using cryogenic
oven. Hold at 10°C for 5 minutes, then program to 160°C at 6°C/min.
0 Column 2' - 30 meter x 0.53 mm ID DB-624 wide-bore capillary, cooling GC oven
to ambient temperatures. Hold at 10°C for 6 minutes, program to 70°C at 10
°C/min, program to 120°C at 5°C/min, then program to 180°C at 8°C/min.
d MDL based on a 25-mL sample volume.
8260B - 34
Revision 2
January 1995
-------
TABLE 2
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON NARROW-BORE CAPILLARY COLUMNS
Compound
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl chloride
Bromome thane
Chloroethane
Tri chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene chloride
trans-l,2-Dichloroethene
1,1-Dichloroethane
cis-l,2-Dichloroethene
2,2-Dichloropropane
Chloroform
Bromochl oromethane
1 , 1 , 1 -Tri chloroethane
1,2-Dichloroethane
1,1-Dichloropropene
Carbon tetrachloride
Benzene
1,2-Dichloropropane
Trichloroethene
Dibromomethane
Bromodi chl oromethane
Toluene
1,1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Tetrachloroethene
1 ,2-Dibromoethane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
Bromoform
o-Xylene
Styrene
1,1,2 , 2-Tetrachl oroethane
1,2,3-Trichloropropane
Isopropyl benzene
Retention Time (minutes)
Column 3a
0.88
0.97
1.04
1.29
1.45
1.77
2.33
2.66
3.54
4.03
5.07
5.31
5.55
5.63
6.76
7.00
7.16
7.41
7.41
8.94
9.02
9.09
9.34
11.51
11.99
12.48
12.80
13.20
13.60
14.33
14.73
14.73
15.30
15.30
15.70
15.78
15.78
15.78
16.26
16.42
MDLb
(M9/L)
0.11
0.05
0.04
0.06
0.02
0.07
0.05
0.09
0.03
0.03
0.06
0.08
0.04
0.09
0.04
0.02
0.12
0.02
0.03
0.02
0.02
0.01
0.03
0.08
0.08
0.08
0.07
0.05
0.10
0.03
0.07
0.03
0.06
0.03
0.20
0.06
0.27
0.20
0.09
0.10
8260B - 35
Revision 2
January 1995
-------
TABLE 2
(Continued)
Compound
Bromobenzene
2-Chlorotoluene
n-Propyl benzene
4-Chlorotoluene
1 , 3 , 5-Tri methyl benzene
tert-Butylbenzene
1,2, 4 -Tri methyl benzene
sec-Butyl benzene
1,3-Dichlorobenzene
p-Isopropyltoluene
1 , 4-Di chl orobenzene
1,2-Dichlorobenzene
n-Butyl benzene
l,2-Dibromo-3-chloropropane
1, 2, 4-Tri chl orobenzene
Naphthalene
Hexachlorobutadiene
1,2, 3 -Tri chl orobenzene
Retention Time (minutes)
Column 3a
16.42
16.74
16.82
16.82
16.99
17.31
17.31
17.47
17.47
17.63
17.63
17.79
17.95
18.03
18.84
19.07
19.24
19.24
MDLb
(M9/L)
0.11
0.08
0.10
0.06
0.06
0.33
0.09
0.12
0.05
0.26
0.04
0.05
0.10
0.50
0.20
0.10
0.10
0.14
a Column 3 - 30 meter x 0.32 mm ID DB-5 capillary with 1 jum film thickness,
b MDL based on a 25-mL sample volume.
8260B - 36
Revision 2
January 1995
-------
TABLE 3
ESTIMATED QUANTITATION LIMITS FOR VOLATILE ANALYTES6
Estimated Quantitation Limits
5-mL Ground Water 25-mL Ground water Low Soil/Sediment6
Purge (/ug/L) Purge (/ng/L)
Estimated Quantitation Limit (EQL) - The lowest concentration that can be
reliably achieved within specified limits of precision and accuracy during
routine laboratory operating conditions. The EQL is generally 5 to 10
times the MDL. However, it may be nominally chosen within these guidelines
to simplify data reporting. For many analytes the EQL analyte concentration
is selected for the lowest non-zero standard in the calibration curve.
Sample EQLs are highly matrix-dependent. The EQLs listed herein are
provided for guidance and may not always be achievable. See the following
footnote for further guidance on matrix-dependent EQLs.
EQLs listed for soil/sediment are based on wet weight. Normally data are
reported on a dry weight basis; therefore, EQLs will be higher, based on
the percent dry weight in each sample.
Other Matrices Factor0
Water miscible liquid waste 50
High concentration soil and sludge 125
Non-water miscible waste 500
c EQL = [EQL for low soil sediment (Table 3)] x [Factor].
For non-aqueous samples, the factor is on a wet-weight basis.
8260B - 37 Revision 2
January 1995
-------
TABLE 4
BFB (4-BROMOFLUOROBENZENE) MASS INTENSITY CRITERIA8
m/z Required Intensity (relative abundance)
50 15 to 40% of m/z 95
75 30 to 60% of m/z 95
95 Base peak, 100% relative abundance
96 5 to 9% of m/z 95
173 Less than 2% of m/z 174
174 Greater than 50% of m/z 95
175 5 to 9% of m/z 174
176 Greater than 95% but less than 101% of m/z 174
177 5 to 9% of m/z 176
Alternate tuning criteria may be used, (e.g. CLP, Method 524.2, or
manufacturers' instructions), provided that method performance is not
adversely affected.
8260B - 38 Revision 2
January 1995
-------
TABLE 5
CHARACTERISTIC MASSES (m/z) FOR PURGEABLE ORGANIC COMPOUNDS
Compound
Primary
Characteristic
Ion
8260B - 39
Secondary
Characteristic
Ion(s)
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Ally! alcohol
Allyl chloride
Benzene
Benzyl chloride
Bromoacetone
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
iso-Butanol
n-Butanol
2-Butanone
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chloroacetonitrile
Chlorobenzene
1-Chlorobutane
Chl orodi bromomethane
Chloroethane
2-Chloroethanol
Bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Chloroprene
3-Chloropropionitrile
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Di bromomethane
1 , 2-Di chl orobenzene
58
41
56
53
57
76
78
91
136
156
128
83
173
94
74
56
72
91
105
119
76
117
82
48
112
56
129
64 (49*)
49
109
63
83
50 (49*)
53
54
91
91
75
129
107
93
146
43
41, 40, 39
55, 58
52, 51
57, 58, 39
76, 41, 39, 78
-
91, 126, 65, 128
43, 136, 138, 93, 95
77, 158
49, 130
85, 127
175, 254
96
43
41
43, 72
92, 134
134
91, 134
78
119
44, 84, 86, 111
75
77, 114
49
208, 206
66 (51*)
49, 44, 43, 51, 80
111, 158, 160
65, 106
85
52 (51*)
53, 88, 90, 51
54, 49, 89, 91
126
126
155, 157
127
109, 188
95, 174
111, 148
Revision 2
January 1995
-------
TABLE 5
(continued)
Compound
Primary
Characteristic
Ion
8260B - 40
Secondary
Characteristic
Ion(s)
l,2-Dich"lorobenzene-d4
1,3-Dichlorobenzene
1,4-Dichlorobenzene
cis-l,4-Dichloro-2-butene
trans -l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans-l,2-Dichloroethene
1,2-Dichloropropane
1 , 3-Di chl oropropane
2,2-Dichloropropane
1, 3-Di chl oro-2-propanol
1,1-Dichloropropene
cis-1, 3-Di chl oropropene
trans- 1, 3-Di chl oropropene
1 , 2 , 3 , 4-Di epoxybutane
Di ethyl ether
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethyl ene oxide
Ethyl methacrylate
Hexachlorobutadiene
Hexachloroethane
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Isopropyl benzene
p-Isopropyl toluene
Malononitrile
Methacrylonitrile
Methyl acrylate
Methyl -t-butyl ether
Methylene chloride
Methyl ethyl ketone
Methyl iodide
Methyl methacrylate
152
146
146
75
53
85
63
62
96
96
96
63
76
77
79
75
75
75
55
74
88
57
31
88
91
44
69
225
201
43
44
142
43
105
119
66
41
55
73
84
72
142
69
115, 150
111, 148
111, 148
75, 53, 77
88, 75
87
65, 83
98
61, 63
61, 98
61, 98
112
78
97
79, 43, 81
110, 77
77, 39
77, 39
55, 57, 56
45, 59
88, 58, 43
57, 49, 62
45, 27, 46
43, 45, 61
106
44, 43, 42
69, 41, 99
223, 227
166, 199,
, 124,
, 49
, 57
, 51
> 86,
203
89
114
58, 57, 100
44, 43, 42
127, 141
43, 41, 42
120
134, 91
66, 39, 65
41, 67, 39
85
57
86, 49
43
142, 127,
, 53
, 74
, 38
, 52,
141
66
69, 41, 100, 39
Revision 2
January 1995
-------
TABLE 5
(continued)
Compound
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
4-Methyl -2-pentanone
Naphthalene
Nitrobenzene
2-Nitropropane
2-Picoline
Pentachloroethane
Propargyl alcohol
6-Propiolactone
Propionitrile (ethyl cyanide)
n-Propylamine
n-Propyl benzene
Pyridine
Styrene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,1,1-Trichl oroethane
1,1, 2 -Trichl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1, 2, 4-Tri methyl benzene
1 , 3 , 5 -Tri methyl benzene
Vinyl acetate
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
100
128
123
46
93
167
55
42
54
59
91
79
104
180
180
131
83
164
92
97
83
95
151
75
105
105
43
62
106
106
106
43, 58, 85
-
51, 77
-
93, 66, 92, 78
167, 130, 132, 165, 169
55, 39, 38, 53
42, 43, 44
54, 52, 55, 40
59, 41, 39
120
52
78
182, 145
182, 145
133, 119
131, 85
129, 131, 166
91
99, 61
97, 85
97, 130, 132
101, 153
77
120
120
86
64
91
91
91
8260B - 41
Revision 2
January 1995
-------
TABLE 5
(continued)
Primary Secondary
Characteristic Characteristic
Internal Standards/Surrogates Ion Ion(s)
Benzene-d6
Bromobenzene-d5
Bromochl oromethane-d2
1,4-Difluorobenzene
Chlorobenzene-d5
l,4-Dichlorobenzene-d4
l,l,2-Trichloroethane-d3
4-Bromof 1 uorobenzene
Chloroform-d.,
Di bromof 1 uoromethane
Dichloroethane-d4
Toluene-d8
Pentaf 1 uorobenzene
FT uorobenzene
84
82
51
114
117
152
100
95
84
113
102
98
168
96
83
162
131
115,
174,
77
150
176
Characteristic ion for an ion trap mass spectrometer (to be used when ion-
molecule reactions are observed).
8260B - 42 Revision 2
January 1995
-------
TABLE 6
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR
VOLATILE ORGANIC COMPOUNDS IN WATER DETERMINED
WITH A WIDE-BORE CAPILLARY COLUMN
Compound
Benzene
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
l,2-Dibromo-3-Chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichlorobenzene
1,2-Dichlorobenzene
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans-l,2-Dichloroethene
1,2-Dichloropropane
1,3-Dichloropropane
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Isopropylbenzene
p-Isopropyltoluene
Methylene chloride
Cone. Number
Range of
(/jg/L) Samples
0.1 -
0.1 -
0.5 -
0.1 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.1 -
0.5 -
0.5 -
0.5 -
0.1 -
0.1 -
0.5 -
0.1 -
0.5 -
0.5 -
0.1 -
0.5 -
0.2 -
0.5 -
0.5 -
0.1 -
0.1 -
0.5 -
0.1 -
0.1 -
0.1 -
0.5 -
0.5 -
0.1 -
0.5 -
0.5 -
0.1 -
0.1 -
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
31
30
24
30
18
18
18
16
18
24
31
24
24
23
31
31
24
31
24
24
31
24
31
18
24
31
34
18
30
30
31
12
18
31
18
16
23
30
%
Recovery3
97
100
90
95
101
95
100
100
102
84
98
89
90
93
90
99
83
92
102
100
93
99
103
90
96
95
94
101
93
97
96
86
98
99
100
101
99
95
Standard
Deviation
of Recovery13 RSD
6.5
5.5
5.7
5.7
6.4
7.8
7.6
7.6
7.4
7.4
5.8
8.0
5.5
8.3
5.6
8.2
16.6
6.5
4.0
5.6
5.8
6.8
6.6
6.9
5.1
5.1
6.3
6.7
5.2
5.9
5.7
14.6
8.7
8.4
6.8
7.7
6.7
5.0
5.7
5.5
6.4
6.1
6.3
8.2
7.6
7.6
7.3
8.8
5.9
9.0
6.1
8.9
6.2
8.3
19.9
7.0
3.9
5.6
6.2
6.9
6.4
7.7
5.3
5.4
6.7
6.7
5.6
6.1
6.0
16.9
8.9
8.6
6.8
7.6
6.7
5.3
8260B - 43
Revision 2
January 1995
-------
TABLE 6
(continued)
Compound
Cone. Number Standard
Range of % Deviation
(jug/L) Samples Recovery8 of Recovery6 RSD
Naphthalene
n-Propylbenzene
Styrene
1,1,1, Z-Tetrachloroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1,1,2-Tri chloroethane
Trichloroethene
Tri chlorof1uoromethane
1,2,3-Trichloropropane
1,2,4-Trimethylbenzene
1,3,5-Trimethyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
0.1 -100
0.1
0.1
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.5
- 10
-100
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 31
- 10
- 10
31
31
39
24
30
24
18
18
18
18
18
24
24
16
18
23
18
18
31
18
104
100
102
90
91
89
102
109
108
98
104
90
89
108
99
92
98
103
97
104
8.6
5.8
7.3
6.1
5.7
6.0
8.1
9.4
9.0
,9
,6
6.5
7.2
15.6
8.0
6.8
6.5
7.4
6.3
8.0
7.
7.
8.2
5.8
7.2
6.8
5.3
6.8
8.0
8.6
8.3
8.1
7.3
7.3
8.1
14.4
8.1
7.4
6.7
7.2
6.5
7.7
Recoveries were calculated using internal standard method. The internal
standard was fluorobenzene.
Standard deviation was calculated by pooling data from three concentrations.
8260B - 44
Revision 2
January 1995
-------
TABLE 7
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR
VOLATILE ORGANIC COMPOUNDS IN WATER DETERMINED
WITH A NARROW-BORE CAPILLARY COLUMN
Compound
Number
Cone. of
(jug/L) Samples
Standard
% Deviation
Recovery8 of Recovery13
RSD
Benzene
Bromobenzene
Bromochl oromethane
Bromodichl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3 -chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans -1, 2-Di chl oroethene
1, 2-Di chl oropropane
1 , 3-Di chl oropropane
2,2-Dichloropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Isopropyl benzene
p-Isopropyltoluene
Methylene chloride
Naphthalene
n-Propyl benzene
0.1
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.1
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
99
97
97
100
101
99
94
110
110
108
91
100
105
101
99
96
92
99
97
93
97
101
106
99
98
100
95
100
98
96
99
99
102
99
100
102
113
97
98
99
6.2
7.4
5.8
4.6
5.4
7.1
6.0
7.1
2.5
6.8
5.8
5.8
3.2
4.7
4.6
7.0
10.0
5.6
5.6
5.6
3.5
6.0
6.5
8.8
6.2
6.3
9.0
3.7
7.2
6.0
5.8
4.9
7.4
5.2
6.7
6.4
13.0
13.0
7.2
6.6
6.3
7.6
6.0
4.6
5.3
7.2
6.4
6.5
2.3
6.3
6.4
5.8
3.0
4.7
4.6
7.3
10.9
5.7
5.8
6.0
3.6
5.9
6.1
8.9
6.3
6.3
9.5
3.7
7.3
6.3
5.9
4.9
7.3
5.3
6.7
6.3
11.5
13.4
7.3
6.7
8260B - 45
Revision 2
January 1995
-------
TABLE 7
(Continued)
Compound
Styrene
1,1,1, 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1, 2, 4-Tri methyl benzene
1,3,5-Trimethylbenzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
Cone.
(M9/L)
0.5
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.5
0.5
0.5
0.1
0.5
0.5
0.5
Number
of
Samples
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
-------
TABLE 8
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Surrogate Compound
4-Bromofl uorobenzene8
Di bromof 1 uoromethane8
Toluene-d80
Dichloroethane-d4a
Water
86-115
86-118
88-110
80-120
Soil/Sediment
74-121
80-120
81-117
80-120
Single laboratory data, for guidance only.
TABLE 9
QUANTITY OF EXTRACT REQUIRED FOR ANALYSIS OF HIGH CONCENTRATION SAMPLES
Approximate Concentration Range
(M9/kg)
500 -
1,000 -
5,000 -
25,000 -
10,000
20,000
100,000
500,000
Volume of Extract8
100 /iL
50 /iL
10 /iL
100 /iL of 1/50 dilution"
Calculate appropriate dilution factor for concentrations exceeding this table.
a The volume of solvent added to 5 mL of water being purged should be kept
constant. Therefore, add to the 5-mL syringe whatever volume of solvent is
necessary to maintain a volume of 100 fj,i added to the syringe.
b Dilute an aliquot of the solvent extract and then take 100 juL for analysis.
8260B - 47
Revision 2
January 1995
-------
TABLE 10
DIRECT INJECTION ANALYSIS OF NEW OIL AT 5 PPM (METHOD 3585)
Compound Recovery (%)
Acetone
Benzene
n-Butanol*,**
iso-Butanol*,**
Carbon tetrachloride
Carbon disulfide**
Chlorobenzene
Chloroform
1 , 4-Di chl orobenzene
1,2-Dichloroethane
1,1-Dichloroethene
Diethyl ether
Ethyl acetate
Ethyl benzene
Hexachloroethane
Methylene chloride
Methyl ethyl ketone
MIBK
Nitrobenzene
Pyridine
Tetrachloroethene
Tri chl orof 1 uoromethane
1 , 1 , 2 -Tri chl orotri f 1 uoroethane
Toluene
Trichloroethene
Vinyl chloride
o-Xylene
m/p-Xylene
91
86
107
95
86
53
81
84
98
101
97
76
113
83
71
98
79
93
89
31
82
76
69
73
66
63
83
84
%RSD
14.8
21.3
27.8
19.5
44.7
22.3
29.3
29.3
24.9
23.1
45.3
24.3
27.4
30.1
30.3
45.3
24.6
31.4
30.3
35.9
27.1
27.6
29.2
21.9
28.0
35.2
29.5
29.5
Blank
(ppm)
1.9
0.1
0.5
0.9
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.2
0.0
0.0
0.4
0.0
0.0
0.0
0.0
0.0
0.0
0.6
0.0
0.0
0.4
0.6
Spike
(ppm)
5.0
0.5
5.0
5.0
0.5
5.0
5.0
6.0
7.5
0.5
0.7
5.0
5.0
5.0
3.0
5.0
5.0
5.0
2.0
5.0
0.7
5.0
5.0
5.0
0.5
0.2
5.0
10.0
* Alternate mass employed
** IS quantitation
Data are taken from Reference 9.
8260B - 48
Revision 2
January 1995
-------
TABLE 11
SINGLE LABORATORY PERFORMANCE
DATA FOR THE DIRECT INJECTION METHOD - USED OIL (METHOD 3585)
Compound Recovery (%)
Acetone**
Benzene
Benzene-d6
n-Butanol**
iso-Butanol*,**
Carbon tetrachloride
Carbon tetrachloride-13C
Carbon disulfide**
Chlorobenzene
Chlorobenzene-d5
Chloroform
Chloroform-d!
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4
1,2-Dichloroethane**
1,1-Dichloroethene*
l,l-Dichloroethene-d2
Diethyl ether**
Ethyl acetate*,**
Ethyl benzene
Ethylbenzene-d10
Hexachloroethane
Hexachloroethane-13C
Methylene chloride**
Methyl ethyl ketone**
4-Methyl-2-pentanone (MIBK)**
Nitrobenzene
Nitrobenzene-d5
Pyridine**
Pyridine-d5
Tetrachl oroethene**
Trichlorofluoromethane**
l,l,2-Cl3F3ethane**
Toluene
Toluene-d8
Trichl oroethene
Trichloroethene-d,
Vinyl chloride**
105
3135
56
100
132
143
99
95
148
60
149
51
142
53
191
155
68
95
126
1298
63
132
54
86
107
100
111
65
68
ND
101
91
81
2881
63
152
55
100
%RSD
54
44
44
71
27
68
44
63
71
44
74
44
72
44
54
51
44
66
39
44
44
72
45
65
64
74
80
53
85
--
73
70
70
44
44
57
44
69
Blank
(ppm)
2.0
14
2.9
12
0
0
5.1
0
0
3.6
0
2.6
0
3.4
0
0
3.4
0
0
54
3.6
0
3.5
0.3
0
0.1
0
4.0
0
0
0
0
0
128
3.6
0
2.8
0
Spike
(ppm)
5.0
0.5
0.5
5.0
5.0
0.5
0.5
5.0
5.0
5.0
6.0
6.0
7.5
7.5
0.5
0.7
0.7
5.0
5.0
5.0
5.0
3.0
3.0
5.0
5.0
5.0
2.0
2.0
5.0
5.0
0.7
5.0
5.0
5.0
5.0
0.5
0.5
0.2
8260B - 49
Revision 2
January 1995
-------
TABLE 11
(Continued)
Compound
o-Xylene
o-Xylene-d10
m-/p-Xylene
p-Xylene-d10
Recovery (%)
2292
76
2583
67
%RSD
44
44
44
44
Blank
(ppm)
105
4.2
253
3.7
Spike
(ppm)
5.0
5.0
10.0
10.0
* Alternate mass employed
** IS quantitation
ND = Not Detected
Data are based on seven measurements and are taken from Reference 9.
8260B - 50 Revision 2
January 1995
-------
TABLE 12
METHOD DETECTION LIMITS (METHOD 5031)
MDL (Mg/L)
Compound
Macro8
Micro"
Concentration Factor
Macro Micro
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Allyl Alcohol
1-Butanol
Crotonaldehyde
1,4-Dioxane
Ethyl Acetate
Isobutyl alcohol
Methanol
Methyl Ethyl Ketone
2-Methyl-l-propanol
n-Nitroso-di-n-butylamine
Paraldehyde
2-Picoline
1-Propanol
Propionitrile
Pyridine
o-Toluidine
31
57
-
16
7
-
12
12
-
7
38
16
-
14
10
7
-
11
4
13
-
4
12
10
-
2
-
7
11
-
8
-
5
-
-
-
7
2
-
-
25-500
25-500
-
25-500
25-500
-
25-500
25-500
-
25-500
25-500
25-500
-
25-500
25-500
25-500
-
25-500
25-500
25-500
-
200
100
100
-
250
-
150
100
-
140
-
250
-
-
-
240
200
-
-
a Produced by analysis of seven aliquots of reagent water spiked at 25 ppb at
the listed compounds; calculations based on internal standard technique and
use of the following equation:
MDL = 3.134 x Std. Dev. of low concentration spike (ppb).
b Data are from seven aliquots of spiked ground water, using Method 8015.
c When a 40-mL sample is used, and the first 100 /iL of distillate are collected.
8260B - 51
Revision 2
January 1995
-------
TABLE 13
TARGET COMPOUNDS, SURROGATES, AND INTERNAL STANDARDS (METHOD 5031)
Target Compound
Surrogate
Internal Standard
Acetone
Acetonitrile
Acrylonitrile
Ally! alcohol
Crotonaldehyde
1,4-Dioxane
Isobutyl alcohol
Methanol
Methyl ethyl ketone
N-Nitroso-di-n-butyl amine
Paraldehyde
2-Picoline
Propionitrile
Pyridine
o-Toluidine
d6-Acetone
d3-Acetonitrile
d8-Isopropyl alcohol
d7-Dimethyl formamide
d8-Isopropyl alcohol
d8-l,4-Dioxane
d7-Dimethyl formamide
d3-Methanol
d8-Isopropyl alcohol
d7-Dimethyl formamide
d7-Dimethyl formamide
d7-Dimethyl formamide
d8-Isopropyl alcohol
d5-Pyridine
d7-Dimethyl formamide
d8-Isopropyl alcohol
d8-Isopropyl alcohol
d7-Dimethyl formamide
dg-Isopropyl alcohol
d7-Dimethyl formamide
8260B - 52
Revision 2
January 1995
-------
TABLE 14
RECOMMENDED CONCENTRATIONS FOR CALIBRATION SOLUTIONS (METHOD 5031}
Compound
Internal Standards
d5-benzyl alcohol
d14-Diglyme
d7-Dimethyl formamide
d8-Isopropyl alcohol
Surrogates
dg-Acetone
d3-Acetonitrile
dg-l,4-Dioxane
d3-Methanol
d5-Pyridine
Target Compounds
Acetone
Acetonitrile
Acrylonitrile
Allyl alcohol
Crotonaldehyde
1,4-Dioxane
Isobutyl alcohol
Methanol
Methyl ethyl ketone
n-Nitroso-di-n-butylamine
Paraldehyde
2-Picoline
Prop ion itrile
Pyridine
o-Toluidine
Concentration(s) (ng/|LiL)
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
.0,
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
10.
o,
o,
o,
o,
o,
o,
o,
o,
o,
o,
o,
o,
0,
o,
o,
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
25.
o,
o,
o,
o,
o,
0,
0,
0,
0,
0,
o,
o,
o,
o,
o,
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
8260B - 53
Revision 2
January 1995
-------
TABLE 15
CHARACTERISTIC IONS AND RETENTION TIMES FOR VOCs (METHOD 5031)
Quantitation
Compound Ion"
Internal Standards
d8-Isopropyl alcohol
d14-Diglyme
d7-Dimethyl formamide
Surrogates
d6-Acetone
d3-Methanol
dg-Acetonitrile
d8-l,4-Dioxane
d5-Pyridine
d5-Phenolc
Target Compounds
Acetone
Methanol
Methyl ethyl ketone
Methacrylonitrile0
Acrylonitrile
Acetonitrile
Methyl isobutyl ketonec
Propionitrile
Crotonaldehyde
1,4-Dioxane
Paraldehyde
Isobutyl alcohol
Allyl alcohol
Pyridine
2-Picoline
n-Nitroso-di -n-butyl amine
Aniline0
o-Toluidine
Phenol0
49
66
50
46
33
44
96
84
99
43
31
43
67
53
41
85
54
41
58
45
43
57
79
93
84
93
106
94
Secondary
Ions
98,64
80
64,42
35,30
42
64,34
56,79
71
58
29
72,57
41
52,51
40,39
100,58
52,55
70
88,57
89
33,42
39
50,52
66
116
66,92
107
66,65
Retention
Time (min)b
1.75
9.07
9.20
1.03
1.75
2.63
3.97
6.73
15.43
1.05
1.52
1.53
2.38
2.53
2.73
2.78
3.13
3.43
4.00
4.75
5.05
5.63
6.70
7.27
12.82
13.23
13.68
15.43
8 These ions were used for quantitation in selected ion monitoring.
b GC column: DB-Wax, 30 meter x 0.53 mm, 1 ^m film thickness.
Oven program: 45°C for 4 min, increased to 220°C at 120C/min.
0 Compound removed from target analyte list due to poor accuracy and precision.
8260B - 54 Revision 2
January 1995
-------
TABLE 16
METHOD ACCURACY AND PRECISION BY MEAN PERCENT RECOVERY AND PERCENT
RELATIVE STANDARD DEVIATION3 (METHOD 5031 - MACRODISTILLATION TECHNIQUE)
(Single Laboratory and Single Operator)
25 ppb
Compound Mean %R
d6-Acetone
d3-Acetonitrile
d8-l,4-Dioxane
d3-Methanol
d5-Pyridine
Acetone
Acetonitrile
Acrylonitrile
Ally! alcohol
Crotonaldehyde
1,4-Dioxane
Isobutyl alcohol
Methanol
Methyl ethyl ketone
n-Nitroso-di-
n-butyl amine
Paraldehyde
Picoline
Propionitrile
Pyridine
o-Toluidine
66
89
56
43
83
67
44
49
69
68
63
66
50
55
57
65
81
67
74
52
Spike
%RSD
24
18
34
29
6.3
45
35
42
13
22
25
14
36
37
21
20
12
22
7.4
31
100 DDb
Mean %R
69
80
58
48
84
63
52
47
70
68
55
66
46
56
61
66
81
69
72
54
Spike
%RSD
14
18
11
19
7.8
14
15
27
9.7
13
16
5.7
22
20
15
11
6.8
13
6.7
15
500 ppb
Mean %R
65
70
61
56
85
60
56
45
73
69
54
65
49
52
72
60
84
68
74
58
Soike
%RSD
16
10
18
14
9.0
14
15
27
10
13
13
7.9
18
19
18
8.9
8.0
13
7.3
12
Data from analysis of seven aliquots of reagent water spiked at each
concentration, using a quadrapole mass spectrometer in the selected ion
monitoring mode.
8260B - 55
Revision 2
January 1995
-------
TABLE 17
RECOVERIES IN SAND SAMPLES FORTIFIED AT 4 /jg/kg (ANALYSIS BY METHOD 5035)
Recovery per Replicate (nq)
Compound
Vinyl chloride
Trichlorofluoromethane
1,1-Dichloroethene
Methylene chloride
trans- 1,2-Dichloroethene
1,2-Dichloroethane
cis- 1,2-Dichloroethene
Bromochloromethane
Chloroform
1,1,1-Trichloroethane
Carbon tetrachloride
Benzene
Trichloroethene
1,2-Dichloropropane
Dibromomethane
Bromodi chl oromethane
Toluene
1, 1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Chlorobenzene
1,1,1,2-Tetrachloroethane
Ethyl benzene
p-Xylene
o-Xylene
Styrene
Bromoform
iso-Propyl benzene
Bromobenzene
1,2,3-Trichloropropane
n-Propyl benzene
2-Chlorotoluene
4-Chlorotoluene
1 ,3, 5-Tri methyl benzene
sec-Butyl benzene
1 , 2 , 4-Tri methyl benzene
1, 3 -Di chlorobenzene
p- i so-Propyl tol uene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
n-Butyl benzene
1,2, 4-Tri chlorobenzene
Hexachlorobutadiene
1,2, 3 -Tri chlorobenzene
Fluorobenzene (Area)
8
13
17
24
22
18
26
24
26
21
23
22
21
24
25
25
28
25
25
26
22
22
25
22
24
23
26
25
19
25
26
23
21
24
25
30
20
21
18
18
13
14
17
14
1
.0
.3
.1
.5
.7
.3
.1
.5
.5
.5
.6
.4
.5
.9
.4
.7
.3
.4
.4
.3
.9
.4
.6
.5
.2
.9
.8
.3
.9
.9
.0
.6
.0
.0
.9
.6
.3
.6
.1
.4
.1
.5
.6
.9
7056
2
7.
16.
16.
22.
23.
18.
23.
25.
26.
23.
24.
23.
20.
26.
26.
26.
25.
24.
24.
26.
22.
27.
25.
22.
23.
21.
25.
25.
21.
23.
23.
23.
19.
22.
25.
39.
20.
22.
21.
22.
20.
14.
22.
15.
5
5
7
7
6
0
1
4
0
0
2
9
5
3
4
7
0
5
2
2
5
7
0
0
1
5
6
1
8
0
8
8
7
1
3
2
6
1
2
5
3
9
5
9
7087
3
6.7
14.9
15.1
19.7
19.4
16.7
22.6
20.9
22.1
23.9
22.6
20.4
19.2
23.1
21.6
24.1
24.8
21.6
22.7
23.7
19.8
25.1
22.1
19.8
21.6
20.9
26.0
24.2
20.0
25.6
22.6
21.3
18.4
22.5
27.8
22.4
18.2
21.6
20.0
22.5
19.5
15.7
21.6
16.5
6794
4
5.4
13.0
14.8
19.4
18.3
15.6
20.3
20.1
18.9
16.7
18.3
17.4
14.4
19.0
20.4
17.9
16.3
17.7
17.0
18.2
14.6
19.4
14.9
13.9
14.0
14.3
20.1
15.4
15.5
15.9
13.9
13.0
12.1
13.8
16.1
18.0
13.0
16.0
13.2
15.2
10.8
8.8
13.2
11.9
7115
5
6.6
10.3
15.6
20.6
20.1
15.9
20.8
20.1
22.1
31.2
23.3
19.2
19.1
23.3
23.6
23.0
23.6
22.1
22.2
23.2
19.4
22.6
24.0
20.3
20.4
20.5
23.5
24.6
19.1
21.4
21.9
21.5
18.3
22.9
28.6
22.7
17.6
22.8
17.4
19.9
18.7
12.3
21.6
13.9
6215
Mean
6
13
15
21
20
16
22
22
23
23
22
20
18
23
23
23
23
22
22
23
19
23
22
19
20
20
24
22
19
22
21
20
17
21
24
26
17
20
18
19
16
13
19
14
.8
.6
.9
.4
.8
.9
.6
.2
.1
.4
.4
.7
.9
.3
.5
.5
.6
.2
.3
.5
.9
.4
.3
.7
.7
.2
.4
.9
.3
.2
.6
.6
.9
.1
.7
.6
.9
.8
.0
.7
.5
.3
.3
.6
RSD
13.0
15.2
5.7
9.1
0.7
6.4
9.0
10.2
12.2
21.2
9.4
11.2
12.7
10.5
9.6
13.1
16.9
12.1
12.8
12.5
15.0
12.0
17.5
15.7
17.3
15.7
9.9
16.6
10.7
15.8
19.0
19.2
17.1
17.6
18.1
28.2
15.2
11.8
15.3
13.9
23.1
18.8
18.2
11.3
Mean
Rec
34.2
68.0
79.2
107
104
84.4
113
111
116
117
112
103
94.6
117
117
117
118
111
112
118
99.3
117
112
98.5
103
101
122
114
96.3
111
106
103
89.5
105
124
133
89.7
104
90.0
96.6
82.4
66.2
96.3
73.1
Data in Tables 17, 18, and 19 are from Reference 11
8260B - 56
Revision 2
January 1995
-------
TABLE 18
RECOVERIES IN C-HORIZON SOILS FORTIFIED AT 4 jug/kg (ANALYSIS BY METHOD 5035)
Recovery per
Compound
Vinyl chloride
Trichlorofl uoromethane
1,1-Dichloroethene
Methylene chloride
trans- 1,2-Dichloroethene
1,1-Dichloroethane
cis- 1,2-Dichloroethene
Bromochl oromethane
Chloroform
1,1,1-Trichloroethane
Carbon tetrachloride
Benzene
Trichloroethene
1,2-Dichloropropane
Dibromomethane
Bromodichl oromethane
Toluene
1,1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
o-Xylene
Styrene
Bromoform
iso-Propyl benzene
Bromobenzene
1,1,2 , 2-Tetrachl oroethane
1,2,3-Trichloropropane
n-Propyl benzene
2-Chlorotoluene
4-Chlorotoluene
1 ,3 , 5-Trimethyl benzene
sec-Butyl benzene
1 , 2 , 4-Tri methyl benzene
1, 3 -Di chlorobenzene
p- i so-Propyl to! uene
1,4-Di chlorobenzene
1,2-Di chlorobenzene
n-Butyl benzene
1 , 2 , 4-Tri chl orobenzene
Hexachl orobutadi ene
Naphthalene
1,2, 3 -Tri chl orobenzene
Fluorobenzene (Area)
33
37
21
20
21
23
21
22
20
16
13
21
19
21
20
20
22
21
21
20
20
19
21
20
20
18
20
21
20
23
18
20
19
19
20
21
20
17
20
18
18
19
15
18
13
14
1
.4
.7
.7
.9
.8
.8
.6
.3
.5
.4
.1
.1
.6
.8
.9
.9
.2
.0
.4
.9
.8
.5
.1
.0
.7
.3
.1
.0
.4
.3
.4
.4
.1
.0
.8
.4
.5
.6
.5
.5
.4
.6
.2
.7
.9
.9
21100
2
31.
20.
33.
19.
18.
21.
18.
19.
17.
11.
11.
19.
16.
19.
17.
18.
17.
16.
17.
18.
18.
19.
18.
17.
17.
15.
15.
18.
16.
17.
14.
18.
17.
15.
18.
18.
18.
15.
17.
13.
15.
15.
17.
16.
11.
15.
0
8
5
4
9
9
8
5
1
9
3
3
4
0
9
0
3
5
3
1
4
0
3
4
2
9
9
1
2
9
6
9
3
5
0
3
6
9
0
8
0
9
2
2
1
2
23300
3
30.9
20.0
39.8
18.7
20.4
21.3
18.5*
19.3
17.3
10.7
13.0
18.7
16.5
18.3
17.9
18.9
18.8
17.2
18.7
19.0
17.6
17.8
18.5
18.2
16.8
16.2
17.1
19.2
17.2
21.2
15.6
17.9
16.1
16.8
17.4
18.9
16.8
15.6
17.1
14.8
15.4
15.9
17.4
15.5
10.2
16.8
23000
Replicate
4
29.7
21.8
30.2
18.3
17.9
21.3
18.2
19.0
16.5
9.5
11.8
18.2
16.5
18.8
17.2
18.2
17.0
17.2
18.6
18.8
16.8
17.2
16.9
16.3
16.2
15.3
17.5
18.4
16.7
18.8
16.1
17.0
16.0
15.9
16.1
17.0
15.3
14.2
15.6
16.7
15.3
14.4
13.6
13.8
10.8
13.7
22300
(nq)
5
28.
20.
32.
18.
17.
20.
18.
19.
15.
9.
11.
16.
15.
16.
18.
17.
15.
16.
16.
16.
14.
16.
15.
14.
14.
13.
16.
15.
15.
16.
15.
14.
14.
13.
14.
14.
13.
14.
13.
14.
13.
18.
12.
16.
11.
12.
6
5
5
4
8
5
2
2
9
4
2
9
5
5
3
3
9
5
7
6
8
5
3
4
8
7
1
6
4
8
6
3
4
6
7
9
7
4
4
9
5
9
1
6
4
7
Mean
30.8
24.1
31.6
19.1
19.4
21.8
19.0
20.0
17.5
11.6
12.1
18.8
16.9
18.9
18.4
18.6
18.2
17.7
18.5
18.7
17.7
18.0
18.0
17.3
17.1
15.9
17.3
18.4
17.2
19.6
16.1
17.7
16.7
16.4
17.4
18.1
17.0
15.6
16.7
15.7
15.5
16.9
15.1
16.1
11.5
14.7
RSD
5.2
28.2
18.5
5.1
7.9
5.2
6.7
6.0
9.2
22.4
6.7
7.4
8.3
9.0
6.9
6.6
12.0
9.6
8.8
7.5
11.2
6.2
10.6
10.9
11.4
9.3
8.6
9.6
10.1
12.1
8.0
11.6
9.2
10.6
11.7
11.8
14.1
7.9
13.9
10.5
10.5
11.7
13.5
10.0
11.0
9.5
Mean
Rec
154
121
158
95.7
96.8
109
95.2
100
87.3
57.8
60.5
94.1
84.5
94.4
92.1
93.2
91.2
88.4
92.6
93.3
88.4
90.0
90.0
86.3
85.7
79.3
86.7
92.2
85.9
96.0
80.3
88.4
83.6
81.8
86.9
90.5
85.0
77.8
83.6
78.7
77.6
84.6
75.4
80.7
57.4
73.2
21200
8260B - 57
Revision 2
January 1995
-------
TABLE 19
RECOVERIES IN GARDEN SOIL FORTIFIED AT 4 jug/kg (ANALYSIS BY METHOD 5035)
Recovery per
Compound
Vinyl chloride
Trichlorofluoromethane
1,1-Dichloroethene
Methylene chloride
trans-l,2-Dichloroethene
1,1-Dichloroethane
cis-l,2-Dichloroethene
Bromochloromethane
Chloroform
1,1,1-Trichloroethane
Carbon tetrachloride
Benzene
Trichloroethene
1,2-Dichloropropane
Dibromomethane
Bromodichloromethane
Toluene
1,1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
o-Xylene
Styrene
Bromoform
iso-Propyl benzene
Bromobenzene
1,1,2 , 2-Tetrachl oroethane
1,2,3-Trichloropropane
n-Propyl benzene
2-Chlorotoluene
4-Chlorotoluene
1, 3, 5-Tri methyl benzene
sec-Butyl benzene
1 , 2, 4-Tri methyl benzene
1, 3 -Di chlorobenzene
p- i so-Propyl tol uene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
n-Butyl benzene
1,2, 4-Tri chlorobenzene
Hexachlorobutadiene
Naphthalene
1,2, 3 -Tri chlorobenzene
Fluorobenzene (Area)
12
33
27
25
2
24
8
11
16
24
19
21
12
19
7
14
42
13
13
14
8
16
22
41
31
0
8
18
5
14
11
13
8
5
31
13
38
3
14
3
3
17
2
4
5
2
1
.7
.7
.7
.4
.8
.1
.3
.1
.7
.6
.4
.4
.4
.0
.3
.9
.6
.9
.3
.5
.4
.7
.1
.4
.7
.6
.1
.1
.0
.0
.4
.3
.1
.3
.5
.7
.6
.7
.0
.6
.4
.8
.8
.5
.2
16500
2
10.9
6.4
20.5
23.9
3.0
26.3
10.2
11.8
16.9
22.8
20.3
22.0
16.5
18.8
8.0
15.9
39.3
15.2
16.7
13.1
10.0
16.7
21.4
38.4
30.8
0
8.9
18.8
5.4
13.5
12.7
13.3
9.0
5.4
27.5
13.4
32.4
3.6
14.1
3.5
4.3
13.8
2.9
4.0
5.1
2.3
15900
9
30
24
24
3
27
8
10
17
22
22
22
14
19
6
15
45
1
11
14
8
15
23
43
34
0
9
9
5
14
11
14
11
5
33
16
40
3
16
3
4
14
3
6
5
2
3
.8
.3
.1
.7
.3
.0
.7
.2
.0
.1
.2
.4
.9
.7
.9
.9
.1
.4
.3
.5
.3
.6
.1
.8
.3
.1
.7
.3
.7
.7
.7
.7
.5
.0
.4
.8
.7
.1
.3
.0
.0
.3
.1
.5
.4
15600
Replicate (nq)
4
8.1
27.8
15.1
22.2
2.2
20.5
5.8
8.8
13.8
16.2
20.0
19.6
9.0
16.0
5.6
12.8
39.9
21.3
10.9
11.9
6.9
15.8
20.1
38.3
30.4
0
7.0
18.3
4.4
15.3
11.7
12.8
8.7
4.8
31.1
13.8
34.1
3.0
13.9
2.6
3.5
18.9
2.6
5.6
4.7
2.2
17300
5
7.2
22.9
13.2
24.2
2.4
21.2
6.4
9.0
15.0
20.9
20.2
20.4
9.9
17.6
6.8
13.9
45.3
14.9
9.5
14.4
7.8
15.7
22.6
44.0
33.2
0
7.7
19.6
4.0
17.1
11.9
13.9
7.9
4.5
33.6
15.4
40.3
3.2
15.1
2.8
3.6
24.0
3.2
6.0
5.6
2.3
15800
Mean
9.7
24.2
20.1
24.1
2.7
23.8
7.9
10.2
15.9
21.3
20.4
21.2
12.5
18.2
6.9
14.7
42.4
15.9
12.3
13.7
8.3
16.1
21.9
41.2
32.1
0
8.3
18.9
4.8
14.9
11.8
13.6
9.1
5.0
31.3
14.5
37.3
3.4
14.8
3.0
3.8
17.6
3.0
5.3
5.3
2.3
RSD
20
39
26
4
15
11
20
11
7
13
4
4
22
7
11
8
5
17
20
7
12
3
4
6
4
0
9
3
11
8
4
4
14
7
6
8
9
8
5
10
8
21
8
15
6
3
.2
.6
.9
.4
.0
.0
.1
.2
.9
.4
.6
.9
.9
.1
.3
.3
.9
.0
.3
.6
.1
.2
.8
.1
.6
.4
.5
.6
.5
.5
.7
.8
.9
.8
.3
.1
.0
.2
.2
.3
.2
.5
.1
.2
.5
Mean
Rec
48.7
121
101
120
13.6
119
39.4
50.9
79.3
107
102
106
62.7
91.0
34.6
73.3
212
79.6
61.7
68.3
41.3
80.4
109
206
160
0
41.4
94.4
24.1
74.5
59.0
68.1
45.6
25.2
157
72.5
186
17.2
73.8
15.0
19.0
88.0
15.0
26.4
26.5
11.4
8260B - 58
Revision 2
January 1995
-------
TABLE 20
VOLATILE ORGANIC ANALYTE RECOVERY FROM SOIL
USING VACUUM DISTILLATION (METHOD 5032)a
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans -1, 2 -Tri chloroethane
cis-l,2-Dichloroethene
Chloroform
Dichloroethane
Butanone
1,1,1 -Tri chl oroethane
Carbon tetrachloride
Vinyl acetate
Bromodi chloromethane
1,1,2 , 2-Tetrachl oroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1,1,2-Trichloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachl oroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
Soil/H20b
Recovery
Mean RSD
61
58
54
46
60
INTe
47
48
61
54
60
104
177
INT
124
172
88
93
96
105
134
98
119
126
99
123
131
155
152
90
94
98
114
106
97
105
20
20
12
10
2
INT
13
9
6
7
4
11
50
36
13
122
11
4
13
8
10
9
8
10
7
12
13
18
20
9
3
7
13
8
9
8
Soil/Oil0
Recovery
Mean RSD
40
47
46
41
65
44
53
47
58
60
72
93
117
38
72
INT
INT
91
50
102
84
99
125
72
CONTf
94
58
164
185
123
CONT
93
CONT
93
CONT
112
18
13
11
8
8
8
10
5
9
7
6
6
8
INT
16
INT
23
12
6
16
10
31
16
CONT
13
18
19
20
14
CONT
18
CONT
18
CONT
12
Soil/Oil/H20
Recovery
Mean RSD
108
74
72
52
76
47
58
61
56
63
114
151
134
104
104
111
107
100
142
97
112
102
173
169
128
112
112
144
68
13
20
14
11
4
3
6
5
8
151,2
222
26
23
7
6
8
5
16
4
9
9
29
18
7
5
5
13
8260B - 59
Revision 2
January 1995
-------
TABLE 20
(continued)
Compound
Soil/H20b
Recovery
Mean RSD
Soil/Oil0
Recovery
Mean RSD
Soil/Oil/H20
Recovery
Mean RSD
Surrogates
1,2-Dichloroethane
Toluene-d8
Bromofluorobenzene
177
96
139
50
6
13
117
79
37
8
12
13
151
82
62
22
6
5
a Results are for 10 min. distillations times, and condenser temperature held at
-10°C. A 30 m x 0.53 mm ID stable wax column with a 1 jum film thickness was
used for chromatography. Standards and samples were replicated and precision
value reflects the propagated errors. Each analyte was spiked at 50 ppb.
Vacuum distillation efficiencies (Method 5032) are modified by internal
standard corrections. Method 8260 internal standards may introduce bias for
some analytes. See Method 5032 to identify alternate internal standards with
similar efficiencies to minimize bias.
b Soil samples spiked with 0.2 mL water containing analytes and then 5 ml water
added to make slurry.
0 Soil sample + 1 g cod liver oil, spiked with 0.2 mL water containing analytes.
d Soil samples + 1 g cod liver oil, spiked as above with 5 mL of water added to
make slurry.
6 Interference by co-eluting compounds prevented accurate measurement of
analyte.
f Contamination of sample matrix by analyte prevented assessment of efficiency.
8260B - 60 Revision 2
January 1995
-------
TABLE 21
VACUUM DISTILLATION EFFICIENCIES FOR VOLATILE ORGANIC ANALYTES
IN FISH TISSUE (METHOD 5032)a
Compound
Efficiency
Mean (%) RSD (%)
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Trichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Tri chloroethane
Carbon tetrachloride
Vinyl acetate
Bromodichloromethane
1,1,2 , 2-Tetrachl oroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1, 1, 2 -Tri chloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
N/Ab
N/Ab
N/Ab
N/Ab
CONTC
CONTC
79
122
126
109
106
111
117
INTd
106
83
INTd
97
67
117
92
98
71
92
129
102
58
INTd
113
66
CONTC
65
74
57
46
83
36
39
35
46
22
32
27
30
34
22
20
23
22
31
19
20
35
24
19
37
20
19
19
14
13
20
8260B - 61
Revision 2
January 1995
-------
TABLE 21
(continued)
Efficiency
Compound Mean (%) RSD (%)
Surrogates
1,2-Dichloroethane 115 27
Toluene-d8 88 24
Bromofluorobenzene 52 15
Results are for 10 min. distillation times and condenser temperature held at
-10°C. Five replicate 10-g aliquots of fish spiked at 25 ppb were analyzed
using GC/MS external standard quantitation. A 30 m x 0.53 mm ID stable wax
column with a 1 MM film thickness was used for chromatography. Standards were
replicated and results reflect 1 sigma propagated standard deviation.
No analyses.
Contamination of sample matrix by analyte prevented accurate assessment of
analyte efficiency.
Interfering by co-eluting compounds prevented accurate measurement of analyte.
8260B - 62 Revision 2
January 1995
-------
TABLE 22
METHOD DETECTION LIMITS (MDL) FOR VOLATILE ORGANIC ANALYTES
IN FISH TISSUE (METHOD 5032)a
Method Detection Limit (ppb)
Compound
Chl oromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Dichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1 , 1 , 1-Tri chloroethane
Carbon tetrachloride
Vinyl acetate
Bromodi chl oromethane
1,1,2 , 2-Tetrachl oroethane
1,2-Dichloropropane
trans -1,3-Di chl oropropene
Trichloroethene
Di bromochl oromethane
1 , 1 , 2-Tr i chl oroethane
Benzene
cis- 1,3-Di chl oropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
External
Standard Method
7.8
9.7
9.5
9.2
CONTb
CONTb
5.4
4.0
4.0
4.4
4.7
5.6
3.3
INTC
1.1
3.2
I NT0
3.2
4.4
3.8
3.4
3.1
3.5
4.4
3.6
3.5
4.9
7.7
7.5
4.3
3.0
3.3
3.6
3.5
3.7
3.3
Internal
Standard Method
7.3
9.8
9.4
10.0
CONTb
CONTb
4.9
5.7
3.5
4.0
4.1
5.0
3.2
INTC
4.2
3.5
INTC
2.8
3.8
3.7
3.0
4.0
3.2
3.3
3.2
3.0
4.0
8.0
8.0
4.0
2.5
2.8
3.5
3.3
3.5
4.7
Footnotes are on the following page.
8260B - 63 Revision 2
January 1995
-------
TABLE 22
(continued)
Values shown are the average MDLs for studies on three non-consecutive days,
involving seven replicate analyses of 10 g of fish tissue spiked a 5 ppb.
Daily MDLs were calculated as three times the standard deviation.
Quantitation was performed by GC/MS Method 8260 and separation with a 30 m x
0.53 mm ID stable wax column with a 1 /urn film thickness.
Contamination of sample by analyte prevented determination.
Interference by co-eluting compounds prevented accurate quantisation.
8260B - 64 Revision 2
January 1995
-------
TABLE 23
VOLATILE ORGANIC ANALYTES RECOVERY FOR WATER
USING VACUUM DISTILLATION (METHOD 5032)a
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methyl ene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Dichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Trichloroethane
Carbon tetrachloride
Vinyl acetate
Bromodichloromethane
1,1,2, 2 -Tetrachl oroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oroethane
1 , 1 , 2-Tri chl oroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachl oroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
5 mL H20b
Recovery
Mean RSD
114
131
131
110
87
83
138
105
118
105
106
114
104
83
118
102
90
104
85
100
105
98
99
98
97
106
93
60
79
101
100
98
100
98
96
96
27
14
13
15
16
22
17
11
10
11
7
6
6
50
9
6
16
3
17
6
8
4
8
7
4
5
16
17
24
3
6
6
3
4
4
7
20 mL H20C
Recovery
Mean RSD
116
121
120
99
105
65
133
89
119
107
99
104
109
106
109
108
99
110
81
103
105
99
99
100
100
105
94
63
63
97
97
98
92
97
94
95
29
14
16
8
15
34
23
4
11
14
5
8
8
31
9
12
7
5
7
2
4
2
6
4
5
4
8
16
14
7
8
4
8
9
8
6
20 mL H20/0il
Recovery
Mean RSD
176
113
116
96
77
119
99
96
103
96
104
107
144
INTC
113
109
72
99
111
104
92
95
90
76
112
98
57
78
68
77
85
88
73
88
60
72
67
21
23
16
6
68
47
18
25
18
23
21
19
23
27
36
5
43
7
4
5
25
12
10
3
21
23
15
14
5
16
13
16
12
14
8260B - 65
Revision 2
January 1995
-------
TABLE 23
(continued)
5 ml H20b 20 ml H20C 20 mL H20/0il
Recovery Recovery Recovery
Compound Mean RSD Mean RSD Mean RSD
Surrogates
1,2-Dichloroethane
Toluene-d8
Bromof 1 uorobenzene
104
104
106
6
5
6
109
102
106
6
2
9
144
76
40
19
7
8
3 Results are for 10 min. distillation times, and condenser temperature held at
-10"C. A 30 m x 0.53 mm ID stable wax column with a 1 p,m film thickness was
used for chromatography. Standards and samples were replicated and precision
values reflect the propagated errors. Concentrations of analytes were 50 ppb
for 5-mL samples and 25 ppb for 20-mL samples. Recovery data generated with
comparison to analyses of standards without the water matrix.
b Sample contained 1 gram cod liver oil and 20 ml water. An emulsion was
created by adding 0.2 ml of water saturated with lecithin.
0 Interference by co-eluting compounds prevented accurate assessment of
recovery.
8260B - 66 Revision 2
January 1995
-------
TABLE 24
METHOD DETECTION LIMITS (MDL) FOR VOLATILE ORGANIC ANALYTES
USING VACUUM DISTILLATION (METHOD 5032) (INTERNAL STANDARD METHOD)'
Compound
Chi oromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-1, 2 -Dichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Tri chloroethane
Carbon tetrachloride
Vinyl acetate
Bromodichl oromethane
1 , 1 ,2,2-Tetrachloroethane
1,2-Dichloropropane
trans-1, 3 -Dichloropropene
Trichloroethene
Di bromochl oromethane
1,1, 2 -Tri chloroethane
Benzene
ci s - 1 , 3-Di chl oropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
Water"
(M9A)
3.2
2.8
3.5
5.9
3.1
5.6
2.5
2.9
2.2
2.2
2.0
2.4
1.7
7.4
1.8
1.4
11.8
1.6
2.5
2.2
1.5
1.6
1.7
2.1
0.5
1.4
1.8
4.6
3.5
1.4
1.0
1.4
1.5
1.4
1.5
1.7
Soil0
8.0
4.9
6.0
6.0
4.0
CONT9
2.0
3.2
2.0
1.4
2.3
1.8
1.5
INTh
1.7
1.5
INTh
1.4
2.1
2.1
1.7
1.7
1.5
1.7
1.5
1.7
1.5
3.6
4.6
1.6
3.3
1.4
2.8
1.4
2.9
3.4
Tissued
7.3
9.8
9.4
10.0
CONT9
CONT9
4.9
5.7
3.5
4.0
4.1
5.0
3.2
INTh
4.2
3.5
INTh
2.8
3.8
3.7
3.0
4.0
3.2
3.3
3.2
3.0
4.0
8.0
8.0
4.0
2.5
2.8
3.5
3.3
3.5
4.7
Oil6
(mg/kg)
N/Af
N/Af
N/Af
N/Af
0.05
0.06
0.18
0.18
0.14
0.10
0.07
0.07
0.06
INTh
0.10
0.13
INTh
0.06
0.02
0.15
0.05
0.04
0.07
0.05
0.05
0.04
0.05
INTh
INTh
0.10
0.05
0.06
0.04
0.18
0.20
0.07
Footnotes are found on the following page.
8260B - 67
Revision 2
January 1995
-------
TABLE 24
(continued)
8 Quantitation was performed using GC/MS Method 8260 and chromatographic
separation with a 30 m x 0.53 mm ID stable wax column with aim film
thickness. Method detection limits are the average MDLs for studies on
three non-consecutive days.
b Method detection limits are the average MDLs for studies of three non-
consecutive days. Daily studies were seven replicated analyses of 5 mL
aliquots of 4 ppb soil. Daily MDLs were three times the standard deviation.
0 Daily studies were seven replicated analyses of 10 g fish tissue spiked at
5 ppb. Daily MDLs were three times the standard deviation. Quantitation
was performed using GC/MS Method 8260 and chromatographic separation with a
30 m x 0.53 mm ID stable wax column with a 1 /xm film thickness.
d Method detection limits are estimated analyzing 1 g of cod liver oil samples
spiked at 250 ppm. Five replicates were analyzed using Method 8260.
e No analyses.
f Contamination of sample by analyte prevented determination.
9 Interference by co-eluting compounds prevented accurate quantisation.
8260B - 68 Revision 2
January 1995
-------
TABLE 25
METHOD DETECTION LIMITS (MDL) FOR VOLATILE ORGANIC ANALYTES
(METHOD 5032) (EXTERNAL STANDARD METHOD)8
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans -1,2-Dichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Trichloroethane
Carbon tetrachloride
Vinyl acetate
Bromodi chloromethane
1,1,2,2-Tetrachloroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1,1,2-Trichloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
Waterb
(M9/L)
3.1
2.5
4.0
6.1
3.1
33. Of
2.5
3.4
2.3
3.0
2.4
2.7
1.6
57. Of
1.6
1.5
23. Of
2.0
3.6
2.9
2.3
2.5
2.1
2.7
1.7
2.1
2.3
4.6
3.8
1.8
1.8
2.4
2.4
2.0
2.3
2.4
Soilc
(M9A9)
8.6f
4.9f
7.1f
7.5f
3.3
CONTh
3.2
3.8
1.7
3.2
2.7
2.6
1.7
INT
2.4
1.7
INT
2.3
3.2
3.7
2.4
3.0
2.9
2.8
2.9
2.5
2.5
4.6
3.9
2.6
4.4
2.6
4.1
2.5
3.9
4.1
Tissued
tug/kg)
7.8
9.7
9.5
9.2
CONTh
CONTh
5.4
4.0
4.0
4.4
4.7
5.6
3.3
INT
1.1
3.2
INT1
3.2
4.4
3.8
3.8
3.1
3.5
4.4
3.6
3.5
4.9
7.7
7.5
4.3
3.0
3.3
3.6
3.5
3.7
3.3
Oil6
(nig/ kg)
N/A9
N/A9
N/A9
N/Afl
0.08
0.12
0.19
0.19
0.13
0.09
0.08
0.06
0.06
INT
0.08
0.15
INT
0.05
0.09
0.12
0.08
0.06
0.04
0.07
0.03
0.06
0.10
INT
INT
0.12
0.09
0.07
0.09
0.16
0.18
0.08
8260B - 69
Revision 2
January 1995
-------
TABLE 25
(continued)
8 Method detection limits are the average MDLs for studies on three non-
consecutive days. Daily studies were seven replicate analyses of 5-mL
aliquots of water spiked at 4 ppb. Daily MDLs were three times the standard
deviation.
b Daily studies were seven replicate analyses of 5-mL aliquots of water spiked
at 4 ppb.
0 These studies were seven replicate analyses of 5-g aliquots of soil spiked
at 4 ppb.
d These studies were seven replicate analyses of 10-g aliquots of fish tissue
spiked at 5 ppb.
e Method detection limits were estimated by analyzing cod liver oil samples
spiked at 250 ppb. Five replicates were analyzed using Method 8260.
f Method detection limits were estimated by analyzing replicate 50 ppb
standards five times over a single day.
8 No analyses.
h Contamination of sample by analyte prevented determination.
' Interference by co-eluting compound prevented accurate quantitation.
8260B - 70 Revision 2
January 1995
-------
TABLE 26
VOLATILE ORGANIC ANALYTE RECOVERY FROM OIL
USING VACUUM DISTILLATION (METHOD 5032)a
Compound
Recovery
Mean (%) RSD (%)
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Trichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Trichloroethane
Carbon tetrachloride
Vinyl acetate
Bromodi chloromethane
1,1,2 , 2-Tetrachl oroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1, 1, 2 -Tri chloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
N/Ab
N/Ab
N/Ab
N/Ab
62
108
98
97
96
86
99
93
138
INTC
89
129
INTC
106
205
107
98
102
168
95
146
98
94
INTC
INTC
117
108
101
96
120
87
90
32
55
46
24
22
23
11
14
31
14
23
14
46
24
13
8
21
7
10
11
18
22
8
12
10
46
23
10
8260B - 71
Revision 2
January 1995
-------
TABLE 26
(continued)
Recovery
Compound Mean (%) RSD (%)
Surrogates
1,2-Dichloroethane 137 30
Toluene-d8 84 6
Bromofluorobenzene 48 2
Results are for 10 min. distillation times and condenser temperature held at -
10°C. Five replicates of 10-g fish aliquots spiked at 25 ppb were analyzed.
Quantitation was performed with a 30 m x 0.53 mm ID stable wax column with a
1 fj.m film thickness. Standards and samples were replicated and precision
value reflects the propagated errors. Vacuum distillation efficiencies (Method
5032) are modified by internal standard corrections. Method 8260 internal
standards may bias for some analytes. See Method 5032 to identify alternate
internal standards with similar efficiencies to minimize bias.
Not analyzed.
Interference by co-evaluating compounds prevented accurate measurement of
analyte.
8260B - 72 Revision 2
January 1995
-------
TABLE 27
METHOD DETECTION LIMITS (MDL) FOR VOLATILE ORGANIC ANALYTES
IN OIL (METHOD 5032)8
Method Detection Limit (ppb)
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Dichloroethene
cis-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
2-Butanone
1,1,1-Trichloroethane
Carbon tetrachloride
Vinyl acetate
Bromodi chl oromethane
1,1,2,2-Tetrachloroethane
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1 , 1 ,2-Trichloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform
2-Hexanone
4-Methyl -2-pentanone
Tetrachloroethene
Toluene
Chlorobenzene
Ethyl benzene
Styrene
p-Xylene
o-Xylene
External
Standard Method
N/Ab
N/Ab
N/Ab
N/Ab
80
120
190
190
130
90
80
60
60
INTC
80
150
INTC
50
90
120
80
60
40
70
30
60
100
INTC
INTC
120
90
70
90
160
180
80
Internal
Standard Method
N/Ab
N/Ab
N/Ab
N/Ab
50
60
180
180
140
100
70
70
60
INTC
100
130
INTC
60
20
150
50
40
70
50
50
40
50
INTC
INTC
100
50
60
40
180
200
70
8260B - 73
Revision 2
January 1995
-------
TABLE 27
(continued)
Method detection limits are estimated as the result of five replicated
analyses of 1 g cod liver oil spiked at 25 ppb. MDLs were calculated as
three times the standard deviation. Quantitation was performed using a 30
m x 0.53 mm ID stable wax column with a 1 jum film thickness.
No analyses.
Interference by co-eluting compounds prevented accurate quantisation.
8260B - 74 Revision 2
January 1995
-------
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-------
TABLE 29
PRECISION AND MDL DETERMINED FOR ANALYSIS OF FORTIFIED SANDa (METHOD 5021)
Compound
Benzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Dibromomethane
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
tran s-1, 2-Di chl oroethene
1,2-Dichloropropane
1,1-Dichloropropene
cis-l,3-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Methylene chloride
Naphthalene
Styrene
1,1,1 , 2-Tetrachl oroethane
1, 1,2, 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,2, 4 -Tri chl orobenzene
1,1, 1 -Tri chl oroethane
1 , 1 , 2 -Tri chl oroethane
Trichloroethene
% RSD
3.0
3.4
2.4
3.9
11.6
3.6
3.2
5.6
3.1
4.1
5.7
3.2
2.8
3.3
3.4
3.7
3.0
4.5
3.0
3.3
3.2
2.6
2.6
3.2
3.4
4.8
4.1
8.2
16.8
7.9
3.6
2.6
9.8
3.5
4.2
2.7
2.6
2.3
MDL (Mg/kg)
0.34
0.27
0.21
0.30
1.3
0.32
0.24
0.51
0.30
3.5b
0.40
0.29
0.20
0.27
0.24
0.30
0.28
0.41
0.24
0.28
0.27
0.22
0.21
0.30
0.27
0.47
0.38
0.62C
3.4C
0.62
0.27
0.20
1.2C
0.38
0.44
0.27
0.20
0.19
8260B- 76
Revision 2
January 1995
-------
TABLE 29
(continued)
Compound % RSD MDL (jug/kg)
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
Vinyl chloride
m-Xylene/p-Xylene
o-Xylene
2.7
1.5
4.8
3.6
3.6
0.31
0.11
0.45
0.37
0.33
Most compounds spiked at 2 ng/g (2
Incorrect ionization due to methanol
Compound detected in unfortified sand at >1 ng
8260B- 77 Revision 2
January 1995
-------
TABLE 30
RECOVERIES IN GARDEN SOIL FORTIFIED AT 20 /^g/kg (ANALYSIS BY METHOD 5021)
Recovery
Compound Sample 1
Benzene
Bromochloromethane
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
l,2-Dibromo-3-chloro-
propane
1,2-Dibromoethane
Dibromomethane
1 , 2-Di chl orobenzene
1, 3 -Di chlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans -1, 2-Di chl oroethene
1 , 2-Di chl oropropane
1,1-Dichloropropene
cis-l,3-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Methylene chloride
Naphthalene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1, 2, 4-Tri chl orobenzene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
37.6
20.5
21.1
23.8
21.4
27.5
25.6
25.0
21.9
21.0
20.8
20.1
22.2
18.0
21.2
20.1
25.3
23.0
20.6
24.8
21.6
22.4
22.8
26.3
20.3
24.7
23.0
26.0
13.8
24.2
21.4
18.6
25.2
28.6
15.0
28.1
20.8
per Replicate (nq)
Sample 2
35.2
19.4
20.3
23.9
19.5
26.6
25.4
24.4
20.9
19.9
20.8
19.5
21.0
17.7
21.0
20.9
24.1
22.0
19.5
23.8
20.0
21.4
22.2
25.7
19.5
24.5
25.3
25.7
12.7
23.3
20.2
17.8
24.8
27.9
14.4
27.2
19.6
Sample 3
38.4
20.0
22.8
25.1
19.7
28.6
26.4
25.3
21.7
21.3
21.0
20.6
22.8
17.1
20.1
19.9
25.4
22.7
19.8
24.4
21.6
22.2
23.4
28.0
21.1
25.5
25.2
26.1
11.8
23.3
21.3
19.0
26.4
30.9
12.9
29.9
21.7
Mean
(ng)
37.1
20.0
21.4
24.3
20.2
27.6
25.8
24.9
21.5
20.7
20.9
20.1
22.0
17.6
20.8
20.3
24.9
22.6
20.0
24.3
21.1
22.0
22.8
26.7
20.3
24.9
24.5
25.9
12.8
23.6
21.0
18.5
25.5
29.1
14.1
28.4
20.7
Recovery
RSD (%)
3.7
2.3
4.9
2.4
4.2
3.0
1.7
1.5
2.0
2.9
0.5
2.2
3.4
2.1
2.3
2.1
2.4
1.9
2.3
1.7
3.6
2.0
2.1
3.7
3.2
1.7
4.3
0.7
6.4
1.8
2.6
2.7
2.7
4.4
6.3
4.0
4.2
185a
100
107
121
101
138
129
125
108
104a
104
100
110
88.0
104
102
125
113
100
122
105
110
114
133
102
125
123
130a
63. 8a
118
105
92.3
127
146a
70.5
142
104
8260B- 78
Revision 2
January 1995
-------
TABLE 30
(continued)
Recovery per Replicate (nq)
Compound
Trichloroethene
Trichlorofl uoromethane
1,2,3-Trichloropropane
Vinyl chloride
m-Xylene/p-Xylene
o-Xylene
Sample
26.3
25.9
18.8
24.8
24.3
23.1
1 Sample 2
24.9
24.8
18.3
23.2
23.9
22.3
Sample 3
26.8
26.5
19.3
23.9
25.3
23.4
Mean
(ng)
26.0
25.7
18.8
24.0
24.5
22.9
Recovery
RSD
3.1
2.7
2.2
2.7
2.4
2.0
(%)
130
129
94.0
120
123
115
Compound found in unfortified garden soil matrix at >5 ng.
8260B- 79
Revision 2
January 1995
-------
TABLE 31
METHOD DETECTION LIMITS AND BOILING POINTS
FOR VOLATILE ORGANICS (ANALYSIS BY METHOD 5041)'
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
1,1,1-Tri chloroethane
Carbon tetrachloride
Bromodi chl oromethane
1,1,2 , 2-Tetrachl oroethane**
1,2-Dichloropropane
trans-l,3-Dichloropropene
Trichloroethene
Di bromochl oromethane
1,1,2-Trichloroethane
Benzene
cis-l,3-Dichloropropene
Bromoform**
Tetrachloroethene
Toluene
Chlorobenzene^
Ethyl benzene**
Styrene**
Tri chl orof 1 uoromethane
lodomethane
Acrylonitrile
Dibromomethane
1,2,3-Trichloropropane**
total Xylenes**
Detection
Limit (ng)
58
26
14
21
9
35
11
14
12
11
11
13
8
8
11
23
12
17
11
21
26
26
27
26
11
15
15
21
46
17
9
13
14
37
22
Boiling
Point (°C)
-24
4
-13
13
40
56
46
32
57
48
62
83
74
77
88
146
95
112
87
122
114
80
112
150
121
111
132
136
145
24
43
78
97
157
138-144
Footnotes are found on the following page.
8260B- 80 Revision 2
January 1995
-------
TABLE 31
(continued)
* The method detection limit (MDL) is defined in Chapter One. The detection
limits cited above were determined according to 40 CFR, Part 136, Appendix
B, using standards spiked onto clean VOST tubes. Since clean VOST tubes
were used, the values cited above represent the best that the methodology
can achieve. The presence of an emissions matrix will affect the ability of
the methodology to perform at its optimum level.
** Boiling Point greater than 130°C. Not appropriate for quantitative sampling
by Method 0030.
8260B- 81 Revision 2
January 1995
-------
TABLE 32
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION (METHOD 5041)
Bromochloromethane
Acetone
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chioromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
Trichloroethene
trans-l,2-Dichloroethene
lodomethane
Methylene chloride
Trichlorofluoromethane
Vinyl chloride
Chlorobenzene-d5
4-Bromofluorobenzene (surrogate)
Chlorobenzene
Ethyl benzene
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
Toluene-d8 (surrogate)
1,2,3-Trichloropropane
Xylenes
1,4-Di f1uorobenzene
Benzene
Bromodi chloromethane
Bromoform
Carbon tetrachloride
Chlorodi bromomethane
Dibromomethane
1,2-Di chloropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
8260B- 82
Revision 2
January 1995
-------
TABLE 33
METHOD 0040 - COMPOUNDS DEMONSTRATED TO BE APPLICABLE TO THE METHOD
Compound
Di chl orodi f 1 uoromethane
Vinyl chloride
1,3-Butadiene
1, 2-Dichloro-l, 1,2,2-
tetrafluoroethane
Methyl bromide
Tr i chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene chloride
1 , 1 , 2-Tri chl oro-tri f 1 uoroethane
Chloroform
1,1,1-Trichloroethane
Carbon tetrachloride
Benzene
Trichloroethene
1 , 2-Di chl oropropane
Toluene
Tetrachloroethene
Boiling
Point
(°C)
-30
-19
-4
4
4
24
31
40
48
61
75
77
80
87
96
111
121
Condensation
Point
at 20 °C (%)
Gas
Gas
Gas
Gas
Gas
88
22
44
37
21
13
11
10
8
5
3
2
Estimated
Detection
Limit3 (ppm)
0.20
0.11
0.90
0.14
0.14
0.18
0.07
0.05
0.13
0.04
0.03
0.03
0.16
0.04
0.05
0.08
0.03
Since this value represents a direct injection (no concentration) from the
Tedlar® bag, these values are directly applicable as stack detection limits.
8260B- 83
Revision 2
January 1995
-------
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METHOD 8260B
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
CAPILLARY COLUMN TECHNIQUE
713-715 Reference
5000 aeries methods
or Method 0031
7 2 Set GC/MS
op«r«tmg conditions.
7.3.1 Tun. GC/MS
system with BFB
7.3.2 Assemble appropriate
• ample introduction
equipment.
7 2 Set GC/MS
operating conditions.
733 Perform analysis
of standards.
732 Assemble
purge-end-trap device
• nd proper* calibration
standarda
733 Perform purga-
and-lrap indytx of
• tandard*.
7 2 S*l GC/MS
op«r«ting oondition
7 3 1 Tun* GC/MS
• V«t«m with Bf B.
733 Ptrform •ncl
of it*ndard«.
7 3.4 Caleulil* RF
r«lativ« to int»rn«l
• nna«tO«.
73$ Calculate mean HF
and vanfy SPCCa for
minirr>um avaraga raaponaa
factor cntana
730 Caiculata tha SO
and RSO of RF valuaa
for CCCa
I
7 4 Perform GC/MS
calibration verification.
7.5 Perform GC/MS
anelyeie.
701 Identify enelytee
by compering that sempie
and stendard meee
spectra, and retention times.
7 7 Calculate the
concentration of each
identified anelyte
i
r
8260B- 88
Revision 2
January 1995
-------
METHOD 8270C
SEMIVOLATILE ORGANIC COMPOUNDS BY
GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/HS): CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8270 is used to determine the concentration of semivolatile
organic compounds in extracts prepared from many types of solid waste matrices,
soils, air sampling media and water samples. Direct injection of a sample may
be used in limited applications. The following compounds can be determined by
this method:
Compounds
CAS No8
Appropriate Preparation Techniques'"
3510 3520
3540/
3541 3550
3580
Acenaphthene
Acenaphthene-d10 (IS)
Acenaphthylene
Acetophenone
2-Acetylaminofluorene
1-Acetyl-2-thiourea
Aldrin
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
3-Amino-9-ethylcarbazole
Anilazine
Aniline
o-Anisidine
Anthracene
Aramite
1016
1221
1232
1242
1248
1254
1260
83-32-9
Aroclor
Aroclor
Aroclor
Aroclor
Aroclor
Aroclor
Aroclor
Azinphos-methyl
Barban
Benzidine
Benzoic acid
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(g,h,i)perylene
Benzo(a)pyrene
208-
98-
53-
591
309-
117-
60-
92-
132-
101-
62-
90-
120-
140-
12674-
11104-
11141-
53469-
12672-
11097-
11096-
86-
101-
92-
65-
56-
205-
207-
191-
50-
96-8
86-2
96-3
08-2
00-2
79-3
09-3
67-1
32-1
05-3
53-3
04-0
12-7
57-8
11-2
28-2
16-5
21-9
29-6
69-1
82-5
50-0
27-9
87-5
85-0
55-3
99-2
08-9
24-2
32-8
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
HS(43)
X
X
X
X
X
X
X
HS(62)
LR
CP
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
ND
X
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
ND
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
CP
X
X
X
X
X
X
8270C - 1
Revision 3
January 1995
-------
Appropriate Preparation
Compounds
p-Benzoquinone
Benzyl alcohol
a-BHC
/3-BHC
S-BHC
7-BHC (Lindane)
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl ) ether
Bis(2-chloroisopropyl ) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Bromoxynil
Butyl benzyl phthalate
Captafol
Captan
Carbaryl
Carbofuran
Carbophenothion
Chlordane (technical)
Chlorfenvinphos
4-Chloroaniline
Chlorobenzilate
5-Chloro-2-methylanil ine
4-Chloro-3 -methyl phenol
3-(Chloromethyl )pyridine
hydrochloride
1-Chloronaphthalene
2-Chloronaphthalene
2-Chlorophenol
4-Chloro-l ,2-phenylenediamine
4-Chloro- 1,3- phenyl enedi ami ne
4-Chlorophenyl phenyl ether
Chrysene
Chrysene-d12 (IS)
Coumaphos
p-Cresidine
Crotoxyphos
2-Cyclohexyl -4, 6-dinitro -phenol
4,4'-DDD
4,4'-DDE
4,4'-DDT
Demeton-0
Demeton-S
Diallate (cis or trans)
2,4-Diaminotoluene
Dibenz(a,j)acridine
CAS Noa
106-51-4
100-51-6
319-84-6
319-85-7
319-86-8
58-89-9
111-91-1
111-44-4
108-60-1
117-81-7
101-55-3
1689-84-5
85-68-7
2425-06-1
133-06-2
63-25-2
1563-66-2
786-19-6
57-74-9
470-90-6
106-47-8
510-15-6
95-79-4
59-50-7
6959-48-4
90-13-1
91-58-7
95-57-8
95-83-0
5131-60-2
7005-72-3
218-01-9
56-72-4
120-71-8
7700-17-6
131-89-5
72-54-8
72-55-9
50-29-3
298-03-3
126-75-0
2303-16-4
95-80-7
224-42-0
3510
OE
X
X
X
X
X
X
X
X
X
X
X
X
HS(55)
HS(40)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
HS(68)
X
X
DC,OE(42)
X
3520
ND
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
ND
3540/
3541
ND
ND
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
ND
ND
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
ND
Techni
3550
ND
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
ND
ND
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
ND
quesb
3580
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
X
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
8270C - 2
Revision 3
January 1995
-------
Appropriate Preparation
Compounds
Dibenz( a, h) anthracene
Dibenzofuran
Dibenzo(a,e)pyrene
1 , 2-Di bromo-3-chl oropropane
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (IS)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Dieldrin
Diethyl phthalate
Diethylstilbestrol
Diethyl sulfate
Dihydrosaffrole
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl aminoazobenzene
7,12-Dimethylbenz(a)-
anthracene
3,3'-Dimethylbenzidine
a , a-Di methyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4,6-Dinitro-2-methylphenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
Dioxathion
Diphenylamine
5,5-Diphenylhydantoin
1,2-Diphenylhydrazine
Di-n-octyl phthalate
Disulfoton
Endosulfan I
Endosulfan II
CAS Noa
53-70-3
132-64-9
192-65-4
96-12-8
84-74-2
117-80-6
95-50-1
541-73-1
106-46-7
91-94-1
120-83-2
87-65-0
62-73-7
141-66-2
60-57-1
84-66-2
56-53-1
64-67-5
56312-13-1
60-51-5
119-90-4
60-11-7
57-97-6
119-93-7
122-09-8
105-67-9
131-11-3
528-29-0
99-65-0
100-25-4
534-52-1
51-28-5
121-14-2
606-20-2
39300-45-3
88-85-7
78-34-2
122-39-4
57-41-0
122-66-7
117-84-0
298-04-4
959-98-8
33213-65-9
3510
X
X
ND
X
X
OE
X
X
X
X
X
X
X
X
X
X
X
AW,OS(67)
LR
ND
HE,HS(31)
X
X
CP(45)
X
ND
X
X
X
X
HE(14)
X
X
X
X
CP,HS(28)
X
ND
X
X
X
X
X
X
X
3520
X
X
ND
X
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
3540/
3541
X
ND
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
Techni
3550
X
X
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
quesb
3580
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
X
LR
X
CP
X
X
X
X
X
X
X
X
X
X
X
CP
X
ND
X
X
X
X
X
X
X
8270C - 3
Revision 3
January 1995
-------
Appropriate Preparation
Compounds
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
EPN
Ethion
Ethyl carbamate
Ethyl methanesulfonate
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr)
2-Fluorophenol (surr)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachlorophene
Hexachl oropropene
Hexamethyl phosphoramide
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3 -Methyl cholanthrene
4,4'-Methylenebis
(2-chloroanil ine)
4,4'-Methylenebis
(N,N-di methyl anil ine)
Methyl methanesulfonate
2-Methyl naphthalene
Methyl parathion
2-Methyl phenol
CAS Noa
1031-07-8
72-20-8
7421-93-4
53494-70-5
2104-64-5
563-12-2
51-79-6
62-50-0
52-85-7
115-90-2
55-38-9
33245-39-5
206-44-0
86-73-7
321-60-8
367-12-4
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
70-30-4
1888-71-7
680-31-9
123-31-9
193-39-5
465-73-6
78-59-1
120-58-1
143-50-0
21609-90-5
121-75-5
108-31-6
72-33-3
91-80-5
72-43-5
56-49-5
101-14-4
101-61-1
66-27-3
91-57-6
298-00-0
95-48-7
3510
X
X
X
X
X
X
DC(28)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
AW,CP(62)
X
X
ND
X
X
X
DC(46)
X
X
HS(5)
HE
X
X
X
X
OE,OS(0)
X
X
X
X
X
3520
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
X
ND
ND
3540/
3541
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
Techniques'3
3550
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
3580
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
X
X
X
X
8270C - 4
Revision 3
January 1995
-------
Appropriate Preparation
Compounds
3 -Methyl phenol
4-Methylphenol
2-Methylpyridine
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
Naphthalene
Naphthalene-d8 (IS)
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
Nitrobenzene-d5 (surr)
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
Nitroqui no! ine-1 -oxide
N-Ni trosodi -n- butyl ami ne
N-Nitrosodiethylamine
N-Ni trosodimethyl ami ne
N-Nitrosomethylethylamine
N-Ni trosodi phenyl ami ne
N-Ni trosodi -n-propyl amine
N-Nitrosomorphol ine
N-Ni trosopi peri dine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachl orobenzene
Pentachloronitrobenzene
Pentachl orophenol
Perylene-d12 (IS)
Phenacetin
Phenanthrene
Phenanthrene-d10 (IS)
CAS Noa
108-39-4
106-44-5
109-06-8
7786-34-7
315-18-4
2385-85-5
6923-22-4
300-76-5
91-20-3
130-15-4
134-32-7
91-59-8
54-11-5
602-87-9
88-74-4
99-09-2
100-01-6
99-59-2
98-95-3
92-93-3
1836-75-5
88-75-5
100-02-7
99-55-8
56-57-5
924-16-3
55-18-5
62-75-9
10595-95-6
86-30-6
621-64-7
59-89-2
100-75-4
930-55-2
152-16-9
101-80-4
56-38-2
608-93-5
82-68-8
87-86-5
62-44-2
85-01-8
3510
X
X
X
X
HE,HS(68)
X
HE
X
X
X
X
OS(44)
X
DE(67)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
LR
X
X
X
X
X
X
X
X
X
3520
ND
ND
X
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
X
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
X
ND
ND
X
X
ND
X
X
3540/
3541
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
Techni
3550
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
quesb
3580
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
8270C - 5
Revision 3
January 1995
-------
Compounds
Appropriate Preparation Techniquesb
3540/
CAS Noa 3510 3520 3541 3550 3580
Phenobarbital 50-06-6
Phenol 108-95-2
Phenol -d6 (surr)
1,4-Phenylenediamine 106-50-3
Phorate 298-02-2
Phosalone 2310-17-0
Phosmet 732-11-6
Phosphamidon 13171-21-6
Phthalic anhydride 85-44-9
2-Picoline 109-06-8
Piperonyl sulfoxide 120-62-7
Pronamide 23950-58-5
Propylthiouracil 51-52-5
Pyrene 129-00-0
Pyridine 110-86-1
Resorcinol 108-46-3
Safrole 94-59-7
Strychnine 60-41-3
Sulfallate 95-06-7
Terbufos 13071-79-9
Terphenyl-d14(surr) 1718-51-0
1,2,4,5-Tetrachlorobenzene 95-94-3
2,3,4,6-Tetrachlorophenol 58-90-2
Tetrachlorvinphos 961-11-5
Tetraethyl dithiopyrophosphate 3689-24-5
Tetraethyl pyrophosphate 107-49-3
Thionazine 297-97-2
Thiophenol (Benzenethiol ) 108-98-5
Toluene diisocyanate 584-84-9
o-Toluidine 95-53-4
Toxaphene 8001-35-2
2,4,6-Tribromophenol (surr)
1,2,4-Trichlorobenzene 120-82-1
2,4,5-Trichlorophenol 95-95-4
2,4,6-Trichlorophenol 88-06-2
Trifluralin 1582-09-8
2,4,5-Trimethylaniline 137-17-7
Trimethyl phosphate 512-56-1
1,3,5-Trinitrobenzene 99-35-4
Tris(2,3-dibromopropyl) phosphate 126-72
Tri-p-tolyl phosphate 78-32-0
0,0,0-Triethyl phosphorothioate 126-68-1
X
DC(28)
DC(28)
X
X
HS(65)
HS(15)
HE(63)
CP,HE(1)
X
X
X
LR
X
ND
DC,OE(10)
X
AW,OS(55)
X
X
X
X
X
X
X
X
X
X
HE(6)
X
X
X
X
X
X
X
X
HE(60)
X
-7 X
X
X
ND
X
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
CP
ND
X
X
LR
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
Footnotes may be found on the following page.
8270C - 6
Revision 3
January 1995
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KEY TO ANALYTE LIST
a Chemical Abstract Service Registry Number
b See Sec. 1.2 for other acceptable preparation methods.
IS = This compound may be used as an internal standard.
surr = This compound may be used as a surrogate.
AW = Adsorption to walls of glassware during extraction and storage.
CP = Nonreproducible chromatographic performance.
DC = Unfavorable distribution coefficient (number in parenthesis is percent
recovery).
HE = Hydrolysis during extraction accelerated by acidic or basic conditions
(number in parenthesis is percent recovery).
HS = Hydrolysis during storage (number in parenthesis is percent stability).
LR = Low response.
ND = Not determined.
OE = Oxidation during extraction accelerated by basic conditions (number in
parenthesis is percent recovery).
OS = Oxidation during storage (number in parenthesis is percent stability).
X = Greater than 70 percent recovery by this technique.
1.2 In addition to the sample preparation methods listed in the above
analyte list, Method 3542 describes sample preparation for semivolatile organic
compounds in air sampled by Method 0010 (Table 11 contains surrogate performance
data), Method 3545 describes an automated solvent extraction device for
semivolatiles in solids (Table 12 contains performance data), and Method 3561
describes a supercritical fluid extraction of solids for PAHs (see Tables 13, 14,
and 15 for performance data).
1.3 Method 8270 can be used to quantitate most neutral, acidic, and basic
organic compounds that are soluble in methylene chloride and capable of being
eluted, without derivatization, as sharp peaks from a gas chromatographic
fused-silica capillary column coated with a slightly polar silicone. Such
compounds include polynuclear aromatic hydrocarbons, chlorinated hydrocarbons and
pesticides, phthalate esters, organophosphate esters, nitrosamines, haloethers,
aldehydes, ethers, ketones, anilines, pyridines, quinolines, aromatic nitro
compounds, and phenols, including nitrophenols. See Table 1 for a list of
compounds and their characteristic ions that have been evaluated on the specified
GC/MS system.
1.3.1 In most cases, Method 8270 is not appropriate for the
quantitation of multicomponent analytes, e.g., Aroclors, Toxaphene,
technical Chlordane, etc., because of limited sensitivity for those
analytes. When these analytes have been identified by another technique,
Method 8270 is appropriate for confirmation of the presence of these
analytes when concentration in the extract permits.
8270C - 7 Revision 3
January 1995
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1.4 The following compounds may require special treatment when being
determined by this method:
1.4.1 Benzidine may be subject to oxidative losses during solvent
concentration and its chromatographic behavior is poor.
1.4.2 Under the alkaline conditions of the extraction step from
aqueous matrices, a-BHC, 7-BHC, Endosulfan I and II, and Endrin are subject
to decomposition. Neutral extraction should be performed if these
compounds are expected.
1.4.3 Hexachlorocyclopentadiene is subject to thermal decomposition
in the inlet of the gas chromatograph, chemical reaction in acetone
solution, and photochemical decomposition.
1.4.4 N-nitrosodimethylamine is difficult to separate from the
solvent under the chromatographic conditions described.
1.4.5 N-nitrosodiphenylamine decomposes in the gas chromatographic
inlet and cannot be separated from diphenylamine.
1.4.6 Pentachlorophenol, 2,4-dinitrophenol, 4-nitrophenol, benzoic
acid, 4,6-dinitro-2-methylphenol, 4-chloro-3-methylphenol, 2-nitroaniline,
3-nitroaniline, 4-chloroaniline, and benzyl alcohol are subject to erratic
chromatographic behavior, especially if the GC system is contaminated with
high boiling material.
1.4.7 Pyridine may perform poorly at the GC injection port
temperatures specified in the method. Lowering the injection port
temperature may reduce the amount of degradation. The analyst needs to use
caution if modifying the injection port temperature as the performance of
other analytes may be adversely affected.
1.4.8 In addition, analytes in the list provided above are flagged
when there are limitations caused by sample preparation and/or
chromatographic problems for.
1.5 The estimated quantitation limit (EQL) of Method 8270 for determining
an individual compound is approximately 1 mg/kg (wet weight) for soil/sediment
samples, 1-200 mg/kg for wastes (dependent on matrix and method of preparation),
and 10 M9/L f°r ground water samples (see Table 2). EQLs will be
proportionately higher for sample extracts that require dilution to avoid
saturation of the detector.
1.6 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate the
ability to generate acceptable results with this method.
8270C - 8 Revision 3
January 1995
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2.0 SUMMARY OF METHOD
2.1 The samples are prepared for analysis by gas chromatography/mass
spectrometry (GC/MS) using the appropriate sample preparation (refer to Method
3500) and, if necessary, sample cleanup procedures (refer to Method 3600).
2.2 The semivolatile compounds are introduced into the GC/MS by injecting
the sample extract into a gas chromatograph (GC) with a narrow-bore fused silica
capillary column. The GC column is temperature-programmed to separate the
analytes, which are then detected with a mass spectrometer (MS) interfaced to the
gas chromatograph.
2.3 Analytes eluted from the capillary column are introduced into the mass
spectrometer via a jet separator or a direct connection. Identification of
target analytes is accomplished by comparing their mass spectra with the electron
impact (or electron impact-like) spectra of authentic standards. Quantitation
is accomplished by comparing the response of a major (quantitation) ion relative
to an internal standard with a five-point calibration curve.
2.4 The method includes specific calibration and quality control steps
that supersede the general requirements provided in Method 8000.
3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, and spikes must be evaluated
for interferences. Determine if the source of interference is in the preparation
and/or cleanup of the samples and take corrective action to eliminate the
problem.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed with solvent between sample injections. Whenever
an unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross-contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system
4.1.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless injection
and all required accessories, including syringes, analytical columns, and
gases. The capillary column should be directly coupled to the source.
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID) 1 //m film thickness
silicone-coated fused-silica capillary column (J&W Scientific DB-5 or
equivalent).
4.1.3 Mass spectrometer
4.1.3.1 Capable of scanning from 35 to 500 amu every 1 sec
or less, using 70 volts (nominal) electron energy in the electron
8270C - 9 Revision 3
January 1995
-------
impact ionization mode. The mass spectrometer must be capable of
producing a mass spectrum for decafluorotriphenylphosphine (DFTPP)
which meets the criteria in Table 3 when 1 /zL of the GC/MS tuning
standard is injected through the GC (50 ng of DFTPP).
4.1.3.2 An ion trap mass spectrometer may be used if it is
capable of axial modulation to reduce ion-molecule reactions and can
produce electron impact-like spectra that match those in the EPA/NIST
Library. The mass spectrometer must be capable of producing a mass
spectrum for DFTPP which meets the criteria in Table 3 when 5 or 50
ng are introduced.
4.1.4 GC/MS interface - Any GC-to-MS interface may be used that
gives acceptable calibration points at 50 ng per injection for each
compound of interest and achieves acceptable tuning performance criteria.
For a narrow-bore capillary column, the interface is usually
capillary-direct into the mass spectrometer source.
4.1.5 Data system - A computer system should be interfaced to the
mass spectrometer. The system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
the duration of the chromatographic program. The computer should have
software that can search any GC/MS data file for ions of a specific mass
and that can plot such ion abundances versus time or scan number. This
type of plot is defined as an Extracted Ion Current Profile (EICP).
Software should also be available that allows integrating the abundances
in any EICP between specified time or scan-number limits. The most recent
version of the EPA/NIST Mass Spectral Library should also be available.
4.1.6 Guard column (optional) - (J&W Deactivated Fused Silica, 0.25
mm ID x 6 m, or equivalent) between the injection port and the analytical
column joined with column joiners (Hewlett-Packard Catalog No. 5062-3556,
or equivalent).
4.2 Syringe - 10-/A.
4.3 Volumetric flasks, Class A - Appropriate sizes with ground-glass
stoppers.
4.4 Balance - Analytical, capable of weighing 0.0001 g.
4.5 Bottles - glass with Teflon®-!ined screw caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
8270C - 10 Revision 3
January 1995
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5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock standard solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 10-mL
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96% or greater, the weight
may be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.3.2 Transfer the stock standard solutions into bottles with
Teflon®-!ined screw-caps. Store at -10°C or less and protect from light.
Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.3.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10,
chrysene-d12, and perylene-d12 (see Table 5). Other compounds may be used as
internal standards as long as the requirements given in Sec. 7.3.2 are met.
5.4.1 Dissolve 0.200 g of each compound with a small volume of
carbon disulfide. Transfer to a 50 ml volumetric flask and dilute to
volume with methylene chloride so that the final solvent is approximately
20% carbon disulfide. Most of the compounds are also soluble in small
volumes of methanol, acetone, or toluene, except for perylene-d12. The
resulting solution will contain each standard at a concentration of 4,000
ng/juL. Each 1 mL sample extract undergoing analysis should be spiked with
10 juL of the internal standard solution, resulting in a concentration of
40 ng//iL of each internal standard. Store at -10°C or less when not in
use.
5.4.2 If a more sensitive mass spectrometer is employed to achieve
lower detection levels, a more dilute internal standard solution may be
required. Area counts of the internal standard peaks should be between
50-200% of the area of the target analytes in the mid-point calibration
analysis.
5.5 GC/MS tuning standard - A methylene chloride solution containing 50
ng//iL of decafluorotriphenylphosphine (DFTPP) should be prepared. The standard
should also contain 50 ng/jLtL each of 4,4'-DDT, pentachlorophenol, and benzidine
to verify injection port inertness and GC column performance. Store at -10°C or
8270C - 11 Revision 3
January 1995
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less when not in use. If a more sensitive mass spectrometer is employed to
achieve lower detection levels, a more dilute tuning solution may be necessary.
5.6 Calibration standards - A minimum of five calibration standards should
be prepared. One of the calibration standards should be at a concentration near,
but above, the method detection limit and the other standards should correspond
to the range of concentrations found in actual samples but should not exceed the
working range of the GC/MS system. Each standard should contain each analyte for
detection by this method.
5.6.1 It is the intent of EPA that all target analytes for a
particular analysis be included in the calibration standard(s). These
target analytes may not include the entire list of analytes (Sec. 1.1) for
which the method has been demonstrated. However, the laboratory shall not
report a quantitative result for a target analyte that was not included in
the calibration standard(s).
5.6.2 Each 1-mL aliquot of calibration standard should be spiked
with 10 juL of the internal standard solution prior to analysis. All
standards should be stored at -10°C or less, and should be freshly prepared
once a year, or sooner if check standards indicate a problem. The
calibration verification standard should be prepared weekly and stored at
4°C.
5.7 Surrogate standards - The recommended surrogates are phenol-de,
2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-d5, 2-fluorobiphenyl, and
p-terphenyl-d14. See Method 3500 for instructions on preparing the surrogate
solutions.
5.7.1 Determine what the appropriate concentration should be for the
blank extracts after all extraction, cleanup, and concentration steps.
Inject this concentration into the GC/MS to determine recovery of surrogate
standards in all blanks, spikes, and sample extracts. Take into account
all dilutions of sample extracts.
NOTE: Method 3561 (SFE Extraction of PAHs) recommends the use of bromobenzene
and p-quaterphenyl to better cover the range of PAHs listed in the method.
5.7.2 If a more sensitive mass spectrometer is employed to achieve
lower detection levels, a more dilute surrogate solution may be necessary.
5.8 Matrix spike and laboratory control standards - See Method 3500 for
instructions on preparing the matrix spike standard. The same standard may be
used as the laboratory control standard.
5.8.1 Determine what concentration should be in the blank extracts
after all extraction, cleanup, and concentration steps. Inject this
concentration into the GC/MS to determine recovery of all matrix and
laboratory control spikes. Take into account all dilutions of sample
extracts.
8270C - 12 Revision 3
January 1995
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5.8.2 If a more sensitive mass spectrometer is employed to achieve
lower detection levels, a more dilute matrix and LCS spiking solution may
be necessary.
5.9 Acetone, hexane, methylene chloride, isooctane, carbon disulfide,
toluene, and other appropriate solvents - All solvents should be pesticide
quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
6.2 Store the sample extracts at -10°C, protected from light, in screw-cap
vials equipped with unpierced Teflon®-lined septa.
7.0 PROCEDURE
7.1 Sample preparation
7.1.1 Samples are normally prepared by one of the following methods
prior to GC/MS analysis.
Matrix
Air
Water
Soil/sediment
Waste
Methods
3542
3510, 3520, 3535
3540, 3541, 3545, 3550, 3560, 3561
3540, 3541, 3545, 3550, 3560, 3561, 3580
7.1.2 In very limited applications direct injection of the sample
into the GC/MS system with a IQ-^L syringe may be appropriate. The
detection limit is very high (approximately 10,000 M9/L). Therefore, it
is only permitted where concentrations in excess of 10,000 /ug/L are
expected.
7.2 Extract cleanup - Extracts may be cleaned up by any of the following
methods prior to GC/MS analysis.
Analytes of interest Methods
Aniline & aniline derivatives
Phenols
Phthalate esters
Nitrosamines
Organochlorine pesticides & PCBs
Nitroaromatics and cyclic ketones
Polynuclear aromatic hydrocarbons
Haloethers
Chlorinated hydrocarbons
Organophosphorus pesticides
8270C - 13
3620
3630, 3640, 8041a
3610, 3620, 3640
3610, 3620, 3640
3610, 3620, 3630, 3660, 3665
3620, 3640
3611, 3630, 3640
3620, 3640
3620, 3640
3620
Revision 3
January 1995
-------
Analytes of interest
Methods
Petroleum waste
All base, neutral, and acid
priority pollutants
3611, 3650
3640
"Method 8041 includes a derivatization technique followed by GC/ECD
analysis, if interferences are encountered on GC/FID.
7.3 Initial calibration
Establish the GC/MS
recommendations as guidance.
operating conditions, using the following
Mass range:
Scan time:
Initial temperature:
Temperature program:
Final temperature:
Injector temperature:
Transfer line temperature:
Source temperature:
Injector:
Injection volume:
Carrier gas:
Ion trap only:
35-500 amu
1 sec/scan
40°C, hold for 4 minutes
40-270°C at 10°C/min
270°C, hold until benzo[g,h,iJperylene elutes
250-300°C
250-300°C
According to manufacturer's specifications
Grob-type, splitless
1-2 ML
Hydrogen at 50 cm/sec or helium at 30 cm/sec
Set axial modulation, manifold temperature,
and emission current to manufacturer's
recommendations
Split injection is allowed if the sensitivity of the mass spectrometer is
sufficient.
7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 3 for a 50 ng injection of DFTPP. Analyses should not begin until
these criteria are met. Background subtraction should be straightforward
and designed only to eliminate column bleed or instrument background ions.
7.3.1.1 The GC/MS tuning standard solution should also be
used to assess GC column performance and injection port inertness.
Degradation of DDT to DDE and ODD should not exceed 20%. (See Sec.
8.0 of Method 8081 for the percent breakdown calculation). Benzidine
and pentachlorophenol should be present at their normal responses, and
no peak tailing should be visible.
7.3.1.2 If degradation is excessive and/or poor
chromatography is noted, the injection port may require cleaning. It
may also be necessary to break off the first 6-12 in. of the capillary
column. The use of a guard column (Sec. 4.1.6) between the injection
port and the analytical column may help prolong analytical column
performance.
7.3.2 The internal standards selected in Sec. 5.4 should permit most
of the components of interest in a chromatogram to have retention times of
8270C - 14
Revision 3
January 1995
-------
0.80-1.20 relative to one of the internal standards. Use the base peak ion
from the specific internal standard as the primary ion for quantitation
(see Table 1). If interferences are noted, use the next most intense ion
as the quantitation
quantitation).
ion (i.e. for 1,4-dichlorobenzene-d4, use 152 m/z for
7.3.3 Analyze 1-2 jxL of each calibration standard (containing
internal standards) and tabulate the area of the primary characteristic ion
against concentration for each compound (as indicated in Table 1). The
injection volume must be the same for all standards and sample extracts.
Figure 1 shows a chromatogram of a calibration standard containing
base/neutral and acid analytes.
Calculate response factors (RFs) for each compound relative to one of
the internal standards as follows:
RF -
where:
A x C
S IS
A x (f
A8 =
C,s =
Peak area (or height) of the analyte or surrogate.
Peak area (or height) of the internal standard.
Concentration of the analyte or surrogate, in /xg/L.
Concentration of the internal standard, in M9/L.
7.3.4 System performacne check compounds (SPCCs)
A system performance check must be performed to ensure that minimum
average RFs are met before the calibration curve is used. For
semivolatiles, the System Performance Check Compounds (SPCCs) are:
N-nitroso-di-n-propylamine; hexachlorocyclopentadiene; 2,4-dinitrophenol;
and 4-nitrophenol.
The minimum acceptable average RF for these compounds is 0.050. These
SPCCs typically have very low RFs (0.1-0.2) and tend to decrease in
response as the chromatographic system begins to deteriorate or the
standard material begins to deteriorate. They are usually the first to
show poor performance. Therefore, they must meet the minimum requirement
when the system is calibrated.
7.3.4.1 Calculate the mean response factor and the relative
standard deviation (RSD) of the response factors for each compound.
The RSD should be less than 15% for each compound. However, the RSD
for each individual Calibration Check Compound (CCC) (see Table 4)
must be less than 30%.
_ ERF,
mean RF = RF = ^ -
SD =
E(RF,-RF):
1=1
n-1
8270C - 15
Revision 3
January 1995
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SD
RSD = ±1 x 100
RF
7.3.4.2 If the RSD of any CCC is 30% or greater, then the
chromatographic system is too reactive for analysis to begin. Clean
or replace the injector liner and/or capillary column, then repeat the
calibration procedure beginning with Section 7.3.
7.3.4.3 The relative retention time (RRT) of each compound
in each calibration standard should agree within 0.06 RRT units.
Late-eluting compounds usually have much better agreement.
7.3.5 Linearity - If the RSD of any compound is 15% or less, then
the relative response factor is assumed to be constant over the calibration
range, and the average relative response factor may be used for
quantitation (Sec. 7.6.2).
7.3.5.1 If the RSD of any compound is greater than 15%, see
Sec. 7.0 in Method 8000 for options on dealing with non-linear
calibrations. One of the options must be applied to GC/MS calibration
in this situation, or a new initial calibration must be performed.
NOTE: Method 8000 specifies a linearity criterion of 20% RSD. That criterion
pertains to GC and HPLC methods other than GC/MS. Method 8270 requires
15% RSD as evidence of sufficient linearity to employ an average response
factor.
7.3.5.2 When the RSD exceeds 15%, the plotting and visual
inspection of a calibration curve can be a useful diagnostic tool.
The inspection may indicate analytical problems, including errors in
standard preparation, the presence of active sites in the
chromatographic system, analytes that exhibit poor chromatographic
behavior, etc.
7.4 GC/MS calibration verification
7.4.1 Prior to the analysis of samples or calibration standards, the
GC/MS tuning standard must be analyzed. A 50-ng injection of DFTPP must
result in a mass spectrum for DFTPP which meets the criteria in Table 3.
These criteria must be demonstrated at the beginning of each 12-hour shift.
7.4.2 A calibration standard(s) at the mid-point concentration of
the initial calibration range containing all semivolatile analytes,
including all required surrogates, must be analyzed once every 12 hours
during which analyses are performed. Compare the response factors from the
standards analyzed every 12 hours with the SPCC (Sec. 7.4.3) and CCC (Sec.
7.4.4) criteria.
7.4.3 System performance check compounds (SPCCs)
7.4.3.1 A system performance check must be made during every
12-hour analytical shift. Each SPCC compound in the calibration
8270C - 16 Revision 3
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verification standard must meet a minimum response factor of 0.050.
This is the same check that is applied during the initial calibration.
If the minimum response factors are not met, the system must be
evaluated, and corrective action must be taken before sample analysis
begins. Possible problems include standard mixture degradation,
injection port inlet contamination, contamination at the front end of
the analytical column, and active sites in the column or
chromatographic system. This check must be met before sample analysis
begins.
7.4.4 Calibration check compounds (CCCs)
7.4.4.1 After the system performance check is met, the CCCs
listed in Table 4 are used to check the validity of the initial
calibration.
RF - RF
% Difference = v- x 100
RF
where:
RF = Mean response factor from the initial calibration
RFV = Response factor for the CCC from the calibration verification
standard
7.4.4.2 If the percent difference for each CCC is less than
or equal to 20%, then the initial calibration is assumed to be valid.
If the criterion is not met (> 20% difference) for any one CCC, then
corrective action must be taken.
7.4.4.3 Problems similar to those listed under SPCCs could
affect the CCCs. If the problem cannot be corrected by other
measures, a new five-point initial calibration must be generated. The
CCC criteria must be met before sample analysis begins. If the CCCs
are not included in the list of analytes for a project, and therefore
not included in the calibration standards, then all analytes must meet
the 20% difference criterion.
7.4.5 The internal standard responses and retention times in the
calibration verification standard must be evaluated immediately after or
during data acquisition. If the retention time for any internal standard
changes by more than 30 seconds from the last calibration verification (12
hours), then the chromatographic system must be inspected for malfunctions
and corrections must be made, as required. If the EICP area for any of the
internal standards changes by a factor of two (-50% to +100%) from the
previous calibration verification standard, the mass spectrometer must be
inspected for malfunctions and corrections must be made, as appropriate.
When corrections are made, reanalysis of samples analyzed while the system
was malfunctioning is required.
7.5 GC/MS analysis of samples
7.5.1 It is highly recommended that sample extracts be screened on
a GC/FID or GC/PID using the same type of capillary column used in the
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GC/MS system. This will minimize contamination of the GC/MS system from
unexpectedly high concentrations of organic compounds.
7.5.2 Allow the sample extract to warm to room temperature. Just
prior to analysis, add 10 n\_ of the internal standard solution to the 1-mL
concentrated sample extract obtained from sample preparation.
7.5.3 Inject a 1-2 /jL aliquot of the sample extract into the GC/MS
system, using the same operating conditions that were used for the
calibration (Sec. 7.3). The volume to be injected should contain 100 ng
of base/neutral and 200 ng of acid surrogates (assuming 100% recovery),
unless a more sensitive GC/MS system is being used and the surrogate
solution is less concentrated then that listed in Sec. 5.7.
7.5.4 If the response for any quantitation ion exceeds the initial
calibration range of the GC/MS system, the sample extract must be diluted
and reanalyzed. Additional internal standard must be added to the diluted
extract to maintain the same concentration as in the calibration standards
(40 ng/juL, unless a more sensitive GC/MS system is being used).
7.5.5 The use of selected ion monitoring (SIM) is acceptable for
applications requiring detection limits below the normal range of electron
impact mass spectrometry. However, SIM may provide a lesser degree of
confidence in the compound identification unless multiple ions are
monitored for each compound.
7.6 Qualitative analysis
7.6.1. The qual itative identification of compounds determined by this
method is based on retention time and on comparison of the sample mass
spectrum, after background correction, with characteristic ions in a
reference mass spectrum. The reference mass spectrum must be generated by
the laboratory using the conditions of this method. The characteristic
ions from the reference mass spectrum are defined as the three ions of
greatest relative intensity, or any ions over 30% relative intensity, if
less than three such ions occur in the reference spectrum. Compounds are
identified when the following criteria are met.
7.6.1.1 The intensities of the characteristic ions of a
compound must maximize in the same scan or within one scan of each
other. Selection of a peak by a data system target compound search
routine where the search is based on the presence of a target
chromatographic peak containing ions specific for the target compound
at a compound-specific retention time will be accepted as meeting this
criterion.
7.6.1.2 The RRT of the sample component is within ± 0.06 RRT
units of the RRT of the standard component.
7.6.1.3 The relative intensities of the characteristic ions
agree within 30% of the relative intensities of these ions in the
reference spectrum. (Example: For an ion with an abundance of 50%
in the reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
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7.6.1.4 Structural isomers that produce very similar mass
spectra should be identified as individual isomers if they have
sufficiently different GC retention times. Sufficient GC resolution
is achieved if the height of the valley between two isomer peaks is
less than 25% of the sum of the two peak heights. Otherwise,
structural isomers are identified as isomeric pairs.
7.6.1.5 Identification is hampered when sample components
are not resolved chromatographically and produce mass spectra
containing ions contributed by more than one analyte. When gas
chromatographic peaks obviously represent more than one sample
component (i.e., a broadened peak with shoulder(s) or a valley between
two or more maxima), appropriate selection of analyte spectra and
background spectra is important.
7.6.1.6 Examination of extracted ion current profiles of
appropriate ions can aid in the selection of spectra, and in
qual itative identification of compounds. When analytes coelute (i.e.,
only one chromatographic peak is apparent), the identification
criteria may be met, but each analyte spectrum will contain extraneous
ions contributed by the coeluting compound.
7.6.2 For samples containing components not associated with the
calibration standards, a library search may be made for the purpose of
tentative identification. The necessity to perform this type of
identification will be determined by the purpose of the analyses being
conducted. Data system library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other.
For example, the RCRA permit or waste delisting requirements may
require the reporting of non-target analytes. Only after visual comparison
of sample spectra with the nearest library searches may the analyst assign
a tentative identification. Guidelines for tentative identification are:
(1) Relative intensities of major ions in the reference spectrum
(ions > 10% of the most abundant ion) should be present in the
sample spectrum.
(2) The relative intensities of the major ions should agree within ±
20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should be
present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination
or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
8270C - 19 Revision 3
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peaks. Data system library reduction programs can sometimes
create these discrepancies.
7.7 Quantitative analysis
7.7.1 Once a compound has been identified, the quantisation of that
compound will be based on the integrated abundance of the primary
characteristic ion from the EICP.
7.7.2 If the RSD of a compound's response factor is 15% or less,
then the concentration in the extract may be determined using the average
response factor (RF) from initial calibration data (Sec. 7.3.4.1). See
Method 8000, Sec. 7.0, for the equations describing internal standard
calibration and either linear or non-linear calibrations.
7.7.3 Where applicable, the concentration of any non-target analytes
identified in the sample (Sec. 7.6.2) should be estimated. The same
formulae should be used with the following modifications: The areas Ax and
Ais should be from the total ion chromatograms, and the RF for the compound
should be assumed to be 1.
7.7.4 The resulting concentration should be reported indicating:
(1) that the value is an estimate, and (2) which internal standard was used
to determine concentration. Use the nearest internal standard free of
interferences.
7.7.5 Quantitation of multicomponent compounds (e.g., Toxaphene,
Aroclors, etc.) is beyond the scope of Method 8270. Normally, quantitation
is performed using a GC/ECD, by Methods 8081 or 8082. However, Method 8270
may be used to confirm the identification of these compounds, when the
concentrations are at least 10 ng//nL in the concentrated sample extract.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quality control procedures to ensure the proper operation of the
various sample preparation and/or sample introduction techniques can be found in
Method 3500. Each laboratory should maintain a formal quality assurance
program. The laboratory should also maintain records to document the quality of
the data generated.
8.2 Quality control procedures necessary to evaluate the GC system
operation are found in Method 8000, Sec. 7.0 and include evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
In addition, instrument QC requirements may be found in the following sections
of Method 8270:
8.2.1 The GC/MS system must be tuned to meet the DFTPP
specifications in Sees. 7.3.1 and 7.4.1.
8.2.2 There must be an initial calibration of the GC/MS system as
described in Sec. 7.3.
8270C - 20 Revision 3
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8.2.3 The GC/MS system must meet the SPCC criteria specified in Sec.
7.4.3 and the CCC criteria in Sec. 7.4.4, each 12 hours.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch and the
addition of surrogates to each field sample and QC sample.
8.4.1 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time
a set of samples is analyzed or there is a change in reagents, a method
blank should be analyzed as a safeguard against chronic laboratory
contamination. The blanks should be carried through all stages of sample
preparation and measurement.
8.4.2 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.3 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.4.4 See Method 8000, Sec. 8.0 for the details on carrying out
sample quality control procedures for preparation and analysis.
8.5 Surrogate recoveries - The laboratory must evaluate surrogate recovery
data from individual samples versus the surrogate control limits developed by the
laboratory. See Method 8000, Sec. 8.0 for information on evaluating surrogate
data and developing and updating surrogate limits.
8.6 The experience of the analyst performing GC/MS analyses is invaluable
to the success of the methods. Each day that analysis is performed, the
8270C - 21 Revision 3
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calibration verification standard should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal? Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still performing acceptably, the
injector is leaking, the injector septum needs replacing, etc. If any changes
are made to the system (e.g., the column changed), recalibration of the system
must take place.
8.7 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Method 8250 (the packed column version of Method 8270) was tested by
15 laboratories using organic-free reagent water, drinking water, surface water,
and industrial wastewaters spiked at six concentrations ranging from 5 to 1,300
jug/L. Single operator accuracy and precision, and method accuracy were found
to be directly related to the concentration of the analyte and essentially
independent of the sample matrix. Linear equations to describe these
relationships are presented in Table 7. These values are presented as guidance
only and are not intended as absolute acceptance criteria. Laboratories should
generate their own acceptance criteria for capillary column method performance.
(See Method 8000)
9.2 Chromatograms from calibration standards analyzed with Day 0 and Day
7 samples were compared to detect possible deterioration of GC performance.
These recoveries (using Method 3510 extraction) are presented in Table 8.
9.3 Method performance data (using Method 3541 Automated Soxhlet
extraction) are presented in Table 9. Single laboratory accuracy and precision
data were obtained for semivolatile organics in a clay soil by spiking at a
concentration of 6 mg/kg for each compound. The spiking solution was mixed into
the soil during addition and then allowed to equilibrate for approximately 1 hour
prior to extraction. The spiked samples were then extracted by Method 3541
(Automated Soxhlet). Three determinations were performed and each extract was
analyzed by gas chromatography/ mass spectrometry following Method 8270. The low
recovery of the more volatile compounds is probably due to volatilization losses
during equilibration. These data are listed in Table 10 and were taken from
Reference 7.
9.4 Surrogate precision and accuracy data are presented in Table 11 from
a field dynamic spiking study based on air sampling by Method 0010. The trapping
media were prepared for analysis by Method 3542 and subsequently analyzed by
Method 8270.
9.5 Single laboratory precision and bias data (using Method 3545
Accelerated Solvent Extraction) for semivolatile organic compounds are presented
in Table 12. The samples were conditioned spiked samples prepared and certified
by a commercial supplier that contained 57 semivolatile organics at three
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concentrations (250, 2500, and 12,500 ug/kg) on three types of soil (clay, loam
and sand). Spiked samples were extracted both by the Dionex Accelerated Solvent
Extraction system and by Perstorp Environmental Soxtec™ (automated Soxhlet). The
data presented in Table 12 represents seven replicate extractions and analyses
for each individual sample and were taken from reference 9. The average
recoveries from the three matrices for all analytes and all replicates relative
to the automated Soxhlet data are as follows: clay 96.8%, loam 98.7% and sand
102.1%. The average recoveries from the three concentrations also relative to
the automated Soxhlet data are as follows: low-101.2%, mid-97.2% and high-99.2%.
9.6 Single laboratory precision and bias data (using Method 3561 SFE
Extraction of PAHs with a variable restrictor and solid trapping material) were
obtained for the method analytes by the extraction of two certified reference
materials (one, EC-1, a lake sediment from Environment Canada and the other,
HS-3, a marine sediment from the National Science and Engineering Research
Council of Canada, both naturally-contaminated with PAHs). The SFE instrument
used for these extractions was a Hewlett-Packard Model 7680. Analysis was by
GC/MS. Average recoveries from six replicate extractions range from 85 to 148%
(overall average of 100%) based on the certified value (or a Soxhlet value if a
certified value was unavailable for a specific analyte) for the lake sediment.
Average recoveries from three replicate extractions range from 73 to 133%
(overall average of 92%) based on the certified value for the marine sediment.
The data are found in Tables 13 and 14 and were taken from Reference 10.
9.7 Single laboratory precision and accuracy data (using Method 3561 SFE
Extraction of PAHs with a fixed restrictor and liquid trapping) were obtained for
twelve of the method analytes by the extraction of a certified reference material
(a soil naturally contaminated with PAHs). The SFE instrument used for these
extractions was a Dionex Model 703-M. Analysis was by GC/MS. Average recoveries
from four replicate extractions range from 60 to 122% (overall average of 89%)
based on the certified value. Following are the instrument conditions that were
utilized to extract a 3.4 g sample: Pressure - 300 atm; Time - 60 min.;
Extraction fluid - C02; Modifier - 10% 1:1 (v/v) methanol/methylene chloride;
Oven temperature - 80°C; Restrictor temperature - 120°C; and, Trapping fluid -
chloroform (methylene chloride has also been used). The data are found in Table
15 and were taken from Reference 11.
10.0 REFERENCES
1. Eichelberger, J.W., Harris, I.E., and Budde, W.L., "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass Spectrometry
Systems", Analytical Chemistry, 47, 995-1000, 1975.
2. "Method Detection Limit for Methods 624 and 625", Olynyk, P., Budde, W.L.,
and Eichelberger, J.W., unpublished report, October 1980.
3. "Interlaboratory Method Study for EPA Method 625-Base/Neutrals, Acids, and
Pesticides", Final Report for EPA Contract 68-03-3102.
4. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis: Some
Practical Aspects", Journal of the Association of Official Analytical
Chemists (AOAC), 48, 1037, 1965.
8270C - 23 Revision 3
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5. Lucas, S.V., Kornfeld, R.A., "GC-MS Suitability Testing of RCRA Appendix
VIII and Michigan List Analytes ", U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
February 20, 1987, Contract No. 68-03-3224.
6. Engel, T.M., Kornfeld, R.A., Warner, J.S., and Andrews, K.D., "Screening
of Semivolatile Organic Compounds for Extractability and Aqueous Stability
by SW-846, Method 3510", U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268, June
5, 1987, Contract 68-03-3224.
7. Lopez-Avila, V. (W. Beckert, Project Officer); "Development of a Soxtec
Extraction Procedure for Extraction of Organic Compounds from Soils and
Sediments"; U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Las Vegas, NV, October 1991; EPA
600/X-91/140.
8. Bursey, J., Merrill, R., McAllister, R., and McGaughey, J., "Laboratory
Validation of VOST and SemiVOST for Halogenated Hydrocarbons from the Clean
Air Act Amendments List", Vol. 1 and 2, U.S. Environmental Protection
Agency, EPA 600/R-93/123a and b, (NTIS PB 93-227163 and 93-27171), Research
Triangle Park, NC, July 1993.
9. Richter, B., Ezzell, J., and Felix, D., "Single Laboratory Method
Validation Report: Extraction of Target Compound List/Priority Pollutant
List BNAs and Pesticides using Accelerated Solvent Extraction (ASE) with
Analytical Validation by GC/MS and GC/ECD", Document 101124, Dionex
Corporation, Salt Lake City, UT, June 16, 1994.
10. Lee, H.B., Peart, T.E., Hong-You, R.L., and Gere, D.R., "Supercritical
Carbon Dioxide Extraction of Polycyclic Aromatic Hydrocarbons from
Sediments", J. Chromatography, A 653 83-91 (1993).
11. Personal communication from Sue Warner, EPA Region 3, Central Regional
Laboratory, 839 Bestgate Road, Annapolis, MD 21401.
8270C - 24 Revision 3
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TABLE 1
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
2-Picoline
Aniline
Phenol
Bis(2-chloroethyl) ether
2-Chlorophenol
1,3-Dichlorobenzene
l,4-Dichlorobenzene-d4 (IS)
1,4-Dichlorobenzene
Benzyl alcohol
1,2-Dichlorobenzene
N-Ni trosomethylethyl ami ne
Bis(2-chloroisopropyl) ether
Ethyl carbamate
Thiophenol (Benzenethiol)
Methyl methanesulfonate
N-Nitrosodi-n-propyl amine
Hexachloroethane
Maleic anhydride
Nitrobenzene
Isophorone
N-Nitrosodiethylamine
2-Nitrophenol
2,4-Dimethylphenol
p-Benzoquinone
Bis(2-chloroethoxy)methane
Benzoic acid
2,4-Dichlorophenol
Trimethyl phosphate
Ethyl methanesulfonate
1,2,4-Trichlorobenzene
Naphtha!ene-d8 (IS)
Naphthalene
Hexachlorobutadiene
Tetraethyl pyrophosphate
Diethyl sulfate
4-Chloro-3-methyl phenol
2-Methylnaphthalene
2-Methylphenol
Hexachloropropene
Hexachlorocyclopentadi ene
N-Nitrosopyrrolidine
Acetophenone
3,
5.
5.
5.
5.
,75a
,68
,77
.82
.97
6.27
6.35
6.40
6.78
6.85
6.97
.22
,27
,42
.48
.55
.65
.65
.87
8.53
8.70
8.75
9.
9.
03
13
9.23
9.38
9.48
9.53
9.62
9.67
9.75
9.82
10.43
11.07
11.37
11.68
11.87
12.40
12.45
12.60
12.65
12.67
8270C - 25
93 66,92
93 66,65
94 65,66
93 63,95
128 64,130
146 148,111
152 150,115
146 148,111
108 79,77
146 148,111
88 42,88,43,56
45 77,121
62 62,44,45,74
110 110,66,109,84
80 80,79,65,95
70 42,101,130
117 201,199
54 54,98,53,44
77 123,65
82 95,138
102 102,42,57,44,56
139 109,65
122 107,121
108 54,108,82,80
93 95,123
122 105,77
162 164,98
110 110,79,95,109,140
79 79,109,97,45,65
180 182,145
136 68
128 129,127
225 223,227
99 99,155,127,81,109
139 139,45,59,99,111,125
107 144,142
142 141
107 107,108,77,79,90
213 213,211,215,117,106,141
237 235,272
100 100,41,42,68,69
105 71,105,51,120
Revision 3
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TABLE 1
(continued)
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
4-Methylphenol
2,4,6-Trichlorophenol
o-Toluidine
3-Methylphenol
2-Chloronaphthalene
N-Nitrosopiperi dine
1,4-Phenylenediamine
1-Chloronaphthalene
2-Nitroaniline
5-Chloro-2-methyl aniline
Dimethyl phthalate
Acenaphthylene
2,6-Dinitrotoluene
Phthalic anhydride
o-Anisidine
3-Nitroaniline
Acenaphthene-d10 (IS)
Acenaphthene
2,4-Dinitrophenol
2,6-Dinitrophenol
4-Chloroaniline
Isosafrole
Dibenzofuran
2,4-Diaminotoluene
2,4-Dinitrotoluene
4-Nitrophenol
2-Naphthylamine
1,4-Naphthoquinone
p-Cresidine
Dichlorovos
Diethyl phthalate
Fluorene
2,4,5-Trimethyl aniline
N-Nitrosodi-n-butyl amine
4-Chlorophenyl phenyl ether
Hydroquinone
4,6-Dinitro-2-methylphenol
Resorcinol
N-Nitrosodiphenylamine
Safrole
Hexamethyl phosphoramide
3-(Chloromethyl)pyridine hydrochloride
Diphenylamine
12.82
12.85
12.87
12.93
13.30
13.55
13.62
13.65"
13.75
14.28
14.48
14.57
14.62
14.62
15.00
15.02
15.05
15.13
15.35
15.47
15.50
15.60
15.63
15.78
15.80
15.80
16.00a
16.23
16.45
16.48
16.70
16.70
16.70
16.73
16.78
16.93
17.05
17.13
17.17
17.23
17.33
17.50
17.54a
107
196
106
107
162
114
108
162
65
106
163
152
165
104
108
138
164
154
184
162
127
162
168
121
165
139
143
158
122
109
149
166
120
84
204
110
198
110
169
162
135
92
169
8270C - 26
107,108,77,79,90
198,200
106,107,77,51,79
107,108,77,79,90
127,164
42,114,55,56,41
108,80,53,54,52
127,164
92,138
106,141,140,77,89
194,164
151,153
63,89
104,76,50,148
80,108,123,52
108,92
162,160
153,152
63,154
162,164,126,98,63
127,129,65,92
162,131,104,77,51
139
121,122,94,77,104
63,89
109,65
115,116
158,104,102,76,50,130
122,94,137,77,93
109,185,79,145
177,150
165,167
120,135,134,91,77
84,57,41,116,158
206,141
110,81,53,55
51,105
110,81,82,53,69
168,167
162,162,104,77,103,135
135,44,179,92,42
92,127,129,65,39
168,167
Revision 3
January 1995
-------
TABLE 1
(continued)
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
1,2,4,5-Tetrachlorobenzene
1-Naphthylamine
1-Acetyl -2-thiourea
4-Bromophenyl phenyl ether
Toluene diisocyanate
2,4,5-Trichlorophenol
Hexachlorobenzene
Nicotine
Pentachlorophenol
5-Nitro-o-toluidine
Thionazine
4-Nitroaniline
Phenanthrene-d10 (IS)
Phenanthrene
Anthracene
1,4-Dinitrobenzene
Mevinphos
Naled
1,3-Dinitrobenzene
Diallate (cis or trans)
1,2-Dinitrobenzene
Diallate (trans or cis)
Pentachlorobenzene
5-Nitro-o-anisidine
Pentachloronitrobenzene
4-Nitroquinoline-l-oxide
Di-n-butyl phthalate
2,3,4,6-Tetrachlorophenol
Dihydrosaffrole
Demeton-0
Fluoranthene
1,3,5-Trinitrobenzene
Dicrotophos
Benzidine
Trifluralin
Bromoxynil
Pyrene
Monocrotophos
Phorate
Sulfall ate
Demeton-S
Phenacetin
Dimethoate
17.97 216
18.20 143
18.22 118
18.27 248
18.42 174
18.47 196
18.65 284
18.70 84
19.25 266
19.27 152
19.35 107
19.37 138
19.55 188
19.62 178
19.77 178
19.83 168
19.90 127
20.03 109
20.18 168
20.57 86
20.58 168
20.78 86
21.35 250
21.50 168
21.72 237
21.73 174
21.78 149
21.88 232
22.42 135
22.72 88
23.33 202
23.68 75
23.82 127
23.87 184
23.88 306
23.90 277
24.02 202
24.08 127
24.10 75
24.23 188
24.30 88
24.33 108
24.70 87
8270C - 27
216,214,179,108,143,218
143,115,89,63
43,118,42,76
250,141
174,145,173,146,132,91
196,198,97,132,99
142,249
84,133,161,162
264,268
77,152,79,106,94
96,107,97,143,79,68
138,65,108,92,80,39
94,80
179,176
176,179
168,75,50,76,92,122
127,192,109,67,164
109,145,147,301,79,189
168,76,50,75,92,122
86,234,43,70
168,50,63,74
86,234,43,70
250,252,108,248,215,254
168,79,52,138,153,77
237,142,214,249,295,265
174,101,128,75,116
150,104
232,131,230,166,234,168
135,64,77
88,89,60,61,115,171
101,203
75,74,213,120,91,63
127,67,72,109,193,237
92,185
306,43,264,41,290
277,279,88,275,168
200,203
127,192,67,97,109
75,121,97,93,260
188,88,72,60,44
88,60,81,89,114,115
180,179,109,137,80
87,93,125,143,229
Revision 3
January 1995
-------
TABLE 1
(continued)
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
Phenobarbital
Carbofuran
Octamethyl pyrophosphoramide
4-Aminobiphenyl
Dioxathion
Terbufos
a, a -Dimethylphenylami ne
Pronamide
Aminoazobenzene
Dichlone
Dinoseb
Disulfoton
Fluchloralin
Mexacarbate
4,4'-Oxydianiline
Butyl benzyl phthalate
4-Nitrobiphenyl
Phosphamidon
2-Cyclohexyl-4,6-Dinitrophenol
Methyl parathion
Carbaryl
Dimethyl ami noazobenzene
Propylthiouracil
Benz(a)anthracene
Chrysene-d12 (IS)
3,3'-Dichlorobenzidine
Chrysene
Malathion
Kepone
Fenthion
Parathion
Anilazine
Bis(2-ethylhexyl) phthalate
3,3'-Dimethylbenzidine
Carbophenothion
5-Nitroacenaphthene
Methapyrilene
Isodrin
Captan
Chlorfenvinphos
Crotoxyphos
Phosmet
EPN
24.70 204
24.90 164
24.95 135
25.08 169
25.25 97
25.35 231
25.43 58
25.48 173
25.72 197
25.77 191
25.83 211
25.83 88
25.88 306
26.02 165
26.08 200
26.43 149
26.55 199
26.85 127
26.87 231
27.03 109
27.17 144
27.50 225
27.68 170
27.83 228
27.88 240
27.88 252
27.97 228
28.08 173
28.18 272
28.37 278
28.40 109
28.47 239
28.47 149
28.55 212
28.58 157
28.73 199
28.77 97
28.95 193
29.47 79
29.53 267
29.73 127
30.03 160
30.11 157
8270C - 28
204,117,232,146,161
164,149,131,122
135,44,199,286,153,243
169,168,170,115
97,125,270,153
231,57,97,153,103
58,91,65,134,42
173,175,145,109,147
92,197,120,65,77
191,163,226,228,135,193
211,163,147,117,240
88,97,89,142,186
306,63,326,328,264,65
165,150,134,164,222
200,108,171,80,65
91,206
199,152,141,169,151
127,264,72,109,138
231,185,41,193,266
109,125,263,79,93
144,115,116,201
225,120,77,105,148,42
170,142,114,83
229,226
120,236
254,126
226,229
173,125,127,93,158
272,274,237,178,143,270
278,125,109,169,153
109,97,291,139,155
239,241,143,178,89
167,279
212,106,196,180
157,97,121,342,159,199
199,152,169,141,115
97,50,191,71
193,66,195,263,265,147
79,149,77,119,117
267,269,323,325,295
127,105,193,166
160,77,93,317,76
157,169,185,141,323
Revision 3
January 1995
-------
TABLE 1
(continued)
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
Tetrachlorvinphos
Di-n-octyl phthalate
2-Aminoanthraquinone
Barban
Aramite
Benzo(b)fluoranthene
Nitrofen
Benzo(k)fluoranthene
Chiorobenzilate
Fensulfothion
Ethion
Diethylstilbestrol
Famphur
Tri-p-tolyl phosphateb
Benzo(a)pyrene
Perylene-d12 (IS)
7,12-Dimethylbenz(a)anthracene
5,5-Diphenylhydantoin
Captafol
Dinocap
Methoxychlor
2-Acetylaminofluorene
4,4'-Methylenebis(2-chloroaniline)
3,3'-Dimethoxybenzidine
3-Methylcholanthrene
Phosalone
Azinphos-methyl
Leptophos
Mi rex
Tris(2,3-dibromopropyl) phosphate
Dibenz(a,j)acridine
Mestranol
Coumaphos
Indeno(l,2,3-cd)pyrene
Dibenz(a,h)anthracene
Benzo(g,h,i)perylene
1,2:4,5-Di benzopyrene
Strychnine
Piperonyl sulfoxide
Hexachlorophene
Aldrin
Aroclor 1016
Aroclor 1221
30.27 329 109,329,331,79,333
30.48 149 167,43
30.63 223 223,167,195
30.83 222 222,51,87,224,257,153
30.92 185 185,191,319,334,197,321
31.45 252 253,125
31.48 283 283,285,202,139,253
31.55 252 253,125
31.77 251 251,139,253,111,141
31.87 293 293,97,308,125,292
32.08 231 231,97,153,125,121
32.15 268 268,145,107,239,121,159
32.67 218 218,125,93,109,217
32.75 368 368,367,107,165,198
32.80 252 253,125
33.05 264 260,265
33.25 256 256,241,239,120
33.40 180 180,104,252,223,209
33.47 79 79,77,80,107
33.47 69 69,41,39
33.55 227 227,228,152,114,274,212
33.58 181 181,180,223,152
34.38 231 231,266,268,140,195
34.47 244 244,201,229
35.07 268 268,252,253,126,134,113
35.23 182 182,184,367,121,379
35.25 160 160,132,93,104,105
35.28 171 171,377,375,77,155,379
35.43 272 272,237,274,270,239,235
35.68 201 137,201,119,217,219,199
36.40 279 279,280,277,250
36.48 277 277,310,174,147,242
37.08 362 362,226,210,364,97,109
39.52 276 138,227
39.82 278 139,279
41.43 276 138,277
41.60 302 302,151,150,300
45.15 334 334,335,333
46.43 162 162,135,105,77
47.98 196 196,198,209,211,406,408
66 263,220
222 260,292
190 224,260
8270C - 29 Revision 3
January 1995
-------
TABLE 1
(continued)
Compound
Retention Primary Secondary
Time (min) Ion Ion(s)
IS = internal standard
surr = surrogate
"Estimated retention times
Substitute for the non-specific mixture, tricresyl phosphate
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1260
a-BHC
0-BHC
<5-BHC
7-BHC (Lindane)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
1 , 2-Di phenyl hydrazi ne
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
2-Fluorobiphenyl (surr)
2-Fluorophenol (surr)
Heptachlor
Heptachlor epoxide
Nitrobenzene-d5 (surr)
N-Nitrosodimethylamine
Phenol -de (surr)
Terphenyl-d14 (surr)
2,4,6-Tribromophenol (surr)
Toxaphene
190
222
292
292
360
183
181
183
183
235
246
235
79
77
195
337
272
263
67
317
172
112
100
353
82
42
99
244
330
159
224,260
256,292
362,326
362,326
362,394
181,109
183,109
181,109
181,109
237,165
248,176
237,165
263,279
105,182
339,341
339,341
387,422
82,81
345,250
67,319
171
64
272,274
355,351
128,54
74,44
42,71
122,212
332,141
231,233
8270C - 30
Revision 3
January 1995
-------
TABLE 2
ESTIMATED QUANTITATION LIMITS (EQLs) FOR SEMIVOLATILE ORGANICS
Compound
Estimated Quantitation Limits8
Ground water Low Soil/Sediment15
8270C - 31
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetylaminofluorene
1-Acetyl -2-thiourea
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
Anilazine
o-Anisidine
Anthracene
Aramite
Azinphos-methyl
Barban
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzoic acid
Benzo(g,h,i)perylene
Benzo(a)pyrene
p-Benzoquinone
Benzyl alcohol
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl ) ether
Bis(2-chloroisopropyl ) ether
4-bromophenyl phenyl ether
Bromoxynil
Butyl benzyl phthalate
Captafol
Captan
Carbaryl
Carbofuran
Carbophenothion
Chlorfenvinphos
4-Chloroanil ine
Chi orobenzi late
5-Chloro-2-methylanil ine
4- Chi oro -3 -methyl phenol
3-(Chloromethyl )pyridine hydrochloride
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenyl phenyl ether
10
10
10
20
1000
20
10
20
100
10
10
20
100
200
10
10
10
50
10
10
10
20
10
10
10
10
10
10
20
50
10
10
10
20
20
10
10
20
100
10
10
10
660
660
ND
ND
ND
ND
ND
ND
ND
ND
660
ND
ND
ND
660
660
660
3300
660
660
ND
1300
660
660
660
660
ND
660
ND
ND
ND
ND
ND
ND
1300
ND
ND
1300
ND
660
660
660
Revision 3
January 1995
-------
TABLE 2
(continued)
Compound
Estimated Quantitation Limits8
Ground water Low Soil/Sedimentb
M9/L /xg/kg
Chrysene
Coumaphos
p-Cresidine
Crotoxyphos
2-Cyclohexyl -4,6-dinitrophenol
Demeton-0
Demeton-S
Diallate (cis or trans)
Diallate (trans or cis)
2,4-Diaminotoluene
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Dibenzo(a,e)pyrene
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Diethyl phthalate
Diethylstilbestrol
Diethyl sulfate
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl ami noazobenzene
7, 1 2 -Di methyl benz( a) anthracene
3,3'-Dimethylbenzidine
a, a-Di methyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
10
40
10
20
100
10
10
10
10
20
10
10
10
10
10
NA
10
10
10
20
10
10
10
10
10
20
100
20
100
10
10
10
ND
10
10
40
20
40
50
50
10
10
100
20
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
660
660
660
1300
660
ND
ND
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
3300
3300
660
660
ND
ND
8270C - 32
Revision 3
January 1995
-------
TABLE 2
(continued)
Estimated Quantitation Limits3
Ground water Low Soil/Sedimentb
Compound M9/L M9/kg
5,5-Diphenylhydantoin
Di-n-octyl phthalate
Disulfoton
EPN
Ethion
Ethyl carbamate
Bis(Z-ethylhexyl) phthalate
Ethyl methanesulfonate
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl orophene
Hexachl oropropene
Hexamethylphosphoramide
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methylenebis(2-chloroanil ine)
Methyl methanesulfonate
2-Methylnaphthalene
Methyl parathion
2-Methylphenol
3-Methylphenol
4-Methyl phenol
Mevinphos
Mexacarbate
Mi rex
20
10
10
10
10
50
10
20
20
40
10
20
10
10
10
10
10
10
50
10
20
ND
10
20
10
10
20
10
50
NA
20
100
10
10
NA
10
10
10
10
10
10
10
20
10
ND
660
ND
ND
ND
ND
660
ND
ND
ND
ND
ND
660
660
660
660
660
660
ND
ND
ND
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
ND
660
ND
660
ND
ND
ND
8270C - 33 Revision 3
January 1995
-------
TABLE 2
(continued)
Compound
Estimated Quantitation Limits3
Ground water Low Soil/Sedimentb
8270C - 34
Monocrotophos
Naled
Naphthalene
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroanil ine
5-Nitro-o-anisidine
Nitrobenzene
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
4-Nitroquinoline-l-oxide
N-Nitrosodi-n -butyl ami ne
N-Nitrosodi ethyl ami ne
N-Nitrosodiphenylamine
N-Nitroso-di-n-propylamine
N-Nitrosopiperidine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianil ine
Parathion
Pentachlorobenzene
Pentachloronitrobenzene
Pentachlorophenol
Phenacetin
Phenanthrene
Phenobarbital
Phenol
1,4-Phenylenediamine
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
40
20
10
10
10
10
20
10
50
50
20
10
10
10
20
10
50
10
40
10
20
10
10
20
40
200
20
10
10
20
50
20
10
10
10
10
10
100
40
100
100
ND
100
10
ND
ND
660
ND
ND
ND
ND
ND
3300
3300
ND
ND
660
ND
ND
660
3300
ND
ND
ND
ND
660
660
ND
ND
ND
ND
ND
ND
ND
3300
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
Revision 3
January 1995
-------
TABLE 2
(continued)
Estimated Quantitation Limits3
Ground water Low Soil/Sedimentb
Compound iLig/L M9/kg
Propylthiouracil 100 ND
Pyrene 10 660
Pyridine ND ND
Resorcinol 100 ND
Safrole 10 ND
Strychnine 40 ND
Sul fall ate 10 ND
Terbufos 20 ND
1,2,4,5-Tetrachlorobenzene 10 ND
2,3,4,6-Tetrachlorophenol 10 ND
Tetrachlorvinphos 20 ND
Tetraethyl pyrophosphate 40 ND
Thionazine 20 ND
Thiophenol (Benzenethiol ) 20 ND
Toluene diisocyanate 100 ND
o-Toluidine 10 ND
1,2,4-Trichlorobenzene 10 660
2,4,5-Trichlorophenol 10 660
2,4,6-Trichlorophenol 10 660
Trifluralin 10 ND
2,4,5-Trimethylaniline 10 ND
Trimethyl phosphate 10 ND
1,3,5-Trinitrobenzene 10 ND
Tris(2,3-dibromopropyl) phosphate 200 ND
Tri-p-tolyl phosphate(h) 10 ND
0,0,0-Triethyl phosphorothioate NT ND
a Sample EQLs are highly matrix-dependent. The EQLs listed here are provided
for guidance and may not always be achievable.
b EQLs listed for soil/sediment are based on wet weight. Normally, data are
reported on a dry weight basis, therefore, EQLs will be higher based on the
% dry weight of each sample. These EQLs are based on a 30-g sample and gel
permeation chromatography cleanup.
ND = Not Determined
NA= Not Applicable
NT= Not Tested
Other Matrices Factor0
High-concentration soil and sludges by sonicator 7.5
Non-water miscible waste 75
CEQL = (EQL for Low Soil/Sediment given above in Table 2) x (Factor)
8270C - 35 Revision 3
January 1995
-------
TABLE 3
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA3'6
Mass Ion Abundance Criteria
51 30-60% of mass 198
68 < 2% of mass 69
70 < 2% of mass 69
127 40-60% of mass 198
197 < 1% of mass 198
198 Base peak, 100% relative abundance
199 5-9% of mass 198
275 10-30% of mass 198
365 > 1% of mass 198
441 Present but less than mass 443
442 > 40% of mass 198
443 17-23% of mass 442
Data taken from Reference 3.
Alternate tuning criteria may be used (e.g., CLP, Method 525, or
manufacturers' instructions), provided that method performance is not
adversely affected.
TABLE 4
CALIBRATION CHECK COMPOUNDS (CCC)
Base/Neutral Fraction Acid Fraction
Acenaphthene 4-Chloro-3-methyl phenol
1,4-Dichlorobenzene 2,4-Dichlorophenol
Hexachlorobutadiene 2-Nitrophenol
N-Nitrosodiphenylamine Phenol
Di-n-octyl phthalate Pentachlorophenol
Fluoranthene 2,4,6-Trichlorophenol
Benzo(a)pyrene
8270C - 36 Revision 3
January 1995
-------
TABLE 5
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
1,4-Dichlorobenzene-cL
Naphtha!ene-da
Acenaphthene-d
10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl)
ether
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propyl-
amine
Phenol
Phenol-de (surr)
2-Picoline
Acetophenone
Benzoic acid
Bi s(2-chloroethoxy)methane
4-Chloroaniline
4-Chioro-3-methyl phenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a,a-Dimethyl-
phenethylamine
2,4-Dimethylphenol
Hexachlorobutadiene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr)
2-Nitrophenol
N-Nitrosodi-n-butylamine
N-Nitrosopiperi dine
1,2,4-Trichlorobenzene
Acenaphthene
Acenaphthylene
1-Chloronaphthalene
2-Chloronaphthalene
4-Chlorophenyl
phenyl ether
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Fluorene
2-Fluorobiphenyl
(surr)
Hexachlorocyclo-
pentadiene
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
4-Nitrophenol
Pentachlorobenzene
1,2,4,5-Tetra-
chlorobenzene
2,3,4,6-Tetra-
chlorophenol
2,4,6-Tribromo-
phenol (surr)
2,4,6-Trichloro-
phenol
2,4,5-Trichloro-
phenol
(surr) = surrogate
8270C - 37
Revision 3
January 1995
-------
TABLE 5
(Continued)
Phenanthrene-d
10
Chrysene-d12
Perylene-d12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl
ether
Di-n-butyl phthalate
4,6-Dinitro-2-methyl-
phenol
Diphenylamine
Fluoranthene
Hexachlorobenzene
N-Nitrosodiphenylamine
Pentachlorophenol
Pentachloronitrobenzene
Phenacetin
Phenanthrene
Pronamide
Benzidine
Benzo(a)anthracene
Bis(2-ethylhexyl)
phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethylaminoazobenzene
Pyrene
Terphenyl-d14 (surr)
7,12-Dimethylbenz-
(a)anthracene
Di-n-octyl phthalate
Indeno(l,2,3-cd)
pyrene
3-Methylchol-
anthrene
Benzo(b)fluor-
anthene
Benzo(k)fluor-
anthene
Benzo(g,h,i)-
perylene
Benzo(a)pyrene
Dibenz(a,j)acridine
Dibenz(a,hj-
anthracene
(surr) = surrogate
8270C - 38
Revision 3
January 1995
-------
TABLE 6
QC ACCEPTANCE CRITERIA3
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Benzo(b)fl uoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Benzo(g,h,i)perylene
Benzyl butyl phthalate
/3-BHC
S-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl ) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3' -Dichl orobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
27.6
40.2
39.0
32.0
27.6
38.8
32.3
39.0
58.9
23.4
31.5
21.6
55.0
34.5
46.3
41.1
23.0
13.0
33.4
48.3
31.0
32.0
61.6
70.0
16.7
30.9
41.7
32.1
71.4
30.7
26.5
23.2
21.8
29.6
31.4
16.7
32.5
32.8
20.7
37.2
54.7
24.9
Range_
for x
(M9/L)
60.1-132.3
53.5-126.0
7.2-152.2
43.4-118.0
41.8-133.0
42.0-140.4
25.2-145.7
31.7-148.0
D-195.0
D-139.9
41.5-130.6
D-100.0
42.9-126.0
49.2-164.7
62.8-138.6
28.9-136.8
64.9-114.4
64.5-113.5
38.4-144.7
44.1-139.9
D-134.5
19.2-119.7
D-170.6
D-199.7
8.4-111.0
48.6-112.0
16.7-153.9
37.3-105.7
8.2-212.5
44.3-119.3
D-100.0
D-100.0
47.5-126.9
68.1-136.7
18.6-131.8
D-103.5
D-188.8
42.9-121.3
71.6-108.4
D-172.2
70.9-109.4
7.8-141.5
Range
P. Ps
(%)
47-145
33-145
D-166
27-133
33-143
24-159
11-162
17-163
D-219
D-152
24-149
D-110
12-158
33-184
36-166
8-158
53-127
60-118
25-158
17-168
D-145
4-136
D-203
D-227
1-118
32-129
D-172
20-124
D-262
29-136
D-114
D-112
39-139
50-158
4-146
D-107
D-209
26-137
59-121
D-192
26.155
D-152
8270C - 39
Revision 3
January 1995
-------
TABLE 6
(continued)
Compound
Test
cone.
(M9/L)
Limit
for s
(M9/L)
Range_
for x
(M9/L)
Range
P> Ps
(%)
Hexachlorobutadiene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi-n-propylamine
Aroclor 1260
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
4-Chloro-3 -methyl phenol
2-Chlorophenol
2,4-Chlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
26.3
24.5
44.6
63.3
30.1
39.3
55.4
54.2
20.6
25.2
28.1
37.2
28.7
26.4
26.1
49.8
93.2
35.2
47.2
48.9
22.6
31.7
37.8-102.2
55.2-100.0
D-150.9
46.6-180.2
35.6-119.6
54.3-157.6
13.6-197.9
19.3-121.0
65.2-108.7
69.6-100.0
57.3-129.2
40.8-127.9
36.2-120.4
52.5-121.7
41.8-109.0
D-172.9
53.0-100.0
45.0-166.7
13.0-106.5
38.1-151.8
16.6-100.0
52.4-129.2
24-116
40-113
D-171
21-196
21-133
35-180
D-230
D-164
54-120
52-115
44-142
22-147
23-134
39-135
32-119
D-191
D-181
29-182
D-132
14-176
5-112
37-144
Standard deviation of four recovery measurements, in jug/L
Average recovery for four recovery measurements, in
Measured percent recovery
s
x
P, Ps =
D = Detected; result must be greater than zero
a
Criteria from 40 CFR Part 136 for Method 625, using a packed GC column.
These criteria are based directly on the method performance data in
Table 7. Where necessary, the limits for recovery have been broadened
to assure applicability of the limits to concentrations below those used
to develop Table 7. These values are for guidance only. Appropriate
derivation of acceptance criteria for capillary columns should result in
much narrower ranges. See Method 8000 for information on developing and
updating acceptance criteria for method performance.
8270C - 40
Revision 3
January 1995
-------
TABLE 7
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION6
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo(g,h,i)perylene
Benzyl butyl phthalate
/3-BHC
5-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl )
ether
Bis(2-ethylhexyl)
phthalate
4-Bromophenyl phenyl
ether
2-Chl oronaphthal ene
4-Chlorophenyl phenyl
ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Accuracy, as
recovery, x'
(M9/L)
0.96C+0.19
0.89C+0.74
0.78C+1.66
0.80C+0.68
0.88C-0.60
0.93C-1.80
0.87C-1.56
0.90C-0.13
0.98C-0.86
0.66C-1.68
0.87C-0.94
0.29C-1.09
0.86C-1.54
1.12C-5.04
1.03C-2.31
0.84C-1.18
0.91C-1.34
0.89C+0.01
0.91C+0.53
0.93C-1.00
0.56C-0.40
0.70C-0.54
0.79C-3.28
0.88C+4.72
0.59C4-0.71
0.80C+0.28
0.86C-0.70
0.73C-1.47
1.23C-12.65
0.82C-0.16
0.43C+1.00
0.20C+1.03
0.92C-4.81
1.06C-3.60
0.76C-0.79
0.39C+0.41
0.76C-3.86
Single analyst
precision, s/
(M9/L)
0.15X-0.12
0.24X-1.06
0.27X-1.28
0.21X-0.32
0.15X+0.93
0.22X+0.43
0.19X+1.03
0.22X+0.48
0.29X+2.40
O.lSx+0.94
0.20X-0.58
0.34X+0.86
0.35x-0.99
0.16x4-1.34
0.24x+0.28
0.26x+0.73
0.13x+0.66
0.07X+0.52
0.20X-0.94
0.28x4-0.13
0.29X-0.32
0.26x-1.17
0.42X+0.19
0.30X+8.51
0.13x4-1.16
0.20x+0.47
0.25x+0.68
0.24x+0.23
0.28x4-7.33
0.20X-0.16
0.28x4-1.44
0.54x4-0.19
0.12X+1.06
0.14x4-1.26
0.21X+1.19
0.12X+2.47
O.lSx+3.91
Overall
precision,
S' (M9/L)
0.21X-0.67
0.26X-0.54
0.43x4-1.13
0.27X-0.64
0.26X-0.21
0.29x4-0.96
0.35X+0.40
0.32x4-1.35
0.51X-0.44
0.53x4-0.92
0.30X+1.94
0.93X-0.17
0.35X+0.10
0.26x4-2.01
0.25x4-1.04
0.36x4-0.67
0.16X+0.66
0.13X+0.34
0.30X-0.46
0.33X-0.09
0.66X-0.96
0.39X-1.04
0.65X-0.58
0.59X+0.25
0.39x4-0.60
0.24X+0.39
0.41x4-0.11
0.29x4-0.36
0.47x4-3.45
0.26X-0.07
0.52x4-0.22
1.05X-0.92
0.21X+1.50
0.19x4-0.35
0.37x4-1.19
0.63X-1.03
0.73X-0.62
8270C - 41
Revision 3
January 1995
-------
TABLE 7
(continued)
Compound
Fluoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi-n-propylamine
Aroclor 1260
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
4 -Chi oro -3 -methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-THchlorophenol
Accuracy, as
recovery, x'
Ug/L)
0.81C+1.10
0.90C-0.00
0.87C-2.97
0.92C-1.87
0.74C+0.66
0.71C-1.01
0.73C-0.83
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.22
0.81C-10.86
0.87C+0.06
0.84C-0.16
0.94C-0.79
0.84C+0.35
0.78C+0.29
0.87C-0.13
0.71C+4.41
0.81C-18.04
1.04C-28.04
0.07C-1.15
0.61C-1.22
0.93C+1.99
0.43C+1.26
0.91C-0.18
Single analyst
precision, s/
(M9/L)
0.22X-0.73
0.12X+0.26
0.24X-0.56
0.33x-0.46
O.lSx-0.10
0.19X+0.92
0.17X+0.67
0.29x+1.46
0.27X+0.77
0.21X-0.41
0.19X+0.92
0.27X+0.68
0.35X+3.61
0.12x+0.57
0.16X+0.06
O.lSx+0.85
0.23x+0.75
O.lSx+1.46
O.lBx+1.25
0.16X+1.21
0.38X+2.36
O.lOx+42.29
O.lSx+1.94
0.38x+2.57
0.24x+3.03
0.26X+0.73
0.16x+2.22
Overall
precision,
S' (M9/L)
0.28X-0.60
0.13X+0.61
O.BOx-0.23
0.28X+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
O.lSx+0.25
0.15X+0.31
0.21X+0.39
0.29X+1.31
0.28X+0.97
0.21X+1.28
0.22X+1.31
0.42X+26.29
0.26X+23.10
0.27X+2.60
0.44X+3.24
0.30X+4.33
0.35X+0.58
0.22X+1.81
x' =
s/ =
S' =
C =
x =
Expected recovery for one or more measurements of a sample containing a
concentration of C, in
Expected single analyst standard deviation of measurements at an average
concentration of x, in /^g/L.
Expected inter! aboratory_standard deviation of measurements at an average
concentration found of x, in /xg/L.
True value for the concentration, in ng/L.
Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8270C - 42
Revision 3
January 1995
-------
TABLE 8
EXTRACTION EFFICIENCY AND AQUEOUS STABILITY RESULTS
Percent Recovery Percent Recovery
Compound on Day 0 on Day 7
Mean RSD Mean RSD
3-Amino-9-ethylcarbazole 80 8 73 3
4-Chloro-l,2-phenylenediamine 91 1 108 4
4-Chloro-l,3-phenylenediamine 84 3 70 3
l,2-Dibromo-3-chloropropane 97 2 98 5
Dinoseb 99 3 97 6
Parathion 100 2 103 4
4,4'-Methylenebis(N,N-dimethylaniline) 108 4 90 4
5-Nitro-o-toluidine 99 10 93 4
2-Picoline 80 4 83 4
Tetraethyl dithiopyrophosphate 92 7 70 1
Data taken from Reference 6.
8270C - 43 Revision 3
January 1995
-------
TABLE 9
MEAN PERCENT RECOVERIES AND PERCENT RSD VALUES FOR SEMIVOLATILE ORGANICS FROM
SPIKED CLAY SOIL AND TOPSOIL BY AUTOMATED SOXHLET EXTRACTION (METHOD 3541)
WITH HEXANE-ACETONE (1:1)'
Compound
Clay Soil
Mean
Recovery RSD
Topsoil
Mean
Recovery RSD
1,3-Dichlorobenzene
1,2-Dichlorobenzene
Nitrobenzene
Benzal chloride
Benzotrichloride
4-Chloro-2-nitrotoluene
Hexachl orocycl opentadi ene
2,4-Dichloronitrobenzene
3,4-Dichloronitrobenzene
Pentachlorobenzene
2,3,4 , 5-Tetrachl oron i trobenzene
Benefin
alpha-BHC
Hexachl orobenzene
delta-BHC
Heptachlor
Aldrin
Isopropalin
Heptachlor epoxide
trans-Chlordane
Endosulfan I
Dieldrin
2,5-Dichlorophenyl -4-nitrophenyl ether
Endrin
Endosulfan II
p,p'-DDT
2,3,6-Trichlorophenyl-
4'-nitrophenyl ether
2,3,4-Trichlorophenyl-
4'-nitrophenyl ether
Mi rex
0
0
0
0
0
0
4.1
35.2
34.9
13.7
55.9
62.6
58.2
26.9
95.8
46.9
97.7
102
90.4
90.1
96.3
129
110
102
104
134
110
112
104
--
--
--
15
7.6
15
7.3
6.7
4.8
7.3
13
4.6
9.2
12
4.3
4.4
4.5
4.4
4.7
4.1
4.5
4.1
2.1
4.8
4.4
5.3
0
0
0
0
0
0
7.8
21.2
20.4
14.8
50.4
62.7
54.8
25.1
99.2
49.1
102
105
93.6
95.0
101
104
112
106
105
111
110
112
108
--
23
15
11
13
6.0
2.9
4.8
5.7
1.3
6.3
7.4
2.3
2.4
2.3
2.2
1.9
2.1
3.7
0.4
2.0
2.8
3.3
2.2
The operating conditions for the Soxtec apparatus were as follows: immersion
time 45 min; extraction time 45 min; the sample size was 10 g; the spiking
concentration was 500 ng/g, except for the surrogate compounds at 1000 ng/g,
2,5-Dichlorophenyl-4-nitrophenyl ether, 2,3,6-Trichlorophenyl-4-nitrophenyl
ether, and 2,3,4-Trichlorophenyl-4-nitrophenyl ether at 1500 ng/g,
Nitrobenzene at 2000 ng/g, and 1,3-Dichlorobenzene and 1,2-Dichlorobenzene at
5000 ng/g.
8270C - 44
Revision 3
January 1995
-------
TABLE 10
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR THE EXTRACTION
OF SEMIVOLATILE ORGANICS FROM SPIKED CLAY BY
AUTOMATED SOXHLET (METHOD 3541)a
Compound
Phenol
Bis(2-chloroethyl )ether
2-Chlorophenol
Benzyl alcohol
2-Methylphenol
Bis(2-chloroisopropyl )ether
4-Methyl phenol
N-Nitroso-di-n-propylamine
Nitrobenzene
Isophorone
2-Nitrophenol
2,4-Dimethylphenol
Benzole acid
Bis(2-chloroethoxy)methane
2,4-Dichlorophenol
1,2,4-Trichlorobenzene
Naphthalene
4-Chloroaniline
4 -Chi oro -3 -methyl phenol
2-Methylnaphthalene
Hexachl orocycl opentadi ene
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
2-Chloronaphthalene
2-Nitroaniline
Dimethyl phthalate
Acenaphthylene
3-Nitroanil ine
Acenaphthene
2,4-Dinitrophenol
4-Nitrophenol
Dibenzofuran
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diethyl phthalate
4-Chlorophenyl-phenyl ether
Fluorene
4-Nitroaniline
4, 6-Dinitro-2-methyl phenol
N-Nitrosodiphenylamine
Mean
Recovery
47.8
25.4
42.7
55.9
17.6
15.0
23.4
41.4
28.2
56.1
36.0
50.1
40.6
44.1
55.6
18.1
26.2
55.7
65.1
47.0
19.3
70.2
26.8
61.2
73.8
74.6
71.6
77.6
79.2
91.9
62.9
82.1
84.2
68.3
74.9
67.2
82.1
79.0
63.4
77.0
RSD
5.6
13
4.3
7.2
6.6
15
6.7
6.2
7.7
4.2
6.5
5.7
7.7
3.0
4.6
31
15
12
5.1
8.6
19
6.3
2.9
6.0
6.0
5.2
5.7
5.3
4.0
8.9
16
5.9
5.4
5.8
5.4
3.2
3.4
7.9
6.8
3.4
8270C - 45
Revision 3
January 1995
-------
TABLE 10
(continued)
Compound
4-Bromophenyl-phenyl ether
Hexachlorobenzene
Pentachlorophenol
Phenanthrene
Anthracene
Di-n-butyl phthalate
Fluoranthene
Pyrene
Butyl benzyl phthalate
3,3'-Dichlorobenzidine
Benzo(a)anthracene
Bis(2-ethylhexyl) phthalate
Chrysene
Di-n-octyl phthalate
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Indeno(l,2,3-cd)pyrene
Dibenzo( a, h) anthracene
Benzo(g,h,i)perylene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachloroethane
Hexachlorobutadiene
Mean
Recovery
62.4
72.6
62.7
83.9
96.3
78.3
87.7
102
66.3
25.2
73.4
77.2
76.2
83.1
82.7
71.7
71.7
72.2
66.7
63.9
0
0
0
0
0
RSD
3.0
3.7
6.1
5.4
3.9
40
6.9
0.8
5.2
11
3.8
4.8
4.4
4.8
5.0
4.1
4.1
4.3
6.3
8.0
--
--
--
--
— ~
a Number of determinations was three. The operating conditions for the Soxtec
apparatus were as follows: immersion time 45 min; extraction time 45 min; the
sample size was 10 g clay soil; the spike concentration was 6 rug/kg per
compound. The sample was allowed to equilibrate 1 hour after spiking.
Data taken from Reference 7.
8270C - 46 Revision 3
January 1995
-------
TABLE 11
PRECISION AND BIAS VALUES FOR METHOD 35421
Compound
2-Fluorophenol
Phenol -d5
Nitrobenzene-d5
2-Fluorobiphenyl
2,4,6-Tribromophenol
Terphenyl-du
Mean
Recovery
74.6
77.8
65.6
75.9
67.0
78.6
Standard
Deviation
28.6
27.7
32.5
30.3
34.0
32.4
Relative Standard
Deviation Percent
38.3
35.6
49.6
39.9
50.7
41.3
1 The surrogate values shown in Table 11 represent mean recoveries for
surrogates in all Method 0010 matrices in a field dynamic spiking study.
8270C - 47
Revision 3
January 1995
-------
TABLE 12
ACCELERATED SOLVENT EXTRACTION (METHOD 3545) RECOVERY VALUES AS % OF SOXTEC"
COMPOUND
Phenol
Bis(2-chloroethyl) ether
2-Chlorophenol
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
1 , 2-Di chl orobenzene
2-Methyl phenol
Bis(2-chloroisopropy1 Jet her
o-Toluidine
N-Ni troso-di -n-propyl ami ne
Hexachloroethane
Nitrobenzene
Isophorone
2 ,4-Di methyl phenol
2-Nitrophenol
Bis(chloroethoxy)methane
2,4-Dichlorophenol
1, 2, 4-Tri chl orobenzene
Naphthalene
4-Chloroanll Ine
Hexachl orobutadi ene
4-Chl oro-3-methyl phenol
2-Methyl naphthalene
Hexachl orocycl opentadi ene
2 ,4,6-Trichlorophenol
2 , 4 , 5-Tri chl orophenol
2-Chl oronaphthal ene
2-N1troan11 Ine
2,6-Dinitrotoluene
Acenaphthylene
3-Ni troanil Ine
Acenaphthene
4-Nitrophenol
2,4-Dinitrotoluene
Dibenzofuran
4-Chl orophenyl -phenyl ether
Ruorene
4-Nitroaniline
N-NI trosodi phenyl ami ne
4-Bromophenyl -phenyl ether
CLAY
LOW
93.3
102.1
100.8
127.7
127.9
116.8
98.9
109.4
100.0
103.0
97.1
104.8
100.0
100.0
80.7
94.4
88.9
98.0
101.7
100.0
101.1
90.4
93.2
100.0
94.6
84.4
100.0
90.0
83.1
104.9
*224.0
102.1
0.0
73.9
89.5
83.0
85.2
77.8
82.6
85.6
MID
78.7
85.1
82.6
129.7
127.0
115.8
82.1
71.5
89.7
79.1
125.1
82.4
86.4
104.5
80.5
80.6
87.8
97.8
97.2
*150.2
98.7
80.2
89.9
100.0
90 0
91.9
91.3
83.4
90.6
95.9
115.6
92.6
93.2
91.9
91.7
94.5
94.9
114.8
96.7
92.9
HIGH
135.9
109.1
115.0
110.0
110.5
101.3
119.7
108.0
117.2
107.7
111.0
106.6
98.2
140.0
107.9
94.7
111.4
98.8
123.6
*162.4
102.2
114.7
94.6
0.0
112.0
109.6
93.6
97.4
91.6
100.5
97.6
97.6
121.5
100.2
109.3
98.7
89.2
94.5
93.8
92.8
LOAM
LOW
73.9
96.0
93.8
*364.2
*365.9
*159.2
87.6
81.8
100.0
83.9
*245.4
86.8
87.1
100.0
91.4
86.5
85.9
123.0
113.2
100.0
124.1
79.0
104.1
100.0
84.2
96.1
97.6
71.3
86.4
99.0
100.0
97.2
18.1
84.7
98.5
95.7
102.0
129.6
92.9
91.1
MID
82.8
88.0
88.9
129.9
127.8
113.4
89.4
81.0
*152.5
88.1
117.1
84.6
87.5
114.4
86.7
84.4
87.6
93.7
102.9
125.5
90.3
85.2
92.2
100.0
91.2
80.7
93.4
88.4
90.6
97.9
111.8
96.9
87.1
93.8
92.2
94.3
95.5
103.6
93.4
107.6
HIGH
124.6
103.6
111.1
119.0
116.4
105.5
111.0
88.6
120.3
96.2
128.1
101.7
109.7
123.1
103.2
99.6
103.5
94.5
129.5
*263.6
98.0
109.8
105.9
6.8
103.6
103.6
98.3
89.9
90.3
108.8
107.8
104.4
116.6
98.9
111.4
94.2
93.8
95.4
116.4
89.4
SAND
LOW
108.8
122.3
115.0
*241.3
*309.6
*189.3
133.2
118.1
100.0
109.9
*566.7
119.7
135.5
100.0
122.1
130.6
123.3
137.0
*174.5
100.0
134.9
131.6
146.2
100.0
101.6
108.9
106.8
112.1
104.3
118.5
0.0
114.2
69.1
100.9
113.8
111.4
121.3
*154.1
97.5
118.0
MID
130.6
119.9
115.3
*163.7
*164.1
134.0
128.0
148.3
*199.5
123.3
147.9
122.1
118.4
*180.6
107.1
110.7
107.0
99.4
114.0
*250.8
96.1
116.2
99.1
100.0
95.9
83.9
93.0
113.3
84.7
97.8
111.7
92.0
90.5
84.3
92.7
87.7
85.7
89.3
110.9
97.5
HIGH
89.7
90.8
91.9
107.1
105.8
100.4
92.1
94.8
102.7
91.4
103.7
93.3
92.7
96.3
87.0
93.2
92.1
95.3
89.8
114.9
96.8
90.1
93.3
*238.3
89.8
87.9
92.0
87.7
90.9
92.0
99.0
89.0
84.5
87.3
90.4
90.3
90.9
87.5
86.7
87.1
AVE
102.0
101.9
101.6
120.6
119.2
112.5
104.7
100.2
110.3
98.1
118.6
100.2
101.7
109.8
96.3
97.2
98.6
104.2
106.1
108.1
104.7
99.7
102.1
75.8
95.9
94.1
96.2
92.6
90.3
101.7
92.9
98.4
75.6
90.7
98.8
94.4
95.4
99.1
96.8
95.8
8270C - 48
Revision 3
January 1995
-------
TABLE 12
(continued)
COMPOUND
Hexachl orobenzene
Pentachlorophenol
Phenanthrene
Anthracene
Carbazole
Fluoranthene
Pyrene
3 , 3 ' -Di chl orobenzi dl ne
Benzo(a)anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)f"luoranthene
Benzo(a)pyrene
Indeno(l,2,3-cd)pyrene
Dibenz (a, h) anthracene
Benzo(g,h,i )perylene
Average
CLAY
LOW
95.4
68.2
92.1
101.6
94.4
109.9
106.5
100.0
98.1
100. 0
106.6
102.4
107.9
95.1
85.0
98.0
95.1
MID
91.7
85.9
93.7
95.0
99.3
101.4
105.8
*492.3
107.0
108.5
109.9
105.2
105.5
105.7
102.6
0.0
94.3
HIGH
92.3
107.7
93.3
93.5
96.6
94.3
107.6
131.4
98.4
100.2
75.6
88.4
80.8
93.8
82.0
81.2
101.0
LOAM
LOW
95.4
53.2
100.0
92.5
105.5
111.6
116.7
100.0
119.3
116.8
121.7
125.5
122.3
126.0
118.8
0.0
95.5
MID
93.6
89.8
97.8
101.8
96.7
96.6
90.7
*217.6
98.6
93.0
100.7
99.4
97.7
105.2
100.7
33.6
96.5
HIGH
83.7
88.1
113.3
118.4
111.4
109.6
127.5
*167.6
104.0
117.0
93.9
95.1
104.6
90.4
91.9
78.6
104.1
SAND
LOW
106.8
96.6
124.4
123.0
115.7
123.2
103.4
100.0
105.0
106.7
106.9
144.7
101.7
133.6
142.3
128.7
113.0
MID
94.3
59.8
101.0
94.5
83.2
85.4
95.5
*748.8
93.4
93.6
81.9
89.2
86.2
82.6
71.0
83.0
100.9
HIGH
90.0
81.3
89.9
90.6
88.9
92.7
93.2
100.0
89.3
90.2
93.6
78.1
92.0
91.9
93.1
94.2
92.5
AVE
93.7
81.2
100.6
101.2
99.1
102.7
105.2
116.5
101.5
102.9
99.0
103.1
99.9
102.7
98.6
66.4
* Values greater than 150% were not used to determine the averages, but the 0% values were
used.
8270C - 49
Revision 3
January 1995
-------
TABLE 13
SINGLE LABORATORY ACCURACY AND PRECISION FOR THE EXTRACTION OF PAHs
FROM A CERTIFIED REFERENCE SEDIMENT EC-1, USING METHOD 3561 (SEE - SOLID TRAP)
Compound
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Indeno(l,2,3-cd)pyrene
Benzo(g,h,i)perylene
Dibenz( a, h) anthracene
Certified
Value
(mg/kg)
(27.9)b
(0.8)
(0.2)
(15.3)
15.8 ± 1.2
(1.3)
23.2 ± 2.0
16.7 ± 2.0
8.7 ± 0.8
(9.2)
7.9 ± 0.9
4.4 ± 0.5
5.3 ± 0.7
5.7 ± 0.6
4.9 ± 0.7
(1.3)
Percent of
SFE Value3 Certified
(mg/kg) Value
41.3 ± 3.6
0.9 ± 0.1
0.2 ± 0.01
15.6 ± 1.8
16.1 ± 1.8
1.1 ± 0.2
24.1 ± 2.1
17.2 ± 1.9
8.8 ± 1.0
7.9 ± 0.9
8.5 ± 1.1
4.1 ± 0.5
5.1 ± 0.6
5.2 ± 0.6
4.3 ± 0.5
1.1 ± 0.2
(148)
(112)
(100)
(102)
102
(88)
104
103
101
(86)
108
91
96
91
88
(85)
SFE
RSD
8.7
11.1
0.05
11.5
11.2
18.2
8.7
11.0
11.4
11.4
12.9
12.2
11.8
11.5
11.6
18.2
a Relative standard deviations for the SFE values are based on six replicate
extractions.
b Values in parentheses were obtained from, or compared to, Soxhlet extraction
results which were not certified.
Data are taken from Reference 10.
8270C - 50
Revision 3
January 1995
-------
TABLE 14
SINGLE LABORATORY ACCURACY AND PRECISION FOR THE EXTRACTION OF PAHs
FROM A CERTIFIED REFERENCE SEDIMENT HS-3, USING METHOD 3561 (SFE - SOLID TRAP)
Compound
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Indeno(l,2,3-cd)pyrene
Benzo(g,h,i)perylene
Dibenz( a, h) anthracene
Certified
Value
(mg/kg)
9.0
0.3
4.5
13.6
85.0
13.4
60.0
39.0
14.6
14.1
7.7
2.8
7.4
5.0
5.4
1.3
+
±
±
±
±
+
±
±
±
±
±
±
±
±
±
±
0
0
1
3
20
0
9
9
2
2
1
2
3
2
1
0
.7
.1
.5
.1
.0
.5
.0
.0
.0
.0
.2
.0
.6
.0
.3
.5
SFE Value8
(mg/kg)
7
0
3
10
86
12
54
32
12
12
8
3
6
4
4
1
.4
.4
.3
.4
.2
.1
.0
.7
.1
.0
.4
.2
.6
.5
.4
.1
+
±
±
+
±
±
±
±
±
±
±
±
±
±
±
±
0.
0.
0.
1.
9.
1.
6.
3.
1.
1.
0.
0.
0.
0.
0.
0.
6
1
3
3
5
5
1
7
3
3
9
5
8
6
6
3
Percent of
Certified SFE
Value RSD
82
133
73
77
101
90
90
84
83
85
109
114
89
90
82
85
8.1
25.0
9.0
12.5
11.0
12.4
11.3
11.3
10.7
10.8
10.7
15.6
12.1
13.3
13.6
27.3
Relative standard deviations for the SFE values are based on three replicate
extractions.
Data are taken from Reference 10.
8270C - 51
Revision 3
January 1995
-------
TABLE 15
SINGLE LABORATORY ACCURACY AND PRECISION FOR THE EXTRACTION OF PAHs
FROM A CERTIFIED REFERENCE SOIL SRS103-100, USING METHOD 3561
(SFE - LIQUID TRAP)
Compound
Naphthalene
2-Methylnaphthalene
Acenaphthene
Dibenzofuran
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene +
Benzo(k)fluoranthene
Benzo(a)pyrene
Certified
Value
(mg/kg)
32.4
62.1
632
307
492
1618
422
1280
1033
252
297
153
97.2
+
±
+
±
±
+
±
±
+
+
±
±
±
8.2
11.5
105
49
78
340
49
220
285
38
26
22
17.1
SFE Value8
(mg/kg)
29.55
76.13
577.28
302.25
427.15
1278.03
400.80
1019.13
911.82
225.50
283.00
130.88
58.28
Percent of
Certified SFE
Value RSD
91
122
91
98
87
79
95
80
88
89
95
86
60
10.5
2.0
2.9
4.1
3.0
3.4
2.6
4.5
3.1
4.8
3.8
10.7
6.5
Relative standard deviations for the SFE values are based on four replicate
extractions.
Data are taken from Reference 11.
8270C - 52
Revision 3
January 1995
-------
FIGURE 1
GAS CHROMATOGRAM OF BASE/NEUTRAL AND ACID CALIBRATION STANDARD
fclC
CUTM: 5lbHieew786 kl
CML!: 51BHStt6e786 13
SCMNS 2t* TO 2780
8to 6:26:89
S**PLE: BASE AC 10 STO, 2UL/lw<.. UL
CCMDS.:
RriHCE: (, 1.27W LP££L: N 6. 4.8 UUnN: H U, l.a J 6 bhi£: U 20 3
133513.
AJL
33:28
25*
41:46
8270C - 53
Revision 3
January 1995
-------
METHOD 8270C
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY
(GC/MS): CAPILLARY COLUMN TECHNIQUE
7.1 Prepare sample
using appropriate
3500 series method.
7.2 If necessary,
cleanup extract using
appropriate 3600
series method.
7.3 Establish GC/MS
operating conditions.
Tune to DFTPP.
Perform initial
calibration.
7.4 Perform daily
calibration verification
with SPCCs and CCCs
prior to analysis of
samples.
7.5.1 Screen extract
on GC/FID or GC/PID
to identify highly
contaminated samples.
Dilute those samples
as needed.
7.5.3 Analyze
extract by GC/MS.
7.5.4
Does any
'response exceed^
initial calibration
curve
range?
INo
Yes
7.5.4 Dilute
extract.
7.6 Identify analyte
by comparing the
sample and standard
mass spectra.
7.7 Calculate
concentration of
each individual analyte
confirmed present.
Report results.
8270C - 54
Revision 3
January 1995
-------
METHOD 8280A
THE ANALYSIS OF POLYCHLORINATED DIBENZO-P-DIOXINS AND
POLYCHLORINATED DIBENZOFURANS BY HIGH RESOLUTION GAS
CHROMATOGRAPHY/LOW RESOLUTION MASS SPECTROMETRY (HRGS/LRMS)
1.0 SCOPE AND APPLICATION
1.1 This method is appropriate for the detection and quantitative
measurement of2,3,7,8-tetrachlorinated dibenzo-p-dioxin (2,3,7,8-TCDD), 2,3,7,8-
tetrachlorinated dibenzofuran (2,3,7,8-TCDF), and the 2,3,7,8-substituted penta-,
hexa-, hepta-, and octachlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans
(PCDFs) (Figure 1) in water (at part-per-billion concentrations), soil, fly ash,
and chemical waste samples, including stillbottoms, fuel oil, and sludge matrices
(at part-per-mi11 ion concentrations). The following compounds can be determined
by this method (see Sec. 1.4 for a discussion of "total" concentrations).
Compound
CAS Registry No.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)
1,2,3,7,8-Pentachlorodibenzo-p-dioxin (PeCDD)
1,2,3,4,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,7,8,9-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,4,6,7,8-Heptachlorodibenzo-p-dioxin (HpCDD)
1,2,3,4,5,6,7,8-Octachlorodibenzo-p-dioxin (OCDD)
2,3,7,8-Tetrachlorodibenzofuran (TCDF)
1,2,3,7,8-Pentachlorodibenzofuran (PeCDF)
2,3,4,7,8-Pentachlorodibenzofuran (PeCDF)
1,2,3 , 4, 7 ,8-Hexachl orodi benzofuran (HxCDF)
1,2,3,6,7,8-Hexachlorodibenzofuran (HxCDF)
1,2,3,7,8,9-Hexachlorodibenzofuran (HxCDF)
2, 3, 4, 6, 7, 8 -Hexachl orodi benzofuran (HxCDF)
1,2,3,4,6,7 , 8-Heptachl orodi benzofuran (HpCDF)
1,2,3,4,7,8,9-Heptachlorodibenzofuran (HpCDF)
1,2,3,4,5,6,7,8-Octachlorodibenzofuran (OCDF)
Total Tetrachlorodibenzo-p-dioxin (TCDD)
Total Pentachlorodibenzo-p-dioxin (PeCDD)
Total Hexachlorodibenzo-p-dioxin (HxCDD)
Total Heptachlorodibenzo-p-dioxin (HpCDD)
Total Tetrachlorodi benzofuran (TCDF)
Total Pentachlorodibenzofuran (PeCDF)
Total Hexachlorodibenzofuran (HxCDF)
Total Heptachlorodibenzofuran (HpCDF)
1746-01-6
40321-76-4
39227-28-6
57653-85-7
19408-74-3
35822-46-9
3268-87-9
51207-31-9
57117-41-6
57117-31-4
70648-26-9
57117-44-9
72918-21-9
60851-34-5
67562-39-4
55673-89-7
39001-02-0
41903-57-5
36088-22-9
34465-46-8
37871-00-4
55722-27-5
30402-15-4
55684-94-1
38998-75-3
8280A - 1
Revision 1
January 1995
-------
1.2 The analytical method requires the use of high resolution gas
chromatography and low resolution mass spectrometry (HRGC/LRMS) on sample
extracts that have been subjected to specified cleanup procedures. The
calibration range is dependent on the compound and the sample size. The sample
size varies by sample matrix. Table 2 lists the quantitation limits for the
various matrices.
1.3 This method requires the calculation of the 2,3,7,8-TCDD toxicity
equivalence according to the procedures given in the U.S. Environmental
Protection Agency "Update of Toxicity Equivalency Factors (TEFs) for Estimating
Risks Associated with Exposures to Mixtures of Chlorinated Dibenzo-p-Dioxins and
Dibenzofurans (CDDs/CDFs)" February 1989 (EPA 625/3-89/016). If the toxicity
equivalence is greater than or equal to 0.7 ppb (soil or fly ash), 7 ppt
(aqueous), or 7 ppb (chemical waste), analysis on a column capable of resolving
all 2,3,7,8-substituted PCDDs/PCDFs is necessary. If the expected concentrations
of the PCDDs and PCDFs are below the quantitation limits in Table 2, use Method
8290.
1.4 This method contains procedures for reporting the total concentration
of all PCDDs/PCDFs in a given level of chlorination (i.e. Total TCDD, Total
PeCDF, etc.), although complete chromatographic separation of all 210 possible
PCDDs/PCDFs is not possible under the instrumental conditions described here.
1.5 This method is restricted for use only by analysts experienced with
residue analysis and skilled in HRGC/LRMS. Each analyst must demonstrate the
ability to generate acceptable results with this method.
1.6 Because of the extreme toxicity of these compounds, the analyst must
take necessary precautions to prevent the exposure of laboratory personnel or
others to materials known or believed to contain PCDDs or PCDFs. Typical
infectious waste incinerators are not satisfactory devices for disposal of
materials highly contaminated with PCDDs or PCDFs. A laboratory planning to use
these compounds should prepare a disposal plan. Additional safety instructions
are outlined in Sec. 11.0.
2.0 SUMMARY OF THE METHOD
2.1 This procedure uses a matrix-specific extraction, analyte-specific
cleanup, and high-resolution capillary column gas chromatography/low resolution
mass spectrometry (HRGC/LRMS) techniques.
2.2 If interferants are encountered, the method provides selected cleanup
procedures to aid the analyst in their elimination. The analysis flow chart is
shown at the end of this procedure.
2.3 A specified amount of water, soil, fly ash, or chemical waste samples
is spiked with internal standards and extracted according to a matrix-specific
extraction procedure. Aqueous samples judged to contain 1% or more solids are
filtered, and solid samples that show an aqueous phase are centrifuged before
extraction. The extraction procedures and solvents are:
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2.3.1 Soil, fly ash, or chemical waste samples are extracted with
the combination of a Dean-Stark water trap and a Soxhlet extractor using
toluene.
2.3.2 Water samples are extracted with a separatory funnel or
liquid-liquid extractor using methylene chloride.
2.4 The extracts are spiked with 37Cl4-2,3,7,8-TCDD and submitted to an
acid-base washing treatment, dried and concentrated. The extracts are cleaned
up by column chromatography on alumina, silica gel, and activated carbon on
Celite 545® and concentrated again.
2.5 An aliquot of the concentrated extract is injected into an HRGC/LRMS
system capable of performing the selected ion monitoring.
2.6 The identification of the target compounds is based on their ordered
elution and comparison to standard solutions (Table 1) from an appropriate GC
column and MS identification. Isomer specificity for all 2,3,7,8-substituted
PCDDs/PCDFs cannot be achieved on a single column. The use of both DB-5 and
SP2331 (or equivalent) columns is advised. No analyses can proceed unless all
the criteria for retention times, peak identification, signal-to-noise and ion
abundance ratios are met by the GC/MS system after the initial calibration and
calibration verification.
2.7 A calculation of the toxicity equivalent concentration (TEQ) of each
sample is made using international consensus toxicity equivalence factors (TEFs),
and the TEQ is used to determine if the concentrations of target compounds in the
sample are high enough to warrant confirmation of the results on a second GC
column.
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines which may cause
misinterpretation of chromatographic data. All of these materials must be
demonstrated to be free from interferents under the conditions of analysis by
running laboratory method blanks.
3.2 The use of high purity reagents and pesticide grade solvents helps to
minimize interference problems. Purification of solvents by distillation, in all
glass systems, may be required.
3.3 Interferants co-extracted from the sample will vary considerably from
source to source, depending upon the industrial process being sampled. PCDDs and
PCDFs are often associated with other interfering chlorinated compounds such as
PCBs and polychlorinated diphenyl ethers (PCDPEs) which may be found at
concentrations several orders of magnitude higher than that of the analytes of
interest. Retention times of target analytes must be verified using reference
standards. While certain cleanup techniques are provided as part of this method,
unique samples may require additional cleanup techniques to achieve the
sensitivity specified in this method.
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3.4 High resolution capillary columns are used to resolve as many isomers
as possible; however, no single column is known to resolve all of the 210
isomers. The columns employed by the laboratory in these analyses must be
capable of resolving all 17 of the 2,3,7,8-substituted PCDDs/PCDFs sufficiently
to meet the method specifications.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system:
4.1.1 Gas chromatograph - An analytical system with a temperature-
programmable gas chromatograph and all necessary accessories including
syringes, analytical columns, and gases. The GC injection port shall be
designed for capillary columns; a splitless or an on-column injection
technique is recommended. A 2-juL injection volume is assumed throughout
this method; however, with some GC injection ports, other volumes may be
more appropriate. A l-/uL injection volume may be used if adequate
sensitivity and precision can be demonstrated.
4.1.2 GC column - Fused silica capillary columns are needed. The
columns shall demonstrate the required separation of all 2,3,7,8-specific
isomers whether a dual column or a single column analysis is chosen.
Column operating conditions shall be evaluated at the beginning and end of
each 12 hour period during which samples or concentration calibration
solutions are analyzed.
4.1.2.1 Isomer specificity for all 2,3,7,8-substituted
PCDDs/PCDFs cannot be achieved on the 60 m DB-5 column. Problems have
been associated with the separation of 2,3,7,8-TCDD from 1,2,3,7-TCDD
and 1,2,6,8-TCDD, and separation of 2,3,7,8-TCDF from 2,3,4,7-TCDF.
Because of the toxicologic concern associated with 2,3,7,8-TCDD and
2,3,7,8-TCDF, additional analyses may be necessary for some samples,
as described in Sec. 7.12. In instances where the toxicity equivalent
concentration (TEQ) is greater than 0.7 ppb (solids), 7 ppt (aqueous),
or 7 ppb (chemical waste), the reanalysis of the sample extract on a
60 m SP-2330 or SP-2331 GC column (or equivalent column) may be
required in order to determine the concentrations of the individual
2,3,7,8-substituted isomers.
4.1.2.2 For any sample analyzed on a DB-5 or equivalent
column in which either 2,3,7,8-TCDD or 2,3,7,8-TCDF is reported as an
Estimated Maximum Possible Concentration (Sec. 7.21), regardless of
TEF-adjusted concentration or matrix, analysis of the extract is
required on a second GC column which provides better specificity for
these two isomers.
4.1.2.3 Analysis on a single column is acceptable if the
required separation of all the 2,3,7,8-specific isomers is
demonstrated, and the minimum acceptance criteria outlined in Sees.
7.4, 7.5, and 7.6 are met. See Sec. 7.2 for the specifications for
the analysis of the 2,3,7,8-specific isomers using both dual columns
and single columns.
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4.2 Mass spectrometer - A low resolution instrument is designated,
utilizing 70 volts (nominal) electron energy in the electron impact ionization
mode. The system must be capable of selected ion monitoring (SIM) for at least
18 ions per cycle, with a cycle time of 1 sec or less. Minimum integration time
for SIM is 25 ms per m/z. The integration time used to analyze samples shall be
identical to the time used to analyze the initial calibration and calibration
verification solutions and quality control samples.
4.2.1 Interfaces - GC/MS interfaces constructed of all glass or
glass-lined materials are necessary. Glass can be deactivated by
silanizing with dichlorodimethylsilane. Inserting a fused silica column
directly into the MS source is recommended. Care must be taken not to
expose the end of the column to the electron beam.
4.2.2 Data system - An interfaced data system is necessary to
acquire, store, reduce and output mass spectral data.
4.3 Miscellaneous equipment
4.3.1 Nitrogen evaporation apparatus (N-Evap* Analytical Evaporator
Model 111, Organomation Association Inc., Northborough, MA, or equivalent).
4.3.2 Balance capable of accurately weighing ±0.01 g.
4.3.3 Water bath - Equipped with concentric ring cover and
temperature controlled within ±2"C.
4.3.4 Stainless steel (or glass) pan large enough to hold contents
of 1 pint sample containers.
4.3.5 Glove box - For use in preparing standards from neat materials
and in handling soil/sediment samples containing fine particulates that may
pose a risk of exposure.
4.3.6 Rotary evaporator, R-110, Buchi/Brinkman - American Scientific
No. E5045-10 or equivalent.
4.3.7 Centrifuge - Capable of operating at 400 x G with a 250-300 ml
capacity.
4.3.8 Drying oven.
4.3.9 Vacuum oven - Capable of drying solvent-washed solid reagents
at 110'C.
4.3.10 Mechanical shaker - A magnetic stirrer, wrist-action or
platform-type shaker that produces vigorous agitation. Used for pre-
treatment of fly ash samples.
4.4 Miscellaneous laboratory glassware
4.4.1 Extraction jars - Amber glass with Teflon®-!ined screw cap;
minimum capacity of approximately 200 ml; must be compatible with
mechanical shaker to be used.
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4.4.2 Kuderna-Danish (K-D) Apparatus - 500-mL evaporating flask, 10-
mL graduated concentrator tubes with ground glass stoppers, three-ball
macro-Synder column.
NOTE: The use of a solvent vapor recovery system (Kontes K-545000-
1006 or K-547300-0000, Ace Glass 6614-30, or equivalent) is
recommended for the purpose of solvent recovery during the
concentration procedures requiring the use of Kuderna-Danish
evaporative concentrators. Incorporation of this apparatus
may be required by State or local municipality regulations
that govern air emissions of volatile organics. EPA
recommends the incorporation of this type of reclamation
system as a method to implement an emissions reduction
program. Solvent recovery is a means to conform with waste
minimization and pollution prevention initiatives.
4.4.3 Disposable Pasteur pipets, 150 mm long x 5 mm ID.
4.4.4 Disposable serological pipets, 10-mL for preparation of the
carbon column described in Sec. 7.10.
4.4.5 Vials - 0.3-mL and 2-mL amber borosilicate glass with conical
shaped reservoir and screw caps lined with Teflon®-faced silicone disks.
4.4.6 Funnels - Glass; appropriate size to accommodate filter paper
(12.5 cm).
4.4.7 Chromatography columns - 300 mm x 10.5 mm glass
chromatographic column fitted with Teflon® stopcock.
4.4.8 Soxhlet apparatus, 500-mL flask, all glass - Complete with
glass extractor body, condenser, glass extraction thimbles, heating mantle,
and variable transformer for heat control.
NOTE: Extraction thimbles must be of sufficient size to hold 100 g of
sand, 5 g of silica gel, and at least 10 g of solid sample, with
room to mix the sand and sample in the thimble.
4.4.9 Dean-Stark water separator apparatus, with a Teflon® stopcock.
Must fit between Soxhlet extractor body and condenser.
4.4.10 Concentrator tubes - 15-mL conical centrifuge tubes.
4.4.11 Separatory funnels - 125-mL and 2-L separatory funnels with
a Teflon® stopcock.
4.4.12 Continuous liquid-liquid extractor - 1-L sample capacity,
suitable for use with heavier than water solvents.
4.4.13 Teflon® boiling chips - wash with hexane prior to use.
4.4.14 Buchner funnel - 15 cm.
4.4.15 Filtration flask - For use with Buchner funnel, 1-L capacity.
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4.5 Filters
4.5.1 Filter paper - Whatman No. 1 or equivalent.
4.5.2 Glass fiber filter - 15 cm, for use with Buchner funnel.
4.5.3 0.7 /xm, Whatman GFF, or equivalent material compatible with
toluene. Rinse with toluene.
4.6 Glass wool, silanized - Extract with methylene chloride and hexane
before use.
4.7 Laboratory glassware cleaning procedures - Reuse of glassware should
be minimized to avoid the risk of using contaminated glassware. All glassware
that is reused shall be scrupulously cleaned as soon as possible after use,
applying the following procedure.
4.7.1 Rinse glassware with the last solvent used in it.
4.7.2 Wash with hot water containing detergent.
4.7.3 Rinse with copious amounts of tap water and several portions
of organic-free reagent water. Drain dry.
4.7.4 Rinse with pesticide grade acetone and hexane.
4.7.5 After glassware is dry, store inverted or capped with aluminum
foil in a clean environment.
4.7.6 Do not bake reusable glassware as a routine part of cleaning.
Baking may be warranted after particularly dirty samples are encountered,
but should be minimized, as repeated baking may cause active sites on the
glass surface that will irreversibly adsorb PCDDs/PCDFs.
CAUTION: The analysis for PCDDs/PCDFs in water samples is for much lower
concentrations than in soil/sediment, fly ash, or chemical waste
samples. Extreme care must be taken to prevent cross-
contamination between soil/sediment, fly ash, chemical waste and
water samples. Therefore, it is strongly recommended that
separate glassware be reserved for analyzing water samples.
4.8 Pre-extraction of glassware - All glassware should be rinsed or pre-
extracted with solvent immediately before use. Soxhlet-Dean-Stark (SDS)
apparatus and continuous liquid-liquid extractors should be pre-extracted for
approximately three hours immediately prior to use, using the same solvent and
extraction conditions that will be employed for sample extractions. The pooled
waste solvent for a set of extractions may be concentrated and analyzed as a
method of demonstrating that the glassware was free of contamination.
It is recommended that each piece of reusable glassware be numbered in such
a fashion that the laboratory can associate all reusable glassware with the
processing of a particular sample. This will assist the laboratory in:
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1) Tracking down possible sources of contamination for individual
samples,
2) Identifying glassware associated with highly contaminated samples that
may require extra cleaning, and
3) Determining when glassware should be discarded.
5.0 REAGENTS
5.1 Solvents - all solvents must be pesticide grade, distilled-in-glass.
5.1.1 Hexane, C6H14
5.1.2 Methanol, CH3OH
5.1.3 Methylene chloride, CH2C12
5.1.4 Toluene, C6H5CH3
5.1.5 Isooctane, (CH3)3CCH2CH(CH3)2
5.1.6 Cyclohexane, C6H12
5.1.7 Acetone, CH3COCH3
5.1.8 Tridecane, CH3(CH2)nCH3
5.1.9 Nonane, C9H20
5.2 White quartz sand - 60/70 mesh, for use in the Soxhlet-Dean-Stark
(SDS) extractor. Bake at 450°C for 4 hours minimum.
5.3 Sodium sulfate (granular, anhydrous), Na2S04 - Purify by heating at
400"C for 4 hours in a shallow tray, or by extracting with methylene chloride.
If, after heating, the sodium sulfate develops a noticeable grayish cast (due to
the presence of carbon in the crystal matrix) that batch of sodium sulfate is not
suitable for use and should be discarded. Extraction with methylene chloride may
produce sodium sulfate that is suitable for use in such instances, but following
extraction, a reagent blank must be analyzed that demonstrates that there is no
interference from the sodium sulfate.
5.4 Potassium hydroxide, KOH - ACS reagent grade, prepare a 20% (w/v)
solution in organic-free reagent water.
5.5 Sulfuric acid, H2S04, concentrated - ACS reagent grade, specific
gravity 1.84.
5.6 Sodium chloride, NaCl - ACS reagent grade, prepare a 5% (w/v) solution
in organic-free reagent water.
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5.7 Hydrochloric acid, HC1, concentrated - ACS reagent grade, specific
gravity 1.17. Prepare a IN solution in organic-free reagent water for
pretreatment of fly ash samples.
5.8 Column chromatography reagents
5.8.1 Alumina, acidic AG4 - BioRad Laboratories (Catalog No.
132-1240) or equivalent. Soxhlet extract with methylene chloride for 21
hours and activate by heating in a foil-covered glass container for 24
hours at 190°C.
5.8.2 Charcoal carbon - Activated carbon, Carbopak C (Supelco) or
equivalent, prewashed with methanol and dried in vacua at 110CC. (Note:
AX-21 [Anderson Development Company] carbon is no longer available, but
existing stocks may be utilized).
5.8.3 Celite 545 (Supelco) or equivalent.
5.8.4 Silica gel - High-purity grade, type 60, 70-230 mesh. Soxhlet
extract with methylene chloride for 21 hours and activate by heating in a
foil covered glass container for 24 hours at 190"C.
5.8.5 Silica gel impregnated with 2% (w/w) sodium hydroxide - Add
one part by weight of 1 M NaOH solution to two parts silica gel (extracted
and activated) in a screw-cap bottle and mix with a glass rod until free
of lumps.
5.8.6 Silica gel impregnated with 40% (w/w) sulfuric acid. Add two
parts by weight concentrated sulfuric acid to three parts silica gel
(extracted and activated), mix with a glass rod until free of lumps, and
store in a screw-cap glass bottle.
5.9 Calibration solutions (Table 1) - Prepare five tridecane (or nonane)
solutions (CC1-CC5) containing 10 unlabeled and 7 carbon-labeled PCDDs/PCDFs at
known concentrations for use in instrument calibration. One of these five
solutions (CCS) is used as the calibration verification solution and contains 7
additional unlabeled 2,3,7,8-isomers. The concentration ranges are homologue-
dependent, with the lowest concentrations associated with tetra- and
pentachlorinated dioxins and furans (0.1 to 2.0 ng/^L), and the higher
concentrations associated with the hexa- through octachlorinated homologues (0.5
to 10.0 ng/juL). Commercially-available standards containing all 17 unlabeled
analytes in each solution may also be utilized.
5.10 Internal standard solution (Table 3) - Prepare a solution containing
the five internal standards in tridecane (or nonane) at the nominal
concentrations listed in Table 3. Mix 10 /zL with 1.0 ml of acetone before
adding to each sample and blank.
5.11 Recovery standard solution (Table 3) - Prepare a solution in hexane
containing the recovery standards, 13C12-1,2,3,4-TCDD and 13C12-l,2,3,7,8,9-HxCDD,
at concentrations of 5.0 ng/|iL, in a solvent other than tridecane or nonane.
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5.12 Calibration verification solution - Prepare a solution containing
standards to be used for identification and quantitation of target analytes
(Table 4).
5.13 Cleanup standard - Prepare a solution containing 37Cl4-2,3,7,8-TCDD at
a concentration of 5 ng/^L (5 /zg/mL) in tridecane (or nonane). Add this
solution to all sample extracts prior to cleanup. The solution may be added at
this concentration, or diluted into a larger volume of solvent. The recovery of
this compound is used to judge the efficiency of the cleanup procedures.
5.14 Matrix spiking standard - Prepare a solution containing ten of the
2,3,7,8-substituted isomers, at the concentrations listed in Table 5 in tridecane
(or nonane). Use this solution to prepare the spiked sample aliquot. Dilute 10
juL of this standard to 1.0 ml with acetone and add to the aliquot chosen for
spiking.
5.15 Window defining mix - Prepare a solution containing the first and last
eluting isomer of each homologue (Table 6). Use this solution to verify that the
switching times between the descriptors have been appropriately set.
5.16 Column performance solution - Prepare a solution containing
2,3,7,8-TCDD and the other TCDD isomers (1,4,7,8-TCDD and the 1,2,3,7/1,2,3,8-
TCDD pair) that elute closest to 2,3,7,8-TCDD on the SP-2331 column. Use this
solution to verify the chromatographic resolution of the SP-2331 column. The
concentrations of these isomers should be approximately 0.5 ng//zL in tridecane
(or nonane}.
If the laboratory employs a column that has a different elution order than
those specified here, the laboratory must ensure that the isomers eluting closest
to 2,3,7,8-TCDD are represented in the column performance solution.
6.0 SAMPLE COLLECTION, HANDLING, AND PRESERVATION
6.1 See the introductory material to this chapter, Organic Analytes.
6.2 Sample collection
6.2.1 Sample collection personnel should, to the extent possible,
homogenize samples in the field before filling the sample containers. This
should minimize or eliminate the necessity for sample homogenization in the
laboratory. The analyst should make a judgment, based on the appearance
of the sample, regarding the necessity for additional mixing. If the
sample is clearly not homogeneous, the entire contents should be
transferred to a glass or stainless steel pan for mixing with a stainless
steel spoon or spatula before removal of a sample portion for analysis.
6.2.2 Grab and composite samples must be collected in glass
containers. Conventional sampling practices must be followed. The bottle
must not be prewashed with sample before collection. Sampling equipment
must be free of potential sources of contamination.
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6.3 Storage and holding times - All samples should be stored at 4'C in the
dark, extracted within 30 days and completely analyzed within 45 days of
extraction. Whenever samples are analyzed after the holding time expiration
date, the results should be considered to be minimum concentrations and should
be identified as such.
NOTE: The holding times listed in Sec. 6.3*are recommendations. PCDDs and
PCDFs are very stable in a variety of matrices, and holding times under
the conditions listed in Sec. 6.3 may be as high as a year for certain
matrices. Sample extracts, however, should always be analyzed within 45
days of extraction.
7.0 PROCEDURE
Four types of extraction procedures are employed in these analyses,
depending on the sample matrix.
1) Chemical waste samples are extracted by refluxing with a Dean-Stark
water separator.
2) Fly ash samples and soil/sediment samples are extracted in a
combination of a Soxhlet extractor and a Dean-Stark water separator.
3) Water samples are filtered and then the filtrate is extracted using
either a separatory funnel procedure or a continuous liquid-liquid
extraction procedure.
4) The filtered particulates are extracted in a combination of a Soxhlet
extractor and a Dean-Stark water separator.
Sec. 7.1 provides general information on the use of the Soxhlet-Dean-Stark
apparatus. The four matrix-specific extraction procedures are described in
Sees. 7.2 - 7.5.
7.1 General considerations for use of the Soxhlet-Dean-Stark (SDS)
apparatus - The following procedures apply to use of the SDS apparatus for
extracting matrices covered by this protocol.
The combination of a Soxhlet extractor and a Dean-Stark trap is used for
the removal of water and extraction of PCDDs/PCDFs from samples of fly ash,
soil/sediment, and the particulate fraction of water samples.
For soil/sediment samples, the results of these analyses are reported based
on the wet weight of the sample. However, use of the SDS allows the water
content of a sample to be determined from the same aliquot of sample that is also
extracted for analysis. The amount of water evolved from the sample during
extraction is used to approximate the percent solids content of the sample. The
percent solids data may be employed by the data user to approximate the dry
weight concentrations. The percent solids determination does not apply to the
extraction of particulates from the filtration of water samples or to the
extraction of fly ash samples which are treated with an HC1 solution prior to
extraction.
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7.1.1 The extraction of soil/sediment, fly ash, and particulates
from water samples will require the use of a Soxhlet thimble. See Sec. 4.6
for a discussion of pre-extraction of glassware such as the SDS. Prior to
pre-extraction, prepare the thimble by adding 5 g of 70/230 mesh silica gel
to the thimble to produce a thin layer in the bottom of the thimble. This
layer will trap fine particles in the thimble. Add 80-100 g of quartz sand
on top of the silica gel, and place the thimble in the extractor.
7.1.2 Pre-extract the SDS for three hours with toluene, then allow
the apparatus to cool and remove the thimble. Mix the appropriate weight
of sample with the sand in the thimble, being careful not to disturb the
silica gel layer.
7.1.3 If the sample aliquot to be extracted contains large lumps,
or is otherwise not easily mixed in the thimble, the sand and sample may
be mixed in another container. Transfer approximately 2/3 of the sand from
the thimble to a clean container, being careful not to disturb the silica
gel layer when transferring the sand. Thoroughly mix the sand with the
sample with a clean spatula, and transfer the sand/sample mixture to the
thimble.
7.1.4 If a sample with particularly high moisture content is to be
extracted, it may be helpful to leave a small conical depression in the
material in the thimble. This will allow the water to drain through the
thimble more quickly during the early hours of the extraction. As the
moisture is removed during the first few hours of extraction, the
depression will collapse, and the sample will be uniformly extracted.
7.2 Chemical waste extraction (including oily sludge/wet fuel oil and
still bottom/oil)
7.2.1 Assemble a flask, a Dean-Stark trap, and a condenser, and pre-
extract with toluene for three hours (see Sec. 4.6). After pre-extraction,
allow the apparatus to cool, and discard the used toluene, or pool it for
later analysis to verify the cleanliness of the glassware.
7.2.2 Weigh about 1 g of the waste sample to two decimal places into
a tared pre-extracted 125-mL flask. Add 1 ml of the acetone-diluted
internal standard solution (Sec. 5.10) to the sample in the flask. Attach
the pre-extracted Dean-Stark water separator and condenser to the flask,
and extract the sample by refluxing it with 50 ml of toluene for at least
three hours.
Continue refluxing the sample until all the water has been removed.
Cool the sample, filter the toluene extract through a rinsed glass fiber
filter into a 100-mL round bottom flask. Rinse the filter with 10 ml of
toluene; combine the extract and rinsate. Concentrate the combined
solution to approximately 10 ml using a K-D or rotary evaporator as
described in Sees. 7.6.1 and 7.6.2. Transfer the concentrated extract to
a 125-mL separatory funnel. Rinse the flask with toluene and add the rinse
to the separatory funnel. Proceed with acid-base washing treatment per
Sec. 7.7, the micro-concentration per Sec. 7.8, the chromatographic
procedures per Sees. 7.9 and 7.10, and a final concentration per Sec. 7.11.
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7.2.3 Prepare an additional two 1-g aliquots of the sample chosen
for spiking. After weighing the sample in a tared pre-extracted flask
(Sec. 7.2.2), add 1.0 ml of the acetone-diluted matrix spiking standard
solution (Sec. 5.14) to each of the two aliquots. After allowing the
matrix spiking solution to equilibrate to approximately 1 hour, add the
internal standard solution and extract the aliquots as described in Sec.
7.2.2.
7.3 Fly ash sample extraction
7.3.1 Weigh about 10 g of the fly ash to two decimal places, and
transfer to an extraction jar. Add 1 ml of the acetone-diluted internal
standard solution to the sample.
7.3.2 Add 150 ml of 1 N HC1 to the fly ash sample in the jar. Seal
the jar with the Teflon®-!ined screw cap, place on a mechanical shaker, and
shake for 3 hours at room temperature.
7.3.3 Rinse a Whatman #1 (or equivalent) filter paper with toluene,
and then filter the sample through the filter paper in a Buchner funnel
into a 1 L receiving flask. Wash the fly ash with approximately 500 ml of
organic-free reagent water.
7.3.4 Mix the fly ash with the sand in the pre-extracted thimble
(Sec. 7.1.2). Place the filter paper from Sec. 7.3.3 on top of the sand.
Place the thimble in a SDS extractor, add 200 ml toluene, and extract for
16 hours. The solvent should cycle completely through the system 5-10 times
per hour. Cool and filter the toluene extract through a rinsed glass fiber
filter into a 500-mL round-bottom flask. Rinse the filter with 10 ml of
toluene. Concentrate the extract as described in Sees. 7.6.1 or 7.6.2.
Transfer the concentrated extract to a 125-mL separatory funnel. Rinse the
flask with toluene and add the rinse to the separatory funnel. Proceed
with acid-base washing treatment per Sec. 7.7, the micro-concentration per
Sec. 7.8, the chromatographic procedures per Sees. 7.9 and 7.10 and a final
concentration per Sec, 7.11.
NOTE: A blank should be analyzed using a piece of filter paper handled in
the same manner as the fly ash sample.
7.3.5 Prepare an additional two 10-g aliquots of the sample chosen
for spiking for use as the matrix spike and matrix spike duplicate.
Transfer each aliquot to a separate extraction jar and add 1.0 ml of the
acetone-diluted matrix spiking standard solution (Sec. 5.14) to each of the
two aliquots. After allowing the matrix spiking solution to equilibrate
to approximately 1 hour, add the internal standard solution and extract the
aliquots as described in Sec. 7.3.1.
7.4 Soil/sediment sample extraction
NOTE: Extremely wet samples may require centrifugation to remove standing
water before extraction.
7.4.1 Weigh about 10 grams of the soil to two decimal places and
transfer to a pre-extracted thimble (Sec. 7.1.2). Mix the sample with the
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quartz sand, and add 1 ml of the acetone-diluted internal standard solution
(Sec. 5.10) to the sample/sand mixture. Add small portions of the solution
at several sites on the surface of the sample/sand mixture.
7.4.2 Place the thimble in the SDS apparatus, add 200 to 250 mL
toluene, and reflux for 16 hours. The solvent should cycle completely
through the system 5-10 times per hour.
7.4.3 Estimate the percent solids content of the soil/sediment
sample by measuring the volume of water evolved during the SDS extraction
procedure. For extremely wet samples, the Dean-Stark trap may need to be
drained one or more times during the 16-hour extraction. Collect the water
from the trap, measure its volume to the nearest 0.1 mL. Assume a density
of 1.0 g/mL, and calculate the percent solids content according to the
formula below:
n . ,.. Wet weight of sample - Weight of water ,.rt
Percent solids = x 100
Wet weight of sample
7.4.4 Concentrate this extract as described in Sees. 7.6.1 or 7.6.2.
Transfer the concentrated extract to a 125 ml separatory funnel. Rinse the
flask with toluene and add the rinse to the separatory funnel. Proceed
with acid-base washing treatment per Sec. 7.7, the micro concentration per
Sec. 7.8, the chromatographic procedures per Sees. 7.9 and 7.10 and a final
concentration per Sec. 7.11.
7.4.5 Prepare an additional two 10-g aliquots of the sample chosen
for spiking for use as the matrix spike and matrix spike duplicate. After
transferring each aliquot to a separate pre-extracted Soxhlet thimble, add
1.0 ml of the acetone-diluted matrix spiking standard solution (Sec. 5.14)
to each of the two aliquots. After allowing the matrix spiking solution
to equilibrate to approximately 1 hour, add the internal standard solution
(Sec. 5.10) and extract the aliquots as described in Sec. 7.4.1.
7.5 Aqueous sample extraction
7.5.1 Allow the sample to come to ambient temperature, then mark the
water meniscus on the side of the 1-L sample bottle for determination of
the exact sample volume.
7.5.2 Add 1 ml of the acetone-diluted internal standard solution
(Sec. 5.10) to the sample bottle. Cap the bottle, and mix the sample by
gently shaking for 30 seconds.
7.5.3 Filter the sample through a 0.7-jum filter that has been rinsed
with toluene. Collect the aqueous filtrate in a clean flask. If the total
dissolved and suspended solids contents are too much to filter through the
0.7-jum filter, centrifuge the sample, decant, and then filter the aqueous
phase. Procedures for extraction of the particulate fraction are given in
Sec. 7.5.4. The aqueous portion may be extracted using either the
separatory funnel technique (Sec. 7.5.5.1) or a pre-extracted continuous
liquid-liquid extractor (Sec. 7.5.5.2).
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NOTE: Organic-free reagent water used as a blank must also be filtered in
a similar fashion, and subjected to the same cleanup and analysis
as the water samples.
7.5.4 Particulate fraction
7.5.4.1 Combine the particulate on the filter and the filter
itself, and if centrifugation was used, the solids from the centrifuge
bottle(s), with the quartz sand in the pre-extracted Soxhlet thimble.
Place the filter on top of the particulate/sand mixture, and place the
thimble into a pre-extracted SDS apparatus.
7.5.4.2 Add 200 to 250 ml of toluene to the SDS apparatus
and reflux for 16 hours. The solvent should cycle completely through
the system 5-10 times per hour.
7.5.4.3 Allow the Soxhlet to cool, remove the toluene and
concentrate this extract as described in Sees. 7.6.1. or 7.6.2.
7.5.5 Aqueous filtrate
The aqueous filtrate may be extracted by either a separatory funnel
procedure (Sec. 7.5.5.1) or a continuous liquid-liquid extraction procedure
(Sec. 7.5.5.2).
7.5.5.1 Separatory funnel extraction - The filtered aqueous
sample is poured into a 2-L separatory funnel. Add 60 ml methylene
chloride to the sample bottle, seal, and shake 60 seconds to rinse the
inner surface. Transfer the solvent to the separatory funnel and
extract the sample by shaking the funnel for 2 minutes with periodic
venting. Allow the organic layer to separate from the water phase for
a minimum of 10 minutes. Drain the methylene chloride extract into
a 500-mL K-D concentrator (mounted with a 10-mL concentrator tube) by
passing the extract through a funnel packed with a glass wool plug and
half-filled with anhydrous sodium sulfate. Extract the water sample
two more times using 60 mL of fresh methylene chloride each time.
Drain each extract through the funnel into the K-D concentrator.
After the third extraction, rinse the sodium sulfate with at least 30
ml of fresh methylene chloride. Concentrate this extract as described
in Sees. 7.6.1 or 7.6.2.
7.5.5.2 Continuous liquid-liquid extraction - A continuous
liquid-liquid extractor may be used in place of a separatory funnel
when experience with a sample from a given source indicates that a
serious emulsion problem will result or an emulsion is encountered
using a separatory funnel. The following procedure is used for a
continuous liquid-liquid extractor.
7.5.5.2.1 Pre-extract the continuous liquid-liquid
extractor for three hours with methylene chloride and reagent
water. Allow the extractor to cool, discard the methylene
chloride and the reagent water, and add the filtered aqueous
sample to the continuous liquid-liquid extractor. Add 60 ml
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of methylene chloride to the sample bottle, seal and shake
for 30 seconds.
7.5.5.2.2 Transfer the solvent to the extractor.
Repeat the sample bottle rinse with an additional 50 to 100
ml portion of methylene chloride and add the rinse to the
extractor. Add 200 to 500 ml methylene chloride to the
distilling flask and sufficient reagent water to ensure
proper operation. Extract for 16 hours. Allow to cool, then
detach the flask and dry the sample by running it through a
rinsed funnel packed with a glass wool plug and 5 g of
anhydrous sodium sulfate into a 500-mL K-D flask.
Concentrate the extract according to Sees. 7.6.1 or 7.6.2.
7.5.6 Combination of extracts - The extracts from both the
particulate fraction (Sec. 7.5.4) and the aqueous filtrate (Sec. 7.5.5}
must be concentrated using the procedures in Sec. 7.6.1 and then combined
together prior to the acid-base washing treatment in Sec. 7.7.
7.5.7 Determine the original aqueous sample volume by refilling the
sample bottle to the mark and transferring the liquid to a 1-L graduated
cylinder. Record the sample volume to the nearest 5 ml.
7.5.8 Prepare an additional two 1-L aliquots of the sample chosen
for spiking for use as the matrix spike and matrix spike duplicate. Add
1.0 mi of the acetone-diluted matrix spiking standard solution (Sec. 5.14)
to each of the two aliquots in the original sample bottles. After allowing
the matrix spiking solution to equilibrate to approximately 1 hour, add the
internal standard solution and filter and extract the aliquots as described
in Sec. 7.5.2.
7.6 Macro-concentration procedures (all matrices)
Prior to cleanup, extracts from all matrices must be concentrated to
approximately 10 ml. In addition, as noted above, the concentrated extracts from
the aqueous filtrate and the filtered particulates must be combined prior to
cleanup. Two procedures may be used for macro-concentration, rotary evaporator,
or Kuderna-Danish (K-D). Concentration of toluene by K-D involves the use of a
heating mantle, as toluene boils above the temperature of a water bath. The two
procedures are described below.
7.6.1 Concentration by K-D
7.6.1.1 Add one or two clean boiling chips to the flask and
attach a three-ball Snyder column. Pre-wet the column by adding
approximately 1 ml of toluene through the top.
7.6.1.2 Attach the solvent recovery system condenser, place
the round bottom flask in a heating mantle and apply heat as required
to complete the concentration in 15-20 minutes. At the proper rate
of distillation, the balls of the column will actively chatter but the
chambers will not flood.
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7.6.1.3 When the apparent volume of liquid reaches 10 ml,
remove the K-D apparatus from the water bath and allow it to drain and
cool for at least 10 minutes.
7.6.2 Concentration by rotary evaporator
7.6.2.1 Assemble the rotary evaporator according to
manufacturer's instructions, and warm the water bath to 45°C. On a
daily basis, preclean the rotary evaporator by concentrating 100 ml
of clean extraction solvent through the system. Archive both the
concentrated solvent and the solvent in the catch flask for
contamination check if necessary. Between samples, three 2-3 ml
aliquots of toluene should be rinsed down the feed tube into a waste
beaker.
7.6.2.2 Attach the round bottom flask containing the sample
extract to the rotary evaporator. Slowly apply vacuum to the system
and begin rotating the sample flask. Lower the sample flask into the
water bath and adjust the speed of rotation to complete the
concentration in 15-20 minutes. At the proper rate of concentration,
the flow of condensed solvent into the receiving flask will be steady,
but no bumping or visible boiling will occur.
7.6.2.3 When the apparent volume of the liquid reaches
10 ml, shut off the vacuum and the rotation. Slowly admit air into
the system, taking care not to splash the extract out of the sample
flask.
7.7 Micro-concentration procedures (all matrices)
When further concentration is required, either a micro-Snyder column
technique or a nitrogen evaporation technique is used to adjust the extract
to the final volume required.
7.7.1 Micro-Snyder column technique
7.7.1.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two-ball micro-Snyder column. Prewet
the column by adding about 0.5 ml of toluene to the top of the column.
7.7.1.2 Place the round bottom flask in a heating mantle and
apply heat 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.
7.7.1.3 When the apparent volume of liquid reaches 0.5 mL,
remove the K-D apparatus from the water bath and allow it to drain and
cool for at least 10 minutes. Remove the Snyder column and rinse the
flask and its lower joints with about 0.2 ml of solvent and add to the
concentrator tube. Adjust the final volume to 1.0 ml with solvent.
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7.7.2 Nitrogen blowdown technique
7.7.2.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the required
level using a gentle stream of clean, dry nitrogen (filtered through
a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon trap and
the sample.
7.7.2.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be positioned
to prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become dry.
7.7.2.3 When the apparent volume of liquid reaches 0.5 ml,
remove the concentrator tube from the water bath. Adjust the final
volume to 1.0 mL with solvent.
7.8 Acid-base cleanup procedure (all matrices)
7.8.1 The concentrated extracts from all matrices are subjected to
a series of cleanup procedures beginning with an acid-base wash, and
continuing on with silica gel chromatography, alumina chromatography, and
carbon chromatography. Begin the cleanup procedures by quantitatively
transferring each concentrated extract to a separate 125-mL separatory
funnel.
7.8.2 Prior to cleanup, all extracts are spiked with the 37C14-
2,3,7,8-TCDD cleanup standard (Sec. 5.13). The recovery of this standard
is used to monitor the efficiency of the cleanup procedures. Spike 5 /zL
of the cleanup standard (or a larger volume of diluted solution containing
25 ng of 37Cl4-2,3,7,8-TCDD) into each separatory funnel containing an
extract, resulting in a concentration of 0.25 ng/juL in the final extract
analyzed by GC/MS.
CAUTION: Concentrated acid and base produce heat when mixed with aqueous
solutions, and may cause solutions to boil or splatter. Perform
the following extractions carefully, allowing the heat and
pressure in the separatory funnel to dissipate before shaking the
stoppered funnel.
7.8.3 Partition the concentrated extract against 40 ml of
concentrated sulfuric acid. Shake for 2 minutes. Remove and discard the
acid layer (bottom). Repeat the acid washing until no color is visible in
the acid layer. (Perform acid washing a maximum of 4 times.)
7.8.4 Partition the concentrated extract against 40 ml of 5 percent
(w/v) sodium chloride. (Caution: Acid entrained in the extract may
produce heat when mixed with the sodium chloride solution). Shake for two
minutes. Remove and discard the aqueous layer (bottom).
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7.8.5 Partition the concentrated extract against 40 mL of 20 percent
(w/v) potassium hydroxide (KOH). (Caution: Allow heat to dissipate before
shaking). Shake for 2 minutes. Remove and discard the base layer
(bottom). Repeat the base washing until color is not visible in the bottom
layer (perform base washing a maximum of four times). Strong base (KOH)
is known to degrade certain PCDDs/PCDFs; therefore, contact time should be
minimized.
7.8.6 Partition the concentrated extract against 40 ml of 5 percent
(w/v) sodium chloride. (Caution: Base entrained in the extract may produce
heat when mixed with the sodium chloride solution). Shake for 2 minutes.
Remove and discard the aqueous layer (bottom). Dry the organic layer by
pouring it through a funnel containing a rinsed filter half-filled with
anhydrous sodium sulfate. Collect the extract in an appropriate size (100-
to 250-mL) round bottom flask. Wash the separatory funnel with two 15-mL
portions of hexane, pour through the funnel and combine the extracts.
7.8.7 Concentrate the extracts of all matrices to 1.0 mL of hexane
using the procedures described in Sec. 7.7. Solvent exchange is
accomplished by concentrating the extract to approximately 100 /ul_, adding
2-3 ml of hexane to the concentrator tube and continuing concentration to
a final volume of 1.0 ml.
7.9 Silica gel and alumina column chromatographic procedures
7.9.1 Silica gel column - Insert a glass wool plug into the bottom
of a gravity column (1 cm x 30 cm glass column) fitted with a Teflon®
stopcock. Add 1 g silica gel and tap the column gently to settle the
silica gel. Add 2 g sodium hydroxide-impregnated silica gel, 1 g silica
gel, 4 g sulfuric acid-impregnated silica gel, and 2 g silica gel (Sec.
5.8). Tap the column gently after each addition. A small positive
pressure (5 psi) of clean nitrogen may be used if needed.
7.9.2 Alumina column - Insert a glass wool plug onto the bottom of
a gravity column (1 cm x 30 cm glass column) fitted with a Teflon®
stopcock. Add 6 g of the activated acid alumina (Sec. 5.8.1). Tap the top
of the column gently.
NOTE; Check each new batch of silica gel and alumina by combining 50 ^L
of the calibration verification solution (CC3) with 950 /nL of
hexane. Process this solution through both columns in the same
manner as a sample extract (Sees. 7.9.5 through 7.9.9). Concentrate
the calibration verification solution to a final volume of 50 y.1.
Proceed to Sec. 7.14. If the recovery of any of the analytes is
less than 80%, the batch of alumina or silica gel must not be used.
7.9.3 Add hexane to each column until the packing is free of air
bubbles. A small positive pressure (5 psi) of clean dry nitrogen may be
used if needed. Check the columns for channeling. If channeling is
present, discard the column. Do not tap a wetted column.
7.9.4 Assemble the two columns such that the eluate from the silica
gel column drains directly into the alumina column.
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7.9.5 Apply the concentrated extract (in hexane) from Sec. 7.8.7 to
the top of the silica gel column. Rinse the vial with enough hexane (1-2
ml) to complete the quantitative transfer of the sample to the surface of
the silica.
7.9.6 Using 90 ml of hexane, elute the extract from Column 1
directly onto Column 2 which contains the alumina. Do not allow the
alumina column to run dry.
7.9.7 Add 20 ml of hexane to Column 2, and elute until the hexane
level is just below the top of the alumina. Do not discard the eluted
hexane, but collect in a separate flask and store it for later use, as it
may be useful in determining where the labeled analytes are being lost if
recoveries are less than 50%.
7.9.8 Add 20 ml of 20% methylene chloride/80% hexane (v/v) to Column
2 and collect the eluate.
7.9.9 Concentrate the extract to approximately 2 to 3 ml using
the procedures in Sec. 7.7.
CAUTION: Do not concentrate the eluate to dryness. The sample is now
ready to be transferred to the carbon column.
7.10 Carbon column chromatographic procedure
7.10.1 Thoroughly mix 9.0 g activated carbon (Carbopak C, Sec. 5.8.2)
and 41.0 g Celite 545 to produce a 18% w/w mixture. Activate the mixture
at 130°C for 6 hours, and store in a desiccator.
NOTE: Check each new batch of the carbon/Celite mixture by adding 50 ;uL
of the calibration verification solution to 950 /zl_ of hexane.
Process the spiked solution in the same manner as a sample extract
(Sees. 7.10.3 through 7.10.5). Concentrate the calibration
verification solution to 50 /j.1 and proceed with Sec. 7.14. If the
recovery of any of the analytes is less than 80%, this batch of
carbon/Celite mixture may not be used.
7.10.2 Prepare a 4-inch long glass column by cutting off each end of
a 10-mL disposable serological pipet. Fire polish both ends and flare if
desired. Insert a glass wool plug at one end of the column, and pack it
with 1 g of the Carbon/Celite mixture. Insert an additional glass wool
plug in the other end.
CAUTION: It is very important that the column be packed properly to ensure
that carbon fines are not carried into the eluate. PCDDs/PCDFs
will adhere to the carbon fines and greatly reduce recovery. If
carbon fines are carried into the eluate in Sec. 7.10.5, filter
the eluate, using a 0.7 jum filter (pre-rinsed with toluene), then
proceed to Sec. 7.11.
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7.10.3 Rinse the column with:
4 ml Toluene
2 mL of Methylene Chloride/Methanol/Toluene (75:20:5 v/v)
4 ml of Cyclohexane/Methylene Chloride (50:50 v/v)
Discard all the column rinsates.
7.10.4 While the column is still wet, transfer the concentrated
eluate from Sec. 7.9.10 to the prepared carbon column. Rinse the eluate
container with two 0.5-mL portions of hexane and transfer the rinses to the
carbon column. Elute the column with the following sequence of solvents.
10 ml of Cyclohexane/Methylene Chloride (50:50 v/v).
5 ml of Methylene Chloride/Methanol/Toluene (75:20:5 v/v).
NOTE: The above two eluates may be collected and combined, and used as a
check on column efficiency.
7.10.5 Once the solvents have eluted through the column, turn the
column over, and elute the PCDD/PCDF fraction with 20 ml of toluene, and
collect the eluate.
7.11 Final concentration
7.11.1 Evaporate the toluene fraction from Sec. 7.10.5 to
approximately 1.0 ml, using the procedures in Sees. 7.6 and 7.7. Transfer
the extract to a 2.0-mL conical vial using a toluene rinse.
CAUTION: Do not evaporate the sample extract to dryness.
7.11.2 Add 100 juL tridecane (or nonane) to the extract and reduce the
volume to 100 /iL using a gentle stream of clean dry nitrogen (Sec. 7.7).
The final extract volume should be 100 juL of tridecane (or nonane). Seal
the vial and store the sample extract in the dark at ambient temperature
until just prior to GC/MS analysis.
7.12 Chromatographic conditions (recommended)
7.12.1 Establish the GC operating conditions necessary to achieve the
resolution and sensitivity required for the analyses, using the following
conditions as guidance for the DB-5 (or equivalent) column:
Helium Linear Velocity 35 - 40 cm/sec at 240°C
Initial Temperature 170'C
Initial Time 10 minutes
Temperature Program increase to 320"C at 8"C/minute
Hold Time until OCDF elutes
Total Time 40-45 minutes
On the DB-5 column,the Chromatographic resolution is evaluated using the
CCS calibration standard during both the initial calibration and the
calibration verification. The Chromatographic peak separation between the
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13C12-2,3,7,8-TCDD peak and the 13C12-1,2,3,4-TCDD peak must be resolved with
a valley of < 25 percent, where:
Valley = (-) x 100
y
y = the peak height of any TCDD isomer
x = measured as shown in Figure 2
The resolution criteria must be evaluated using measurements made on
the selected ion current profile (SICP) for the appropriate ions for each
isomer. Measurements are not made from total ion current profiles.
Optimize the operating conditions for sensitivity and resolution, and
employ the same conditions for both calibration and sample analyses.
7.12.2 For analyses on a SP-2331 (or equivalent) GC column, the
chromatographic resolution is evaluated before the analysis of any
calibration standards by the analysis of a commercially-available column
performance mixture (Sec. 5.16) that contains the TCDD isomers that elute
most closely with 2,3,7,8-TCDD on this GC column (1,4,7,8-TCDD and the
1,2,3,7/1,2,3,8-TCDD pair). Analyze a 2-/iL aliquot of this solution, using
the column operating conditions and descriptor switching times previously
established. The GC operating conditions for this column should be
modified from those for the DB-5 (or equivalent) column, focusing on
resolution of the closely-eluting TCDD and TCDF isomers.
NOTE; The column performance mixture may be combined with the window
defining mix into a single analysis, provided that the combined
solution contains the isomers needed to determine that criteria for
both analyses can be met.
The chromatographic peak separation between unlabeled 2,3,7,8-TCDD and
the peaks representing all other unlabeled TCDD isomers should be resolved
with a valley of < 25 percent, where:
Valley = (-) x 100
y
y = the peak height of any TCDD isomer
x = measured as shown in Figure 2
The resolution criteria must be evaluated using measurements made on
the selected ion current profile (SICP) for the appropriate ions for each
isomer. Measurements are not made from total ion current profiles.
Further analyses may not proceed until the GC resolution criteria have
been met.
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7.13 GC/MS Calibration
Calibration of the GC/MS system involves three separate procedures, mass
calibration of the MS, establishment of GC retention time windows, and
calibration of the target analytes. These three procedures are described in
Sees. 7.13.1 to 7.13.3. Samples should not be analyzed until acceptable
descriptor switching times, chromatographic resolution, and calibrations are
achieved and documented. The sequence of analyses is shown in Figure 3.
NOTE: The injection volume for all sample extracts, blanks, quality control
samples and calibration solutions must be the same.
7.13.1 Mass calibration - Mass calibration of the MS is recommended
prior to analyzing the calibration solutions, blanks, samples and QC
samples. It is recommended that the instrument be tuned to greater
sensitivity in the high mass range in order to achieve better response for
the later eluting compounds. Optimum results using FC-43 for mass
calibration may be achieved by scanning from 222-510 amu every 1 second or
less, utilizing 70 volts (nominal) electron energy in the electron
ionization mode. Under these conditions, m/z 414 and m/z 502 should be SO-
SO0/, of m/z 264 (base peak).
7.13.2 Retention time windows - Prior to the calibration of the
target analytes, it is necessary to establish the appropriate switching
times for the SIM descriptors (Table 7). The switching times are
determined by the analysis of the Window Defining Mix, containing the first
and last eluting isomers in each homologue (Table 8). Mixes are available
for various columns.
The ions in each of the four recommended descriptors are arranged so
that there is overlap between the descriptors. The ions for the TCDD,
TCDF, PeCDD, and PeCDF isomers are in the first descriptor, the ions for
the PeCDD, PeCDF, HxCDD and HxCDF isomers are in the second descriptor, the
ions for the HxCDD, HxCDF, HpCDD and HpCDF isomers are in the third, and
the ions for the HpCDD, HpCDF, OCDD and OCDF isomers are in the fourth
descriptor. The descriptor switching times are set such that the isomers
that elute from the GC during a given retention time window will also be
those isomers for which the ions are monitored. For the homologues that
overlap between descriptors, the laboratory may use discretion in setting
the switching times. However, do not set descriptor switching times such
that a change in descriptors occurs at or near the expected retention time
of any of the 2,3,7,8-substituted isomers.
7.13.3 Calibration of target analytes - Two types of calibration
procedures, initial calibration and calibration verification, are necessary
(Sees. 7.13.3.1 and 7.13.3.2). The initial calibration is needed before
any samples are analyzed for PCDDs/PCDFs, and intermittently throughout
sample analysis, as dictated by the results of the calibration
verification. The calibration verification is necessary at the beginning
of each 12-hour time period during which sample are analyzed.
7.13.3.1 Initial Calibration - Once the Window Defining Mix
has been analyzed and the descriptor switching times have been
verified (and after the analysis of the column performance solution,
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if using a GC column other than DB-5), analyze the five concentration
calibration solutions (CC1-CC5), described in Table 1, prior to any
sample analysis.
7.13.3.1.1 The relative ion abundance criteria for
PCDDs/PCDFs presented in Table 9 should be met for all
PCDD/PCDF peaks, including the labeled internal and recovery
standards, in all solutions. The lower and upper limits of
the ion abundance ratios represent a ±15% window around the
theoretical abundance ratio for each pair of selected ions.
The 37Cl4-2,3,7,8-TCDD cleanup standard contains no 35C1, thus
the ion abundance ratio criterion does not apply to this
compound.
7.13.3.1.2 If the laboratory uses a GC column other
than those described here, the laboratory must ensure that
the isomers eluting closest to 2,3,7,8-TCDD on that column
are used to evaluate GC column resolution
7.13.3.2 Calculate the relative response factors (RFs) for
the seventeen unlabeled target analytes relative to their appropriate
internal standards (RFJ (Table 10), according to the formulae below.
For the seven unlabeled analytes and the 37Cl4-2,3,7,8-TCDD cleanup
standard that are found only in the CC3 solution, only one RF is
calculated for each analyte. For the other 10 unlabeled analytes,
calculate the RF of each analyte in each calibration standard.
Calculate the RFs for the five labeled internal standards and
the cleanup standard relative to the appropriate recovery standard
(RFjs) (Table 10), in each calibration standard, according to the
following formulae:
(A 1 + A2) x Q.
pp_n n |s
n ~ (A.1 + A2) x Q
v is is ' ^n
(A. 1 + A. 2) x Q
RF. = 1S " ' '
(A 1 + A2) x Q.
v rs rs' MS
where:
A,,1 and An2 = integrated areas of the two quantitation ions of the
isomer of interest (Table 8)
AJ81 and Ais2 = integrated areas of the two quantitation ions of the
appropriate internal standard (Table 8)
Ars1 and Ars2 = integrated areas of the two quantitation ions of the
appropriate recovery standard (Table 8)
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Qn = nanograms of unlabeled target analyte injected
Qis = nanograms of appropriate internal standard injected
Qrs = nanograms of appropriate recovery standard injected.
There is only one quantitation ion for the 37C1 cleanup
standard. Calculate the relative response factor as described for
RFis, using one area for the cleanup standard, and the sum of the areas
of the ions from the recovery standard.
The RFn and xRFis are dimensionless quantities; therefore, the
units used to express the Qn, Qis, and Qrs must be the same.
7.13.3.3 Calculate the relative response factors for the
unlabeled PCDDs/PCDFs relative to the recovery standards (RFrs), where:
RF = RF x RF.
rs n is
This relative response factor is necessary when the sample is diluted
to the extent that the MS response of the internal standard is less
than 10% of its MS response in the calibration verification standard
(Sec. 7.15.3).
7.13.3.4 Relative Response Factor Criteria - Calculate the
mean RF and percent relative standard deviation (%RSD) of the five RFs
(CC1 to CCS) for each unlabeled PCDD/PCDF and labeled internal
standards present in all five concentration calibration solutions.
No mean RF or %RSD calculations are possible for the 2,3,7,8-
substituted isomers or the cleanup standard found only in the CC3
solution.
%RSD = Standard deviation x
Mean RF
The %RSD of the five RFs (CC1-CC5) for the unlabeled PCDDs/PCDFs and
the internal standards should not exceed 15.0%.
7.13.3.5 The response factors to be used for determining the
total homologue concentrations are described in Sec. 7.15.2.
7.13.3.6 Calibration Verification - The calibration
verification consists of two parts: evaluation of the chromatographic
resolution, and verification of the RF values to be used for
quantitation. At the beginning of each 12-hour period, the
chromatographic resolution is verified in the same fashion as in the
initial calibration, through the analysis of the CCS solution on the
DB-5 (or equivalent) column, or through the analysis of the column
performance solution on the SP-2331 (or equivalent) column.
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Prepare the CCS solution by combining the volumes of the
solutions listed in Table 4 to yield a final volume of 1.0 ml at the
concentrations listed for the CCS solution in Table 1. Alternatively,
use a commercially-prepared solution that contains the target analytes
at the CCS concentrations listed in Table 1.
For the DB-5 (or equivalent) column, begin the 12-hour period
by analyzing the CCS solution. Inject a 2-p.l aliquot of the
calibration verification solution (CCS) into the GC/MS. The identical
GC/MS/DS conditions used for the analysis of the initial calibration
solutions must be used for the calibration verification solution.
Evaluate the chromatographic resolution using the QC criteria in Sec.
7.12.1.
For the SP-2331 (or equivalent) column, or other columns with
different elution orders, begin the 12-hour period with the analysis
of a 2-/iL aliquot of the appropriate column performance solution.
Evaluate the chromatographic resolution using the QC criteria in Sec.
7.12.2. If this solution meets the QC criteria, proceed with the
analysis of a 2-/xL aliquot of the CCS solution. The identical
GC/MS/DS conditions used for the analysis of the initial calibration
solutions must be used for the calibration verification solution.
Calculate the RFs for the seventeen unlabeled target analytes
relative to their appropriate internal standards (RFJ and the
response factors for the five labeled internal standards and the
cleanup standard relative to the appropriate recovery standard (RFis),
according to the formulae in Sec. 7.13.3.2.
Calculate the RFs for the unlabeled PCDDs/PCDFs relative to
the recovery standards (RFrs), using the formula in Sec. 7.13.3.3.
Do not proceed with sample analyses until the calibration
verification criteria have been met for:
1) GC Column Resolution Criteria - The chromatographic
resolution on the DB-5 (or equivalent) and /or the SP-2331 (or
equivalent) column must meet the QC criteria in Sec. 7.12. In
addition, the chromatographic peak separation between the 1,2,3,4,7,8-
HxCDD and the 1,2,3,6,7,8-HxCDD in the CCS solution shall be resolved
with a valley of < 50 percent (Figure 2).
2) Ion Abundance Criteria - The relative ion abundances
listed in Table 9 must be met for all PCDD/PCDF peaks, including the
labeled internal and recovery standards.
3) Instrument Sensitivity Criteria - For the CC3 solution,
the signal-to-noise (S/N) ratio shall be greater than 2.5 for the
unlabeled PCDD/PCDF ions, and greater than 10.0 for the labeled
internal and recovery standards.
4) Response Factor Criteria - The measured RFs of each analyte
and internal standard in the CCS solution must be within ±30.0% of the
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mean RFs established during initial calibration for the analytes in
all five calibration standards, and within ±30.0% of the single-point
RFs established during initial calibration for those analytes present
in only the CCS standard (see Sec. 7.13.3.2).
(RF. - RF )
% Difference = —_! c~ x 100
RF.
where:
RFj = Relative response factor established during initial
calibration.
RFC = Relative response factor established during calibration
verification.
7.14 GC/MS analysis of samples
7.14.1 Remove the extract of the sample or blank from storage.
Gently swirl the solvent on the lower portion of the vial to ensure
complete dissolution of the PCDDs/PCDFs.
7.14.2 Transfer a 50-juL aliquot of the extract to a 0.3-mL vial, and
add sufficient recovery standard solution to yield a concentration of
0.5 ng//iL (1.0 ng/juL for 13C12-OCDD). Reduce the volume of the extract
back down to 50 /xL using a gentle stream of dry nitrogen.
7.14.3 Inject a 2-juL aliquot of the extract into the GC/MS
instrument. Reseal the vial containing the original concentrated extract.
Analyze the extract by GC/MS, and monitor all of the ions listed in Table
7. The same MS parameters used to analyze the calibration solutions must
be used for the sample extracts.
7.14.4 Dilution of the sample extract is necessary if the
concentration of any PCDD/PCDF in the sample has exceeded the calibration
range, or the detector has been saturated. An appropriate dilution will
result in the largest peak in the diluted sample falling between the mid-
point and high-point of the calibration range.
7.14.4.1 Dilutions are performed using an aliquot of the
original extract, of which approximately 50 /nL remain from Sec.
7.14.2. Remove an appropriate size aliquot from the vial and add it
to a sufficient volume of tridecane (or nonane) in a clean 0.3-mL
conical vial. Add sufficient recovery standard solution to yield a
concentration of 0.5 ng/juL (1.0 ng/^L for 13C-OCDD). Reduce the
volume of the extract back down to 50 juL using a gentle stream of dry
nitrogen.
7.14.4.2 The dilution factor is defined as the total volume
of the sample aliquot and clean solvent divided by the volume of the
sample aliquot that was diluted.
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7.14.4.3 Inject 2 piL of the diluted sample extract into the
GC/MS, and analyze according to Sees. 7.14.1 through 7.14.3.
7.14.4.4 Diluted samples in which the MS response of any
internal standard is greater than or equal to 10% of the MS response
of that internal standard in the most recent calibration verification
standard are quantitated using the internal standards.
Diluted samples in which the MS response of any internal
standard is less than 10% of the MS response of that internal
standard in the most recent calibration verification standard
are quantitated using the recovery standards (see Sec. 7.15.3).
7.14.5 Identification Criteria - For a gas chromatographic peak to
be unambiguously identified as a PCDD or PCDF, it must meet all of the
following criteria.
7.14.5.1 Retention times - In order to make a positive
identification of the 2,3,7,8-substituted isomers for which an
isotopically labeled internal or recovery standard is present in the
sample extract, the absolute retention time (RT) at the maximum peak
height of the analyte must be within -1 to +3 seconds of the retention
time of the corresponding labeled standard.
In order to make a positive identification of the 2,3,7,8-
substituted isomers for which a labeled standard is not available, the
relative retention time (RRT) of the analyte must be within 0.05 RRT
units of the RRT established by the calibration verification. The RRT
is calculated as follows:
RRT _ retention time of the analyte
retention time of the corresponding internal standard
For non-2,3,7,8-substituted compounds (tetra through hepta),
the retention time must be within the retention time windows
established by the window defining mix for the corresponding homologue
(Sec. 7.13.2).
In order to assure that retention time shifts do not adversely
affect the identification of PCDDs/PCDFs, the absolute retention times
of the two recovery standards added to every sample extract
immediately prior to analysis may not shift by more than ±10 seconds
from their retention times in the calibration verification standard.
7.14.5.2 Peak identification - All of the ions listed in
Table 8 for each PCDD/PCDF homologue and labeled standards must be
present in the SICP. The ion current response for the two
quantitation ions and the M-[COCL]+ ions for the analytes must
maximize simultaneously (±2 seconds). This requirement also applies
to the internal standards and recovery standards. For the cleanup
standard, only one ion is monitored.
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7.14.5.3 Signal-to-noise ratio - The integrated ion current
for each analyte ion listed in Table 8 must be at least 2.5 times
background noise and must not have saturated the detector (Figure 4).
The internal standard ions must be at least 10.0 times background
noise and must not have saturated the detector. However, if the M-
[COCL]+ ion does not meet the 2.5 times S/N requirement but meets all
the other criteria listed in Sec. 7.14.5 and, in the judgement of the
GC/MS Interpretation Specialist the peak is a PCDD/PCDF, the peak may
be reported as positive and the data flagged on the report form.
7.14.5.4 Ion abundance ratios - The relative ion abundance
criteria listed in Table 9 for unlabeled analytes and internal
standards must be met using peak areas to calculate ratios.
7.14.5.4.1 If interferences are present, and ion
abundance ratios are not met using peak areas, but all other
qualitative identification criteria are met (RT, S/N,
presence of all 3 ions), then use peak heights to evaluate
the ion ratio.
7.14.5.4.2 If, in the judgement of the analyst, the
peak is a PCDD/PCDF, then report the ion abundance ratios
determined using peak heights, quantitate the peaks using
peak heights rather than areas for both the target analyte
and the internal standard, and flag the result on the report
form.
7.14.5.5 Polychlorinated diphenyl ether (PCDPE) interferences
The identification of a GC peak as a PCDF cannot be made if
a signal having S/N greater than 2.5 is detected at the same retention
time (±2 seconds) in the corresponding PCDPE channel (Table 8). If
a PCDPE is detected, an Estimated Maximum Possible Concentration
(EMPC) should be calculated for this GC peak according to Sec. 7.15.7,
regardless of the ion abundance ratio, and reported.
7.15 Calculations
7.15.1 For GC peaks that have met all the identification criteria
outlined in Sec. 7.14.5, calculate the concentration of the individual PCDD
or PCDF isomers using the formulae:
ALL MATRICES OTHER THAN WATER:
Q. x (A1 + A2)
C (ug/kg) n n
W x (A.1 + A.2) x RF
^ is is' n
WATER:
Q. x (A1 + A2)
Cn (ng/L) = M" "'
V x (A.1 + A.2) x RF
x IR is * n
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where:
An1 and An2 = integrated ion abundances (peak areas) of the
quantitation ions of the isomer of interest (Table 8).
Ais1 and Ais2 = integrated ion abundances (peak areas) of the
quantitation ions of the appropriate internal standard
(Table 8).
Cn = concentration of unlabeled PCDD/PCDF found in the sample.
W = weight of sample extracted, in grams.
V = volume of sample extracted, in liters.
Qis = nanograms of the appropriate internal standard added to
the sample prior to extraction.
RFn = calculated relative response factor from calibration
verification (see Sec. 7.13.3.6).
NOTE: In instances where peak heights are used to evaluate ion abundance
ratios due to interferences (Sec. 7.14.5.4), substitute peak heights
for areas in the formulae above.
For solid matrices, the units of ng/g that result from the formula
above are equivalent to Atg/kg. Using isotope dilution techniques for
quantitation the concentration data are recovery corrected, and therefore,
the volume of the final extract and the injection volume are implicit in
the value of Qis.
7.15.1.1 For homologues that contain only one 2,3,7,8-
substituted isomer (TCDD, PeCDD, HpCDD, and TCDF), the RF of the
2,3,7,8-substituted isomer from the calibration verification will be
used to quantitate both the 2,3,7,8-substituted isomers and the non-
2,3, 7,8-isomers.
7.15.1.2 For homologues that contain more than one 2,3,7,8-
substituted isomer (HxCDD, PeCDF, HxCDF, and HpCDF), the RF used to
calculate the concentration of each 2,3,7,8-substituted isomers will
be the RF determined for that isomer during the calibration
verification.
7.15.1.3 For homologues that contain one or more non-2,3,7,8-
substituted isomer, the RF used to calculate the concentration of
these isomers will be the lowest of the RFs determined during the
calibration verification for the 2,3,7,8-substituted isomers in that
homologue. This RF will yield the highest possible concentration for
the non-2,3,7,8-substituted isomers.
NOTE: The relative response factors of given isomers within any
homologue may be different. However, for the purposes of these
calculations, it will be assumed that every non-2,3,7,8-
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substituted isomer for a given homologue has the same relative
response factor. In order to minimize the effect of this
assumption on risk assessment, the 2,3,7,8-substituted isomer
with the lowest RF was chosen as representative of each
homologue. All relative response factor calculations for the
non-2,3,7,a—substituted isomers in a given homologue are based
on that isomer.
7.15.2 In addition to the concentrations of specific isomers, the
total homologue concentrations are also reported. Calculate the total
concentration of each homologue of PCDDs/PCDFs as follows:
Total concentration = sum of the concentrations of every positively
identified isomer of each PCDD/PCDF homologue.
The total must include the non-2,3,7,8-substituted isomers as well as
the 2,3,7,8-substituted isomers that are also reported separately. The
total number of GC peaks included in the total homologue concentration
should be reported.
7.15.3 If the area of any internal standard in a diluted sample is
less than 10% of the area of that internal standard in the calibration
verification standard, then the unlabeled PCDD/PCDF concentrations in the
sample shall be estimated using the recovery standard, using the formulae
that follow. The purpose is to ensure that there is an adequate MS
response for quantitation in a diluted sample. While use of a smaller
aliquot of the sample might require smaller dilutions and therefore yield
a larger area for the internal standard in the diluted extract, this
practice leads to other concerns about the homogeneity of the sample and
the representativeness of the aliquot taken for extraction.
ALL MATRICES OTHER THAN WATER
Q x (A1 + A2) x D
nrs » n n '
Cn (ug/kg) =
W x (A1 + A2) x RF
» rs rs'
WATER:
Q x (A1 + A2) x D
"rs » n n *
C (ng/L) =
n n
V x (Ar; * A2) x RF
where:
D = the dilution factor (Sec. 7.14.4.2).
An\ An2, Ars1, Ars2, Qrs, RFrs, W, and V are defined in Sees. 7.13.3.2 and
7.15.1.
7.15.4 Report results for soil/sediment, fly ash, and chemical waste
samples in micrograms per kilogram (ju9/kg) and water samples in nanograms
per liter (ng/L).
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7.15.5 Calculate the percent recovery, Ris, for each internal standard
and the cleanup standard in the sample extract, using the formula:
(A.1 + A2) x Q
R (%) = - -^ - i: - !!= — x 100
where:
Ai.1» A»2» A,.1, Ars2, Qis, Qrs, and RFi8 are defined in Sees. 7.13.3.2 and
7.15.1.
NOTE: When calculating the recovery of the 37Cl4-2,3,7,8-TCDD cleanup
standard, only one m/z is monitored for this standard; therefore,
only one peak area will be used in the numerator of this formula.
Use both peak areas of the 13C12-1,2,3,4-TCDD recovery standard in
the denominator.
7.15.5.1 The 13C12-1,2,3,4-TCDD is used to quantitate the TCDD
and TCDF internal standards and the cleanup standard, and the 13C12-
1,2,3,7,8,9-HxCDD is used to quantitate the HxCDD, HpCDF and OCDD
internal standards (Table 10).
7.15.5.2 If the original sample, prior to any dilutions, has
any internal standard with a percent recovery of less than 25% or
greater than 150%, re-extraction and reanalysis of that sample is
necessary.
7.15.6 Sample specific estimated detection limit - The sample
specific estimated detection limit (EDL) is the estimate made by the
laboratory of the concentration of a given analyte required to produce a
signal with a peak height of at least 2.5 times the background signal
level. The estimate is specific to a particular analysis of the sample,
and will be affected by sample size, dilution, etc.
7.15.6.1 An EDL is calculated for each 2,3,7,8-substituted
isomer that is not identified, regardless of whether or not non-
2,3, 7, 8-substituted isomers in that homologue are present. The EDL
is also calculated for 2,3,7,8-substituted isomers giving responses
for both the quantitation ions that are less than 2.5 times the
background level .
7.15.6.2 Use the formulae below to calculate an EDL for each
absent 2,3,7,8-substituted PCDD/PCDF. The background level (HJ is
determined by measuring the height of the noise at the expected
retention times of both the quantitation ions of the particular
2,3,7,8-substituted isomer.
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ALL MATRICES OTHER THAN WATER:
WATER:
2.5 x Q. x (H1 + H2) x D
EDL (ug/kg) = ^1 Ls 11
W x (HJ + His2) x RFn
2.5 x Q. x (H1 + H2) x D
EDL (ng/L) = -^—-^ =-1
V x (H.1 + H.2) x RF
v is is' n
where:
Hn1 and Hn2 = The peak heights of the noise for both of the
quantitation ions of the 2,3,7,8-substituted isomer
of interest
Hi81and His2 = The peak heights of both the quantitation ions of the
appropriate internal standards
D = dilution factor (Sec. 7.14.4.2).
Qis, RFi8, W and V are defined in Sees. 7.13.3.2 and 7.15.1.
7.15.7 Estimated maximum possible concentration - An estimated
maximum possible concentration (EMPC) is calculated for 2,3,7,8-substituted
isomers that are characterized by a response with a S/N of at least 2.5 for
both the quantitation ions, but that do not meet all the identification
criteria in Sec. 7.15.5. Calculate the EMPC according to the following
formulae:
ALL MATRICES OTHER THAN WATER:
Q. x (A1 + A2) x D
EMPC (ug/kg) = —= —2 11
n W x (A.1 + A.2) x RF
v is is ' n
WATER:
Q. x (A1 + A2) x D
EMPC (ng/L) = -^ -^ -11
n V x (A.1 + A2) x RF
1 is is' n
where:
Ax1 and Ax2 = Areas of both the quantitation ions.
Ais1> Ais2, Qis, RF, D, W, and V are defined in Sees. 7.13.3.2 and 7.15.1.
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7.15.8 Toxic equivalent concentration (TEQ) calculation - The
2,3,7,8-TCDD toxic equivalent concentration of PCDDs/PCDFs present in the
sample is calculated according to the method recommended by the Chlorinated
Dioxins Workgroup (CDWG) of the EPA and the Centers for Disease Control
(CDC). This method assigns a 2,3,7,8-TCDD toxicity equivalency factor
(TEF) to each of the seventeen 2,3,7,8-substituted PCDDs/PCDFs shown in
Table 11 ("Update of Toxicity Equivalency Factors [TEFs] for Estimating
Risks Associated with Exposures to Mixtures of Chlorinated Dibenzo-p-
Dioxins and -Dibenzofurans [CDDs/CDFs]" March 1989 [EPA 625/3-89/016]).
7.15.8.1 The 2,3,7,8-TCDD TEQ of the PCDDs/PCDFs present in
the sample is calculated by summing the product of the concentration
for each of the compounds listed in Table 11 and the TEF for each
compound. The principal purpose of making this calculation is to
provide the data user with a single value, normalized to the toxicity
of 2,3,7,8-TCDD, that can more readily be used in decisions related
to mixtures of these highly toxic compounds.
7.15.8.1.1 The exclusion of homologues such as
mono-, di-, tri- and the non-2,3,7,8-substituted isomers in
the higher homologues does not mean that they are not toxic.
Their toxicity, as estimated at this time, is much less than
the toxicity of the compounds listed in Table 11. Hence,
only the 2,3,7,8-substituted isomers are included in the TEF
calculations. The procedure for calculating the 2,3,7,8-TCDD
toxic equivalence cited above is not claimed by the CDWG to
be based on a thoroughly established scientific foundation.
Rather, the procedure represents a "consensus recommendation
on science policy."
7.15.8.1.2 When calculating the TEQ of a sample,
include only those 2,3,7,8-substituted isomers that were
detected in the sample and met all of the qualitative
identification criteria in Sec. 7.14.5. Do not include EMPC
or EDL values in the TEQ calculation.
7.15.8.2 The TEQ of a sample is also used in this analytical
procedure to determine when second column confirmation may be
necessary. The need for second column confirmation is based on the
known difficulties in separating all 17 of the 2,3,7,8-substituted
PCDDs/PCDFs. Historical problems have been associated with the
separation of 2,3,7,8-TCDD from 1,2,3,7-TCDD and 1,2,6,8-TCDD, and
separation of 2,3,7,8-TCDF from 2,3,4,7-TCDF. Because of the
toxicological concern associated with 2,3,7,8-TCDD and 2,3,7,8-TCDF,
additional analyses may be required for some samples, as described
below.
7.15.8.2.1 If the TEQ calculated in Sec. 7.15.8.1
is greater than 0.7 ppb for soil/sediment or fly ash, 7 ppb
for chemical waste, or 7 ppt for an aqueous sample, then
better isomer specificity may be required than can be
achieved on the DB-5 column. The TEQ values listed here for
the various matrices are equivalent to 70% of the historical
"Action Level" set by the CDC for soil concentrations of
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2,3,7,8-TCDD at Superfund sites. As such, it provides a
conservative mechanism for determining when the additional
specificity provided by a second column confirmation is
required.
7.15.8.2.2 The sample extract may be reanalyzed on
a 60 m SP-2330 or SP-2331 GC column (or equivalent) in order
to achieve better GC resolution, and therefore, better
identification and quantitation of the individual 2,3,7,8-
substituted isomers. Other columns that provide better
specificity for 2,3,7,8-TCDD and 2,3,7,8-TCDF than the DB-5
column may also be used.
7.15.8.2.3 Regardless of the GC column used, for
a gas chromatographic peak to be identified as a 2,3,7,8-
substituted PCDD/PCDF isomer during the second column
confirmation, it must meet the ion abundance, signal-to-
noise, and retention time criteria listed in Sec. 7.14.5.
7.15.8.2.4 For any sample analyzed on a DB-5
or equivalent column in which either 2,3,7,8-TCDD or
2,3,7,8-TCDF is reported as an EMPC, regardless of the
TEQ or matrix, analysis of the extract is necessary on
a second GC column which provides clearer specificity
for these two isomers of greatest toxicological concern.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Each laboratory should maintain a formal quality assurance program.
The labortory should also maintain records to document the quality of the data
generated.
8.2 Quality control procedures necessary to evaluate the GC/MS system
operation include evaluation of chromatographic resolution, retention time
windows, calibration verification and chromatographic analysis of samples.
Performance criteria are given in the following sections of Method 8280A:
8.2.1 GC resolution criteria for the DB-5 or equivalent column are
given in Sec. 7.12.1.
8.2.2 GC resolution criteria for SP-2331 or equivalent column are
given in Sec. 7.12.2.
8.2.3 Initial calibration criteria are given in Sec. 7.13.3.1.
8.2.4 Relative response factor criteria for the initial calibration
criteria are given in Sec. 7.13.3.4.
8.2.5 Calibration verification criteria are given in Sec. 7.13.3.6.
8.2.6 Ion abundance criteria are given in Sees. 7.13.3.1, 7.13.3.6,
and 7.14.5.4.
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8.2.7 Instrument sensitivity criteria are given in Sec. 7.13.3.6.
8.2.8 Relative response factor criteria for the calibration
verification are given in Sec. 7,13.3,6.
8.2.9 Identification criteria are given in Sec. 7.14.5.
8.2.10 Criteria for Isotopic Ratio Measurements for PCDDs/PCDFs are
given in 7.13.3.1, 7.13.3.6, and Table 9.
8.3 Initial Demonstration of Proficiency - Each laboratory must
demonstrate initial proficiency with each sample preparation and determinative
method combination it utilizes, by generating data of acceptable accuracy and
precision for target analytes in a clean matrix. The laboratory must also repeat
the following operations whenever new staff are trained or significant changes
in instrumentation are made. See Method 8000, Sec. 8.0 for information on how
to accomplish this demonstration.
8.4 Sample Quality Control for Preparation and Analysis - The laboratory
must also have procedures for documenting the effect of the matrix on method
performance (precision, accuracy, and detection limit). At a minimum, this
includes the analysis of QC samples including a method blank, a matrix spike, a
duplicate, and a laboratory control sample (LCS) in each analytical batch.
8.4.1 Documenting the effect of the matrix should include the
analysis of at least one matrix spike and one duplicate unspiked sample or
one matrix spike/matrix spike duplicate pair. The decision on whether to
prepare and analyze duplicate samples or a matrix spike/matrix spike
duplicate must be based on a knowledge of the samples in the sample batch.
If samples are expected to contain target analytes, then laboratories may
use one matrix spike and a duplicate analysis of an unspiked field sample.
If samples are not expected to contain target analytes, laboratories should
use a matrix spike and matrix spike duplicate pair.
8.4.2 A Laboratory Control Sample (LCS) should be included with each
analytical batch. The LCS consists of an aliquot of a clean (control)
matrix similar to the sample matrix and of the same weight or volume. The
LCS is spiked with the same analytes at the same concentrations as the
matrix spike. When the results of the matrix spike analysis indicate a
potential problem due to the sample matrix itself, the LCS results are used
to verify that the laboratory can perform the analysis in a clean matrix.
8.5 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. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
Method performance data are currently not available.
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10.0 REFERENCES
1. "Update of Toxicity Equivalency Factors (TEFs) for Estimating Risks
Associated with Exposures to Mixtures of Chlorinated Dibenzo-p-Dioxins and
Dibenzofurans (CDDs/CDFs)", March 1989 (EPA 6251/3-89/016).
2. "Method 8290: Polychlorinated Dibenzodioxins (PCDDs) and Polychlorinated
Dibenzofurans (PCDFs) by High Resolution Gas Chromatography/High Resolution
Mass Spectrometry (HRGC/HRMS)", Test Methods for Evaluating Solid Waste,
Physical/Chemical Methods (EPA OSW SW-846).
3. "Statement of Work for Analysis of Polychlorinated Dibenzo-p-dioxins (PCDD)
and Polychlorinated Dibenzofurans, Multi-Media, Multi-Concentration,
DFLM01.1", September 1991.
4. Method 613: 2,3,7,8-Tetrachlorodibenzo-p-Dioxin, 40 CFR Part 136,
Guidelines Establishing Test Procedures for the Analysis of Pollutants
Under the Clean Water Act, October 26, 1984.
11.0 RECOMMENDED SAFETY AND HANDLING PROCEDURES FOR PCDDs/PCDFs
11.1 The following safety practices are excerpts from EPA Method 613,
Sec. 4 (July 1982 version) and amended for use in conjunction with this method.
The 2,3,7,8-TCDD isomer has been found to be acnegenic, carcinogenic, and
teratogenic in laboratory animal studies. Other PCDDs and PCDFs containing
chlorine atoms in positions 2,3,7,8 are known to have toxicities comparable to
that of 2,3,7,8-TCDD. The analyst should note that finely divided dry soils
contaminated with PCDDs and PCDFs are particularly hazardous because of the
potential for inhalation and ingestion. It is recommended that such samples be
processed in a confined environment, such as a hood or a glove box. Laboratory
personnel handling these types of samples should wear masks fitted with charcoal
filters to prevent inhalation of dust.
11.2 The toxicity or carcinogenicity of each reagent used in this method
is not precisely defined; however, each chemical compound should be treated as
a potential health hazard. From this viewpoint, exposure to these chemicals must
be kept to a minimum. The laboratory is responsible for maintaining a current
awareness file of OSHA regulations regarding the safe handling of the chemicals
listed in this method. A reference file of material safety data sheets should
be made available to all personnel involved in the chemical analysis of samples
suspected to contain PCDDs and/or PCDFs.
11.3 Each laboratory must develop a strict safety program for the handling
of PCDDs and PCDFs. The laboratory practices listed below are recommended.
11.3.1 Contamination of the laboratory will be minimized by
conducting most of the manipulations in a hood.
11.3.2 The effluents of sample splitters for the gas chromatograph
and roughing pumps on the HRGC/HRMS system should pass through either a
column of activated charcoal or be bubbled through a trap containing oil
or high boiling alcohols.
8280A - 37 Revision 1
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11.3.3 Liquid waste should be dissolved in methanol or ethanol and
irradiated with ultraviolet light at a wavelength less than 290 nm for
several days (use F 40 BL lamps, or equivalent). Using this analytical
method, analyze the irradiated liquid wastes and dispose of the solutions
when 2,3,7,8-TCDD and -TCDF congeners can no longer be detected.
11.4 The following precautions were issued by Dow Chemical U.S.A. for safe
handling of 2,3,7,8-TCDD in the laboratory and amended for use in conjunction
with this method. The following statements on safe handling are as complete as
possible on the basis of available toxicological information. The precautions
for safe handling and use are necessarily general in nature since detailed,
specific recommendations can be made only for the particular exposure and
circumstances of each individual use. Assistance in evaluating the health
hazards of particular plant conditions may be obtained from certain consulting
laboratories and from State Departments of Health or of Labor, many of which have
an industrial health service. The 2,3,7,8-TCDD isomer is extremely toxic to
certain kinds of laboratory animals. However, it has been handled for years
without injury in analytical and biological laboratories. Many techniques used
in handling radioactive and infectious materials are applicable to 2,3,7,8-TCDD.
11.4.1 Protective Equipment - Disposable plastic gloves, apron or lab
coat, safety glasses and laboratory hood adequate for radioactive work.
However, PVC gloves should not be used.
11.4.2 Training - Workers must be trained in the proper method of
removing contaminated gloves and clothing without contacting the exterior
surfaces.
11.4.3 Personal Hygiene - Thorough washing of hands and forearms
after each manipulation and before breaks (coffee, lunch, and shift).
11.4.4 Confinement - Isolated work area, posted with signs,
segregated glassware and tools, plastic backed absorbent paper on bench
tops.
11.4.5 Waste - Good technique includes minimizing contaminated waste.
Plastic bag liners should be used in waste cans.
11.4.6 Disposal of Hazardous Wastes - Refer to the November 7, 1986
issue of the Federal Register on Land Ban Rulings for details concerning
the handling of dioxin containing wastes.
11.4.7 Decontamination of Personnel - apply a mild soap with plenty
of scrubbing action. Glassware, tools and surfaces - Chlorothene NU
Solvent (Trademark of the Dow Chemical Company) is the least toxic solvent
shown to be effective. Satisfactory cleaning may be accomplished by
rinsing with Chlorothene, then washing with a detergent and water. Dish
water may be disposed to the sewer after percolation through a charcoal bed
filter. It is prudent to minimize solvent wastes because they require
special disposal through commercial services that are expensive.
11.4.8 Laundry - Clothing known to be contaminated should be disposed
with the precautions described under "Disposal of Hazardous Wastes".
Laboratory coats or other clothing worn in 2,3,7,8-TCDD work area may be
8280A - 38 Revision 1
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laundered. Clothing should be collected in plastic bags. Persons who
convey the bags and launder the clothing should be advised of the hazard
and trained in proper handling. The clothing may be put into a washer
without contact if the launderer knows the problem. The washer should be
run through one full cycle before being used again for other clothing.
11.4.9 Wipe Tests - A useful method for determining cleanliness of
work surfaces and tools is to wipe the surface with a piece of filter
paper, extract the filter paper and analyze the extract.
11.4.10 Inhalation - Any procedure that may generate airborne
contamination must be carried out with good ventilation. Gross losses to
a ventilation system must not be allowed. Handling of the dilute solutions
normally used in analytical and animal work presents no significant
inhalation hazards except in case of an accident.
11.4.11 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.
11.5 It is recommended that personnel working in laboratories where
PCDD/PCDF are handled be given periodic physical examinations (at least
annually). Such examinations should include specialized tests, such as those for
urinary porphyrins and for certain blood parameters which, based upon published
clinical observations, are appropriate for persons who may be exposed to
PCDDs/PCDFs. Periodic facial photographs to document the onset of dermatologic
problems are also advisable.
8280A - 39 Revision 1
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Tables for 8280
Table 1 Concentration Calibration Solutions
Table 2 Quantitation Limits for Target Compounds
Table 3 Internal Standard, Recovery Standard, and Cleanup Standard solutions
Table 4 Calibration Verification Solution
Table 5 Matrix Spiking Solution
Table 6 PCDD/PCDF Isomers in the Window Defining Mix for a 60 m DB-5 Column
Table 7 Recommended Selected Ion Monitoring Descriptors
Table 8 Ions Specified for Selected Ion Monitoring for PCDDs/PCDDs
Table 9 Criteria for Isotopic Ratio Measurements for PCDDs/PCDFs
Table 10 Relationship of Internal Standards to analytes, and Recovery Standards
to Internal Standards, Cleanup Standard, and Analytes
Table 11 2,3,7,8-TCDD Toxicity Equivalency Factors (TEFs) for the
Polychlorinated Dibenzodioxins and Dibenzofurans
Figures for 8280
Figure 1 General structures of PCDDs (top) and PCDFs (bottom).
Figure 2 Valley between 2,3,7,8-TCDD and other closely eluting isomers on a
DB-5 GC column.
Figure 3 Example of the Analytical Sequence for calibrating a SP-2331 Column.
Figure 4 Measurement of the signal-to-noise ratio.
8280A - 40
Revision 1
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TABLE 1
CALIBRATION SOLUTIONS
Concentration
Analyte
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,7,8-PeCDD
* 2,3,4,7,8-PeCDF
* 1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
* 1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
* 1,2,3,7,8,9-HxCDD
* 2,3,4,6,7,8-HxCDF
* 1,2,3,7,8,9-HxCDF
* 1,2,3,4,7,8,9-HpCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,6,7,8-HpCDD
OCDD
OCDF
13C12-2,3,7,8-TCDD
13C12-2,3,7,8-TCDF
13C12-l,2,3,6,7,8-HxCDD
13C12-l,2,3,4,6,7,8-HpCDF
13C12-OCDD
13C12-1234-TCDD
13C12-l23789-HxCDD
37Cl4-2378-TCDD
CC1
0.1
0.1
0.1
0.1
0.25
0.25
0.25
0.25
0.5
0.5
0.5
0.5
0.5
1.0
1.0
0.5
0.5
CC2
0.25
0.25
0.25
0.25
0.625
0.625
0.625
0.625
1.25
1.25
0.5
0.5
0.5
1.0
1.0
0.5
0.5
of Standard in ng/jxL
CC3
0.5
0.5
0.5
0.5
0.5
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
2.5
2.5
0.5
0.5
0.5
1.0
1.0
0.5
0.5
0.25
CC4
1.0
1.0
1.0
1.0
2,5
2.5
2.5
2.5
5.0
5.0
0.5
0.5
0.5
1.0
1.0
0.5
0.5
CC5
2.0
2.0
2.0
2.0
5.0
5.0
5.0
5.0
10.0
10.0
0.5
0.5
0.5
1.0
1.0
0.5
0.5
These compounds are only required in the CC3 solution.
perform % RSD calculations on these analytes.
Therefore, do not
8280A - 41
Revision 1
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TABLE 2
QUANTITATION LIMITS FOR TARGET COMPOUNDS
Analyte
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,7,8-PeCDD
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
2,3,4,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
CAS Number
1746-01-6
51207-31-9
57117-41-6
40321-76-4
57117-31-4
70648-26-9
57117-44-9
39227-28-6
57653-85-7
19408-74-3
60851-34-5
72918-21-9
67562-39-4
35822-46-9
55673-89-7
3268-87-9
39001-02-0
Water
(ng/L)
10
10
25
25
25
25
25
25
25
25
25
25
25
25
25
50
50
Soil
1.0
1.0
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0
Fly
Ash
1.0
1.0
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0
Chemical
Waste*
(Mg/kg)
10
10
25
25
25
25
25
25
25
25
25
25
25
25
25
50
50
Chemical Waste includes the matrices of oils, still bottoms, oily sludge, wet
fuel oil, oil-laced soil, and surface water heavily contaminated with these
matrices.
8280A - 42
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TABLE 3
INTERNAL STANDARD, RECOVERY STANDARD, AND CLEANUP STANDARD SOLUTIONS
INTERNAL STANDARD SOLUTION
Internal Standards Concentration
13Cl2-2,3,7,8-TCDD 5 ng/^L
13Cl2-2,3,7,8-TCDF 5
13C,2-l,2,3,6,7,8-HxCDD 5
13C12-l,2,3,4,6,7,8-HpCDF 10
13CI2-OCDD 10
RECOVERY STANDARD SOLUTION
Recovery Standards Concentration
13CI2-1,2,3,4-TCDD 5 ng/^L
13Cl2-l,2,3,7,8,9-HxCDD 5 ng/jul
CLEANUP STANDARD SOLUTION
Cleanup Standards Concentration
37Cl4-2,3,7,8-TCDD 5 ng/VL
8280A - 43 Revision 1
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TABLE 4
CALIBRATION VERIFICATION SOLUTION
Volume
Solution
500
125
50
50
50
225
CC4 (Table 1)
Supplemental Calibration solution (below)
Internal Standard solution (Table 3)
Recovery Standard solution (Table 3)
Cleanup Standard solution (Table 3)
Tridecane (or nonane)
This solution will yield a final volume of 1.0 mL at the concentrations specified
for the CCS solution in Table 1.
Supplemental Calibration Solution Prepared from Commercially-Available Materials
Analyte
Concentration (ng/ML)
2,3,4,7,8-PeCDF
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,7,8,9-HpCDF
4
10
10
10
10
10
10
8280A - 44
Revision 1
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TABLE 5
MATRIX SPIKING SOLUTION
Analyte
Concentration (ng/,uL)
1
1
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDF
1,2,3,6,7,8-HxCDD
,2,3,4,6,7,8-HpCDF
,2,3,4,6,7,8-HpCDD
OCDD
OCDF
2.5
2.5
6.25
6.25
6.25
6.25
6.25
6.25
12.5
12.5
This solution is prepared in tridecane (or nonane) and diluted with acetone prior
to use (see Sec. 5.16).
8280A - 45
Revision 1
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TABLE 6
PCDD/PCDF ISOMERS IN THE WINDOW DEFINING MIX FOR A 60 m DB-5 COLUMN
Homologue
First
Eluted
Last
Eluted
Approximate
Concentration
TCDD 1,3,6,8-
TCDF 1,3,6,8-
PeCDD 1,2,4,7,9-
PeCDF 1,3,4,6,8-
HxCDD 1,2,4,6,7,9-
HxCDF 1,2,3,4,6,8-
HpCDD 1,2,3,4,6,7,9-
HpCDF 1,2,3,4,6,7,8-
1,2,8,9-
1,2,8,9-
1,2,3,8,9-
1,2,3,8,9-
1,2,3,4,6,7-
1,2,3,4,8,9-
1,2,3,4,6,7,8-
1,2,3,4,7,8,9-
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
8280A - 46
Revision 1
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TABLE 7
RECOMMENDED SELECTED ION MONITORING DESCRIPTORS
Descriptor 1 Descriptor 2 Descriptor 3 Descriptor 4
243
259
277
293
304
306
316
318
320
322
328
332
334
340
342
356
358
376
277
293
311
327
338
340
342
354
356
358
374
376
390
392
402
404
410
446
311
327
345
361
374
376
390
392
402
404
408
410
420
422
424
426
446
480
345
361
379
395
408
410
420
422
424
426
442
444
458
460
470
472
480
514
The ions at m/z 376 (HxCDPE), 410 (HpCDPE), 446 (OCDPE), 480 (NCDPE) and 514
(DCDPE) represent the polychlorinated diphenyl ethers.
The ions in each of the four recommended descriptors are arranged so that there is
overlap between the descriptors. The ions for the TCDD, TCDF, PeCDD, and PeCDF
isomers are in the first descriptor, the ions for the PeCDD, PeCDF, HxCDD and HxCDF
isomers are in the second descriptor, the ions for the HxCDD, HxCDF, HpCDD and
HpCDF isomers are in the third, and the ions for the HpCDD, HpCDF, OCDD and OCDF
isomers are in the fourth descriptor.
8280A - 47 Revision 1
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TABLE 8
IONS SPECIFIED FOR SELECTED ION MONITORING FOR PCDDs/PCDFs
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
Internal Standards
13Cl2-2,3,7,8-TCDD
13Cl2-l,2,3,6,7,8-HxCDD
13CI2-OCDD
13Cl2-2,3,7,8-TCDF
13CI2-1,2,3,4,6,7,8-HPCDF
Recovery Standards
13CI2-1,2,3,4-TCDD
13Cl2-l,2,3,7,8,9-HxCDD
Cleanup Standard
37C14-2,3,7,8-TCDD
Polychlorinated diphenyl
HxCDPE
HpCDPE
OCDPE
NCDPE
DCDPE
Quantitation
320
356
390
424
458
304
340
374
408
442
332
402
470
316
420
332
402
328
ethers
376
410
446
480
514
Ions
322
358
392
426
460
306
342
376
410
444
334
404
472
318
422
334
404
(1)
—
—
—
—
_ _ ..
M-[COC1]+
259
293
327
361
395
243
277
311
345
379
—
—
—
—
—
—
—
265
_ _ _
—
—
—
— — _
(1) There is only one quantitation ion monitored for the cleanup standard,
8280A - 48
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TABLE 9
CRITERIA FOR ISOTOPIC RATIO MEASUREMENTS FOR PCDDs/PCDFs
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
Internal Standards
13C,2-1,2,3,4-TCDD
13C,2-l,2,3,6,7,8-HxCDD
13CI2-OCDD
13C,2-2,3,7,8-TCDF
13CI2-1,2,3,4,6,7,8-HPCDF
Recovery Standards
13CI2-,1,2,3,4-TCDD
13C,2-l,2,3,7,8,9-HxCDD
Selected
Ions
320/322
356/358
390/392
424/426
458/460
304/306
340/342
374/376
408/410
442/444
332/334
402/404
470/472
316/318
420/422
332/334
402/404
Theoretical
Ion
Abundance
0.77
1.55
1.24
1.04
0.89
0.77
1.55
1.24
1.04
0.89
0.77
1.24
0.89
0.77
1.04
0.77
1.24
Control
Limits
0.65
1.24
1.05
0.88
0.76
0.65
1.24
1.05
0.88
0.76
0.65
1.05
0.76
0.65
0.88
0.65
1.05
- 0.89
- 1.86
- 1.43
- 1.20
- 1.02
- 0.89
- 1.86
- 1.43
- 1.20
- 1.02
- 0.89
- 1.43
- 1.01
- 0.89
- 1.20
- 0.89
- 1.43
8280A - 49
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TABLE 10
RELATIONSHIP OF INTERNAL STANDARDS TO ANALYTES, AND RECOVERY
STANDARDS TO INTERNAL STANDARDS, CLEANUP STANDARD, AND ANALYTES
INTERNAL STANDARDS VS. ANALYTES
Internal Standard
Analyte
13,
C12-TCDD
13C12-HxCDD
13,
C12-OCDD
13,
C12-TCDF
13,
C12-HpCDF
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8,9-OCDD
1,2,3,4,6,7,8,9-OCDF
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,5,8,9-HpCDF
1,2,3,4,7,8,9-HpCDF
8280A - 50
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TABLE 10 (continued)
RELATIONSHIP OF INTERNAL STANDARDS TO ANALYTES, AND RECOVERY
STANDARDS TO INTERNAL STANDARDS, CLEANUP STANDARD, AND ANALYTES
RECOVERY STANDARDS VS. ANALYTES, INTERNAL STANDARDS, AND CLEANUP STANDARD
Recovery Standard Analyte, Internal Standard
13C12-1,2,3,4-TCDD 2,3,7,8-TCDD
1,2,3,7,8-PeCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
13C12-2,3,7,8-TCDD
13
37
C12-2378-TCDF
Cl4-2378-TCDD
13C12-l,2,3,7,8,9-HxCDD 1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,5,8,9-HpCDF
1,2,3,4,7,8,9-HpCDF
1,2,3,4,6,7,8,9-OCDD
1,2,3,4,6,7,8,9-OCDF
13C12-l,2,3,6,7,8-HxCDD
l3C12-l,2,3,4,6,7,8-HpCDF
13
C19-OCDD
'12
8280A - 51 Revision 1
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TABLE 11
2,3,7,8-TCDD TOXICITY EQUIVALENCY FACTORS (TEFs) FOR THE PCDDs/PCDFs
Compound
Toxicity Equivalency Factor (TEF)
Mono-, di-, and trichloro dibenzo-p-dioxins
2,3,7,8-tetrachloro-dibenzo-p-dioxin
All other tetrachloro-dibenzo-p-dioxins
1,2,3,7,8-pentachloro-dibenzo-p-dioxin
All other pentachloro-dibenzo-p-dioxins
1,2,3,4,7,8-hexachloro-dibenzo-p-dioxin
1,2,3,6,7,8-hexachloro-dibenzo-p-dioxin
1,2,3,7,8,9-hexachloro-dibenzo-p-dioxin
All other hexachloro-dibenzo-p-dioxins
1,2,3,4,6,7,8-heptachloro-dibenzo-p-dioxin
All other heptachloro-dibenzo-p-dioxins
Octachloro-di benzo-p-di oxi n
All mono-, di-, and trichloro dibenzofurans
2,3,7,8-tetrachlorodibenzofuran
All other tetrachlorodibenzofurans
1,2,3,7,8-pentachlorodibenzofuran
2,3,4,7,8-pentachlorodi benzofuran
All other pentachlorodibenzofurans
1,2,3,4,7,8-hexachlorodi benzofuran
1,2,3,6,7,8-hexachlorodibenzofuran
1,2,3,7,8,9-hexachlorodibenzofuran
2,3,4,6,7,8-hexachlorodibenzofuran
All other hexachlorodibenzofurans
1,2,3,4,6,7,8-heptachlorodibenzofuran
1,2,3,4,7,8,9-heptachlorodibenzofuran
All other heptachlorodibenzofurans
Octachlorod i benzofuran
0.0
1.0
0.0
0.5
0.0
0.1
0.1
0.1
0.0
0.01
0.0
0.001
0.0
0.1
0.0
0.05
0.5
0.0
0.1
0.1
0.1
0.1
0.0
0.01
0.01
0.0
0.001
8280A - 52
Revision 1
January 1995
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FIGURE 1
GENERAL STRUCTURES OF PCDDs (top) AND PCDFs (bottom)
8
o
o
8
O
1
w ^
O
8280A - 53
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January 1995
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FIGURE 2
VALLEY BETWEEN 2,3,7,8-TCDD AND OTHER CLOSELY ELUTING
ISOWERS ON AN OB-5 GC COLUMN
100i
GO
«
n
M
c
c
0)
s
DC
(L)
.« >• • A A_J
22:30
I
24:00
Time
I
25:30
27:00
Selected ion current profile for m/z 322 (TCDDs) produced by MS analysis of
GC performance check solution on a 60 m x 0.32 mm DB-5 fused silica capillary
column with 0.25 urn film thickness.
Injector temp:
Starting temp:
Splitless valve time:
Total time: 60 min
270'C
200*C for 2 min
200 to 220*C 9 5'/min and held for 16 min
220 to 235'C 9 5'/min and held for 7 min
235 to 330'C @ 5*/min and held for 5 min
45 sec
8280A - 54
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FIGURE 3
EXAMPLE OF THE ANALYTICAL SEQUENCE FOR CALIBRATING A SP-2331 COLUMN
Time Analysis
Hour 0 Window Defining Mix
Column Performance Solution (SP-2331)
CC3
CC1 (Initial Calibration)
CC2
CC4
CC5
Blanks and Samples
Hour 12 CC1
Column Performance Solution (SP-2331)
CCS
Blanks and Samples
Hour 24 CC1
Column Performance Solution (SP-2331)
CC3
Blanks and Samples
etc.
CC1 (at completion of all scheduled analyses)
NOTE: CC# represents calibration standards
8280A - 55 Revision 1
January 1995
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FIGURE 4
MEASUREMENT OF THE SIGNAL-TO-NOISE RATIO
100-,
90-
80-
70-
60-
50-
40-
30-
20-
10-
E,
_L
20:00
22:00
24:00
26:00
28.00
30:00
Manual Determination of S/N
The peak height (S) is measured between the mean noise (lines C and D).
These mean signal values are obtained by tracing the line between the
baseline average noise extremes, El and E2, and between the apex average
noise extremes, E3 and E4, at the apex of the signal.
NOTE: It is imperative that the instrument interface amplifier electronic
zero offset be set high enough so that negative going baseline noise
is recorded.
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METHOD 8280A
FLOWCHART FOR THE ANALYSIS OF POLYCHLORINATED DIBENZO-P-DIOXINS AND
POLYCHLORINATED DIBENZOFURANS BY HIGH RESOLUTION GAS
CHROMATOGRAPHY/LOW RESOLUTION MASS SPECTROMETRY (HRGS/LRMS)
Complex Waste Sample
(I) Add Internal Standards
(2) Perform matrix-specific extraction,
Sample Extract
(1) Wash with cone. H2S04
(2) Wash with 5% NaCl
(3) Wash with 20% KOH
(4) Wash with 5% NaCl
(5) Dry extract
(6) Solvent exchange
(7) Silica Gel column
(8) Alumina column
60% CH2Cl2/hexane fraction
(1) Concentrate eluate
(2) Perform carbon column cleanup
(3) Add recovery standard(s)
Analyze by GC/MS
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.3 HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHODS
The following methods are included in this section:
Method 8310:
Method 8315A:
Appendix A:
Method 8316:
Method 8318:
Method 8321A:
Method 8325:
Method 8330:
Method 8331:
Method 8332:
Polynuclear Aromatic Hydrocarbons
Determination of Carbonyl Compounds by High
Performance Liquid Chromatography (HPLC)
Recrystallization of 2,4-
Dinitrophenylhydrazine (DNPH)
Acrylamide, Acrylonitrile and Acrolein by High
Performance Liquid Chromatography (HPLC)
N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC)
Solvent Extractable Non-Volatile Compounds by
High Performance Liquid
Chromatography/Thermospray/Mass Spectrometry
(HPLC/TS/MS) or Ultraviolet (UV) Detection
Solvent Extractable Non-Volatile Compounds by
High Performance Liquid Chromatography/Particle
Beam/Mass Spectrometry (HPLC/PB/MS)
Nitroaromatics and Nitramines by High Performance
Liquid Chromatography (HPLC)
Tetrazene by Reverse Phase High Performance
Liquid Chromatography (HPLC)
Nitroglycerine by High Performance Liquid
Chromatography
FOUR - 12
Revision 3
January 1995
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