S-CUBED
A Division of Maxwell Laboratories, Inc.
CONSOLIDATED GC METHOD
FOR THE DETERMINATION
OF ITD/RCRA PESTICIDES USING
SELECTIVE GC DETECTORS
SSS-R-86-8070
S-CUBED Reference No.: 32145-01
S-CUBED Document No.: R70
Prepared by
P.J. Marsden
Task Manager
Submitted to
W.A. Telliard
Chief, Energy and Mining Industries Branch
Industrial Technology Division
USEPA
401 M Street, Southwest
Washington, D.C. 20460
July, 1986
P.O. Box 1620, La Jolla, California 92038-1620 3398 Carmel Mountain Road, San Diego, California 92121-1095
Tel: (619) 453-0060 TWX: 910-337-1253 Telecopier: (619) 755-0474
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TABLE OF CONTENTS
SECTION PACE
1.0 SUMMARY 1
2.0 REAGENTS AND EQUIPMENT 3
3.0 EXTRACTIONS-APOLAR 10
4.0 EXTRACT CLEANUP-APOLAR 16
5.0 CC ANALYSIS-APOLAR 26
6.0 EXTRACTION-PFIENOXYACID 33
7.0 DERIVATIZATION-PHENOXYACID 39
8.0 CC ANALYSIS-PHENOXYACID 41
9.0 QUALITY CONTROL 46
10.0 REFERENCES 49
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1.0 SUMMARY
1.1 This method is a consolidation of methods 608, 614, 615, 617,
622 and 701 that can be used for the analysis of water, solid
and multimedia samples. The method has been validated for the
analysis of organochiorine and organophosphorous pesticides and
industrial chemicals listed as analyzable by “GC/EC” and
“CC/NPD” in the ITD/RCRA list of analytes (Appendix I).
1.2 Two separate extraction and sample preparation schemes are re-
quired in this method. One for apolar organochiorine (Table 1)
and organophosphorous analytes (Table 2). The other for
phenoxyacid herbicides and their esters (Table 3). The scheme
for the preparation and analyses of all of the method.
1.3 The sample is initially split into separate aliquots for work up
by the different procedures. Apolar analytes are extracted from
liquid samples with methylene chloride using a continuous liquid
extractor or are extracted from solid samples with methylene
chloride/acetone using a sonicator. The apolar extracts are
cleaned using gel permeation chromatography (CPC) followed by
adsorbtion chromatography prior to CC analysis. Phenoxyacid
herbicides and their esters are extracted from acidified liquid
samples with methylene chloride using a continuous liquid ex-
tractor or are extracted from acidified solid samples with
methylene chloride/acetone using a sonicator. The extracted
phenoxyacids are partitioned into aqueous base and their esters
are hydrolyzed to the free acids. The acid fractions are
combined and derivatized with diazomethane then cleaned using
adsorbtion chromatography prior to CC analysis.
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1.4 CC analysis is accomplished on “megabore” capillary columns.
Apolar analytes are analyzed using both electron capture
detection (ECD) (for organochlorine analytes), and flame photo-
metric detection (FPD) (for organophosphates). Derivatized
phenoxyacid herbicides are analyzed by using ECD. The primary
column for all analytes is the DB—5 (or equivalent), the confir-
mation column is the SPB-608.
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2.0 REACENTS AND EQUIPMENT
Sodium Sulfate — anhydrous reagent
four hours, or 120°C for 16 hours,
stored in a glass bottle. Baker
No. 3375 or equivalent.
Methylene chloride, hexane, ethyl
(optional), and methanol: pesticide
is strongly suggested that each
analyzed to demonstrate that it
before use.
grade, heated at 400°C for
cooled in a desiccator, and
anhydrous granular, catalog
ether, acetone, isooctane
quality or equivalent. It
lot of solvent used be
is free of interferences
2.1.3 Primary pesticide and PCB standards will be obtained from the
EPA Quality Assurance Materials Bank, Pesticides and Indus-
trial Chemicals Repository, R.T.P., NC for the purpose of
traceability. Comercial standard should be used for working
solutions but must be shown to be traceable (quantitatively
and qualitatively) to EPA standards.
2.1.4
2.1.5
2.1.6
2.1.7
Mercury (optional)
Copper powder (optional), bright and nonoxidized
Concentrated Sodium hydroxide solution (6 N). Dissolve 24g
NaOH in reagent water and dilute to 100 rnL.
Sulfuric acid solution (1:1 v/v). Slowly add 50 mL H 2 S0 4 (Sp.
gr. 1.84) to ’50 mL of reagent water.
2.1 REAGENTS
2.1.1
2.1.2
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2.1.8 Potassium hydroxide solution (37% w/v). Dissolve 37g of po-
tassium hydroxide in 100 mL of distilled water.
2.1.9 Diol bonded silica 500—mg or 1-g cartridges with stainless
steel frits (Analytichem, Harbor City, CA) or equivalent.
2.1.10 Florisil 500mg cartridges (J.T. Baker), or equivalent.
2.1.11 2,4,5—trichlorophenol for Diol cartridge performance check,
Prepare a 0.1 pg/mL solution in acetone.
2.1.12 Reagent water — Reagent water is defined as a water in which
no interferent is observed at the method detection limit of
any parameter when 1 liter of the reagent water is extracted
and prepared using the sample workup procedures for environ-
mental waters.
2.1.13 N-methyl (N-nitroso-p-.toluenesulfanamide) (Diazald ) fresh and
high purity, Aldrich Chemical Co.
2.1.14 Silicic Acid: 100 mesh powder (optional).
2.1.15 Ten percent acetone in hexane (v/v), prepare by adding 10 mL
of acetone to 90 mL of hexane.
2.1.16 GPC calibration solutions:
2.1.16.1 Bis(2—ethylhexyl)phthalate/pentachlorophenol solution 4 mg/mL
each in methylene chloride.
2.1.16.2 Corn oil - 200 mg/mL in methylene chloride.
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2.1.17 Acidified sodium sulfate. Add 0.5 mL H 2 S0 4 to 100 grams of
sodium sulfate and 30—mL ethyl ether. Completely evaporate
the ether and store at 110°C. (Caution: Transfer the acidi-
fied sodium sulfate into a different vessel before placing it
in the oven. This is required to ensure no flammable ether
residue is placed into a hot oven.)
2.1.18 Dilute sodium hydroxide solution (0.1M). Dissolve 4g NaOH in
reagent water and dilute to 1.OL.
2.2 APPARATUS AND MATERIALS
2.2.1 Apparatus for determining percent moisture.
2.2.1.1 Oven, drying.
2.2.1.2 Desiccator.
2.2.1.3 Crucibles, porcelain (optional).
2.2.1.4 Aluminum weighing pans (optional).
2.2.2 Sonic cell disruptor, Heat Systems - Ultrasonics, Inc. Model
375C (or equivalent) (375 watt with pulsing capability 1/2” or
3/4” disruptor horn).
2.2.3 Sonabox (or equivalent) for use with disrupter.
2.2.4 Beakers, 400-mL.
2.2.5 Kuderna—Danish (K—D) apparatus.
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2.2.5.1 Concentrator tube — 10-mL, graduated (Kontes K-570040-1029, or
equivalent).
2.2.5.2 Evaporative flask - 500-mL (Kontes K-470001-0500, or
equivalent).
2.2.5.3 Snyder column — three—ball macro (Kontes K—503000—0121, or
equivalent).
2.2.6 Powder funnels, 10—cm diameter, for filtration/drying.
2.2.7 Boiling chips
2.2.7.1 Silicon carbide boiling chips (optional) — approximately 10/40
mesh. Heat to 400°C for 30 minutes or solvent rinse before
use.
2.2.7.2 Teflon boiling chips (optional). Solvent rinse before use.
2.2.8 Water bath — heated, with concentric ring cover, capable of
temperature control. The bath should be used in a hood.
2.2.9 Top loading balance, capable of accurately weighing to
0.01 g.
2.2.10 Balance—Analytical, capable of accurately weighing to
aO.0001 g.
2.2.11 Nitrogen evaporation device equipped with a heated bath that
can be maintained at 35-40°C, N—Evap by Organomation Associ-
ates, Inc., South Berlin, MA (or equivalent).
2.2.12 Vials and caps, 2—mL for GC auto sampler.
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2.2.13 Gel permeation chromatography cleanup device: automated
system. Gel permeation chromatograph (GPC) Analytical Bio-
chemical Labs, Inc., GPC Autoprep 1002 (or equivalent) includ-
ing:
2.2.13.1 25-mm ID x 600 - 700-mm glass column packed with 70 g of Bio-
Beads SX—3, Bio—Rad Laboratories (or equivalent).
2.2.13.2 Syringe, 10-mL with luer lock fitting.
2.2.13.3 Syringe filter holder, stainless steel, and filters — TFE
(Celman 4310 or equivalent) or glass fiber.
2.2.13.4 IN detection (optional) Type 6, 254-m 4 u Isco, Inc., Lincoln, NB
(or equivalent).
2.2.14 Vacuum systeni for eluting multiple cleanup cartridges.
2.2.14.1 Vac Elute Manifold (Analytichem International, Harbor City,
CA, J.T. Baker or Supelco) or equivalent.
2.2.14.2 Vacuum trap made from a 500—mL sidearm flask fitted with a 1
hole stopper and glass tubing.
2.2.14.3 Vacuum pressure gauge.
2.2.14.4 Rack for holding 10—mL volumetric flasks in the manifold.
2.2.15 Pyrex glass wool.
2.2.16 Bottle or test tube, 50-mL with Teflon lined screw cap for
sulfur removal.
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2.2.17 Separatory funnels - 1000-mL, 500-mL and 250-mL with Teflon
stopcocks.
2.2.18 Drying column — Chromatographic column approximately 400—mm
long x 19—mm ID, with coarse frit. (Substitution of a small
pad of disposable Pyrex glass wool for the frit will help
prevent cross—contamination of sample extracts.)
2.2.19 Continuous liquid—liquid extractors for use with methylene
chloride with Teflon or glass connecting lines. Hershberg—
Wolf Extractor, Ace Class Company, Vineland, NH P/N 6841—10
(or equivalent).
2.2.20 Glass scintillation vials, at least 20—mL, with screw cap and
Teflon or aluminum foil liner.
•2221 Spatula. Stainless steel or Teflon.
2.2.22 pH Paper. Wide range, Hydrion Papers, Microessential Labora-
tory, Brooklyn, N.Y. (or equivalent).
2.2.23 Pipet, Volumetric 1.0O-mL (optional).
2.2.24 Syringe, 1.00-mL (optional).
2.2.25 Flask, Volumetric 10.0O-mL.
2.2.26 Vials, 10-mL, with screw cap and teflon liner (optional).
2.2.27 Tube, centrifuge, 12 to 15—mL with 19—mm ground glass joint,
(optional).
2.2.28 Snyder Column, micro with a 19-mm ground glass joint.
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2.2.29 Centrifuge, table top (optional).
2.2.30 Centrifuge bottle: 500-mL (Pyrex 1260 or equivalent).
2.2.31 Diazald kit with clear seal joints for generation of diazo—
methane, Aldrich Chemical Co., catalog No. Z10, 025—0.
2.2.32 Flask, filter, 1—L.
2.2.33 Funnel, Buchner, 15-cm.
2.2.34 Paper, filter (Whatman #1, or equivalent), 15—cm.
2.2.35 Gas chromatographic system
detection makeup gas, an
flame photometric detector.
equipped with an integrator
chart recorder.
including two 0.25—inch injectors,
electron capture detector, and a
It is recommended that the GC be
or data system rather than a strip
2.3.36 pH Meter with a combination glass electrode.
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3.0 EXTRACTIONS-APOLAR
3.1 OVERVIEW
3.1.1 Aqueous samples are extracted using methylene chloride in a
continuous liquid—liquid extractor. Extracts are then dried
and concentrated by Kuderna—Danish techniques in preparation
for cleanup (Section 4) and analysis (Section 5).
3.1.2 Soil or sediment samples are mixed with sodium sulfate and
extracted using a 1:1 acetone:methylene chloride solvent by a
sonication technique. Extracts are then filtered, dried,
concentrated by Kuderna—Danish, and solvent exchanged to
methylene chloride in preparation for GPC cleanup (4.2).
3.1.3 Most sludge samples are treated as liquid samples (3.1.1)
after 30g of sludge is added to 1-L of reagent water.
3.2 APOLAR PROTOCOL FOR LIQUIDS
3.2.1 Summary of Sample Preparation Method
3.2.1.1 A 1 L volume of a water sample or a mixture of 30 g of sludge
and 1 L of reagent water is extracted with methylene chloride
using a continuous extractor. The methylene chloride extract
is dried, concentrated, exchanged to hexane, and adjusted to a
volume of 10.0 mL and a 1.0 mL aliquot is cleaned up on a Diol
cartridge prior to CC analysis. Optional CPC (4.2) and sulfur
cleanup (4.4) techniques are also allowed.
3.2.2 Liquid Sample Extraction.
3.2.2.1 Liquid samples must be extracted using continuous extraction.
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3.2.2.2 It is strongly suggested that 30g of sludge be added to 1 L of
reagent water and extracted using the continuous extractor.
With some samples, it may be necessary to place a layer of
glass wool between the methylene chloride and water layers in
the extractor to prevent precipitation of suspended solids
into the methylene chloride during extraction. Any sample in
which solids precipitate through the glass wool must be treat-
ed as solids (3.3).
3.2.2.3 The percent weight loss of volatiles from sludge must be
determined and reported according to the procedures described
in 3.3.1.3.
3.2.2.4 After the sample has been transferred to the extractor,
measure and record the pH with wide range pH paper and adjust
to between pH 5 and 9 with 6 N sodium hydroxide or 1:1
sulfuric acid solution, if required. Record and report which
samples require pH adjustment.
3.2.2.5 Add sufficient methylene chloride to the distilling flask to
ensure proper solvent cycling during operation and extract for
18 hours.
3.2.3 Extract Drying and Concentration
3.2.3.1 Assemble a Kuderna-Danish (K—D) concentrator by attaching a
10-mL concentrator tube to a 500-mL evaporative flask. Pour
the combined extract through anhydrous sodium sulfate and
collect the extract in the K—D concentrator (the sodium sul-
fate can be held in a drying column filled to a height of
about 20—cm or in a powder funnel plugged with glass wool
filled to a height of 4—5-cm [ 3.3.2.5]). Rinse the Erlenmeyer
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flask and column with at least one additional 20 to 30 mL
portion of methylene chloride to complete the quantitative
transfer.
3.2.3.2 Add one or two clean boiling chips to the evaporative flask
and attach a three—ball Snyder column. Pre—wet the Snyder
column by adding about 1 mL methylene chloride to the top:
P lace the K-D apparatus on a hot water bath (60-80°C) so that
the concentrator tube is partially immersed in the hot water
and the entire lower rounded surface of the flask is bathed
with hot vapor. Adjust the vertical position of the apparatus
and the water temperature as required to complete the concen-
tration in 10 to 15 minutes. At the proper rate of distilla-
tion, the balls of the column will actively chatter but the
chambers will not flood with condensed solvent. When the
apparent volume of liquid reaches 3—5 mL, remove the K—D appa-
ratus. Allow it to drain and cool for at least 10 minutes.
Do not allow the evaporator to go dry.
3.2.3.3 If GPC cleanup is to be used, remove the Snyder column, rinse
the flask and its lower joint into the concentrator tube,
adjust the volume to 10.0 mL with methylene chloride and
proceed to Section 4. If no GPC cleanup is required, proceed
with the hexane exchange described below.
3.2.4 Exchange into Hexane
3.2.4.1 Momentarily remove the Snyder column, add 50 mL of hexane and
a new boiling chip and re-attach the Snyder column. Pre—wet
the column by adding about 1 mL of hexane to the top. Concen-
trate the solvent extract as before. When the apparent volume
of liquid reaches 3—5 mL, remove the K-D apparatus and allow
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it to drain and cool at least 10 minutes. Do not allow the
evaporator to go dry.
3.2.4.2 Remove the Snyder column, rinse the flask and its lower joint
into the concentrator tube with 1 to 2 mL of hexane and pro-
ceed to Section 4 for Diol cartridge cleanup.
3.3 APOLAR PROTOCOL FOR SOLIDS
3.3.1 Sample Preparation
3.3.1.1 Mix samples thoroughly, especially composited samples. Dis-
card any foreign objects such as sticks, leaves, and rocks.
Also, decant and discard any standing aqueous phase. The lab
must estimate to the nearest 10 percent (by weight) the amount
of water decanted (and discarded) from the sample and report
that value with the data.
3.3.1.2 Transfer 30 g of sample to 100-mL beaker. Add 50 mL of water
and stir for 1 hour. Determine pH of sample using a glass
electrode and pH meter while the sample is stirring. Report
pH value with the data, but do not attempt to adjust the pH of
the sample before extraction. Discard the portion of the
sample used for pH determination.
3.3.1.3 Transfer 5 to 10 g of the sediment into a tarred crucible or
aluminum weighing pan and weigh to the nearest 0.01 g. Trans-
fer the sediment and weighing pan into an oven monitored at
105°C and dry overnight. Allow the sample and weighing pan to
cool in a desiccator before weighing. Concentrations of in-
dividual analytes will be reported relative to the nonvola—
tiles in the sample. (Caution: Gases volatilized from some
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soil/sediment samples may require that this drying procedure
be carried out in a hood.)
wt of sample — wt of heated sample
X 100 = % weight loss (3.1)
wt of sample
3.3.2 Extraction with Sonic Agitation
3.3.2.1 Weigh approximately 30 g of sample (to the nearest 0.1 g) into
a 400—mL beaker and add 60 g of anhydrous sodium sulfate.
3.3.2.2 Immediately add 80 mL of 1:1 methylene chloride:acetone mix-
ture to the sample.
3.3.2.3 Place the sonicator probe about 1/2 inch below the surface of
the solvent but above the sediment layer.
3.3.2.4 Sonicate for 3 mm., using the 3/4 inch horn at full power
with pulse set at 50 percent. Do not use a microtip.
3.3.2.5 Prepare a filtration/drying bed by placing a plug of glass
wool in the neck of a 10—cm powder funnel and filling the
funnel to approximately half its depth (4 or 5 cm) with anhy—
drous sodium sulfate. Decant the extract through the packed
funnel and collect it in a 500—mL evaporation (K—D) flask.
3.3.2.6 Repeat the extraction two more times with fresh 80 mL portions
of the 1:1 methylene chloride:acetone mixture. Decant the
extraction solvent after each sonication. After the final
sonication, pour the entire sample into the funnel and rinse
with 60—mL portion of 1:1 methylene chloride:acetone mixture.
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3.3.2.7 Some samples may require additional clarification before
evaporation. These should be vacuum filtered through Whatman
#1 paper using a Buchner funnel then transferred to a K—D
apparatus.
3.3.2.8 Add one or two clean boiling chips to the evaporative flask
and attach a three—ball Snyder column. Pre—wet the Snyder
column by adding about 1 mL methylene chloride to the top.
Place the K—D apparatus on a hot water bath (60 to 80°C) so
that the concentrator tube is partially immersed in the hot
water and the entire lower rounded surface of the flask is
bathed with hot vapor. Adjust the vertical position of the
appaiatus and the water temperature as required to complete
the concentration in 10 — 15 minutes. At the proper rate of
distillation the balls of the column will actively chatter but
the chambers will not flood with condensed solvent. In order
to remove as much acetone as possible, reduce the apparent
volume of liquid to less than 3 mL, but do not take to
dryness. Remove the K—D apparatus and allow it to drain and
cool for at least 10 minutes. Make up a 10 mL final volume
with methylene chloride.
Proceed to Part 4.1 for mandatory GPC cleanup.
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4.0 EXTRACT CLEANUP-APOLAR
4.1 REQUIREMENTS
4.1.1 GPC cleanup is mandatory for all apolar analyses of soil/sedi-
ment and sludge extracts. It can also be used for water
samples, if needed. GPC removes many higher molecular weight
contaminants which would otherwise accelerate degradation of
gas chromatography columns and thus reduce instrument perform-
ance. The GPC must meet monthly performance checks.
4.1.2 Diol cartridge cleanup is required for all extracts. It
removes polar organic molecules such as phenols. Each lot
number of Diol cartridges must pass a cartridge performance
check.
4.1.3 Sulfur can be removed by one of two methods, according to
laboratory preference. Chromatogram interference due to sul-
fur is not acceptable.
4.2 EXTRACT CLEANUP BY GEL PERMEATION CHROMATOGRAPHY (GPC)
4.2.1 GPC Setup, Operation and Initial Calibration
4.2.1.1 Packing the column — Place 70 g of Bio Beads SX—3 in a 400—mL
beaker. Allow the beads to swell overnight in methylene
chloride before packing the column. Transfer the swelled
beads to the column and begin pumping solvent through the
column, from bottom to top, at a rate of 5.0 mL/min. After
approximately 1 hour, adjust the pressure on the column to
between 5 and 10 psi and pump an additional 4 hours to remove
air from the column. Adjust the column pressure periodically
as required to maintain 5 to 10 psi.
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4.2.1.2 NOTE: The description of solvent flow rate and column pres-
sure applies only to the ABC GPC apparatus. Laboratories
using equivalent equipment must develop the parameters for
their apparatus which give acceptable performance (described
in Section 4.2.2).
4.2.1.3 The SX—3 Bio Beads column may be used for several months, even
if discoloration occurs. System calibration usually remains
constant over this period of time if column flow rate remains
constant. The calibration must be checked periodically, not
less than once every 30 days, using standard pesticide and PCB
mixtures.
4.2.1.4 NOTE : Some samples should be centrifuged and/or filtered
through an inert filter held in a 25—mm stainless steel holder
before loading onto the CPC in order to remove particulate
matter.
4.2.1.5 Load the 5.0-mL sample loop using 7.5 mL of the concentrate of
the extract of each soil sample (described in 3.3.2.) and of
those water samples (described in 3.2.2 or 3.2.3.) to be
cleaned up by CPC.
4.2.1.6 NOTE : Some samples may have to be loaded into two or more
loops in order to prevent overloading the GPC column. All
highly viscous samples or samples known to contain > 1 g of
nonvolatile residue should be divided and loaded into two or
more loops.
4.2.1.7 Set the instrument terminal for the appropriate number of
samples; set the length of the dump, collect and wash cycles
for the number of minutes determined by either of the calibra—
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tion procedures described below in 4.2.1.9 or 4.2.1.10. Set
up and label one collection flask for each loop by placing the
appropriate Teflon effluent line into the flask, then covering
the flask with aluminum foil. Begin pumping methylene
chloride through the column at 5 mL/min. (5 to 10 psi), allow
the flow to stabilize for at least 15 minutes, then begin the
automatic cleanup sequence by pressing the “auto start”
button.
4.2.1.8 After the appropriate CPC fraction has been collected for each
sample, evaporate the methylene chloride and exchange to
hexane as described in 3.2.5.
4.2.1.9 Two alternate CPC calibration procedures are described below,
one of these two must be used. The first is based on a
combination of gravimetric and CC analysis of collected GPC
fractions, the second is based on monitoring the elution of
standards with a UV detector connected to the GPC column. If
the UV technique is chosen, care must be taken to account for
any difference in volume (elution time) between the CC column
and the detector and the CPC column and the collection vial.
4.2.2 Cravimetric GPC Calibration Procedure
4.2.2.1 Set the “dump”, “collect” and “wash” times for 00, 02, and 00
minutes, respectively. Set the terminal for 20 samples.
4.2.2.2 Under each of the first twenty receiver lines, place a tared
20—mL scintillation vial.
4.2.2.3 Into Sample No. 1 load 5 mL of 200-mg/mL corn oil solution.
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4.2.2.4 Start the auto sequence, collect twenty 10-mL fractions (40
mm.).
4.2.2.5 Passively evaporate each fraction to dryness in a fume hood
(overnight).
4.2.2.6 Reweigh the vials and record the net weight gain.
4.2.2.7 Plot weight of the elute vs. time to determine the elution
profile.
4.2.2.8 Reset the sampling position to 00.
4.2.2.9 Into Sample Loop No. 1, load 5 mL of 0.4-mg/mL PCP/Phthalate
Solution.
4.2.2.10 Set the “collect” time to 03 minutes.
4.2.2.11 Under each of the first twenty receiver lines place a clean
20—mL scintillation vial. ( NOTE : The vials need not be
tared.)
4.2.2.12 Start the auto sequence, collect twenty 15-mL fractions (1
hour).
4.2.2.13 Analyze each fraction by CC to determine the amount of PCP and
phtha late.
4.2.2.14 Plot the amounts of PCP and phthalate to determine the
elution profiles.
4.2.2.15 Choose the “DUMP” time such that 85+ percent of phthalate is
recovered and the corn oil is discarded.
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4.2.2.16 Choose the “COLLECT” time to extend 10 minutes after the
elution of PCP and set a “WASH” time of 10 minutes.
4.2.3 IN Detector Calibration Procedure
4.2.3.1 Connect the end of the column to a IN detector (254 nm).
4.2.3.2 Zero the detector and start the strip chart recorder.
4.2.3.3 Into Sample Loop No. 1 load 5 mL of 20-mg/mL corn oil solution
and into Sample Loop No. 2 load 5 mL of 0.4-mg/mL
PCP/phthalate. (NOTE: The corn oil concentration used in
this UV procedure is a 10—fold dilution of that used in the
gravimetric method.)
4.2.3.4 Inject Sample Loop No. 1, note the injection on the strip
chart recorder.
4.2.3.5 After the corn oil elutes, allow the recorder to continue
for a few minutes to again establish a baseline.
4.2.3.6 Inject Sample Loop No. 2 and note the injection on the strip
chart recorder.
4.2.3.7 Determine the elution times for the corn oil, PCP and
phtha late.
4.2.3.8 Choose a “DUMP” time which removes the corn oil yet recovers
>85 percent of the phthalate. Choose a “COLLECT” time which
continues 10 minutes beyond the elution of pentachlorophenol
and a “WASH” time of 10 minutes.
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4.2.3.9 NOTE : The DUMP and collect
sate for the difference in
detector and the collection
times must be adjusted to compen—
volume of the lines between the
f I ask.
4.2.4
Continuing CPC calibration requirement.
4.2.4.1 At least once ever 30 days, the calibration
verified by determining the recovery of
(Tables 1 and 2).
of the GPC must be
10 method analytes
4.2.4.2 The ten analytes used are chosen by each lab. The choice is
based on which analytes that were most often detected in their
samples during the previous month.
4.2.4.3 The CPC calibration solution is prepared so that the concen-
tration of each analyte is between 0.5 and 5 pg/mL.
4.2.4.5 The collected CRC
tatively to a K-D apparatus
ide is reduced (3.2.3).
exchanged to hexane (3.2.4)
10.0 mL and the sample is
procedures in Section 5.
4.2.4.6 The CRC performance is acceptable
recovered at 85—110 percent and
used. If the recovery is less
110 percent, the column must
brated before more samples are run
loaded into a CPC
established using
4.2.3).
if all of the analytes are
the column can continue to be
than 85 percent, or more than
be repacked (4.2.1) and recali—
4.2.4.4 Using 7.5 mL of the calibration solution
loop and a fraction using the CPC program
either of the described procedures (4.2.2 or
calibration fraction is transferred quanti—
and the volume of methylene chlor—
After cooling, the solvent is
The final volume is adjusted to
analyzed by CC according to the
21
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4.3 DIOL CARTRIDGE CLEANUP
4.3.1 Cartridge Performance Check
Every lot number of Diol cartridges must be tested. A lot of
Diol cartridges is demonstrated as acceptable if all apolar
analytes are recovered at 80—110 percent and if
trichiorophenol is not detected when the compounds are eluted
through a cartridge using the method described in 4.3.4.
4.3.2 Nitrogen Blowdown Technique (Taken from ASTM Method D 3086)
4.3.2.1 Place the concentrator tube in a heating bath (35C) and evap-
orate the solvent to the final volume using a gentle stream of
clean, dry nitrogen (filtered through a column of activated
carbon). The extract must never be allowed to become dry.
4.3.2.2 CAUTION : New plastic tubing must not be used between the
carbon trap and the sample, as it may introduce interferences.
The internal wall of new tubing must be rinsed several times
with hexane then dried prior to use.
4.3.3 Extract Preparation
4.3.3.1 For samples which have been run through the GPC cleanup
solvent, exchange to hexane (3.2.4) and adjust the hexane
extract volume to 5.0 mL using nitrogen blowdown described in
4.3.2. For those aqueous samples not passed through the GPC
step, adjust to 10.0 mL. The different final volumes are
required because only half of the methylene chloride
concentrates are recovered from the GPC cleanup.
4.3.4 Diol Cartridge Cleanup
22
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4.3.4.1 Attach the Vac—Elute vacuum manifold to a water aspirator or a
vacuum pump with a trap installed between the manifold and the
vacuum source. Adjust the vacuum pressure in the manifold to
between 5 and 10 pounds of vacuum.
4.3.4.2 Most liquid samples can be cleaned using a 500-mg Diol
cartridge, the cleanup of soil and sludge extracts must be
accomplished using 1 g cartridges.
4.3.4.3 Prior to cleanup of samples, the cartridges must be washed
with hexane/acetone (9:1). This is accomplished by placing
the cartridge in the vacuum manifold, pulling a vacuum and
passing 5 mL of the hexane/acetone solution through the
cartridge.
4.3.4.4 After the cartridges in the manifold are washed, the vacuum is
released and a rack containing labeled 10—mL volumetric flasks
is placed inside the manifold. Care must be taken to ensure
that the solvent line from each cartridge is placed inside of
the appropriate volumetric flask as the manifold top is re-
placed.
4.3.4.5 After the volumetric flasks are in place, vacuum to the
manifold is restored and a volume of 1.0 mL from each sample,
blank or matrix spike extract to be analyzed is transferred to
the top frit of the appropriate Diol cartridge.
4.3.4.6 NOTE : Because the volumes marked on concentrator tubes are
not necessarily accurate, the use of an 1.00—mL syringe or a
volumetric pipet is required.
23
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4.3.4.7 The analytes in the extract concentrates are then eluted
through the column with 9 mL of hexane/acetone (9:1) and col-
lected into the 10-mL volumetric flasks held in the rack in-
side the vacuum manifold.
4.3.4.8 Transfer the eluate in each volumetric flask to a clean cen-
trifuge tube or 10—mL vial. Use two additional 1 mL hexane
rinses to ensure quantitative transfer of the cartridge
e I uate.
4.3.4.9 Concentrate the extract to 1.0 mL using nitrogen blow-down
(described in 4.3.2).
4.3.4.10 If crystals of sulfur are evident or sulfur is suspected to be
present, proceed to Section 4.4.
4.3.4.11 If the sulfur is not expected to be a problem, transfer the
1.0—mL concentrate to a CC vial and label the vial. The ex-
tract is ready for CC analysis, proceed to Section 5. Store
the extracts at 4C in the dark until analyses are performed.
4.4 SULFUR REMOVAL
4.4.1 Two options are available for the removal of sulfur from
samples. The mercury technique appears to be the most reli-
able, but requires the use of small volumes of mercury in the
laboratory.
4.4.2 CAUTION : Mercury containing waste should be segregated and
disposed of properly.
4.4.3 Mercury Technique
24
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4.4.3.1 Add 1 to 3 drops of mercury for each 1-mL hexane extract in a
clean vial. Seal the vial and agitate for 30 seconds. Filter
or centrifuge and decant to remove all solid precipitate and
liquid mercury. Analyze the extract if the mercury appears
shiny. If the mercury turns black, repeat as necessary.
Dispose of the mercury waste properly.
4.4.3.2 If only a partial set of samples requires sulfur cleanup, an
additional reagent blank is not required.
4.4.4 Copper Technique
4.4.4.1 Bright (non—oxidized) granular copper can be used in place of
mercury in the procedure described in Section 4.4.3.1.
25
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5.0 CC ANALYSIS-APOLAR
5.1 SUMMARY
5.1.1 Sample extracts are analyzed by both GC/ECD for organochlorine
parameters and by GC/FPD for organophosphorous parameters.
5.1.2 The primary column for both organochlorine and organophosphate
analysis is a 15 mx 53mm id DB-5 megabore capillary column
(or equivalent).
5.1.3 The secondary column for both organochlorine and organophos—
phorous analyses is a 15 m x 0.53 mm id SPB-608 megabore ca-
pillary column (or equivalent).
5.2 CC CONDITIONS
5.2.1 Megabore capillary columns can be installed in standard
0.25—inch packed column injector and detector ports using
suitable glass adapters and graphite ferrules. Because the
column flow used is 5 mL/minute, it is necessary to supply a
suitable makeup gas to the detector.
5.2.2 Electron capture detectors should be plumbed with helium or
hydrogen carrier and P-b (Argon/methane) as a detector makeup
gas, the detector temperature should be 275C.
5.2.3 Flame photometric detectors should be plumbed with helium or
hydrogen carrier and nitrogen as a detector makeup, the
detector temperature should be 250°C.
5.2.4 The temperature program for organochlorine analysis is:
26
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T = 50°C
Initial time one minute
Initial temperature ramp 20°/mm to 150°C
Second temperature ramp 8°/mm to 180°C
Third temperature ramp 3°/mm to 250°C
Final hold 15 minutes
Note: It may be necessary to adjust this temperature program
for individual gas chromatographs.
5.2.5 The temperature program for organophosphorous analysis is:
= 50C
Initial time one minute
Initial temperature ramp 5°/mm to 140°C, hold 10 minutes
Second temperature ramp 10°/mm to 250°C
l ive minute final hold.
Note: It may be necessary to adjust this temperature program
for individual gas chromatographs.
5.2.6 Injectors should be set at 260°C.
5.3 CALIBRATION
5.3.1 After establishing the appropriate GC operating conditions
(5.2) for the method parameters, calibrate the CC system using
the external standard technique.
5.3.2 For each parameter of interest, prepare working standards at
three concentration levels over a range of a least two orders
of magnitude. The low concentration should be near to, but
27
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above, the method detection limit (Section 5.7). The medium
concentration must be at least 10 times the low concentration
and the high concentration must be at least 10 times the
medium concentration (at least 100 times the low
concentration). These concentrations define the calibration
range in which analytes can be quantitated.
5.3.3 The peak height, or the peak area are plotted against the mass
injected and used as a calibration curve for each parameter.
Alternatively, the ratio of the response to the mass injected,
defined as the calibration factor (CF), can be calculated for
each parameter at each standard concentration. If the rela-
tive standard deviation of the calibration factor for the
three concentrations is less than 10% detector linearity
through the origin can be assumed and the average calibration
factor can be used in place of a calibration curve.
5.3.4 The working calibration curve or calibration factor must be
verified on each working day by the measurement of one or more
calibration standards. If the response for any parameter
varies from the predicted response by more than 10%, a new
calibration curve or calibration factor must be prepared for
that parameter.
5.4 QUANTITATION OF ANALYSES
5.4.1 Analytes can be quantitated using either manual measurement of
onscale chromatograms, with a modern electronic integrator, or
with a laboratory data system. The analyst can use either
peak height or peak area as the basis for quantitation. The
use of electronic integration or a laboratory data system is
strongly recommended.
28
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5.4.2 If manual quantitation is used, all peaks used for
calibration and for sample analysis must be onscale and give
at least 20 percent deflection from baseline at maximum
height. Guidance in the manual quantitation of peaks is given
in the EPA MANUAL OF ANALYTICAL METHODS FOR THE ANALYSIS OF
PESTICIDES IN HUMANS AND ENVIRONMENTAL SAMPLES , EPA-600/8-80-
038, or in the FDA Pesticide Analytical Manual.
5.4.3 If electronic integration is used, it is the responsibility of
the analyst to be sure that the integration parameters are set
properly and that off—scale chromatograms are within the dyna-
mic range of the device. The analyst should also check for
data flags that indicate improper quantitation of peaks prior
to reporting data to the EPA. In addition, the peak width of
identified analytes must not be more than 2.0 times the width
of the high concentration calibration peak(s) for the same
analyte.
5.4.4 The detector response (peak area or peak height) to all of the
analytes must lie between the response of the low and high
concentrations in the three-point initial calibration in order
to be quantitated.
5.4.5 The concentration of the single component analytes are
calculated using the following equations:
5.4.5.1 Water
(A ) (Vt)
concentration jsg/L = (CF)(v )(V ) (5.10)
Where:
Ax = Response for the parameter to be measured.
CF = Calibration factor for the external standard (5.3.3)..
29
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Vt = Volume of total extract (pL) (take into account any
dilution).
V , = Volume of extract injected (pL).
V = Volume of water extracted (mL).
5.4.5.2 Sediment/Soil
(A ) (Vt)
Concentration pg/kg = (cF)(V 1 )(w 5 )(D) (5.10)
(Dry weight basis)
Where:
A , CF, V = same as given above in 5.3.4.1
100 - percent weight loss
D = 100 (percent weight loss from Section 3.3)
W 5 = Weight of sample extracted (g).
5.4.6 The quantitation of multicomponent pesticide/PCBs must be
accomplished by comparing the sum of the heights or the areas
of at least three major peaks of the multicomponent analyte in
the sample compared with the same peaks in the standard. The
concentration of multicomponent analytes are also calculated
using equations 5.10 and 5.11 where Ax is the sum of the major
peaks of the multicoinponent analyte.
5.4.7 The identification and quantitation of multicomponent analytes
may be complicated by the environmental alteration of the peak
pattern of multicomponent pesticides/PCBs, and by the presence
of coeluting analytes and/or matrix interference.
30
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5.4.8 If more than one multicomponent is observed in a sample, the
laboratory must chose separate peaks to quantitate the differ-
ent multicomponent analytes. A peak common to both analytes
present in the sample must not be used to quantitate both
compounds in the same sample.
5.5 Scheme of Analysis.
5.5.1 A flow scheme for the analysis of samples by this procedure
are given in Figure 1.
5.6 Sample chromatograms.
5.6.1 Sample chromatograms of mixtures containing all single
component analytes and of all multicomponent analytes for both
columns are presented in Figures 2—23.
5.7 Method Detection limits.
5.7.1 Any method is dependent on the complexity of the sample matrix
analyzed, but this procedure can be used to determine all of
the single component apolar organochlorine analytes (Table 1),
except diallate to a detection limit of <50 ppb. If greater
sensitivity is required, the 9 mL of extract not subjected to
Diol cleanup (4.3) can be blown down to 1 mL and cleaned on a
1 g Diol column. This concentrate can be used to give a
method detection limit of <5 ppb.
5.7.2 The procedure can be used to determine all of the single
component apolar organophosphate analytes (Table 2) to a
detection limit of <500 ppm. If greater sensitivity is
required, the 9 mL of extract not subjected to Diol cleanup
31
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(4.3) can be blown down to 1 mL and cleaned on a 1 g Diol
column. This concentrate can be used to give a method
detection limit of <50 ppb.
32
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6.0 EXTRACTION-PHENOXYACIDS
6.1 OVERVIEW
6.1.1 Phenoxyacids and their esters are extracted from acidified aque-
ous samples using methylene chloride using a continuous liquid
extractor.
6.1.2 Acids and esters are extracted from acidified solid samples
using methlene chloride/acetone and a sonicator.
6.1.3 Most sludge samples are treated as liquid samples (6.1.1) after
30g of sludge is added to 1 L of reagent wate r.
6.1.4 The sample extracts are reduced in volume using a K-D apparatus,
the free phenoxyacid herbicides are partitioned into aqueous
base and the esters into methylene chloride. The esters are
hydrolyzed using aqueous potassium hydroxide and neutral organ—
ics are partitioned into methylene chloride and discarded. The
two basic aqueous fractions containing phenoxyacids are combin-
ed, acidified, and the acids are extracted into methylene
chloride. The phenoxyacids are then derivitized with diazometh—
ane, cleaned on adsorption cartridges and analyzed by GC/ECD.
6.2 LIQUID SAMPLE EXTRACTION-PHENOXYACID
6.2.1 Liquid samples must be extracted using continuous extraction.
Using a 1 liter graduated cylinder, measure out a 1 liter sample
aliquot and place it into the continuous extractor.
6.2.2 It is strongly suggested that 30 g of sludge be added to 1 L of
reagent water and extracted using the continuous extrator. With
33
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some samples, it may be necessary to place a layer of glass wool
between the methylene chloride and water layers in the extractor
to prevent precipitation of suspended solids into the methylene
chloride during extraction. Any sample in which solids precipi-
tate through the glass wool must be treated as solids (6.3).
6.2.3 The percent weight loss of volatiles from sludge must be
determined and reported according to the procedure described in
Section 6.3.1.3.
6.2.4 After the sample has been transferred to the extractor, measure
and record the pH of the sample with wide range pH paper and
adjust to below 2 with 1:1 sulfuric acid, if required (Caution:
some samples should be acidified in the hood because of the
potential for generating hydrogen sulfide).
6.2.5 Add sufficient methylene chloride to the distilling flask to
ensure proper cycling during operation and extract for 18 hours.
6.2.6 If solids are present in the rnethylene chloride, after
extraction, filter the organic phase through Watman #1 paper.
6.2.7 Transfer the methylene chloride fraction to a 500 mL separatory
funnel and proceed to the separation of the phenoxyacids and
phenoxyesters (6.4).
6.3 SOLID SAMPLE EXTRACTION-PHENOXYACID
6.3.1 Sample Preparation
6.3.1.1 Mix samples thoroughly, especially composited samples. Discard
any foreign objects such as sticks, leaves, and rocks. Also,
34
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decant and discard any standing aqueous phase. The lab must
estimate to the nearest 10 percent (by weight) the amount of
water decanted (and discarded) from the sample and report that
value on the data sheet.
6.3.1.2 Transfer 30g of sample to a 100 mL beaker. Add 50 mL of water
and stir for 1 hour. Determine pH of sample using a glass
electrode and pH meter while the sample is stirring. Report pH
value with the data, then determine the amount of 1:1 sulfuric
acid/water required to reduce the pH to <2. Discard the portion
of the sample used for pH determination.
6.3.1.3 Transfer 5 to 10 g of the sediment into a tarred crucible or
aluminum weighing pan and weigh to the nearest 0.01 g. Transfer
the sediment and weighing pan into an over monitored at 105°C
and dry overnight. Allow the sample and weighing pan to cool in
a desiccator before weighing. Concentrations of individual
analytes will be reported relative to the dry weight of
sediment. (Caution: Cases volatilized from some soil/sediment
samples may require that this drying procedure be carried out in
a hood.)
wt of sample sample x 100 = % weight loss (6.1)
6.3.2 Extraction with Sonic Agitation
6.3.2.1 Weigh approximately 30 g of sample (to the nearest 0.1-g) into
a 400-mL beaker.
6.3.2.2 Add the amount of 1:1 sulfuric acid/water determined in
6.3.1.2 (Caution: some samples may evolve toxic gases on the
addition 0 f acid).
35
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6.3.3.3 Add 60 g of acidified sodium sulfate.
5.3.2.4 Immediately add 80 mL
mixture to the sample.
of 1:1 methylene chloride:acetone
6.3.2.5 Place the sonicator probe about 1/2 inch below the surface of
the solvent but above the sediment layer.
6.3.2.6 Sonicate for 3 mm., using the 3/4 inch horn at full power
with pulse set at 50 percent. Do not use a microtip.
6.3.2.7 Caution — the sonicator horn must be cleaned in 5% aqueous
sodium bicarbonate and methanol between samples. If acid is
allowed to remain on the horn, it will be damaged.
6.3.2.9 Repeat the extraction two more times with fresh 80 mL portions
of the 1:1 methylene chloride:acetone mixture. Decant the
extraction solvent after each sonication. After the final
sonication, pour the entire sample into the funnel and rinse
with a 60-mL portion of the 1:1 methylene chloride:acetone
mixture.
6.3.2.10 If required, centrifuge the combined extract for 10 minutes to
settle fine particles then filter the extracts through Whatman
#1 filter paper into a 500 mL K-D flask.
6.3.2.8 Prepare a filtration/drying bed by placing a plug of glass 1
wool in the neck of a 10—cm powder funnel and filling the
funnel to approximately half its depth (4 or 5 cm) with
acidified sodium sulfate. Decant the extract through the
packed funnel and collect it in a 500-mL evaporation (K—D)
flask, or a 500 mL centrifuge bottle.
36
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6.3.2.11 Transfer the methlene chloride fraction to a 500 mL separatory
funnel prior to the separation of the phenoxyacids and
phenoxyesters.
6.4 ESTER HYDROLYSIS
6.4.1 Extract the organic phase of the phenoxyacid preparation two
times with 100 mL portions of 0.1 N aqueous sodium hydroxide.
Combine the aqueous (top) layers containing the free acid
herbicides in a beaker and save. The organic (bottom) layer
contains the herbicide esters, which must be hydrolyzed.
6.4.2 Transfer the methylene chloride layer into 500-mL Kuderna-
Danish flask. Add boiling chips to the K—D and fit it with a
three—ball Snyder columns. Wet the Snyder column with 1 mL of
methylene chloride and reduce the methylene chloride to a
volume of approximately 25 mL on the water bath.
6.4.3 Remove the flask from the water bath and allow it to cool.
Then add 5 mL of 37% aqueous potassium hydroxide, 30 mL of
distilled water and 40 mL of methanol to the flask.
6.4.4 Add additional boiling chips to the flask. Ref lux the mixture
in the K—D on the water bath for 2 hours. Remove the flask
from the water bath and cool to room temperature.
6.4.5 Transfer hydrolysate from the K—D flask to a 250 mL separatory
funnel. Extract the aqueous residue with 50 mL of methylene
chloride and discard the methylene chloride (bottom layer)
which contains neutral interferents.
37
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6.4.6 At this point the two aqueous solutions containing the free
herbicide salts from 6.4.1 and 6.4.5 can be combined (they can
be analyzed separately, if required).
6.4.7 Add sulfuric acid to the basic aqueous solutions to adjust the
pH to 1.
6.4.8 Transfer the acidified aqueous solution into a 500 mL
separatory funnel and extract the solution three times with
100 niL portions of methylene chloride.
6.4.9 Combine the organic extracts in 500 mL Kuderna-Danish flasks.
Add boiling chips to the extracts in the flasks and fit them
with three—ball Snyder columns.
6.4.10 Evaporate the methylene chloride to approximately 5 niL on a
hot water bath (the samples may be stored at this stage).
6.4.11 Remove the flasks from water bath. Evaporate the extracts
just to dryness under a stream of nitrogen.
6.4.12 Reconstitute with 1 niL of iso-octane and 0.5 mL of methanol.
dilute to a volume of 4 mL with ether. The sample is now
ready for methylation with diazomethane (the sample should not
be stored overnight at this step).
38
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7.0 ESTERIFICATION
7.1 The diazomethane derivatization procedure described below will
produce an efficient reaction with all of the chlorinated
herbicides described in this method. It should be used only
by an experienced analyst, due to the potential hazards
associated with its use.
7.2 Diazomethane is a carcinogen and can explode under certain
conditions. The following precautions should be taken:
• Use a safety screen.
• Use mechanical pipetting aides to reduce potential contact
with diazomethane.
• 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 power, calcium chloride,
and boiling chips.
7.3 Specific instructions for preparing diazomethane are provided
with the generator kit. They must be followed precisely.
7.4 Add 2 mL of diazomethane solution and let the sample stand for
10 ninutes with occasional swirling. The yellow color of
39
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passed, add an
Colored or highly
of diazomethane
pheoxyacid herbicides.
7.5 After 15 minutes rinse
hundred pL of ethyl ether.
1 mL under a stream of
diazomethane by reacting it
7.6 Transfer all of the volume
cleaned Diol cartridge in
Section 4.3.4) using < 1 mL
the transfer.
inside wall of ampule with several
Reduce the sample to approximately
nitrogen. Remove any excess
with 10 mg silicic acid.
of derivatized phenoxyacid to a
a vacuum manifold (described in
of additional hexane to complete
7.7
Elute the transferred solvent through the cartridge and
discard.
7.8 Elute the derivatized analytes
10 mL of 90:10 hexane/acetone.
flask.
7.9 Reduce the final sample
dry nitrogen (described
through the cartridge with
Collect in a 10 mL volumetric
diazomethane should be evident and should persist for this
period. If the yellow color disappears before 10 minutes has
additional 2 mL of diazomethane solution.
coupler samples will require at least 4 mL
to ensure quantitative reaction of all
volume to 1.00 mL under a stream of
in 4.3.2) prior to CC analysis.
40
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8.0 CC ANALYSIS-PHENOXYACIDS
8.1 SUMMARY
8.1.1 Extracts are analyzed by GC/ECD for methyl esters of
phenoxyacid herbicides.
8.1.2 The primary column for analysis is a 15 m x 0.53 mm ID DB—5
megabore capillary column (or equivalent).
8.1.3 The secondary column for analyses is a 15 m x 0.53 mm ID SPB-
608 megabore capillary column (or equivalent).
8.2 CC CONDITIONS
8.2.1 Megabore capillary columns can be installed in 0.25—inch
packed column injector and detector ports using suitable glass
adapters and graphite ferrules. Because the column flow used
will be approximately 5 mL/minute, it is necessary to supply a
suitable makeup gas to the detector.
8.2.2 ECD’s should be plumbed with helium or hydrogen carrier gas
and P—b as a detector makeup gas.
8.2.3 The initial temperature program to use for phenoxyacid
herbicide analysis on both columns is:
T = 50°C
Initial time one minute
Initial temperature ramp 20°/mm to 150°C
Second temperature ramp 8°/mm to 180C
Third temperature ramp 5°/mm to 250°C
41
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it may be necessary to adjust the temperature program for
individual gas chromatographs.
8.2.4 CC injectors should be set at 260°C.
8.3 CALIBRATION
8.3.1 After establishing the appropriate CC operating conditions
(8.2) for the method parameters, calibrate the CC system using
the external standard technique.
8.3.2 For each parameter of interest, prepare working standards at
three concentration levels over a range of at least two orders
of magnitude by reacting solutions of phenoxyacid standards
with diazomethane using the procedure described in Section 7.0
(quantitative yield of methyl esters from standards and
samples is assumed). The low concencentration should be near
to, but not above, the method detection limit (8.7). The
medium concentration must be at least 10 times the low
concentration and the high concentration must be at least 10
times the medium concentration (at least 100 times the low
concentration). These concentrations define the calibration
range in which analytes can be quantitated.
8.3.3 The peak height, or the peak area are plotted against the mass
injected and used as a calibration curve for each parameter.
Alternatively, the ratio of the response to the mass injected,
defined as the calibration factor (CF), can be calculated for
each parameter at each standard concentration. If the
relative standard deviation of the calibration factor for the
three concentrations is less than 10%, detector linearity
through the origin can be assumed and the average calibration
factor can be used in place of a calibration curve.
42
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8.3.4 The working calibration curve or calibration factor must be
verified on each working day by the measurement of one or more
calibration standards. If the response for any parameter
varies from the predicted response by more than 10%, a new
calibration curve or calibration factor must be prepared for
that parameter.
8.4 QUANTITATION OF ANALYSES
8.4.1 Analytes can be quantitated using either manual measurement of
onscale chromatograms, with a modern electronic integrator, or
with a laboratory data system. The analyst can use either
peak height or peak area as the basis for quantitation. The
use of manual integration is strongly discouraged.
8.4.2 If manual quantitation is used, all peaks used for
calibration and for sample analysis must be onscale and give
at least 20 percent deflection from baseline at maximum
height. Guidance in the manual quantitation of peaks is given
in the EPA MANUAL OF ANALYTICAL METHODS FOR ThE ANALYSIS OF
PESTICIDES IN HUMANS AND ENVIRONMENTAL SAMPLES , EPA-600/8-80-
038, or in the FDA Pesticide Analytical Manual.
8.4.3 If electronic integration is used, it is the responsibility of
the analyst to be sure that the integration parameters are set
properly and that off—scale chromatograms are within the
dynamic range of the device. The analyst should also check
for data flags that indicate improper quantitation of peaks
prior to reporting data to the EPA. In addition, the peak
width of identified analytes must not be more than 2.0 times
the width of the high concentration calibration peak(s) for
the same analyte.
43
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8.4.4 The detector response (peak area or peak height) to all of the
analytes must lie between the response of the low and high
concentrations in the three—point initial calibration in order
to be quantitated.
8.4.5 The concentration of the surrogate and of the single component
analytes are calculated using the following equations:
8.4.5.1 Water
(A ) (Vt)
concentration jsg/L = ( CF (V)(V ) (5.10)
Where:
A = Response for the parameter to be measured.
CF = Calibration factor for the external standard.
Vt = Volume of total extract (pL) (take into account any
dilution).
V = Volume of extract injected (p1.).
= Volume of water extracted (mL).
8.4.5.2 Sediment/Soil
(A ) (Vt)
Concentration pg/kg = (CF)(V ) (W 5 ) (D) (5.10)
(Dry weight basis)
Where:
Ax, CF, V = same as given above in 5.3.4.1
44
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D = 100 - percent weight loss (percent weight loss from Section 3.3)
W = Weight of sample extracted (g)
8.5 FLOW SCHEME
8.5.1 A flow scheme for the analysis of samples by this procedure
are given in Figure 1.
8.6 SAMPLE CHROMATOCRAMS
8.6.1 Sample chromatograms of mixtures containing all single
component analytes and of all multi-component analytes are
presented in Figures 2-23.
8.7 METHOD DETECTION LIMIT
8.7.1 Any method is dependent on the complexity of the sample matrix
analyzed, but this procedure can be used to determine all of
the phenoxyacid analytes (Table 3) to a detection limit of
<5 ppb.
45
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9.0 QUALITY CONTROL
9.1 Each time a set of samples is extracted or there is a change
in reagents, a method blank should be processed to demonstrate
that all glassware and reagents are interference—free. Method
interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware;
these contaminants lead to discrete artifacts, and/or elevated
baselines, in gas chromatograms. Interferences by phthalate
esters especially can pose a major problem in pesticide
analysis when using the electron capture detector. Common
flexible plastics contain varying amounts of phthalates which
are easily extracted during laboratory operations. Cross-
contamination of clean glassware routinely occurs when
plastics are handled. Interferences from phthalates can best
be minimized by avoiding the use of such plastics in the
laboratory. Exhaustive cleanup of reagents and glassware may
be required to eliminate background phthalate contamination.
9.2 Standard quality assurance practices should be used with this
method. Field replicates should be collected to validate the
precision of the sampling technique. Laboratory replicates
should be analyzed to validate the precision of the analysis.
It is suggested that the response of the external standards be
plotted daily as a quality control check. Where doubt exists
over the identification of a peak on the chromatogram,
confirmatory techniques such as CC/MS should be used.
9.3 Waste samples spiked with selected analytes must be analyzed
for at least 10% of the samples to validate the accuracy of
the analysis. The results of the recovery of spiked analytes
must be reported with sample data.
46
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9.4 At the option of the I.T.D., the requirement for spiking at
least 10% of the samples with selected herbicides may be
replaced with a requirement for analyzing a matrix
spike/matrix spike duplicate (MS/MSD) pair for every 20
samples analyzed.
9.5 The MS/MSD pair are prepared by spiking duplicate sample
aliquots with the particular matrix spiking solutions
described below. The MS/MSD pair as well as the unspiked
sample are analyzed by the procedures described in this
protocol. The recoveries of the matrix spike compounds are
reported with the sample data using the formula below.
Matrix spike recovery = SS SR x 100
SSR = Sample spike results
SR = Sample result
SA = Spike added
9.5.1 The apolar MS/MSD samples are each spiked with two solutions.
One for organochlorine compounds and the other for
organophosphates. The spiking solution for organochlorines is
the same one as is used in the CL I ’ analysis of organochlorine
pesticides and is listed below.
Pesticide pg/1.0 mL
gamma-BHC 0.5
Heptachlor 0.5
Aldrin 0.5
Dieldrin 1.0
Endrin 1.0
4,4’-DDT 1.0
47
-------
The spiking solution for organophosphate pesticides is listed
be I ow.
Pesticide j g/1.0 mL
Chlorpyriophos 100
Fenth ion 100
Malathion 100
Parathion, ethyl 100
Carbofenthion 100
The solutions are prepared in acetone and must be allowed to
equilibrate at room temperature before they are spiked.
9.5.2 The phenoxyacid MS/MSD samples are spiked with the mixture
listed below.
Herbicide / sg/1.0 mL
2,4-D 0.5
24,5-T 0.5
The solution is prepared in acetone and must be allowed to
equilibrate at room temperature before it is spiked.
9.6 It is critical that analysts become proficient with capillary
OC analysis before using this method to generate quantitative
resu Its.
9.7 Store stock solutions of standards at 4°C and protect from
light. All such solutions must be checked frequently for
signs of degradation or evaporation. Organophosphorous
standards are particularly prone to hydrolysis during storage.
48
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10.0 REFERENCES
10.1 Statement of Work - CC Methods for Environment Samples,
Pesticides, PCP’s (proposed), Prepared for J. Fisk, Office of
Remedial and Emergency Response, USEPA, by P. Marsden and D.
Bottrel I.
10.2 Single-Laboratory Validation of EPA Method 8150 for the Analysis
of Chlorinated Herbicides in Hazardous Waste, EPA 600/4-85-060,
prepared for D.F. Curka, QAD, EMSL-LV, by F. Shore, E.N. Arnick,
and S.T. Pan.
10.3 Single Laboratory Validation of EPA Method 8140 for the Analysis
of Organophosphorous Pesticides in Hazardous Waste (in
manuscript), prepared for D. Betowski, QAD, EMSL-LV, by V.
Taylor, D. Hickey, and P. Marsden.
10.4 Methods for Organic Chemical Analysis of Municipal and Industrial
Waste Water, EPA 600/4-82-057, by J.E. Longbottom and J.J.
Li chtenberg.
49
-------
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RE L TED COKI- I IUNDS
I2OMFTt ND ME FI-IOL’ Dh—b sr h--e.)C’El
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Oh oruvrophos 1)8 622 i -id rd
frotcciyphos n c .4 ,06)
Di ch I. or-f siil:h tori 70 I 9, o 7 , 91
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RE: lENT [ U I -i I [ Fl :
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( N 4_’ IL S ( .Ui’ ThLJt I Lfl TL:i) HF I ( iL
c i )NIFO( ‘ND Hi:t H OD Di3—-:3 srB —é,ñu
Dtnos&b . 5 1 5
LiD
;: ,4 5 -T o lD
— , —ir. - IL-
ir
E I iF L ) jtiFO IJNDS
f’,
-------
Figure 1. Flow Scheme for the Analysis of Samples.
53
-------
I;”
3.
Dichioran
Br4C 1 rna
BHC,detta
—
L . Heptachlo,- epoxide
Captari
N itr o 4 efl
Chleroberi:yl ata
________ DDD
Endrin Aldehyde
Carboph.nthi on
Endouul4an aulfate
___________________ Enørjri ketone
Hx abromabenz en.
Figure 2A. Sample Chromatogram of Selected Apolar
Organochlorine Pesticides on the DB—5.
54
-------
PCNB
BHC,atpha
H. p CO L jr
- - 1 odrin
_________________________________ Endo u1f an
DDE Dteldrin
L ___ Endrin
Endo5ul4an II
,— - ,
Kepon.
,
_____________ DDT
CaptO 4 Ol
Methexychtor
Figure 2B. Sample Chromatogram of Selected Apolar
Organochiorine Pesticides on the DB—5.
I
I
55
-------
,, .
4
DBCP
BHC ,a1ph
F- 1_NB
BHC,beta
I pL3rhIOr
Srdr in
End uLf I
DDE
Di el dr 1 I
Enciosul4afl II
Figure 3A.
Sample Chromatogram of Selected Apolar Organochlorine
Pesticides on the SPB-608.
S ..
ia. 4’
Endr i ri
DDT
Captof 01 -
e
Mire
56
-------
-lept ach tar
epz’ ide
Endrin ldehyde
Figure 3B. Sample Chromatogram of Selected Apolar Organoch lorine
Pesticides on the SPB .-608.
57
1 •
4 P0
Fr i f 1 jr at in
Di c hi or an
B IIC ,qamma
delta
— I r 1(1
Captan
Ni tro-f en
DOD
Endosuifan quifato
o
L p 4 brjmrbente”t
-------
.4 . )
L
0
U-
>-
4- I
wu
ED
-c
‘no.
.40
N
z
U-
La.:
Li
U
U I ! i i
Figure 4A. Sample Chromatogram of Selected Apolar
Organophosphorous Pesticides on the DB-5.
58
r.j
tO
0
-C
a
0
>,.o
c i
to
0
-C
a
-D
L
0
-I-
-------
-l
— —
C>.
a. .êJ C
o
.4. ’ -,
o
1- U- :
I..J
LL
0
0.2
-I
C
w
C
>
0
L
0
C
U
‘ 0
L
C.
00
I ., 0
1
Figure 4B. Sample Chromatogram of Selected Apolar
Organophosphorous Pesticides on the DB—5.
59
C
C
.1
ft
1,—I
1’•’
I .)
•1i
I—,—,
-4
‘ -4
-------
It
- -
- ac
—
Figure 5A. Sample Chromatogram of Selected Apolar
Organophosphorous Pesticides on the SPB-608.
60
a
QC
I
C
0’ —
-4
-4
C-
LI,
-------
F -
(n - . 4
r
Figure 5B. Sample Chromatogram of Selected Apolar
Organophosphorous Pesticides on the SPB—608.
C
0
r
LC 4J
,- 0
—
to
I.— C
0
to
C
r
-S
0
-I
LI
L i ’
‘-4
-4
Li
I-.-
I L
C L
l. I
CL
(
to
0
>
0
-4
U
.4
61
-------
Figure 6. Sample Chromatogram of Phenoxyacids on the DB-5.
62
-------
Figure 7. Sample Chromatogram of Phenoxyacids on the SPB—608.
63
-------
I,
II
—.
In.
I I
:,
Figure 8. Toxaphene on the DB-5.
64
1(1
II•I J 0
0
I ,
•1
.1.
• II
II
-------
Figure 9.
Toxaphene on the SPB-608.
tf ;
U )-
iI
I I,
I 1n •
65
-------
Figure 10.
Chiordane on the DB-5.
66
• I
4 ,
0
-t
-7 .
U,
4.-, 7’- •
‘I
I
4 .AJ
-------
Figure 11. Chlordane on the SPB—608.
67
r
LO Y
• •• [ I I IIIO I .
(•J• •fl I c m.
• . nfl I
‘a
C,
uI .
.1
0
-t
I I I
-------
Figure 12.
Aroclor 1016/1260 on the DB—5.
68
N.
I,
0
Lk I
7 3
I .2.
I.
-I ‘I
I.
27.
U,
U
-------
I I.I . II,
I I •
•t II I I) Ill II, I )
ID I
lI I It’ll,
•IJ II I I, 4 II
Figure 13.
Aroclor 1016/1260 on the SPB-608.
69
I ,’,
1 TI
I I
T I)
It
‘II
I l
I l I t, I’
T I I
In..
-------
Figure 14. Aroclor 1221 on the DB—5.
70
t .
— Ii —
Iii,
‘C.
‘I II
C’
-------
Figure 15.
Aroclor 1221 on the SPB-608.
a
.0
I
II
I -
C,
71
-------
Figure 16. Aroclor 1232 on the DB—5.
72
-------
Figure 17.
Aroclor 1232 on the SPB-608.
. , I. f
‘
. :
I
‘A
0
73
-------
I
I
Figure 18.
Aroctor 1242 on the DB-5.
P .-
f )
-I
S
I
U-,
‘I
74
-------
Figure 19.
Aroclor 1242 on the SPB—608.
75
,1 jF,JJ! 0 . 7 0
• t • . ;•‘•‘‘ • ‘.
• . I
I ..
I,
( ‘I
C.
-------
Figure 20.
Aroclor 1248 on the DB-5.
. (, ..- .J
_.v,o 7-
.).-• . .7
r , F. . -r
‘7., ‘
.7
.7
‘.7
I
76
-------
Figure 21.
Aroclor 1248 on the SPB-608.
77
•11 r)
I,,
I
I,
-t
a
(I
-------
Figure 22.
Aroclor 1254 on the DB—5.
78
Ire.
I 0
U.
I,
I —
F -)
Is
.7
I
-r
•.0
7-
I !
U,
I,
II,
-------
Figure 23.
Arocior 1254 on the SPB-608.
79
‘
•1
4
I. ,, I .
I . ‘ 3’
.-•r• I ‘i
I I •‘I tUJ I
‘I
-t
II
-j
I-
-I
r
0
U.
“.3 ’
‘I.
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